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EP3634401A1 - Antibody construct conjugates - Google Patents

Antibody construct conjugates

Info

Publication number
EP3634401A1
EP3634401A1 EP18814341.6A EP18814341A EP3634401A1 EP 3634401 A1 EP3634401 A1 EP 3634401A1 EP 18814341 A EP18814341 A EP 18814341A EP 3634401 A1 EP3634401 A1 EP 3634401A1
Authority
EP
European Patent Office
Prior art keywords
immune
amino acid
seq
conjugate
domain
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Withdrawn
Application number
EP18814341.6A
Other languages
German (de)
French (fr)
Inventor
Peter Armstrong Thompson
Badreddin EDRIS
Craig Alan Coburn
Peter Robert Baum
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
ARS Pharmaceuticals Inc
Original Assignee
Silverback Therapeutics Inc
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Silverback Therapeutics Inc filed Critical Silverback Therapeutics Inc
Publication of EP3634401A1 publication Critical patent/EP3634401A1/en
Withdrawn legal-status Critical Current

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Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D417/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00
    • C07D417/14—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing three or more hetero rings
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/68—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment
    • A61K47/6835—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment the modifying agent being an antibody or an immunoglobulin bearing at least one antigen-binding site
    • A61K47/6849—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment the modifying agent being an antibody or an immunoglobulin bearing at least one antigen-binding site the antibody targeting a receptor, a cell surface antigen or a cell surface determinant
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/68—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment
    • A61K47/6891—Pre-targeting systems involving an antibody for targeting specific cells
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
    • C07D401/14—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing three or more hetero rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
    • C07K16/30—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants from tumour cells
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/32—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against translation products of oncogenes
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/70—Immunoglobulins specific features characterized by effect upon binding to a cell or to an antigen
    • C07K2317/71—Decreased effector function due to an Fc-modification
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/95—Fusion polypeptide containing a motif/fusion for degradation (ubiquitin fusions, PEST sequence)

Definitions

  • cancer One of the leading causes of death in the United States is cancer.
  • the conventional methods of cancer treatment like chemotherapy, surgery, or radiation therapy, tend to be either highly toxic or nonspecific to a cancer, or both, resulting in limited efficacy and harmful side effects.
  • the immune system has the potential to be a powerful, specific tool in fighting cancers.
  • tumors can specifically express genes whose products are required for inducing or maintaining the malignant state. These proteins may serve as antigen markers for the development and establishment of more specific anti-cancer immune response.
  • the immune response may include the recruitment of immune cells that target tumors expressing these antigen markers.
  • the immune cells may express genes whose products are important to proper immune function and may serve as markers for specific types of immune cells.
  • the boosting of this specific immune response has the potential to be a powerful anti-cancer treatment that can be more effective than conventional methods of cancer treatment and can have fewer side effects.
  • an immune-modulatory conjugate comprising: (a) an antibody construct comprising an antigen binding domain and an Fc domain, wherein the antigen binding domain binds to a first antigen; (b) a proteolysis targeting module, comprising: (i) a protein targeting moiety that binds to a target protein; (ii) an E3 ubiquitin ligase binding moiety; and (iii) a spacer S that is covalently bound to the protein targeting moiety and to the E3 ubiquitin ligase binding moiety, wherein the spacer is optionally has 1-25 consecutive non-hydrogen atoms; and (c) a linker L that is covalently attached to the antibody construct and to the proteolysis targeting module.
  • the conjugate can be represented by one of the following formulae:
  • the Fc domain of the conjugate is an Fc null.
  • the E3 ubiquitin ligase is selected from the group consisting of
  • VHL Von Hippel-Lindaue E3 ubiquitin ligase
  • HERC5 HUWE1, HYD, ITCH, LNX1, mahogunin, MARCH-I, MARCH-II, MARCH-III,
  • RNF8 RNF19, RNF190, RNF20, RNF34, RNF40, RNF125, RNF128, RNF138, RNF168,
  • the E3 ubiquitin ligase binding moiety binds to VHL, cereblon or MDM2.
  • the protein targeting moiety binds to a protein selected from one of the groups consisting of: a) an aryl hydrocarbon receptor, androgen receptor, estrogen receptor, FK506-binding protein 12, fibroblast growth factor receptor substrate 2,
  • phosphatidylinositol -4,5-biphosphate 3-kinase SMAD family member 3, bromodomain and extra-territorial family of proteins (BET), bromodomain-containing protein 4 member of the BET family, Abelson tyrosine kinase, receptor- interacting serine/threonine-protein kinase 1, estrogen- related receptor, and transforming growth factor beta (TGFP); or b) ALK, Bcr-Abl, BRAF, BTK, c-KIT, EGFR, ErbB2, JAK, MEK, MET, PDGFRB, RET, ROS 1, Syk, SRC, TNIK, TNKS, TNKS2, TrkA, TrkB, TrkC, VEGFR1, VEGFR2, VEGFR3, FGFR1, FGFR2, FGFR3, FGFR4, CSF1R, RON/MS T1R, TYR03, MERTK, AXL, PI3K,
  • the first antigen is a tumor antigen.
  • the tumor antigen can be MUC16, CD5, CD19, CD20, CD25, CD37, CD30, CD33, CD45, CAMPATH-1, BCMA, CS-1, PD-L1, B7-H3, B7-DC, HLD-DR, carcinoembryonic antigen (CEA), TAG-72, EpCAM, MUC1, fo late-binding protein, A33, G250, pro state- specific membrane antigen (PSMA), ferritin, GD2, GD3, GM2, Ley, CA-125, CA19-9, epidermal growth factor, pl85HER2, IL-2 receptor, fibroblast activation protein (FAP), tenascin, a metalloproteinase, endosialin, vascular endothelial growth factor, v 3, WT1, LMP2, HPV E6, HPV E7, EGFRvIII, Her-2/neu, MAGE A3, p53 nonmutant,
  • CEA carcino
  • the first antigen is an immune cell antigen, and can be an antigen present on the surface of an antigen presenting cell such as a dendritic cell or a macrophage.
  • the immune cell antigen is selected from the group consisting of CD40, DEC-205, PD-Ll, PD-1, CD36 mannose scavenger receptor 1, CLEC9A, DC-SIGN, CLEC12A, BDCA-2, OX40L, 41BBL, CD204, MARCO, CLEC5A, Dectin 1, Dectin 2, CLECIOA, CD206, CD64, CD32A, CD 16 A, HVEM, and CD32B.
  • the protein targeting moiety can be a peptide or a non- proteinaceous molecule.
  • the proteolysis targeting module is selected from compounds 1-1, 1-2, 1-3, 1-4, 1-5, 1-6 and 2-1, as disclosed herein.
  • the linker L is a cleavable linker. In some embodiments, the linker L is a non-cleavable linker.
  • the linker L is covalently bound to the antibody construct at a cysteine residue, an engineered cysteine residue, a lysine residue, a glutamate residue, or a glutamine residue of the antibody construct or is covalently bound to the antibody construct using a Sortase linker.
  • the spacer S is an optionally substituted Ci-25 alkylene or optionally substituted Ci- 25 heteroalkylene, wherein the hetero alkylene is a Ci-24 alkylene chain interspersed with one or more groups independently selected from: -0-, -S-, -NH 2 -, and - C(0)NH-; and optionally substituted with a reactive group, R , that can form a functional group selected from an amide bond, an ester bond, an ether bond, a carbonate bond, a carbamate bond, or a thioether bond.
  • the spacer can be optionally substituted with Ci-C 8 alkyl, C 2 -C 8 alkenyl, C 2 - C 8 alkynyl, -(CH 2 0) n iH, -(CH 2 CH 2 0) n iH, -(CH 2 0) n iCH 3 , -C(0)OH or -NH 2 , wherein nl is from 1 to 8.
  • the spacer is substituted with R x .
  • the linker is unsubstituted.
  • the antibody construct further comprises a second binding domain.
  • the second binding domain can specifically bind to an antigen on an immune cell.
  • the immune cell antigen is selected from the group consisting of CD40, PD-1, PD-Ll, DEC-205, CD36 mannose scavenger receptor 1, CLEC9A, DC-SIGN, CLEC12A, BDCA-2, OX40L, 41BBL, CD204, MARCO, CLEC5A, Dectin 1, Dectin 2, CLECIOA, CD206, CD64, CD32A, CD 16 A, HVEM, and CD32B.
  • the second binding domain is attached to the antibody construct at a C-terminal end of the Fc domain.
  • the Fc domain is an Fc domain variant comprising at least one amino acid residue change as compared to a wild type sequence of the Fc domain. In some embodiments, the Fc domain is an Fc domain variant that decreases binding of the Fc domain to an Fc receptor.
  • the at least one amino acid residue change in the Fc domain is: a) F243L, R292P, Y300L, L235V, and P396L, wherein numbering of amino acid residues in the Fc domain is relative to SEQ ID NO: 898; b) S239D and I332E, wherein numbering of amino acid residues in the Fc domain is relative to SEQ ID NO: 898; or c) S298A, E333A, and K334A, wherein numbering of amino acid residues in the Fc domain is relative to SEQ ID NO: 898.
  • the at least one amino acid residue change in the Fc domain is: a) N297A, N297G, N297Q, N297D as in Eu index of Kabat numbering and relative to SEQ ID NO: 898; or b) K322A/L234A/L235A N296A as in the EU index of Kabat numbering and relative to SEQ ID NO: 898; or c) L234F/L235E/P331S N296A as in the EU index of Kabat numbering and relative to SEQ ID NO: 898; or d) P329G/L234A/L235A as in the EU index of Kabat numbering and relative to SEQ ID NO: 898.
  • a pharmaceutical composition comprising a conjugate as described herein along with a pharmaceutically acceptable excipient.
  • the average ratio of proteolysis targeting modules to antibody construct in the conjugate can be from 2 to 6, from 3 to 5 or 1 to 3.
  • the pharmaceutical composition can be lyophilized.
  • kits for treating cancer comprising administering a conjugate as described herein, or a pharmaceutical composition of a conjugate as described herein, to a subject in need thereof.
  • the cancer is a solid tumor, such as breast cancer, pancreatic cancer, colorectal cancer, renal cell cancer, gastric cancer, or lung cancer.
  • the pharmaceutical composition is administered parenterally.
  • the pharmaceutical composition is administered intravenously.
  • the disclosure also provides immune- stimulatory conjugates comprising: (a) an immune- stimulatory compound that binds to a target protein to stimulate an immune response by stimulating the activity of the target protein, reducing immune inhibition by the target protein or increasing degradation of the target protein; (b) an antibody construct comprising an antigen binding domain and an Fc domain, wherein the antigen binding domain binds to at least a first antigen and wherein the Fc domain binds to an Fc receptor; and (c) a linker, wherein the linker is covalently bound to the antibody construct and the linker is covalently bound to the immune- stimulatory compound.
  • the dissociation constant (K d ) for binding of the Fc domain of the conjugate to an Fc receptor is equal to, or up to no greater than about 100 times the K d for binding of a control antibody construct to the Fc receptor, wherein the control antibody construct is the unconjugated antibody construct; and wherein the K d for binding of the immune- stimulatory compound of the conjugate (i.e, attached to the conjugate) to the target protein is equal to, or up to no greater than 100 times the K d for binding of a control compound to the target protein active site or wherein the EC 50 or IC 50 of the immune- stimulatory compound of the conjugate is no greater than 300-fold the EC 50 or IC 50 of a control compound, wherein the control compound is the unbound immune- stimulatory compound.
  • the dissociation constant (K d ) for binding of the Fc domain of the conjugate to an Fc receptor is greater than about 100 times the K d for binding of a control antibody construct to the Fc receptor, wherein the control antibody construct is the unconjugated antibody construct; and wherein the K d for binding of the immune- stimulatory compound of the conjugate (i.e, attached to the conjugate) to the target protein is equal to, or up to no greater than 100 times the K d for binding of a control compound to the target protein active site or wherein the EC 50 or IC 50 of the immune- stimulatory compound of the conjugate is no greater than 300- fold the EC 50 or IC 50 of a control compound, wherein the control compound is the unbound immune- stimulatory compound.
  • the Fc domain is an Fc null.
  • the disclosure provides an immune- stimulatory conjugate comprising: (a) an immune- stimulatory compound that binds to a protein active site to stimulate an immune response; (b) an antibody construct comprising an antigen binding domain and an Fc domain, wherein the antigen binding domain binds to at least a first antigen and wherein the Fc domain binds to an Fc receptor; and (c) a linker, wherein the linker is covalently bound to the antibody construct and the linker is covalently bound to the immune- stimulatory compound.
  • the dissociation constant (K d ) for binding of the Fc domain of the conjugate to an Fc receptor is equal to, or up to no greater than about 100 times the K d for binding of a control antibody construct to the Fc receptor, wherein the control antibody construct is the unconjugated antibody construct; and wherein the K d for binding of immune- stimulatory compound of the conjugate to the protein active site is equal to, or up to no greater than 100 times the K d for binding of a control compound to the protein active site or wherein the EC 50 or IC 50 of the immune- stimulatory compound of the conjugate (attached to the conjugate )is no greater than 300-fold the EC 50 or IC 50 of a control compound, wherein the control compound is the unbound immune- stimulatory compound.
  • the dissociation constant (K d ) for binding of the Fc domain of the conjugate to an Fc receptor is greater than about 100 times the K d for binding of a control antibody construct to the Fc receptor, wherein the control antibody construct is the unconjugated antibody construct; and wherein the K d for binding of immune- stimulatory compound of the conjugate to the protein active site is equal to, or up to no greater than 100 times the K d for binding of a control compound to the protein active site or wherein the EC 50 or IC 50 of the immune- stimulatory compound of the conjugate (attached to the conjugate) is no greater than 300-fold the EC 50 or IC 50 of a control compound, wherein the control compound is the unbound immune- stimulatory compound.
  • the disclosure provides an immune- stimulatory conjugate comprising: (a) an immune- stimulatory compound that binds to a protein active site of a binding protein to stimulate an immune response by inhibition of the activity of the binding protein; (b) an antibody construct comprising an antigen binding domain and an Fc domain, wherein the antigen binding domain binds to at least a first antigen and wherein the Fc domain binds to an Fc receptor; and (c) a linker, wherein the linker is covalently bound to the antibody construct and the linker is covalently bound to the immune- stimulatory compound.
  • the dissociation constant (K d ) for binding of the Fc domain of the conjugate to an Fc receptor is equal to, or up to no greater than about 100 times the K d for binding of a control antibody construct to the Fc receptor, wherein the control antibody construct is the unconjugated antibody construct; and wherein the K d for binding of the immune- stimulatory compound of the conjugate (attached to the conjugate) to the protein active site is equal to, or up to no greater than 100 times the K d for binding of a control compound to the protein active site or wherein the IC 50 of the immune- stimulatory compound of the conjugate (attached to the conjugate) is no greater than 300-fold the IC 50 of a control compound, wherein the control compound is the unbound immune- stimulatory compound.
  • the dissociation constant (K d ) for binding of the Fc domain of the conjugate to an Fc receptor is greater than about 100 times the K d for binding of a control antibody construct to the Fc receptor, wherein the control antibody construct is the unconjugated antibody construct; and wherein the K d for binding of the immune- stimulatory compound of the conjugate (attached to the conjugate) to the protein active site is equal to, or up to no greater than 100 times the K d for binding of a control compound to the protein active site or wherein the IC 50 of the immune- stimulatory compound of the conjugate (attached to the conjugate) is no greater than 300-fold the IC 50 of a control compound, wherein the control compound is the unbound immune- stimulatory compound.
  • the Fc domain is an Fc null.
  • the disclosure provides an immune- stimulatory conjugate comprising: (a) an immune- stimulatory compound that binds to a protein active site of a binding protein to stimulate an immune response by increasing degradation of the binding protein; (b) an antibody construct comprising an antigen binding domain and an Fc domain, wherein the antigen binding domain binds to at least a first antigen and wherein the Fc domain binds to an Fc receptor; and (c) a linker, wherein the linker is covalently bound to the antibody construct and the linker is covalently bound to the immune- stimulatory compound.
  • the dissociation constant (K d ) for binding of the Fc domain of the conjugate to an Fc receptor is equal to, or up to no greater than about 100 times the K d for binding of a control antibody construct to the Fc receptor, wherein the control antibody construct is the unconjugated antibody construct; and wherein the K d for binding of the immune- stimulatory compound of the conjugate
  • the IC 50 of the immune- stimulatory compound of the conjugate is no greater than 300-fold the IC 50 of a control compound, wherein the control compound is the unbound immune- stimulatory compound.
  • the dissociation constant (K d ) for binding of the Fc domain of the conjugate to an Fc receptor is greater than about 100 times the K d for binding of a control antibody construct to the Fc receptor, wherein the control antibody construct is the unconjugated antibody construct; and wherein the K d for binding of the immune- stimulatory compound of the conjugate (attached to the conjugate) to the protein active site is equal to, or up to no greater than 100 times the K d for binding of a control compound to the protein active site or wherein the IC 50 of the immune- stimulatory compound of the conjugate is no greater than 300-fold the IC 50 of a control compound, wherein the control compound is the unbound immune- stimulatory compound.
  • the Fc domain is an Fc null.
  • the EC 50 or IC 50 of the immune- stimulatory compound of the conjugate is no greater than 100-fold the EC 50 or IC 50 of a control compound, wherein the control compound is the unbound immune- stimulatory compound.
  • the EC 50 or IC 50 of the immune- stimulatory compound of the conjugate may be no greater than 10-fold the EC 50 or IC 50 of a control compound, wherein the control compound is the unbound immune- stimulatory compound.
  • the EC 50 or IC 50 of the immune- stimulatory compound of the conjugate may be equivalent to or less than the EC 50 or IC 50 of a control compound, wherein the control compound is the unbound immune- stimulatory compound.
  • the EC 50 or IC 50 of the immune stimulatory compound on an antigen bearing cell is equivalent to or less than the EC 50 or IC 50 of a control compound on the antigen bearing cell and the EC 50 or IC 50 of the immune- stimulatory conjugate is 5-fold greater or more than the EC 50 or IC 50 of the control compound for a non-antigen bearing cell.
  • the immune- stimulatory compound of the conjugate may have a K d for binding to the protein active site of no greater than 50 times the K d for binding of a control compound to the protein active site, wherein the control compound is the unbound immune- stimulatory
  • the immune- stimulatory compound of the conjugate may have a K d for binding to the protein active site of no greater than 10 times the K d for binding of a control compound to the protein active site, wherein the control compound is the unbound immune- stimulatory
  • the immune- stimulatory compound of the conjugate may have a K d for binding to the protein active site of equivalent to or less than the K d for binding of a control compound to the protein active site, wherein the control compound is the unbound immune- stimulatory compound.
  • an immune- stimulatory conjugate comprising: (a) an immune- stimulatory compound that binds to a protein active site to stimulate an immune response; (b) an antibody construct comprising an antigen binding domain and an Fc domain, wherein the antigen binding domain binds to a first antigen and wherein the Fc domain binds to an Fc receptor; and (c) a linker comprising 5- 100 consecutive atoms attachment sites, wherein one attachment site of the linker is covalently bound to the antibody construct and another attachment site of the linker is covalently bound to the immune- stimulatory compound.
  • the dissociation constant (K d ) for binding of the Fc domain of the conjugate to an Fc receptor is equal to, or up to no greater than about 100 times the K d for binding of a control antibody construct to the Fc receptor, wherein the control antibody construct is the unconjugated antibody construct; and wherein the K d for binding of the immune- stimulatory compound when bound to a the linker to the protein active site is no greater than 100 times the K d for binding of a control compound to the protein active site or wherein the EC 50 or IC 50 of the immune- stimulatory compound of the conjugate is no greater than 300-fold the EC 50 or IC 50 of a control compound, wherein the control compound is the unbound immune- stimulatory compound.
  • the dissociation constant (K d ) for binding of the Fc domain of the conjugate to an Fc receptor is greater than about 100 times the K d for binding of a control antibody construct to the Fc receptor, wherein the control antibody construct is the unconjugated antibody construct; and wherein the K d for binding of the immune- stimulatory compound when bound to a the linker to the protein active site is no greater than 100 times the K d for binding of a control compound to the protein active site or wherein the EC 50 or IC 50 of the immune- stimulatory compound of the conjugate is no greater than 300-fold the EC 50 or IC 50 of a control compound, wherein the control compound is the unbound immune- stimulatory compound.
  • the Fc domain is an Fc null.
  • the EC 50 or IC 50 of the immune- stimulatory compound when bound to the 5- 100 atom linker is no greater than 100-fold the EC 50 or IC 50 of a control compound, wherein the control compound is the unbound immune- stimulatory compound.
  • the EC 50 or IC 50 of the immune- stimulatory compound when bound to the 5- 100 atom linker is no greater than 10-fold the EC 50 or IC 50 of a control compound, wherein the control compound is the unbound immune- stimulatory compound.
  • the EC 50 or IC 50 of the immune- stimulatory compound when bound to the 5- 100 atom linker may be equivalent to or less than the EC 50 or IC 50 of a control compound, wherein the control compound is the unbound immune- stimulatory compound.
  • the K d for binding of the immune- stimulatory compound when bound to the 5- 100 atom linker to the protein active site may be no greater than 50 times the K d for binding of a control compound to the protein active site, wherein the control compound is the unbound immune- stimulatory compound.
  • the K d for binding of the of the immune- stimulatory compound when bound to a 5-100 atom linker to the protein active site may be no greater than 10 times the K d for binding of a control compound to the protein active site, wherein the control compound is the unbound immune- stimulatory compound.
  • the K d for binding of the immune- stimulatory compound when bound to a 5-100 atom linker to the protein active site may be equivalent to or less than the K d for binding of a control compound to the protein active site wherein the control compound is the unbound immune- stimulatory compound.
  • the immune- stimulatory conjugate described herein comprises moiety that binds to the binding protein and an E3 ubiquitin ligase binding moiety.
  • the E3 ubiquitin ligase binding moiety may bind to, for example, VHL, cereblon or MDM2.
  • the E3 ubiquitin ligase binding moiety may be selected from among E3 targeting compounds.
  • the E3 ubiquitin ligase binding moiety may be attached to the linker or may be part of the linker.
  • the E3 ubiquitin ligase binding moiety attached to the linker wherein the E3 ubiquitin ligase binding moiety is bound through a first linker having 5-100 consecutive atoms between attachment sites to the immune- stimulatory compound and the E3 ubiquitin ligase binding moiety is bound through a second linker having 5-100 consecutive atoms between attachment points to the antibody construct.
  • the immune- stimulatory compound is a kinase inhibitor and the protein active site is a kinase active site.
  • the linker is covalently bound to the kinase inhibitor at a position on the kinase inhibitor that is at or near the solvent interface of the kinase active site as determined by modeling of the kinase inhibitor in the kinase active site.
  • the linker is covalently bound to the kinase inhibitor at a position on the kinase inhibitor such that when the kinase inhibitor is bound to the active site, the linker extends out from the kinase active site into the solvent, as determined by modeling of the kinase inhibitor in the kinase active site.
  • the kinase inhibitor may be selected from an inhibitor of ALK, Bcr-Abl, BRAF, BTK, c-KIT, EGFR, ErbB2, JAK, MEK, MET, PDGFRB, RET, ROS 1, Syk, SRC, TGFpR2, TNIK, TNKS, TNKS2, TrkA, TrkB, TrkC, VEGF, VEGFR1, VEGFR2, VEGFR3, FGFR1, FGFR2, FGFR3, FGFR4, CSF1R, RON/MS T1R, TYR03, MERTK, AXL, PDKdelta PDKgamma, MAP4K1, PERK, and combinations thereof.
  • the immune- stimulatory compound is selected from a to 11- like receptor agonist, STING agonist, or RIG-I agonist.
  • the immune- stimulatory compound may be a toll-like receptor (TLR) agonist selected from a TLR1 agonist, a TLR2 agonist, a TLR3 agonist, a TLR4 agonist, a TLR5 agonist, a TLR6 agonist, a TLR7 agonist, a TLR8 agonist, a TLR9 agonist, or a TLR10 agonist.
  • TLR toll-like receptor
  • the immune- stimulatory compound may be selected from a pyrimidine, a purine, a guanine nucleoside, an 8-oxoadenine, an imidazoquinoline, a
  • the immune- stimulatory compound may bind to a G protein-coupled receptor (GCPR), an ion channel, a membrane transporter, a phosphatase or an endoplasmic reticulum (ER) protein.
  • GCPR G protein-coupled receptor
  • ER endoplasmic reticulum
  • the immune- stimulatory compound is an antagonist of the GPCR A2aR, the sphingosine 1-phosphate receptor 1, prostaglandin receptor EP3, prostanglandin receptor E2, Frizzled, CXCR4 or an LPA receptor.
  • the immune- stimulatory compound may be an ion channel agonist for CRAC, Kvl.3 or KCa3.1.
  • the immune- stimulatory compound may be an inhibitor of HSP90 or AAA-ATPase p97.
  • the immune- stimulatory conjugate has immune- stimulatory activity while attached to the conjugate or the linker and without undergoing cell processing such as by endosomal or lysosomal degradation, e.g., to release the immune- stimulatory compound or a modified form of the compound from the conjugate or linker.
  • the linker is a non-cleavable linker. The linker may be selected from the group consisting of
  • the linker may comprise one or more carbamate or amide linkages when attached to an immune- stimulatory compound.
  • the linker may be selected from a linker is
  • R x is a reactive moiety and wherein R is selected from the group consisting of hydrogen, alkyl (e.g., d- C 8 alkyl), sulfonate and methyl sulfonate.
  • the linker may be attached to the antibody construct at a cysteine or lysine residue of the antibody construct.
  • the first antigen is a tumor antigen.
  • the first antigen may be at least 80% or 100% homologous to CD5, CD19, CD20, CD25, CD37, CD30, CD33, CD45, CAMPATH-1, BCMA, CS-1, PD-L1, B7-H3, B7-DC (PD-L2), HLD-DR, carcinoembryonic antigen (CEA), TAG-72, EpCAM, MUC1, MUC15, MUC16, folate-binding protein, A33, G250, pro state- specific membrane antigen (PSMA), ferritin, GD2, GD3, GM2, Le y , CA-125, CA19-9, epidermal growth factor, pl85HER2, IL-2 receptor, EGFRvIII (de2-7 EGFR), fibroblast activation protein (FAP), tenascin, a metalloproteinase, endosialin, vascular endothelial growth factor, ⁇ 3, WT1,
  • the antibody construct further comprises a second target binding domain.
  • the target binding domain specifically binds an antigen on an immune cell.
  • the target binding domain may be attached (e.g., conjugated or linked) to the antibody construct at a C-terminal end of the Fc domain.
  • the antigen binding domain may be from an antibody or non-antibody scaffold.
  • the antigen binding domain may be at least 97% homologous to an antigen binding domain from an antibody or non-antibody scaffold.
  • the antibody construct may be a human antibody or a humanized antibody.
  • the Fc domain may be an Fc domain variant comprising at least one amino acid residue change as compared to a wild type sequence of the Fc domain.
  • the Fc domain may be at least about 80% homologous to an Fc domain from an antibody, wherein the Fc domain from an antibody comprises amino acid residues 216 to 447 of an IgGl (within SEQ ID NO: 898), amino acid residues 216 to 443 of an IgG2 (within SEQ ID NO: 899), or amino acid residues 216 to 444 of an IgG4 (within SEQ ID NO: 900).
  • the Fc domain may have at least one amino acid residue change as compared to a wildtype Fc domain. In certain embodiments, the Fc domain may have at least one amino acid residue change as compared to a wildtype Fc domain, wherein the Fc domain is at least 80% homologous to SEQ ID NO: 898.
  • the at least one amino acid residue change may be in an IgG Fc domain: a) F243L, R292P, Y300L, L235V, and P396L, wherein numbering of amino acid residues in the Fc domain is relative to SEQ ID NO:898; b) S239D and I332E, wherein numbering of amino acid residues in the Fc domain is relative to SEQ ID NO: 898; or c)
  • the Fc domain may have at least one amino acid residue change as compared to wildtype, wherein the Fc domain is at comprises at least 80% homologous to SEQ ID NO: 898, and wherein the at least one amino acid residue change is: (a) N296A wherein numbering of amino acid residues in the Fc domain is relative to SEQ ID NO: 898; or (b) N296G N296A wherein numbering of amino acid residues in the Fc domain is relative to SEQ ID NO: 898; or (c) K322A/L234A/L235A N296A wherein numbering of amino acid residues in the Fc domain is relative to SEQ ID NO: 898; or (d) L234F/L235E/P331S N296A wherein numbering of amino acid residues in the Fc domain is relative to SEQ ID NO:
  • the IQ for binding of the antigen binding domain to the first antigen in the presence of the immune- stimulatory compound is less than about 100 nM and is equal to, or up to no greater than about 10 times the IQ of the binding of the antigen binding domain to the first antigen in the absence of the immune- stimulatory compound; and the IQ for binding of the Fc domain to the Fc receptor in the presence of the immune- stimulatory compound is equal to, or up to no greater than about 10 times the IQ for the binding of the Fc domain to the Fc receptor in the absence of the immune- stimulatory compound.
  • the molar ratio of immune- stimulatory compound to antibody in a conjugate is less than 5. In certain embodiments, the average molar ratio of immune- stimulatory compound to antibody construct in a composition of conjugates is less than 5. In certain embodiments, the average molar ratio of immune- stimulatory compound to antibody construct in a composition of conjugates from 1 to 3, 3 to 5, or about 2.
  • the linker may be bound to the antibody construct at an amino acid residue that does not interfere with the Fc domain binding to the Fc receptor.
  • the linker may not be attached to an amino acid residue of an IgG Fc domain selected from a group consisting of: 221, 224, 227, 228, 230, 231, 223, 233, 234, 235, 236, 237, 238, 239, 240, 241, 243, 244, 245, 247, 249, 250, 258, 262, 263, 264, 265, 266, 267, 268, 269, 270, 271, 272, 273, 275, 276, 278, 280, 281, 283, 285, 286, 291, 292, 293, 294, 295, 296, 297, 298, 299, 300, 302, 305, 313, 318, 323, 324, 325, 327, 328, 329, 330, 331, 332, 333, 335, 336, 396, or 428, wherein numbering of amino acid residues in the Fc domain is
  • the linker is bound to the antibody construct at an amino acid of an IgG Fc domain selected from a group consisting of: 221, 224, 227, 228, 230, 231, 223, 233, 234, 235, 236, 237, 238, 239, 240, 241, 243, 244, 245, 247, 249, 250, 258, 262, 263, 264, 265, 266, 267, 268, 269, 270, 271, 272, 273, 275, 276, 278, 280, 281, 283, 285, 286, 291, 292, 293, 294, 295, 296, 297, 298, 299, 300, 302, 305, 313, 318, 323, 324, 325, 327, 328, 329, 330, 331, 332, 333, 335, 336, 396, or 428, wherein numbering of amino acid residues in the Fc domain is according to the EU index as in Kabat.
  • the Fc domain is an IgG Fc domain selected from a group consisting of a human IgGl Fc domain, a human IgG2 Fc domain, a human IgG3 Fc domain, and a human IgG4 Fc domain.
  • the immune- stimulatory conjugate induces the secretion of cytokines by an antigen presenting cell.
  • the disclosure provides a pharmaceutical composition comprising immune- stimulatory conjugates as described herein and a pharmaceutically acceptable excipient.
  • the disclosure provides a method of treating cancer, comprising administering to a subject in need thereof an immune- stimulatory conjugate described herein.
  • the disclosure provides a method of treating cancer, comprising administering to a subject in need thereof a pharmaceutical composition comprising immune- stimulatory conjugates as described herein.
  • FIGURE 1 illustrates a schematic of a conjugate comprising an antibody and a second binding domain.
  • the antibody contains two heavy chains as shown in gray and two light chains as shown in light gray. A portion of the heavy chains contain Fc domains (705 and 720).
  • the antibody comprises a binding domain comprising two antigen binding sites (710 and 715).
  • the second binding domain is attached to the antibody (780 and 785), for example, at the C-terminus of the heavy chains.
  • FIGURE 2 illustrates a schematic of an exemplary conjugate.
  • the conjugate comprises an antibody, which contains two heavy chains as shown in gray and two light chains as shown in light gray.
  • the antibody comprises a binding domain comprising two antigen binding sites (910 and 915), and a portion of the heavy chains contain Fc domains (905 and 920).
  • the immune- stimulatory compounds (930 and 940) are attached to the antibody by linkers (960 and 970).
  • a second binding domain is attached to the antibody (980 and 985), for example, at the C-terminus of the heavy chains.
  • FIGURE 3 illustrates a schematic of an exemplary conjugate.
  • the conjugate comprises the Fc region of an antibody with the heavy chains shown in gray, and two scaffolds as shown in light gray.
  • the conjugate comprises a first binding domain comprising two antigen binding sites (1110 and 1115) in the scaffolds, and a portion of the heavy chains contain Fc domains (1105 and 1120).
  • the immune- stimulatory compounds (1130 and 1140) are conjugated to the Fc domains by linkers (1160 and 1170).
  • a second binding domain is attached to the conjugate (1180 and 1185), for example, at the C-terminus of the heavy chains.
  • FIGURE 4 illustrates a schematic of an exemplary conjugate.
  • the conjugate comprises the F(ab') 2 region of an antibody with the Fab portions of heavy chains shown in gray and light chains shown in light gray, and two scaffolds as shown in dark gray.
  • the conjugate comprises a first binding domain comprising two antigen binding sites (1310 and 1315), and a portion of two scaffolds contain Fc domains (1340 and 1345).
  • the immune- stimulatory compounds (1330 and 1340) are attached to the conjugate by linkers (1360 and 1370).
  • a second binding domain is attached to the Fc domains (1380 and 1385).
  • FIGURE 5 illustrates a schematic of an exemplary conjugate.
  • the conjugate contains two scaffolds as shown in light gray and two scaffolds as shown in dark gray.
  • the conjugate comprises a first binding domain comprising two antigen binding sites (1510 and 1515), and a portion of the two dark gray scaffolds contain Fc domains (1540 and 1545).
  • the immune- stimulatory compounds (1530 and 1540) are conjugated to the conjugate by linkers (1560 and 1570).
  • a second binding domain is attached to the conjugate (1580 and 1585).
  • FIGURE 6 illustrates a schematic of a conjugate comprising an antibody and a second binding domain.
  • the antibody contains two heavy chains as shown in gray and two light chains as shown in light gray. A portion of the heavy chains contain Fc domains (1705 and 1720).
  • the antibody comprises a binding domain comprising two antigen binding sites (1710 and 1715).
  • the second binding domain is attached to the antibody (1780 and 1785), for example, at the C- terminus of the light chains.
  • FIGURE 7 illustrates a schematic of an exemplary conjugate.
  • the conjugate comprises an antibody, which contains two heavy chains as shown in gray and two light chains as shown in light gray.
  • the antibody comprises a binding domain comprising two antigen binding sites (1910 and 1915), and a portion of the heavy chains contain Fc domains (1905 and 1920).
  • the immune- stimulatory compounds (1930 and 1940) are conjugated to the antibody by linkers (1960 and 1970).
  • a second binding domain is attached to the antibody (1980 and 1985), for example, at the C-terminus of the light chains.
  • FIGURE 8 illustrates a schematic of an exemplary conjugate.
  • the conjugate comprises an Fc region of an antibody shown in gray, and two scaffolds as shown in light gray.
  • the conjugate comprises a first binding domain comprising two antigen binding sites (2110 and 2115) in the scaffolds, and a portion containing Fc domains (2105 and 2120).
  • the immune- stimulatory compounds (2130 and 2140) are conjugated to the antibody construct by linkers (2160 and 2170).
  • a second binding domain is attached to the antibody (2180 and 2185).
  • FIGURE 9 illustrates a schematic of an exemplary conjugate.
  • the conjugate comprises the F(ab')2 region of an antibody with heavy chains shown in gray and light chains shown in light gray, and two scaffolds as shown in dark gray.
  • the conjugate comprises a first binding domain comprising two antigen binding sites (2310 and 2315), and a portion of two scaffolds contain Fc domains (2340 and 2345).
  • the immune- stimulatory compounds (2330 and 2340) are conjugated to the antibody by linkers (2360 and 2370).
  • a second binding domain is attached to the antibody (2380 and 2385), for example, at the C-terminus of the light chains.
  • FIGURE 10 illustrates a schematic of an exemplary conjugate.
  • the conjugate contains two scaffolds as shown in light gray and two scaffolds as shown in dark gray.
  • the conjugate comprises a first binding domain comprising two antigen binding sites (2510 and 2515), and a portion of the two dark gray scaffolds contain Fc domains (2540 and 2545).
  • the immune- stimulatory compounds (2530 and 2540) are conjugated to the antibody construct by linkers (2560 and 2570).
  • a second binding domain is attached to the conjugate (2580 and 2585).
  • FIGURE 11 illustrates a schematic of an antibody construct comprising an antibody.
  • the antibody contains two heavy chains and two light chains. A portion of the heavy chains contain Fc domains (2705 and 2720). The antibody comprises a binding domain comprising two antigen binding sites shown in black (2710 and 2715).
  • FIGURE 12 illustrates a schematic of an antibody construct comprising an antibody.
  • the antibody contains two heavy chains and two light chains. A portion of the heavy chains contain Fc domains (2925 and 2930).
  • the antibody comprises a first binding domain comprising two antigen binding sites shown in black (2910 and 2915).
  • the antibody comprises a second binding domain comprising two single chain variable fragments (2905 and 2920) attached to a C- terminus of the light chains.
  • a single chain variable fragment can be attached to a light chain chain at a heavy chain variable domain of the single chain variable fragment.
  • a single chain variable fragment can be attached to a light chain at a light chain variable domain of the single chain variable fragment.
  • FIGURE 13 illustrates a schematic of an antibody construct comprising an antibody.
  • the antibody contains two heavy chains and two light chains. A portion of the heavy chains contain Fc domains (3120 and 3125).
  • the antibody comprises a first binding domain comprising two antigen binding sites shown in black (3110 and 3115).
  • the antibody comprises a second binding domain comprising two single chain variable fragments (3130 and 3135) attached to a C- terminus of the heavy chains.
  • a single chain variable fragment can be attached to a heavy chain chain at a heavy chain variable domain of the single chain variable fragment.
  • a single chain variable fragment can be attached to a heavy chain at a light chain variable domain of the single chain variable fragment.
  • FIGURE 14 illustrates a schematic of an antibody construct comprising an antibody.
  • the antibody contains two heavy chains and two light chains. A portion of the heavy chains contain Fc domains (3330 and 3335).
  • the antibody comprises a first binding domain comprising two antigen binding sites shown in black (3310 and 3315).
  • the antibody comprises a second binding domain comprising two single chain variable fragments (3320 and 3325) attached to a C- terminus of the light chains.
  • a single chain variable fragment can be attached to a light chain chain at a heavy chain variable domain of the single chain variable fragment.
  • a single chain variable fragment can be attached to a light chain at a light chain variable domain of the single chain variable fragment.
  • the antibody comprises a third binding domain comprising two single chain variable fragments (3340 and 3345) attached to a C-terminus of the heavy chains.
  • a single chain variable fragment can be attached to a heavy chain chain at a heavy chain variable domain of the single chain variable fragment.
  • a single chain variable fragment can be attached to a heavy chain at a light chain variable domain of the single chain variable fragment.
  • FIGURE 15A sets forth a structure of an immune- stimulatory compound (left) and an immune- stimulatory compound with an amine handle (right top).
  • FIGURE 15B shows the x-ray crystal structure and binding orientation (pdb code 5D7A) of the immune- stimulatory compound as described in FIGURE 15A in a TNIK (TRAF2 and NCK- interacting protein kinase) active site.
  • TNIK TNIK
  • Specific interactions between the inhibitor and the TNIK active site are described in Masuda et al., TNIK inhibition abrogates colorectal cancer sternness, (2016) Nat Commun 7: 12586-12586.
  • FIGURE 15C shows a close up of the binding orientation of the immune- stimulatory compound as described in FIGURE 15A in a TNIK active site, where the linker and antibody portions are pointing away and sitting outside of the active site.
  • FIGURE 16 sets forth a structure of an immune- stimulatory compound (left) and an immune- stimulatory compound with an amine handle and linker surrogate (right).
  • the structure on the right illustrates that the immune- stimulatory compound is predicted to sit in the enzyme active site, whereas the amine handle and linker surrogate are predicted to sit outside of the enzyme active site, in the solvent.
  • FIGURE 17A sets forth a structure of an immune- stimulatory compound (left) and an immune- stimulatory compound with a linker surrogate (left side of molecule on right).
  • FIGURE 17B shows the x-ray crystal structure and binding orientation (pdb code 5E91) of the immune- stimulatory compound as described in FIGURE 17 A in a TGFPR2 (transforming growth factor, beta receptor II) active site.
  • TGFPR2 transforming growth factor, beta receptor II
  • Specific interactions between the inhibitor and the TGFpRII active site are described in Tebben et al., Crystal structures of apo and inhibitor-bound TGFbetaR2 kinase domain: insights into TGFbetaR isoform selectivity, (2016) Acta Crystallogr., Sect.D 72: 658-674.
  • FIGURE 17C shows a close up of the binding orientation of the immune- stimulatory compound as described in FIGURE 17 A in a TGFPR2 active site, where the linker and antibody construct portions are pointing away and sitting outside of the active site.
  • FIGURE 18A sets forth a structure of an immune- stimulatory compound (left) and an immune- stimulatory compound with an amine handle (upper left in molecule on right).
  • FIGURE 18B shows the x-ray crystal structure and binding orientation (pdb code 3KR8) of the immune- stimulatory compound as described in FIGURE 18A in a TNKS (tankyrase) active site.
  • FIGURE 18C shows a close up of the binding orientation of the immune- stimulatory compound as described in FIGURE 18A in a TNKS active site, where the linker and antibody construct portions are pointing away and sitting outside of the active site.
  • Specific interactions between the inhibitor and the human tankyrase 2 - catalytic PARP domain active site are described in Karlberg et al., Structural basis for the interaction between tankyrase-2 and a potent Wnt-signaling inhibitor, (2010) J.Med.Chem. 53: 5352-5355.
  • FIGURE 19A shows a structure of an immune- stimulatory compound (left) and an immune- stimulatory compound with two amine handles (on right side of molecule on the right).
  • FIGURE 19B shows the x-ray crystal structure and binding orientation (pdb code 4191) of the immune- stimulatory compound as described in FIGURE 19A with dual site binding in the TNKS (tankyrase) active site.
  • TNKS tumoryrase
  • FIGURE 19C shows a close up of the binding orientation of the immune- stimulatory compound as described in FIGURE 19A in a TNKS active site, where the linker and antibody portions are pointing away and sitting outside of the active site.
  • FIGURE 20A illustrates a schematic of an exemplary conjugate and its molecular target.
  • the conjugate comprises an antibody (3405) attached to a linker (3410) that is attached to a drug (3415) at the opposite end of the antibody (3405).
  • the molecular target (3420) has an active site (3425) that is complementary to the drug (3415).
  • FIGURE 20B illustrates a schematic of an active exemplary conjugate that is bound to the the molecular target's active site.
  • the drug (3415) sits within the active site of the molecular target (3420).
  • the linker (3410) and antibody (3405) sit outside of the active site (3430).
  • FIGURE 20C illustrates a schematic of an active drug (3415) and linker (3410) that is bound to the molecular target (3420).
  • FIGURE 20D illustrates a schematic of an active drug (3415) that is bound to the molecular target (3420).
  • FIGURE 21A, FIGURE 21B, and FIGURE 21C show the results of an assay for degradation of TFGpR2 by a TGFpR2-VHL PROTAC anti-HER2 antibody conjugate.
  • FIGURE 22A and FIGURE 22B show the results of an assay for antigen targeted degradation of TGFPR2 by an antibody conjugate with a PROTAC having VHL or Cereblon E3 binding moieties.
  • FIGURE 23A and FIGURE 23B show the results of an assay for cellular levels of TGFpR2 and TGFpRl in the presence of a TGFpR2/TGFpRl-VHL PROTAC with or without the addition of a proteasome inhibitor.
  • potency generally refers to measured bioactivity and may be quantified as an EC50 or IC50. Potency refers to the amount of a compound or conjugate needed to give an effect. For example, the potency of an immune- stimulatory compound which requires a lower amount of the immune- stimulatory compound compared with a different immune- stimulatory compound can be considered to have greater potency. Furthermore, the different immune- stimulatory compound requires a greater amount of the different immune- stimulatory compound to generate a response, and can therefore be considered lower potency. Potencies of bioactive compounds or conjugates may be measured over a concentration range and can be reported as those molar concentrations required to elicit or inhibit a percentage of the measured bioresponse.
  • a concentration required to stimulate 50% of observed maximal activity in the assay may be reported as an effective concentration 50 (EC50), to stimulate 90% activity as an EC90, or to stimulate 10% activity as an EC 10.
  • a concentration of an antagonist required to give 50% maximal inhibition of a biological activity may be reported as an inhibitory concentration 50 (IC50), to inhibit 90% as an IC90, or to inhibit 10% as an ICIO. This may allow for a comparison of the potencies of bioactive compounds on a molar basis by comparison of their EC or IC values for a given bioassay.
  • an immune- stimulatory conjugate or an immune- stimulatory compound with an EC50 or IC50 that is greater than 300 times the EC50 or IC50 of a control requires 300-fold higher, or more than 300-fold higher, concentration compared to the control to achieve a 50% bioresponse and has a potency weaker than the control by at least 300-fold.
  • an immune- stimulatory compound or conjugate that has an EC50 or IC50 not greater than about 300 times the EC50 or IC50 of a control compound may require no more than a 300-fold higher concentration than the control compound to achieve a 50% maximal bioresponse, no greater than 100 times the EC50 or IC50 requires no more than 100-fold higher concentration and no greater than 10 times the EC50 or IC50 requires no more than 10 times the concentration of the control.
  • the potency of the immune- stimulatory compound or conjugate may be within 300-fold or better, 100-fold or better, or 10-fold or better the potency of the control.
  • control compound refers to, in a case of an immune- stimulatory compound, the compound before attachment to a linker and antibody construct; in the case of an antibody construct, the construct before attachment of the linker and immune- stimulatory compound; or, in the case of a conjugate including an E3 ubiquitin ligase binding moiety, control compound refers to the immune- stimulatory compound attached to a linker that is attached to an E3 ubiquitin ligase binding moiety.
  • an "immune- stimulatory compound” as described herein also referred to herein as an immune- stimulatory agent or molecule, refers to a molecule that stimulates the immune system by inducing activation or increasing activity of any of its components.
  • an immune stimulatory compound may bind directly to a component of the immune system and activate or increase an activity of the immune system or a component thereof.
  • An immune stimulatory compound may also bind to an inhibitory component of the immune system and inactivate or decrease the activity of that component, thereby increasing an activity of the immune system.
  • Immune- stimulatory compounds may have or cause antigenic specificity.
  • an immune- stimulatory compound activates macrophages.
  • an immune- stimulatory compound induces the secretion of cytokines by an antigen presenting cell.
  • the increased activity or activation of the immune system may be evaluated using methods known in the field for evaluating an immune response (see, for example, J. Gratama, Cytometry A. 2008 Nov: 73(11): 971-4, the contents of which are incorporated herein in its entirety).
  • a "protein active site” as described herein refers to a region of a protein target where a molecule (such as a substrate molecule or allosteric effector) binds to the target protein.
  • the molecule may trigger a response upon binding to the the binding protein, e.g., inhibition of the activity of the binding protein or degradation of the binding protein.
  • the protein active site includes the residues that interact with, e.g., bind covalently or non-covalently, a molecule such as an immune- stimulatory compound or a portion thereof.
  • the protein active site may define a groove or pocket in the target protein wherein the interface between the groove or pocket and surrounding solvent is the solvent/active site interface of the protein active site.
  • the phrase "stimulate an immune response” refers to stimulating a response from immune cells that increases the activity or responsiveness of the immune system or a part thereof, such as, for example, inducing secretion of cytokines and/or chemokines, activating immune cells, e.g., T cells, B cells, macrophages, dendritic cells, etc., enhancing antigen presenting cell presentation of antigen, inducing antibody-dependent cell-mediated phagocytosis (ADCP), inducing antibody-dependent cell-mediated cytotoxicity (ADCC), NK cell or CD8+ T cell cytolytic activity, and/or blocking immune suppression.
  • cytokines and/or chemokines activating immune cells, e.g., T cells, B cells, macrophages, dendritic cells, etc.
  • enhancing antigen presenting cell presentation of antigen e.g., inducing antibody-dependent cell-mediated phagocytosis (ADCP), inducing antibody-dependent cell-mediated cytotoxicity
  • homologous or “homology” refer to the similarity or identity between a DNA, RNA, nucleotide, amino acid, or protein sequence to another DNA, RNA, nucleotide, amino acid, or protein sequence, respectively. Homology can be expressed in terms of a percentage of sequence identity of a first sequence to a second sequence. Percent (%) sequence identity with respect to a reference DNA sequence is the percentage of DNA nucleotides in a candidate sequence that are identical with the DNA nucleotides in the reference DNA sequence after aligning the sequences and introducing gaps, as necessary.
  • Percent (%) sequence identity with respect to a reference amino acid sequence is the percentage of amino acid residues in a candidate sequence that are identical with the amino acid residues in the reference amino acid sequence after aligning the sequences and introducing gaps, if necessary, to achieve the maximum percent sequence identity, and not considering any conservative substitutions as part of the sequence identity.
  • X can indicate any amino acid.
  • X can be asparagine (N), glutamine (Q), histidine
  • antibody refers to an immunoglobulin molecule that specifically binds to, or is immunologically reactive toward, a specific antigen.
  • the term antibody can include, for example, polyclonal, monoclonal, genetically engineered, and antigen binding fragments thereof.
  • An antibody can be, for example, murine, chimeric, humanized, bispecific, a heteroconjugate, a diabody, a triabody, a tetrabody or a hcAb (heavy chain antibody).
  • An antigen binding fragment can include, for example, a Fab', F(ab') 2 , Fab, Fv, rlgG, scFv, a single domain antibody, V HH , V NAR , sdAb, or nanobody.
  • the term "antigen" refers to a molecule that can be bound by an antibody or a antibody construct.
  • An antigen may elicit an immune response.
  • An antigen can be a protein, polysaccharide, lipid, glycolipid or the like, which can be recognized by an antibody or an immune cell, such as a T cell or a B cell.
  • Exposure of immune cells to one or more of these antigens can elicit a rapid cell division and differentiation response resulting in the formation of clones of the exposed T cells and B cells.
  • B cells can differentiate into plasma cells which in turn can produce antibodies which selectively bind to the antigens.
  • the terms “recognize,” “bind,” and “specifically bind” refer to the specific association or specific binding between an antigen binding domain and a corresponding antigen, or an immune stimulatory compound and a protein, as compared to the non-specific association or non-specific binding of the antigen binding domain or immune- stimulatory compound with a non-target antigen or protein.
  • tumor antigen is an antigen that can be expressed by or is present on, a cancer cell, a neoplastic tumor cell and/or within a tumor microenvironment.
  • an "antibody construct” refers to a construct that contains an antigen binding domain and an Fc domain.
  • an "antigen binding domain” refers to an antigen binding domain from an antibody or from a non-antibody that can specifically bind to the antigen.
  • Antigen binding domains can be numbered when there is more than one antigen binding domain in a given conjugate or construct (e.g., first binding domain, second antigen binding domain, third antigen binding domain, etc.).
  • Different antigen binding domains in the same conjugate or construct can target the same antigen or different antigens (e.g., first binding domain that can bind a tumor antigen, second antigen binding domain that can bind to a tumor antigen or an antigen presenting cell (APC) antigen, and third antigen binding domain that can bind an APC antigen).
  • APC antigen presenting cell
  • a "Fc domain” is an Fc domain from an antibody or from a non-antibody that can bind to an Fc receptor.
  • an "Fc null” refers to an Fc domain that exhibits weak to no binding to any of the Fcgamma receptors.
  • an Fc null domain or region exhibits a reduction in binding affinity (e.g., increase in Kd) to Fc gamma receptors of at least 1000-fold.
  • a "target binding domain” refers to a construct that contains an antigen binding domain from an antibody or from a non-antibody that can bind to the antigen.
  • salt or “pharmaceutically acceptable salt” refers to salts derived from a variety of organic and inorganic counter ions well known in the art.
  • Pharmaceutically acceptable acid addition salts can be formed with inorganic acids and organic acids.
  • Inorganic acids from which salts can be derived include, for example, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like.
  • Organic acids from which salts can be derived include, for example, acetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, /?-toluenesulfonic acid, salicylic acid, and the like.
  • Pharmaceutically acceptable base addition salts can be formed with inorganic and organic bases.
  • Inorganic bases from which salts can be derived include, for example, sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, aluminum, and the like.
  • Organic bases from which salts can be derived include, for example, primary, secondary, and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines, basic ion exchange resins, and the like, specifically such as
  • the pharmaceutically acceptable base addition salt is chosen from ammonium, potassium, sodium, calcium, and magnesium salts.
  • Agonism is the binding of a chemical to a receptor to induce a biological response.
  • a chemical can be, for example, a small molecule, a compound, or a protein.
  • An agonist causes a response
  • an antagonist can block the action of an agonist
  • an inverse agonist can cause a response that is opposite to that of the agonist.
  • a receptor can be activated by either endogenous or exogenous agonists.
  • C x-y when used in conjunction with a chemical moiety, such as alkyl, alkenyl, or alkynyl is meant to include groups that contain from x to y carbons in the chain.
  • C x - y alkyl refers to substituted or unsubstituted saturated hydrocarbon groups, including straight-chain alkyl and branched-chain alkyl groups that contain from x to y carbons in the chain, including haloalkyl groups such as trifluoromethyl and 2,2,2-trifluoroethyl, etc.
  • C x - y alkenyl and “C x - y alkynyl” refer to substituted or unsubstituted
  • unsaturated aliphatic groups analogous in length and possible substitution to the alkyls described above, but that contain at least one double or triple bond, respectively.
  • Carbocycle refers to a saturated, unsaturated or aromatic ring in which each atom of the ring is carbon.
  • Carbocycle includes 3- to 10-membered monocyclic rings, 6- to 12-membered bicyclic rings, and 6- to 12-membered bridged rings.
  • Each ring of a bicyclic carbocycle may be selected from saturated, unsaturated, and aromatic rings.
  • an aromatic ring e.g., phenyl
  • Any combination of saturated, unsaturated and aromatic bicyclic rings, as valence permits, is included in the definition of carbocyclic.
  • Exemplary carbocycles include cyclopentyl, cyclohexyl, cyclohexenyl, adamantyl, phenyl, indanyl, and naphthyl.
  • heterocycle refers to a saturated, unsaturated or aromatic ring comprising one or more heteroatoms.
  • Exemplary heteroatoms include N, O, Si, P, B, and S atoms.
  • Heterocycles include 3- to 10-membered monocyclic rings, 6- to 12-membered bicyclic rings, and 6- to 12-membered bridged rings.
  • Each ring of a bicyclic heterocycle may be selected from saturated, unsaturated, and aromatic rings wherein at least one of the rings includes a heteroatom.
  • an aromatic ring e.g., pyridyl
  • heteroaryl includes aromatic single ring structures, preferably 5- to 7- membered rings, more preferably 5- to 6-membered rings, whose ring structures include at least one heteroatom, preferably one to four heteroatoms, more preferably one or two heteroatoms.
  • heteroaryl also include polycyclic ring systems having two or more cyclic rings in which two or more carbons are common to two adjoining rings wherein at least one of the rings is heteroaromatic, e.g., the other cyclic rings can be aromatic or non-aromatic carbocyclic, or heterocyclic.
  • Heteroaryl groups include, for example, pyrrole, furan, thiophene, imidazole, oxazole, thiazole, pyrazole, pyridine, pyrazine, pyridazine, and pyrimidine, and the like.
  • substituted refers to moieties having substituents replacing a hydrogen on one or more carbons or substitutable heteroatoms, e.g., NH, of the structure. It will be understood that “substitution” or “substituted with” includes the implicit proviso that such substitution is in accordance with permitted valence of the substituted atom and the substituent, and that the substitution results in a stable compound, i.e., a compound which does not spontaneously undergo transformation such as by rearrangement, cyclization, elimination, etc.
  • substituted refers to moieties having substituents replacing two hydrogen atoms on the same carbon atom, such as substituting the two hydrogen atoms on a single carbon with an oxo, imino or thioxo group.
  • substituted is contemplated to include all permissible substituents of organic compounds.
  • the permissible substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and non-aromatic substituents of organic compounds.
  • the permissible substituents can be one or more and the same or different for appropriate organic compounds.
  • the heteroatoms such as nitrogen may have hydrogen substituents and/or any permissible substituents of organic compounds described herein which satisfy the valences of the heteroatoms.
  • Chemical entities having carbon-carbon double bonds or carbon-nitrogen double bonds may exist in Z- or E- form (or cis- or trans- form). Furthermore, some chemical entities may exist in various tautomeric forms. Unless otherwise specified, chemical entities described herein are intended to include all Z-, E- and tautomeric forms as well.
  • a "tautomer” refers to a molecule wherein a proton shift from one atom of a molecule to another atom of the same molecule is possible.
  • the compounds, constructs and conjugates disclosed herein, in some embodiments, are used in different enriched isotopic forms, e.g., enriched in the content of 2 H, 3 H, 11 C, 13 C and/or 14 C.
  • the compound is deuterated in at least one position.
  • deuterated forms can be made by the procedure described in U.S. Patent Nos. 5,846,514 and 6,334,997, or as otherwise known in the art.
  • deuteration can improve the metabolic stability and or efficacy, thus increasing the duration of action of drugs.
  • structures depicted herein are intended to include compounds which differ only in the presence of one or more isotopically enriched atoms.
  • compounds having the present structures except for the replacement of a hydrogen by a deuterium or tritium, or the replacement of a carbon by 13 C- or 14 C-enriched carbon are within the scope of the present disclosure.
  • the compounds of the present disclosure optionally contain unnatural proportions of atomic isotopes at one or more atoms that constitute such compounds.
  • the compounds may be labeled with isotopes, such as for example, deuterium ( 2 H), tritium ( 3 H), iodine- 125 ( 125 I) or carbon- 14 ( 14 C).
  • isotopes such as for example, deuterium ( 2 H), tritium ( 3 H), iodine- 125 ( 125 I) or carbon- 14 ( 14 C).
  • Isotopic substitution with 2 H, U C, 13 C, 14 C, 15 C, 12 N, 13 N, 15 N, 16 N, 16 0, 17 0, 14 F, 15 F, 16 F, 17 F, 18 F, 33 S, 34 S, 35 S, 36 S, 35 C1, 37 C1, 79 Br, 81 Br, and 125 I are all contemplated. All isotopic variations of the compounds of the present invention, whether radioactive or not, are encompassed within the scope of the
  • the compounds disclosed herein have some or all of the 1H atoms replaced with H atoms.
  • the methods of synthesis for deuterium-containing compounds are known in the art and include, by way of non-limiting example only, the following synthetic methods.
  • Deuterated starting materials are readily available and are subjected to the synthetic methods described herein or otherwise known to provide for the synthesis of deuterium- containing compounds. Large numbers of deuterium-containing reagents and building blocks are available commercially from chemical vendors, such as Aldrich Chemical Co.
  • Compounds also include crystalline and amorphous forms of those compounds, pharmaceutically acceptable salts, and active metabolites of these compounds having the same type of activity, including, for example, polymorphs, pseudopolymorphs, solvates, hydrates, unsolvated polymorphs (including anhydrates), conformational polymorphs, and amorphous forms of the compounds, as well as mixtures thereof.
  • parenteral administration and “administered parenterally” as used herein means modes of administration other than enteral and topical administration, usually by injection, and includes, without limitation, intravenous, intramuscular, intraarterial, intrathecal,
  • intracapsular intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intraarticular, subcapsular, subarachnoid, intraspinal and intrasternal injection and infusion.
  • phrases "pharmaceutically acceptable excipient” or “pharmaceutically acceptable carrier” as used herein means a pharmaceutically acceptable material, composition or vehicle, such as a liquid or solid filler, diluent, excipient, solvent or encapsulating material. Each carrier must be “acceptable” in the sense of being compatible with the other ingredients of the formulation and not injurious to the patient.
  • materials which can serve as pharmaceutically acceptable carriers include: (1) sugars, such as lactose, glucose and sucrose; (2) starches, such as corn starch and potato starch; (3) cellulose, and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; (4) powdered tragacanth; (5) malt; (6) gelatin; (7) talc; (8) excipients, such as cocoa butter and suppository waxes; (9) oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; (10) glycols, such as propylene glycol; (11) polyols, such as glycerin, sorbitol, mannitol and polyethylene glycol; (12) esters, such as ethyl oleate and ethyl laurate; (13) agar; (14) buffering agents, such as magnesium hydroxide and aluminum hydroxide;
  • attached refers to a covalent bond, between two or more groups.
  • attached may refer to the connection of two or more groups via a linker, e.g., a linker connecting a second binding domain to an antibody construct.
  • a fusion may refer to a nucleic acid sequence of two separate domains being expressed in frame.
  • a binding domain can be attached as a fusion or by attachment (e.g., conjugation) via a linker to an antibody construct or other portion of a conjugate.
  • an antibody can be fused with an additional binding domain to create an antibody construct containing a fusion of the antibody and the additional binding domain.
  • the antibody construct can be the result of the nucleic acid sequence of the binding domain being expressed in frame with the nucleic acid sequence of the antibody construct.
  • the fusion can be the result of an in- frame nucleotide sequence encoding the antibody construct with the binding domain.
  • an additional binding domain can be attached to an antibody construct via a linker, wherein the linker is attached (i.e., conjugated) to the binding domain and the linker is attached to the antibody construct.
  • the binding domain can be linked to the linker by a chemical conjugation and the antibody construct can be linked to the linker by a chemical conjugation.
  • the additional binding domain can be a second binding domain and/or a third binding domain as described herein.
  • a binding domain can be a first binding domain attached to an Fc domain to produce the antibody construct as described herein, which may produce the first binding domain as a fusion with the Fc domain or a conjugate wherein the first binding domain can be linked to a linker and the linker can be linked to the Fc domain.
  • conjugates of antibody constructs, immune- stimulatory compounds and linkers referred to as antibody construct- immune- stimulatory compound conjugates (also referred to as immune- stimulatory compound-conjugates, antibody conjugates or conjugates).
  • the conjugates may be used in the treatment of various diseases and disorders, including cancers.
  • immune- stimulatory compounds are attached either directly or through a linker to an antibody construct to form antibody construct-immune- stimulatory compound conjugates.
  • the immune- stimulatory compound stimulates the immune system, or a component thereof. In some embodiments, the immune- stimulatory compound has an immune-modulatory activity. In some embodiments, the immune- stimulatory compound has an inhibitory effect on a component of the immune system, thereby stimulating an immune- modulatory activity.
  • conjugates are represented by the following formula:
  • A is an antibody construct
  • L is a linker
  • D is an immune- stimulatory compound
  • x may be from 1 to 20 (wherein each x denotes a separate compound)
  • n may be from 1 to 20
  • z may be from 1 to 20.
  • L is a cleavable linker.
  • L is a non-cleavable linker.
  • x is 1, n is 1 and z may be from 1 to 10, such as from 1 to 9, such as from 1 to 8, such as from 2 to 8, such as from 1 to 6, such as from 3 to 5 or such as about 2.
  • x is 1, n is 2, and z may be from 1 to 10, such as from 1 to 9, such as from 1 to 8, such as from 2 to 8, such as from 1 to 6 or such as from 3 to 5. In certain embodiments, z is 4.
  • x may be from 1-20
  • n may be from 1-20
  • z may be from 1 to
  • conjugates are represented by the following formula:
  • A is an antibody construct
  • L is a linker having the structure -A a -W w -Y y -, where A is a spacer, a is 0 or 1, W is a cleavable unit, w may be from 0 to 10, Y is a stretcher, y may be from 0 to 3, D is an immunomodulatory compound, x may be from 1 to 20 (wherein each x denotes a distinct compound), n may be from 1 to 20, and z may be from 1 to 20. [0120] In some embodiments, a is 1, w is 0, y is 0, x is 1, n is 1, and z may be from 1 to 20, 1 to 10, 1 to 9, 1 to 8, 2 to 8, 1 to 6 or 3 to 5. In certain embodiments, z is 4.
  • a isl, w is 1, y is from 1, x is 1, n is 1 and z may be from 1 to 10, 1 to 9, 1 to 8, 2 to 8, 1 to 6, 3 to 5, 2 or 4.
  • a is 0 or 1
  • w is from 0 to 10
  • y is from 0-3, where at least one of a, w or y is present
  • x may be from 1 to 20
  • n may be from 1-20
  • z may be from 1 to 20.
  • the immune- stimulatory conjugates include a linker (L) that may comprise from 5 to 100 linear non-hydrogen atoms that is covalently attached to an antibody construct (A) and may be:
  • an immune- stimulatory compound CO which itself may be covalently attached to a spacer (S) comprising from 5 to 100 linear no n- hydrogen atoms covalently attached to a second immune- stimulatory compound (C 2 ) as in the following formula:
  • an immune- stimulatory compound C 2
  • a spacer S
  • C 2 an immune- stimulatory compound that may be covalently attached to a spacer (S) comprising from 5 to 100 linear non-hydrogen atoms covalently attached to a formula: , or
  • a spacer comprising from 5 to 100 linear non- hydrogen atoms that is covalently attached immune- stimulatory compounds (Ci and C 2 ) as in the following formula:
  • an immune- stimulatory conjugate includes a proteolysis targeting module (PTM; also referred to as a proteolysis-targetting chimera or PROTAC) that includes an immune-modulatory compound (IMC) that is covalently attached to an E3 ubiquitin ligase binding moiety (ULM) through a spacer (S) and wherein a linker (L) is covalently attached to the protein targeting molecule and to the antibody construct as
  • PTM proteolysis targeting module
  • PROTAC proteolysis-targetting chimera
  • L is a cleavable linker.
  • S is a non-cleavable spacer.
  • Ci is a target protein targeting moiety, such as an immodulatory compound
  • C2 is an E3 ubiquitin ligase binding moiety such that together C 1 -S-C 2 may form a PTM.
  • an immune-modulatory compound or other protein targeting moiety is covalently attached to an E3 ubiquitin ligase binding moiety (ULM) through a spacer (S) and a linker (L) is covalently attached to the spacer, n is from 1-20 and z is from 1 to 20 as represented by the for
  • L is a cleavable linker. In some embodiments, L is a non- cleavable linker. In some embodiments, S is non-cleavable.
  • an immune-modulatory compound or other protein targeting moiety is covalently attached to an E3 ubiquitin ligase binding moiety (ULM) through a spacer (S) and a linker (L) is covalently attached to the IMC, n is from 1-20 and z is from 1 to 20 as represented by the formula:
  • L is a cleavable linker. In some embodiments, L is a non- cleavable linker.
  • an immune-modulatory compound or other protein targeting moiety is covalently attached to an E3 ubiquitin ligase binding moiety (ULM) through a spacer (S) and linker L is covalently attached to the ubiquitin E3 ligase moiety (ULM), n is from 1-20 and z is from 1 to 20 as represented by the formula:
  • L is a cleavable linker. In some embodiments, L is a non- cleavable linker. In some embodiments, S is non-cleavable.
  • tumor antigens there are several general groups of tumor antigens, including but not limited to: (i) viral tumor antigens which can be identical for any viral tumor of this type, (ii) carcinogenic tumor antigens which can be specific for patients and for the tumors, (iii) isoantigens of the transplantation type or tumor-specific transplantation antigens which can be different in all individual types of tumor but can be the same in different tumors caused by the same inciting biological event; and (iv) embryonic antigens.
  • viral tumor antigens which can be identical for any viral tumor of this type
  • carcinogenic tumor antigens which can be specific for patients and for the tumors
  • isoantigens of the transplantation type or tumor-specific transplantation antigens which can be different in all individual types of tumor but can be the same in different tumors caused by the same inciting biological event
  • embryonic antigens embryonic antigens.
  • tumor antigens include the following: CD5, CD19, CD20, CD25, CD37, CD30, CD33, CD45, CAMPATH-1, BCMA, CS-1, PD-L1, B7-H3, B7-DC (PD-L2), HLD-DR, carcinoembryonic antigen (CEA), TAG-72, EpCAM, MUC1, MUC15, MUC16, fo late-binding protein, A33, G250, prostate- specific membrane antigen (PSMA), ferritin, GD2, GD3, GM2, Le y , CA-125, CA19-9, epidermal growth factor, pl85HER2, IL-2 receptor, EGFRvIII (de2-7 EGFR), fibroblast activation protein (FAP), tenascin, a metalloproteina
  • antibodies that target immune cells to boost the immune response have also been developed, including the following: an anti-CD40 antibody, an anti-CD47 antibody, anti-TNFR2 antibody, an anti-DEC205 antibody, an anti-CD36 mannose scavenger receptor 1 antibody, an anti- CLEC9A antibody, an anti-CLEC12A antibody, an anti-DC-SIGN antibody, an anti-BDCA-2 antibody, an anti-OX40L antibody, an anti-41BBL antibody, an anti-CD204 antibody, an anti- MARCO antibody, an anti-CLEC5A antibody, an anti-Dectin 1 antibody, and anti-Dectin 2 antibody, an anti-CLEClOA antibody, an anti-CD206 antibody, an anti-CD64 antibody, an anti- CD32A antibody, an anti-CD 16A antibody, an anti-HVEM antibody, an anti-CD38 antibody, an anti-PD-Ll antibody, an anti-TREM2 antibody, an anti-
  • CD40 Cluster of Differentiation 40
  • TNF-R Tumor Necrosis Factor Receptor
  • CD40 can be a 50 kDa cell surface glycoprotein that can be constitutively expressed in normal cells, such as monocytes, macrophages, B lymphocytes, dendritic cells, endothelial cells, smooth muscle cells, fibroblasts and epithelium, and in tumor cells, including B-cell lymphomas and many types of solid tumors.
  • Expression of CD40 can be increased in antigen presenting cells in response to IL- ⁇ , IFN- ⁇ , GM-CSF, and LPS induced signaling events.
  • agonistic CD40 monoclonal antibodies examples include CP-870,893, dacetuzumab, Chi Lob 7/4, SEA-CD40, ADC- 1013, 3C3, or 3G5.
  • Cluster of Differentiation 205 is a member of the C-type multilectin family of endocytic receptors, which can include the macrophage mannose receptor (MMR) and the phospholipase A2 receptor (PLA 2 R).
  • DEC-205 can be a 205 kDa endocytic receptor highly expressed in cortical thymic epithelial cells, thymic medullary dendritic cells (CDl lc + CD8 + ), subpopulations of peripheral dendritic cells (CDl lc + CD8 + ).
  • the DEC-205 + CDl lc + CD8 + dendritic cells can function in cross-presentation of antigens derived from apoptotic cells. Additionally, DEC-205 can be significantly upregulated during DC maturation. DEC-205 can also be expressed at moderate levels in B cells and low levels in macrophages and T cells.
  • the receptor- antigen complex can be internalized whereupon the antigen can be processed and be presented on the DC surface by a major histocompatibility complex class II (MHC II) or MHC class I.
  • MHC II major histocompatibility complex class II
  • DEC-205 can deliver antigen to DCs for antigen presentation on MHC class II and cross-presentation on MHC class I.
  • DEC-205 mediated antigen delivery for antigen presentation in DCs without an inflammatory stimulus can result in tolerance.
  • DEC-205 mediated antigen delivery in DCs in the presence of a maturational stimulus e.g., a CD40 agonist
  • a maturational stimulus e.g., a CD40 agonist
  • CD36 mannose scavenger receptor 1 is an oxidized LDL receptor with two
  • transmembrane domains located in the caveolae of the plasma membrane can be classified as a Class B scavenger receptor, which can be characterized by involvement in the removal of foreign substances and waste materials. This receptor can also be involved in cell adhesion, phagocytosis of apoptotic cells, and metabolism of long-chain fatty acids.
  • TNFR2 tumor necrosis factor receptor 2
  • TNFRSF1B tumor necrosis factor receptor super family IB
  • CD 120b CD 120b
  • soluble TNFR2 can be generated via two distinct mechanisms: (1) shedding via proteolytic processing of the full membrane anchored from, and (2) translation from an alternatively spliced message encoding the extracellular domains of TNFR2.
  • TNFR2 is the receptor with high affinity for TNF-alpha and approximately 5-fold lower affinity for homotrimeric lymphotoxin-alpha.
  • TNFR2 Tumor Necrosis Factor Receptor Type II
  • TNF-receptor 1 Tumor Necrosis Factor Receptor Type II
  • c-IAPl can potentiate TNF- induced apoptosis by the ubiquitination and degradation of TNF-receptor-associated factor 2, which mediates anti- apoptotic signals.
  • Knockout studies in mice suggest a role of TNFR2 in protecting neurons from apoptosis by stimulating antioxidative pathways.
  • CLEC9A is a group V C-type lectin receptor. This receptor can be expressed as on myeloid lineage cells, and can be characterized as an activation receptor.
  • CLEC12A is a member of the C-type lectin/C-type lectin like domain super family that can be a negative regulator of granulocyte and monocyte function. It can also be involved in cell adhesion, cell-cell signaling, and glycoprotein turnover, and can play a role in the inflammatory response.
  • DC- SIGN Dendritic cell-specific inter cellular adhesion molecule-3-grabbing non-integrin
  • CD209 is a C-type lectin receptor that can be expressed on the surface of macrophages and dendritic cells. This receptor can recognize and bind to mannose type carbohydrates and be involved in activating phagocytosis, can mediate dendritic cell rolling, and can be involved in CD4+ T cell activation.
  • BDCA-2 is a C-type lectin that is a membrane protein of plasmacytoid dendritic cells. It can be involved in plasmacytoid dendritic cell function, such as ligand internalization and presentation.
  • OX40L which is also referred to as CD252, is the ligand for CD 134 that can be expressed on dendritic cells. It can be involved in T cell activation.
  • 41BBL which is also referred to as CD137L, is a member of the TNF superfamily, and can be expressed on B cells, dendritic cells, activated T cells, and macrophages. It can provide co-stimulatory signal for T cell activation and expansion.
  • CD204 which is also referred to as macrophage scavenger receptor 1
  • the gene for CD204 can encode three different class A macrophage scavenger receptor isoforms.
  • the type 1 and type 2 isoforms can be involved in binding, internalizing, and processing negatively charged macromolecules, such as low density lipoproteins.
  • the type 3 isoform can undergo altered intracellular processing in which it can be retained within the endoplasmic reticulum, and has been shown to have a dominant negative effect on the type 1 and type 2 isoforms.
  • Macrophage receptor with collagenous structure which is also referred to as SCARA2
  • SCARA2 is a class A scavenger receptor with collagen-like and cysteine-rich domains. It can be expressed in macrophages, and can bind to modified low density lipoproteins. It can be involved in the removal of foreign substances and waste materials.
  • C-type lectin domain family 5 member A (CLEC5A) is a C-type lectin. It can be involved in the myeloid lineage activating pathway.
  • Dendritic cell-associated c-type lectin- 1 (Dectin 1), which is also referred to as CLEC7A, is member of the C-type lectin/C-type lectin- like super family. It can be expressed by myeloid dendritic cells, monocytes, macrophages, and B cells, and can be involved in antifungal immunity.
  • Dendritic cell-associated c-type lectin-2 (Dectin 2), which is also referred to as CLEC6A, is member of the C-type lectin/C-type lectin- like super family. It can be expressed by dendritic cells, macrophages, monocytes and neutrophils. It can be involved in antifungal immunity.
  • CLECIOA which is also referred to as CD301, is member of the C-type lectin/C-type lectin-like super family. It can be expressed by dendritic cells, monocytes, and CD33+ myeloid cells, and can be involved in macrophage adhesion and migration.
  • CD206 which is also referred to as macrophage mannose receptor, is a C-type lectin type I membrane glycoprotein. It can be expressed on dendritic cells, macrophages and endothelial cells, and can act as a pattern recognition receptor and bind high-mannose structures of viruses, bacteria, and fungi.
  • CD64 which is also referred to as FcyRI, is a high affinity Fc receptor for IgG. It can be expressed by monocytes and macrophages. It can be involved in mediating phagocytosis, antigen capture, and antibody dependent cell-mediated cytoxicity.
  • CD32A which is also referred to as FcyRIIa, is a low affinity Fc receptor. It can be expressed on monocytes, granulocytes, B cells, and eosinophils. It can be involved in
  • CD 16 A which is also referred to as FcyRIIIa, is low affinity Fc receptor. It can be expressed on NK cells, and can be involved in phagocytosis and antibody dependent cell- mediated cytotoxicity.
  • HVEM Herpesvirus entry mediator
  • CD270 Herpesvirus entry mediator
  • CD32B which is also referred to as FcyRIIb, is a low affinity Fc receptor. It can be expressed on B cells and myeloid dendritic cells. It can be involved in inhibiting maturation and cell activation of dendritic cells.
  • the HER2/neu human epidermal growth factor receptor 2/receptor tyro sine-protein kinase erbB-2
  • the HER2/neu protein can function as a receptor tyrosine kinase and autophosphorylates upon dimerization with binding partners.
  • HER2/neu can activate several signaling pathways including, for example, mitogen-activated protein kinase, phosphoinositide 3-kinase,
  • phospholipase Cy phospholipase Cy
  • protein kinase C protein kinase C
  • signal transducer and activator of transcription STAT
  • Examples of antibodies that can target and inhibit HER2/neu can include trastuzumab and pertuzumab.
  • EGFR epidermal growth factor receptor
  • Mutations that can lead to EGFR overexpression or over activity can be associated with a number of cancers, including squamous cell carcinoma and glioblastomas.
  • EGFR can function as a receptor tyrosine kinase and ligand binding can trigger dimerization with binding partners and autophosphorylation.
  • the phosphorylated EGFR can then activate several downstream signaling pathways including mitogen-activated protein kinase, phosphoinositide 3- kinase, phospholipase Cy, protein kinase C, and signal transducer and activator of transcription (STAT).
  • Examples of antibodies that can target and inhibit EGFR can include cetuximab, panutumumab, nimotuzumab, and zalutumumab.
  • One mutant variant of EGFR is EGFRvIII (epidermal growth factor receptor variant III, also referred to as de2-7EGFR).
  • EGFRvIII is the result of an EGFR gene rearrangement in which exons 2-7 of the extracellular domain are deleted. This mutation can result in a mutant receptor incapable of binding to any known ligand. The resulting receptor can engage in a constitutive low-level signaling and can be implicated in tumor progression.
  • Examples of antibodies that can target EGFRvIII can include AMG595 and ABT806.
  • C-Met hepatocyte growth factor receptor
  • C-Met encodes a member of the receptor tyrosine kinase family of proteins.
  • C-Met overexpression and over activity can be implicated in various cancers including lung adenocarcinomas, and high c-Met levels can be associated with poor patient outcome. Binding of hepatocyte growth factor can induce dimerization and
  • the c-Met receptor can activate various downstream signaling pathways including mitogen-activated protein kinase, phosphoinositide 3-kinase, and protein kinase C pathways.
  • the antibody onartuzumab can target and inhibit c-Met.
  • HER3 human epidermal growth factor receptor 3 encodes a member of the human epidermal growth factor receptor family. Ligand binding can induce dimerization and autophosphorylation of cytoplasmic tyrosine residues that then can recruit signaling proteins for downstream signaling pathway activation including mitogen-activated protein kinase and phosphoinoside 3-kinase pathways. HER3 can play an active role in cell proliferation and survival, and can be overexpressed, overactive, and/or mutated in various cancers. For example, HER3 can be overexpressed in breast, ovarian, prostate, colon, pancreas, stomach, oral, and lung cancers. The antibody patritumab can target and inhibit HER3.
  • MUC1 (mucin 1, cell surface associated) encodes a member of the mucin family of glycosylated proteins that can play an important role in cell adhesion and forming protective mucosal layers on epithelial surfaces.
  • MUC1 can be proteolytically cleaved into alpha and beta subunits that form a heterodimeric complex with the N-terminal alpha subunit providing cell- adhesion functionality and the C-terminal beta subunit modulating cell signaling pathways including the mitogen activated map kinase pathway.
  • MUC1 can play a role in cancer progression, for example, by regulating TP53-mediated transcription.
  • MUC1 overexpression, aberrant intracellular localization, and glycosylation changes can all be associated with carcinomas including pancreatic cancer cells.
  • the antibody clivatuzumab can target MUC1.
  • MUC16 (mucin 16, cell surface associated) encodes the largest member of the mucin family of glycosylated proteins that can play an important role in cell adhesion and forming protective mucosal layers on epithelial surfaces.
  • MUC16 can be a highly glycosylated 2.5MDa transmembrane protein that can provide a hydrophilic lubricating barrier on epithelial cells.
  • the cytoplasmic tail of MUC16 can be involved with various signaling pathways including the JAK2-STAT3 and Src kinase pathways.
  • a peptide epitope of MUC16 can be used as biomarker for detecting ovarian cancer. Elevated expression of MUC16 can be present in advanced ovarian cancers and pancreatic cancers.
  • the antibody sofituzumab can target MUC16.
  • EPCAM epidermal cell adhesion molecule
  • EPCAM epidermal cell adhesion molecule
  • EPCAM can also be a pluripotent stem cell marker.
  • EPCAM can modulate a variety of pathways including cell-cell adhesion, cellular proliferation, migration, invasion, maintenance of a pluripotent state, and differentiation in the context of tumor cells.
  • the antibodies edrecolomab and adecatumumab can target EPCAM.
  • MSLN (mesothelin) encodes a 40 kDa cell GPI-anchored membrane surface protein believed to function in cell adhesion. MSLN is overexpressed in mesothelioma and certain types of pancreatic, lung, and ovarian cancers. MSLN-related peptides that circulate in serum of patients suffering from pleural mesothelioma are used as biomarkers for monitoring the disease. MSLN may promote metastasis by inducing matrix metalloproteinase 7 and 9 expression. The monoclonal antibody anetumab has been developed to target MSLN.
  • CA6 carbonic anhydrase VI encodes one of several isozymes of carbonic anhydrase.
  • CA6 is found in salivary glands and may play a role in the reversible hydration of carbon dioxide.
  • CA6 is expressed in human serous ovarian adenocarcinomas.
  • the monoclonal antibody huDS6 has been developed to target CA6.
  • NAPI2B sodium/phosphate cotransporter 2B encodes a type II sodium-phosphate cotransporter. NAPI2B is highly expressed on the tumor surface in lung, ovarian, and thyroid cancers as well as in normal lung pneumocytes. The monoclonal antibody lifastuzumab has been developed to target NAPI2B.
  • TROP2 trophoblast antigen 2 encodes a transmembrane glycoprotein that acts as an intracellular calcium signal transducer. TROP2 binds to multiple factors such as IGF-1, claudin- 1, claudin-7, cyclin Dl, and PKC. TROP2 including intracellular calcium signaling and the mitogen activated protein kinase pathway. TROP 2 plays a role in cell self-renewal, proliferation, invasion, and survival. Discovered first in trophoblast cells that have the ability to invade uterine decidua during placental implantation, TROP2 overexpression has been shown to be capable of stimulating cancer growth.
  • TROP2 overexpression has been observed in breast, cervix, colorectal, esophagus, lung, non-Hodgkin' s lymphoma, chronic lymphocytic lymphoma, Raji Burkitt lymphoma, oral squamous cell, ovarian, pancreatic, prostate, stomach, thyroid, urinary bladder, and uterine carcinomas.
  • the monoclonal antibody sacituzumab has been developed to target TROP2.
  • CEA carcinomaembryonic antigen
  • CEACAM5 a member of a family of related glycoproteins involved in cell adhesion.
  • CEA is a biomarker for gastrointestinal cancers and may promote tumor development by means of its cell adhesion function.
  • CEA levels have been found to be elevated in serum of individuals with colorectal carcinoma.
  • CEA levels have also been found to be elevated in gastric carcinoma, pancreatic carcinoma, lung carcinoma, breast carcinoma, and medullary thyroid carcinoma.
  • the monoclonal antibodies PR1A3 and Ab2-3 have been developed to target CEA.
  • CLDN18.2 (claudin 18) encodes a member of the claudin family of integral membrane proteins.
  • CLDN18.2 is a component of tight junctions that create a physical barrier to prevent diffusion of solutes and water through the paracellular space between epithelial cells.
  • CLDN18.2 is overexpressed in infiltrating ductal adenocarcinomas, but is reduced in some gastric carcinomas.
  • the monoclonal antibody claudiximab has been developed to target CLDN18.2.
  • FAP fibroblast activation protein, alpha
  • FAP fibroblast activation protein, alpha
  • FAP fibroblast activation protein, alpha
  • FAP is believed to play a role in many processes including tissue remodeling, fibrosis, wound healing, inflammation, and tumor growth.
  • FAP enhances tumor growth and invasion by promoting angiogenesis, collagen fiber degradation and apoptosis, and by downregulating the immune response.
  • FAP is selectively expressed on fibroblasts within the tumor stroma.
  • the monoclonal antibody sibrotuzumab has been developed to target FAP.
  • EphA2 (EPH Receptor A2) encodes a member of the ephrin receptor subfamily of the protein- tyro sine kinase family. EphA2 binds to ephrin-A ligands. Activation of EphA2 receptor upon ligand binding can result in modulation of migration, integrin- mediated adhesion, proliferation, and differentiation. EphA2 is overexpressed in various cancers including breast, prostate, urinary bladder, skin, lung, ovarian, and brain cancers. High EphA2 expression is also correlated with poor prognosis. The monoclonal antibodies DS-8895a optl, DS-8895 opt2, and the 1C1 antibody in MEDI-547 have been developed to target EphA2.
  • RON macrophage stimulating 1 receptor encodes a cell surface receptor for macrophage stimulating protein (MSP) with tyrosine kinase activity and belongs to the MET proto-oncogene family.
  • MSP macrophage stimulating protein
  • RON plays a significant role in KRAS oncogene addiction and has also been shown to be overexpressed in pancreatic cancers.
  • Altered Ron expression and activation has been associated with decreased survival and cancer progression in various cancers including gastric, colon, breast, bladder, renal cell, ovarian, and hepatocellular cancers.
  • the monoclonal antibody narnatumab has been developed to target RON.
  • LY6E lymphocyte antigen 6 complex, locus E encodes an interferon alpha-inducible GPI-anchored cell membrane protein. LY6E is overexpressed in numerous cancers including lung, gastric, ovarian, breast, kidney, pancreatic, and head and neck carcinomas. The monoclonal antibody in RG7841 has been developed to target LY6E.
  • FRA farletuzumab and mirvetuximab have been developed to target FRA.
  • PSMA prote specific membrane antigen
  • M28 peptidase family is a type II transmembrane glycoprotein belonging to the M28 peptidase family that is expressed in all types of prostate tissues. PSMA is upregulated in cancer cells within the prostate and is used as a marker for prostate cancer. PSMA expression may also serve as a predictor of disease recurrence in prostate cancer patients.
  • the monoclonal antibodies J591 variant 1 and J591 variant 2 have been developed to target PSMA.
  • DLL3 (delta-like 3) encodes a ligand in the Notch signaling pathway that is associated with neuroendocrine cancer. DLL3 is most highly expressed in the fetal brain and is involved in somitogenesis in the paraxial mesoderm. DLL3 is expressed on tumor cell surfaces but not in normal tissues. The monoclonal antibody rovalpituzumab has been developed to target DLL3.
  • PTK7 tyrosine protein kinase-like 7 encodes a receptor tyrosine kinase that lacks catalytic tyrosine kinase activity but is nevertheless capable of signal transduction. PTK7 interacts with the WNT signaling pathway, which itself has important roles in epithelial mesenchymal transition and various cancers such as breast cancer. PTK7 overexpression has been associated with patient prognosis depending on the cancer type.
  • the monoclonal antibodies in PF-06647020 and the anti-PTK7 antibody described by SEQ ID NOs: 440 and 445 have been developed to target PTK7.
  • LIVl (LIV-1 protein, estrogen regulated) encodes a member of the LIV-1 subfamily of ZIP (Zrt-, Irt-like proteins) zinc transporters.
  • LIVl is an estrogen regulated protein that transports zinc and/or other ions across the cell membrane. Elevated levels of LIVl have been shown in estrogen receptor positive breast cancers, and LIVl is used as a marker of ER-positive cancers. LIVl has also been implicated as a downstream target of the STAT3 transcription factor and as playing an essential role in the nuclear localization of the Snail transcription factor that modulates epithelial-to-mesenchymal transition. The monoclonal antibody in SGN-LIV1A has been developed to target LIV1.
  • RORl receptor tyrosine kinase-like orphan receptor 1 encodes a member of the ROR family of orphan receptors. RORl has been found to bind Wnt5a, a non-canonical Wnt via a Frizzled domain (FZD), and plays an important role in skeletal, cardiorespiratory, and
  • RORl expression is upregulated in B-Cell chronic lymphocytic leukemia, acute lymphocytic leukemia, non-Hodgkin lymphoma, and myeloid malignancies.
  • the monoclonal antibody cirmtuzumab has been developed to target RORl.
  • MAGE-A3 (melanoma-associated antigen 3) encodes a member of the melanoma- associated antigen gene family.
  • the function of MAGE-A3 is not known, but its elevated expression has been observed in various cancers including melanoma, non-small cell lung cancer, and in putative cancer stem cell populations in bladder cancer.
  • the monoclonal antibody described by SEQ ID NOs: 479 and 484 has been developed to target MAGE- A3.
  • NY-ESO-1 (New York esophageal squamous cell carcinoma 1) encodes a member of the cancer-testis family of proteins. Cancer-testis antigen expression is normally restricted to testicular germ cells in adult tissues, but has been found to be aberrantly expressed in various tumors including soft tissue sarcomas, melanoma, epithelial cancers, and myxoid and round cell liposarcomas. The monoclonal antibody described by SEQ ID Nos: 492 and 497 has been developed to target NY-ESO-1.
  • an antigen can be a cell surface marker on target cells associated with a disease or condition.
  • An antigen can be a peptide or fragment thereof.
  • An antigen can be expressed on an immune cell.
  • An antigen can be expressed on an antigen-presenting cell.
  • An antigen can be expressed on a dendritic cell, a macrophage, or a B cell.
  • An antigen on an antigen presenting cell can be a cell lineage marker or a cell surface protein expressed preferentially on antigen presenting cells or a subset of antigen presenting cells.
  • An antigen can be a peptide presented in a major histocompatibility complex by cell.
  • a cell surface marker recognized by the antigen binding domain can include macromolecules associated with viral and bacterial diseases or infections, autoimmune diseases and cancerous diseases.
  • An antigen can be a tumor antigen or fragment thereof.
  • a tumor antigen can be any antigen listed on tumor antigen databases, such as TANTIGEN, or peptide databases for T cell-defined tumor antigens, such as the Cancer Immunity Peptide database.
  • a tumor antigen can also be any antigen listed in the review by Chen (Chen, Cancer Immun 2004 [updated 2004 Mar 10; cited 2004 Apr 1]).
  • An antigen can be or can be at least 80% homologous CD5, CD19, CD20, CD25, CD37, CD30, CD33, CD45, CAMPATH-1, BCMA, CS-1, PD-L1, B7-H3, B7-DC (PD-L2), HLD-DR, carcinoembryonic antigen (CEA), TAG-72, EpCAM, MUC1, MUC15, MUC16, fo late-binding protein, A33, G250, pro state- specific membrane antigen (PSMA), ferritin, GD2, GD3, GM2, Le y , CA-125, CA19-9, epidermal growth factor, pl85HER2, IL-2 receptor, EGFRvIII (de2-7 EGFR), fibroblast activation protein (FAP), tenascin, a metalloproteinase, endosialin, vascular endothelial growth factor, ⁇ 3, WT1, LMP2, HPV E6, HPV E7, Her-2/neu, p53
  • Claudin-16 Claudin-16
  • CLDN18.2 RON, LY6E, FRA, DLL3, PTK7, Uroplakin-IB (UPK1B), LIV1, ROR1, STRA6, TMPRSS3, TMPRSS4, TMEM238, Clorfl86, Fos-related antigen 1, VEGFR1, endoglin, VTCN1 (B7-H4), VISTA, or gpNMB.
  • an antigen binding domain recognizes a single antigen. In some embodiments, an antigen binding domain recognizes two or more different antigens.
  • An antibody construct can include one or more binding domains.
  • an antibody construct can comprise a first binding domain.
  • An antibody construct can also comprise a second binding domain.
  • a binding domain can specifically bind to an antigen on a cell surface.
  • a binding domain can specifically bind to an antigen on a cell surface, for example, of a tumor or cancer cell, an immune cell, or of an antigen presenting cell, such as a dendritic cell or macrophage.
  • a binding domain can be a cell surface receptor agonist.
  • a binding domain can be an antigen binding domain.
  • An antigen binding domain can be a cell surface receptor agonist.
  • An antigen binding domain can be a domain that can specifically bind to an antigen.
  • An antigen binding domain can specifically bind to a tumor antigen.
  • An antigen binding domain can be an antigen-binding portion of an antibody or an antigen binding fragment thereof.
  • a binding domain can recognize a single antigen.
  • An antibody construct can include, for example, one, two, three, four, five, six, seven, eight, nine, ten, or more antigen binding domains.
  • An antibody construct, such as an antibody can include two antigen binding domains in which each antigen binding domain can recognize the same antigen.
  • An antibody construct, such as a bi- specific antibody can include two antigen binding domains in which each antigen binding domain can recognize different antigens.
  • An antibody construct can include three antigen binding domains in which each antigen binding domain can recognize different antigens.
  • An antibody construct can include three antigen binding domains in which two of the antigen binding domains can recognize the same antigen.
  • An antigen binding domain can be in a scaffold, in which a scaffold is a supporting framework for the antigen binding domain.
  • An antigen binding domain can be in a non-antibody scaffold.
  • An antigen binding domain can be in an antibody scaffold.
  • An antibody construct can comprise an antigen binding domain in a scaffold.
  • the antibody construct can further comprise an Fc fusion protein product.
  • the antibody construct is an Fc fusion protein.
  • an antibody construct can comprise an antigen binding domain that can specifically bind to a tumor antigen.
  • a tumor antigen also referred to as a tumor associated antigen, is an antigen that can be expressed by a cancer cell, a neoplastic tumor cell and/or within a tumor microenvironment. It is preferably not expressed or expressed at low levels on normal (non-cancerous) cells.
  • a tumor antigen can be an antigen expressed on a cell associated within a tumor, such as a neoplastic cell, or a tumor associated cell such as a stromal cell, endothelial cell, fibroblast, or tumor- infiltrating immune cell.
  • the tumor antigen Her2/Neu can be overexpressed by certain types of breast and ovarian cancer.
  • a tumor antigen can also be ectopically expressed by a tumor and contribute to deregulation of the cell cycle, reduced apoptosis, metastasis, or escape from immune surveillance.
  • Tumor antigens can generally be proteins or polypeptides derived therefrom, but also can be glycans, lipids, or other small organic molecules.
  • a tumor antigen can arise through increases or decreases in post-translational processing exhibited by a cancer cell compared to a normal cell, for example, protein glycosylation, protein lipidation, protein phosphorylation, or protein acetylation.
  • a binding domain specifically can bind to a tumor associated antigen, such as CD5, CD19, CD20, CD25, CD37, CD30, CD33, CD45, CAMPATH- 1 antigen, BCMA, CS-1, PD-L1, B7-H3, B7-DC (PD-L2), HLD-DR, carcinoembryonic antigen (CEA), TAG-72, EpCAM, MUC1, MUC15, MUC16, fo late-binding protein, A33, G250, prostate- specific membrane antigen (PSMA), ferritin, GD2, GD3, GM2, Le y , CA-125, CA19-9, epidermal growth factor, pl85HER2, IL-2 receptor, EGFRvIII (de2-7 EGFR), fibroblast activation protein (FAP), tenascin, a metalloproteinase, endosialin, vascular endothelial growth factor, ⁇ 3, WT1, LMP2, HPV E6,
  • a tumor associated antigen
  • a binding domain specifically can bind to a tumor associated antigen having at least 80%, 90%, 95%, 97%, 98%, 99% or 100% sequence identity to CD5, CD19, CD20, CD25, CD37, CD30, CD33, CD45, CAMPATH- 1 antigen, BCMA, CS-1, PD-L1, B7-H3, B7-DC (PD-L2), HLD-DR, carcinoembryonic antigen (CEA), TAG-72, EpCAM, MUCl, MUC15, MUC16, fo late-binding protein, A33, G250, pro state- specific membrane antigen
  • PSMA ferritin
  • CA-125 CA19-9
  • epidermal growth factor pl85HER2
  • IL-2 receptor IL-2 receptor
  • EGFRvIII (de2-7 EGFR), fibroblast activation protein (FAP), tenascin, a metalloproteinase, endosialin, vascular endothelial growth factor, ⁇ 3, WTl, LMP2, HPV E6, HPV E7, Her-2/neu, p53 nonmutant, NY-ESO-1, MelanA/M ART 1 , Ras mutant, gplOO, p53 mutant, PR1, bcr-abl, tyrosinase, survivin, PSA, hTERT, a Sarcoma translocation breakpoint protein, EphA2, PAP, ML-IAP, AFP, ERG, NA17, PAX3, ALK, androgen receptor, cyclin B l, MYCN, RhoC, TRP-2, mesothelin (MSLN), PSCA, MAGE Al, MAGE A3, CYP1B 1, PLAV1, BORIS, GloboH, ETV6-
  • a binding domain specifically binds to an antigen selected from the group consisting of comprising Her2/Neu (CD340), EGFR, CMET, HER3, MUCl, MUCl 6,
  • EPCAM EPCAM
  • MSLN CA6, NAPI2B, TROP2, CEA, CLDN18.2, EGFRvIII, FAP, EphA2, RON,
  • a binding domain of an antibody construct can be selected from any domain that specifically binds to an antigen, including but not limited to, from a monoclonal antibody, a polyclonal antibody, a recombinant antibody, or a functional fragment thereof, for example, a heavy chain variable domain (V H ) and a light chain variable domain (V L ), or from a non-antibody such as a DARPin, an affimer, an avimer, a knottin, a monobody, an affinity clamp, an ectodomain, a receptor ectodomain, a receptor, a cytokine, a ligand, an immunocytokine, a T-cell receptor, a VNAR, an anticalin, or a recombinant T-cell receptor.
  • V H heavy chain variable domain
  • V L light chain variable domain
  • a non-antibody such as a DARPin, an affimer, an avimer, a knottin, a mono
  • a binding domain of an antibody construct is from a monoclonal antibody, a polyclonal antibody, a recombinant antibody, or a functional fragment thereof, for example, a heavy chain variable domain (V H ) and a light chain variable domain (V L ).
  • the antigen binding domain of an antibody construct can be at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% homologous to an antigen binding domain selected from, but not limited to, a monoclonal antibody, a polyclonal antibody, a recombinant antibody, or a functional fragment thereof, for example, a heavy chain variable domain (V H ) and a light chain variable domain (V L ), or from a non-antibody, such as a DARPin, an affimer, an avimer, a knottin, a monobody, an affinity clamp, an ectodomain, a receptor ectodomain, a receptor, a cytokine, a ligand, an immunocytokine, a T-cell receptor, a VNAR, an anticalin, or a
  • a binding domain for example an antigen binding domain from a monoclonal antibody, can comprise a light chain and a heavy chain.
  • the monoclonal antibody binds to an antigen present on the surface of an antigen presenting cell (APC antigen) and comprises the light chain of an anti-APC antigen antibody and the heavy chain of an anti-APC antigen antibody, which bind an APC antigen.
  • the monoclonal antibody binds to a tumor antigen comprises the light chain of a tumor antigen (anti-tumor) antibody and the heavy chain of a tumor antigen (anti-tumor) antibody, which bind to the tumor antigen.
  • An antibody construct can be an antibody.
  • An antibody molecule can include of two identical light protein chains (light chains) and two identical heavy protein chains (heavy chains), all held together covalently by interchain disulfide linkages. Structurally, various functions of an antibody can be confined to discrete protein regions or domains.
  • the sites that can recognize and can bind to antigen consist of three complementarity determining regions (CDRs; also referred to as hyper- variable regions) that lie within the variable heavy chain regions and within the variable light chain regions at the N-terminal ends of the two heavy and two light chains.
  • CDRs complementarity determining regions
  • the constant domains can provide the general framework of the antibody and may not be involved directly in binding the antibody to an antigen, but can be involved in various effector functions, such as participation of the antibody in antibody-dependent cellular cytotoxicity (ADCC).
  • ADCC antibody-dependent cellular cytotoxicity
  • variable domains of natural light and heavy chains can have the same general structures, and each domain can comprise four framework regions, whose sequences can be somewhat conserved, connected by three CDRs.
  • the four framework regions can largely adopt a ⁇ -sheet conformation and the CDRs can form loops connecting, and in some aspects forming part of, the ⁇ -sheet structure.
  • the CDRs in each chain can be held in close proximity by the framework regions and, with the CDRs from the other chain, can contribute to the formation of the antigen binding site.
  • An antibody can include an antibody of any type, which can be assigned to different classes of immunoglobins, e.g., IgA, IgD, IgE, IgG, and IgM. Several different classes can be further divided into isotypes, e.g., IgGl, IgG2, IgG3, IgG4, IgAl, and IgA2. Exemplary heavy chain sequences of reference antibodies can be used to identify residue variants and mutants.
  • An exemplary heavy chain sequence for human IgGl heavy chain is that of the human IgGl antibody, and can comprise:
  • An exemplary heavy chain reference sequence for human IgG2 heavy chain can comprise:
  • An exemplary heavy chain reference sequence for human IgG4 heavy chain can comprise:
  • the heavy-chain constant regions (Fc) that corresponds to the different classes of immunoglobulins can be ⁇ , ⁇ , ⁇ , ⁇ , and ⁇ .
  • the light chains can be one of either kappa or ⁇ and lambda or ⁇ , based on the amino acid sequences of the constant domains.
  • the Fc region can contain an Fc domain.
  • An Fc receptor can bind an Fc domain.
  • An Fc domain can comprise amino acid residues 216 to 447 of an IgGl, which are part of SEQ ID NO: 898.
  • An Fc domain can comprise amino acid residues 216 to 442 of an IgG2, which are part of SEQ ID NO: 899.
  • An Fc domain can comprise amino acid residues 216 to 444 of an IgG4, which are part of SEQ ID NO: 900.
  • Antibody constructs can also include any fragment or recombinant forms thereof (e.g., scFVs and domain antibodies).
  • Antibody constructs can also include any fragment or
  • non-antibody scaffolds including but not limited to anti-calins, affibodies, affilins, atrimers, avimers, bicyclic peptides, centyrins, Cys-knots, Darpins, fibronections, Kunitz domains, O-bodies, or peptibodies.
  • An antibody construct can comprise an antigen binding domain of an antibody.
  • An antigen binding domain of an antibody can comprise one or more light chain CDRs (LCDRs) and one or more heavy chain CDRs (HCDRs), or one or more LCDRs or one or more HCDRs.
  • LCDRs light chain CDRs
  • HCDRs heavy chain CDRs
  • an antibody binding domain of an antibody construct can comprise one or more of the following: a light chain complementary determining region 1 (LCDR1), a light chain
  • an antibody binding domain can comprise one or more of the following: a heavy chain complementary determining region 1 (HCDR1), a heavy chain complementary determining region 2 (HCDR2), or a heavy chain complementary determining region 3 (HCDR3).
  • an antigen binding domain comprises LCDR1, LCDR2, LCDR3, HCDR1, HCDR2 and HCDR3.
  • the CDRs described herein can be defined according to the IMGT (the international ImMunoGeneTics information system).
  • An antigen binding domain can comprise only the heavy chain of an antibody.
  • An antigen binding domain can comprise only the variable domain of the heavy chain of an antibody.
  • an antigen binding domain can comprise only the light chain of an antibody.
  • An antigen binding domain can comprise only the variable light chain of an antibody.
  • An antibody construct can comprise an antibody fragment.
  • An antibody fragment can include (i) a Fab fragment, a monovalent fragment consisting of the V L , V H, C L and C HI domains; (ii) a F(ab') 2 fragment, a bivalent fragment comprising two Fab fragments linked by a disulfide bridge at the hinge region; and (iii) a Fv fragment consisting of the V L and V H domains of a single arm of an antibody.
  • the two domains of the Fv fragment, V L and V H can be coded for by separate genes, they can be linked by a synthetic linker to be made as a single protein chain in which the V L and V H regions pair to form monovalent molecules.
  • F(ab') 2 and Fab' moieties can be produced recombinantly.
  • An Fv can be the minimum antibody fragment which contains a complete antigen- recognition and antigen-binding site. This region can consist of a dimer of one heavy chain and one light chain variable domain in tight, non-covalent association. In this configuration the three hypervariable regions of each variable domain can interact to define an antigen-binding site on the surface of the V H -V L dimer. A single variable domain (or half of an Fv comprising only CDRs specific for an antigen) can recognize and bind antigen, although at a lower affinity than the entire binding site.
  • An antibody used herein can be chimeric or "humanized.” Chimeric and humanized forms of non-human (e.g., murine) antibodies can be chimeric immunoglobulins,
  • the humanized antibody can comprise substantially all of at least one, and typically two, variable domains, in which all or substantially all of the CDR regions correspond to those of a non-human immunoglobulin and all or substantially all of the frame work regions (FR) are those of a human immunoglobulin sequence.
  • the humanized antibody can also comprise at least a portion of an immunoglobulin constant region (Fc), typically that of a human immunoglobulin consensus sequence.
  • an antibody described herein can be a human antibody.
  • human antibodies can include antibodies having, for example, the amino acid sequence of a human immunoglobulin and include antibodies isolated from human immunoglobulin libraries or from animals transgenic for one or more human immunoglobulins that do not express endogenous immunoglobulins. Human antibodies can be produced using transgenic mice which are incapable of expressing functional endogenous immunoglobulins, but which can express human immunoglobulin genes. Completely human antibodies that recognize a selected epitope can be generated using guided selection. In this approach, a selected non-human monoclonal antibody, e.g., a mouse antibody, is used to guide the selection of a completely human antibody recognizing the same epitope
  • An antibody described herein can be a bispecific antibody or a dual variable domain antibody (DVD).
  • Bispecific and DVD antibodies are monoclonal, often human or humanized, antibodies that have binding specificities for at least two different antigens.
  • An antibody described herein can be a derivatized antibody.
  • derivatized antibodies can be modified by glycosylation, deglycosylation, defucosylation, acetylation, pegylation, phosphorylation, amidation, derivatization by known protecting/blocking groups, proteolytic cleavage, or linkage to a cellular ligand or other protein.
  • An antibody described herein can have a sequence that has been modified to alter at least one constant region-mediated biological effector function relative to the corresponding wild type sequence.
  • the antibody can be modified to reduce at least one constant region-mediated biological effector function relative to an unmodified antibody, e.g., reduced binding or increased binding to the Fc receptor (FcR).
  • FcR binding can be reduced or increased by, for example, mutating the immunoglobulin constant region segment of the antibody at particular regions necessary for FcR interactions.
  • An antibody described herein can be modified to acquire or improve at least one constant region-mediated biological effector function relative to an unmodified antibody, e.g., to enhance FcyR interactions.
  • an antibody with a constant region that binds FcyRIIA, FcyRIIB and/or FcyRIIIA with greater affinity than the corresponding wild type constant region can be produced according to the methods described herein.
  • An antibody described herein can bind to tumor cells, such as an antibody against a cell surface receptor or a tumor antigen.
  • An antibody construct can comprise a first binding domain.
  • An antibody construct can comprise a first binding domain that specifically binds an antigen.
  • An antibody construct can comprise a first binding domain that specifically binds a tumor antigen.
  • a first binding domain can specifically bind a tumor antigen, wherein the tumor antigen is selected from the group consisting CD5, CD19, CD20, CD25, CD37, CD30, CD33, CD45, CAMPATH-1, BCMA, CS-1, PD-Ll, B7-H3, B7-DC (PD-L2), HLD-DR, carcinoembryonic antigen (CEA), TAG-72, EpCAM, MUC1, MUC15, MUC16, fo late-binding protein, A33, G250, pro state- specific membrane antigen (PSMA), ferritin, GD2, GD3, GM2, Le y , CA-125, CA19-9, epidermal growth factor, pl85HER2, IL-2 receptor, EGFRvIII (de2-7 EGFR),
  • HMWMAA HMWMAA
  • AKAP-4 SSX2
  • XAGE 1 Legumain
  • Tie Page4
  • VEGFR2 VEGFR2
  • MAD-CT-1 MAD-CT-1
  • PDGFR-B MAD-CT-2, ROR2, TRAIL 1, MAGE A4, MAGE C2, GAGE, EGFR, CMET, HER3, EPCAM, CA6, NAPI2B, TROP2, Claudin-6 (CLDN6), Claudin-16 (CLDN16),
  • An antibody construct can comprise a first binding domain that specifically binds a tumor antigen on a tumor cell, a tumor fragment, an immune cell or an antigen presenting cell.
  • An antibody construct can comprise a first binding domain that specifically binds a tumor antigen.
  • the construct can comprise a first binding domain comprising one or more CDRs, typically a set of six CDRs.
  • a first binding domain can comprise at least 80% sequence identity to any sequence in TABLE 1.
  • An antibody construct can comprise a first binding domain that binds a tumor antigen, wherein the first binding domain comprises at least 80% sequence identity to: a) HCDR1 comprising an amino acid sequence of SEQ ID NO: 13, HCDR2 comprising an amino acid sequence of SEQ ID NO: 14, HCDR3 comprising an amino acid sequence of SEQ ID NO: 15, LCDR1 comprising an amino acid sequence of SEQ ID NO: 18, LCDR2 comprising an amino acid sequence of SEQ ID NO: 19, and LCDR3 comprising an amino acid sequence of SEQ ID NO: 20; b) HCDR1 comprising an amino acid sequence of SEQ ID NO: 26, HCDR2 comprising an amino acid sequence of SEQ ID NO: 27, HCDR3 comprising an amino acid sequence of SEQ ID NO: 28, LCDR1 comprising an amino acid sequence of SEQ ID NO: 31, LCDR2 comprising an amino acid sequence of SEQ ID NO: 32, and LCDR3 comprising an amino acid sequence of SEQ ID NO: 33; c) HCDR1 comprising an
  • HCDR2 comprising an amino acid sequence of SEQ ID NO: 209
  • HCDR3 comprising an amino acid sequence of SEQ ID NO: 210
  • LCDR1 comprising an amino acid sequence of SEQ ID NO: 213
  • LCDR2 comprising an amino acid sequence of SEQ ID NO: 214
  • LCDR3 comprising an amino acid sequence of SEQ ID NO: 215
  • HCDRl comprising an amino acid sequence of SEQ ID NO: 805
  • HCDR2 comprising an amino acid sequence of SEQ
  • HCDR3 comprising an amino acid sequence of SEQ ID NO: 807
  • LCDR1 comprising an amino acid sequence of SEQ ID NO: 808
  • LCDR2 comprising an amino acid sequence of SEQ ID NO: 809
  • LCDR3 comprising an amino acid sequence of SEQ ID NO:
  • HCDRl comprising an amino acid sequence of SEQ ID NO: 823
  • HCDR2 comprising an amino acid sequence of SEQ ID NO: 824
  • HCDR3 comprising an amino acid sequence of SEQ
  • LCDR1 comprising an amino acid sequence of SEQ ID NO: 826
  • LCDR2 comprising an amino acid sequence of SEQ ID NO: 827
  • LCDR3 comprising an amino acid sequence of SEQ ID NO: 828
  • HCDRl comprising an amino acid sequence of SEQ ID NO:
  • HCDR2 comprising an amino acid sequence of SEQ ID NO: 222
  • HCDR3 comprising an amino acid sequence of SEQ ID NO: 223
  • LCDR1 comprising an amino acid sequence of SEQ
  • LCDR2 comprising an amino acid sequence of SEQ ID NO: 227
  • LCDR3 comprising an amino acid sequence of SEQ ID NO: 228
  • HCDRl comprising an amino acid sequence of SEQ ID NO: 260
  • HCDR2 comprising an amino acid sequence of SEQ ID NO: 261
  • HCDR3 comprising an amino acid sequence of SEQ ID NO: 262
  • LCDR1 comprising an amino acid sequence of SEQ ID NO: 265
  • LCDR2 comprising an amino acid sequence of SEQ ID NO:
  • HCDRl comprising an amino acid sequence of SEQ ID NO: 273, HCDR2 comprising an amino acid sequence of SEQ ID NO: 274, HCDR3 comprising an amino acid sequence of SEQ ID NO: 275,
  • LCDR1 comprising an amino acid sequence of SEQ ID NO: 278, LCDR2 comprising an amino acid sequence of SEQ ID NO: 279, and LCDR3 comprising an amino acid sequence of SEQ ID NO:
  • HCDRl comprising an amino acid sequence of SEQ ID NO: 286, HCDR2 comprising an amino acid sequence of SEQ ID NO: 287, HCDR3 comprising an amino acid sequence of SEQ ID NO: 288, LCDR1 comprising an amino acid sequence of SEQ ID NO: 291,
  • LCDR2 comprising an amino acid sequence of SEQ ID NO: 292
  • LCDR3 comprising an amino acid sequence of SEQ ID NO: 293
  • HCDRl comprising an amino acid sequence of
  • HCDR2 comprising an amino acid sequence of SEQ ID NO: 300, HCDR3 comprising an amino acid sequence of SEQ ID NO: 301, LCDR1 comprising an amino acid sequence of SEQ ID NO: 304, LCDR2 comprising an amino acid sequence of SEQ ID NO: 305, and LCDR3 comprising an amino acid sequence of SEQ ID NO: 306; x) HCDRl comprising an amino acid sequence of SEQ ID NO: 312, HCDR2 comprising an amino acid sequence of SEQ ID NO: 313, HCDR3 comprising an amino acid sequence of SEQ ID NO: 314, LCDR1 comprising an amino acid sequence of SEQ ID NO: 317, LCDR2 comprising an amino acid sequence of SEQ ID NO: 318, and LCDR3 comprising an amino acid sequence of SEQ ID NO: 319; y) HCDRl comprising an amino acid sequence of SEQ ID NO: 325, HCDR2 comprising an amino acid sequence of SEQ ID NO: 326, HC
  • LCDR1 comprising an amino acid sequence of SEQ ID NO: 421, LCDR2 comprising an amino acid sequence of SEQ ID NO: 422, and LCDR3 comprising an amino acid sequence of SEQ ID NO: 423; gg) HCDRl comprising an amino acid sequence of SEQ ID NO:
  • HCDR2 comprising an amino acid sequence of SEQ ID NO: 430
  • HCDR3 comprising an amino acid sequence of SEQ ID NO: 431
  • LCDR1 comprising an amino acid sequence of SEQ
  • HCDRl comprising an amino acid sequence of SEQ ID NO: 442
  • HCDR2 comprising an amino acid sequence of SEQ ID NO: 443
  • HCDR3 comprising an amino acid sequence of SEQ ID NO: 444
  • LCDR1 comprising an amino acid sequence of SEQ ID NO: 447
  • LCDR2 comprising an amino acid sequence of SEQ ID NO:
  • HCDRl comprising an amino acid sequence of SEQ ID NO: 455, HCDR2 comprising an amino acid sequence of SEQ ID NO: 456, HCDR3 comprising an amino acid sequence of SEQ ID NO: 457,
  • LCDR1 comprising an amino acid sequence of SEQ ID NO: 460
  • LCDR2 comprising an amino acid sequence of SEQ ID NO: 461
  • LCDR3 comprising an amino acid sequence of SEQ ID NO:
  • HCDRl comprising an amino acid sequence of SEQ ID NO: 468
  • HCDR2 comprising an amino acid sequence of SEQ ID NO: 469
  • HCDR3 comprising an amino acid sequence of SEQ ID NO: 470
  • LCDR1 comprising an amino acid sequence of SEQ ID NO: 473
  • LCDR2 comprising an amino acid sequence of SEQ ID NO: 474
  • LCDR3 comprising an amino acid sequence of SEQ ID NO: 475
  • HCDRl comprising an amino acid sequence of
  • HCDR1 comprising an amino acid sequence of SEQ ID NO: 486, LCDR2 comprising an amino acid sequence of SEQ ID NO: 487, and LCDR3 comprising an amino acid sequence of SEQ ID NO: 488; 11) HCDRl comprising an amino acid sequence of SEQ ID NO: 494, HCDR2 comprising an amino acid sequence of SEQ ID NO:
  • HCDR3 comprising an amino acid sequence of SEQ ID NO: 496
  • LCDR1 comprising an amino acid sequence of SEQ ID NO: 499
  • LCDR2 comprising an amino acid sequence of SEQ ID NO: 500
  • LCDR3 comprising an amino acid sequence of SEQ ID NO:
  • HCDRl comprising an amino acid sequence of SEQ ID NO: 673
  • HCDR2 comprising an amino acid sequence of SEQ ID NO: 674
  • HCDR3 comprising an amino acid sequence of
  • An antibody construct can comprise a first binding domain that specifically binds a tumor antigen.
  • An antibody construct can comprise a first binding domain comprising one or more variable domains.
  • An antibody construct can comprise a first binding domain comprising a light chain variable domain (V L domain).
  • a first binding domain can have at least 80% or 100% sequence identity to any V L sequence in TABLE 2.
  • An antibody construct can comprise a first binding domain comprising a heavy chain variable domain (V H domain).
  • a first binding domain can comprise at least 80% or 100% sequence identity to any V H sequence in TABLE 2.
  • a first binding domain can comprise at least 80% sequence identity to any sequence in TABLE 2.
  • a first binding domain can have a pair of V H and V L regions, having sequence selected from the pairs in TABLE 2.
  • An antibody construct can comprise a first binding domain that specifically binds a tumor antigen, wherein the first binding domain comprises: a) a V H sequence having at least 80% or 100% sequence identity to an amino acid sequence of SEQ ID NO: 12, and a V L sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 17; b) a V H sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 25, and a V L sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 30; c) a V H sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 38, and a V L sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 43; d) a V H sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 51, and a V L sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 56; e) a V H sequence having
  • V L sequence having at least 80% sequence identity to an amino acid sequence of SEQ
  • V L sequence having at least 80% sequence identity to an amino acid sequence of SEQ
  • An antibody construct can comprise a first binding domain and an Fc domain, wherein the first binding domain and the Fc domain comprise an antibody.
  • a first binding domain can specifically bind a tumor antigen.
  • An antibody construct can comprise an antibody light chain wherein the antibody construct specifically binds to an antigen.
  • An antibody construct can comprise a light chain comprising at least 80% or 100% sequence identity to a light chain sequence in TABLE 3, wherein the antibody construct specifically binds to an antigen.
  • An antibody construct can comprise an antibody heavy chain, wherein the antibody construct specifically binds to an antigen.
  • An antibody construct can comprise a heavy chain comprising at least 80% or 100% sequence identity to a heavy chain sequence in TABLE 3, wherein the antibody construct specifically binds to an antigen.
  • An antibody construct can comprise at least 80% sequence identity to any sequence in TABLE 3, wherein the antibody construct specifically binds to an antigen.
  • An antibody construct can have a pair of heavy and light chains having sequences selected from the pairs of sequences in TABLE 3, wherein the antibody construct specifically binds to an antigen.
  • An antibody construct can comprise an anti-tumor antibody, wherein the antibody comprises: a) a heavy chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 11, and a light chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 16; b) a heavy chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 24, and a light chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 29; c) a heavy chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 37, and a light chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 42; d) a heavy chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 50, and a light chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 55; e) a heavy chain sequence having at least 80% sequence identity to an amino amino acid sequence
  • An antibody construct can comprise a second binding domain that specifically binds to an antigen.
  • An antibody construct can comprise a second binding domain that specifically binds to an antigen on an antigen presenting cell.
  • An antigen presenting cell can be a dendritic cell or a macrophage.
  • a second binding domain can specifically bind to an antigen on an immune cell such as an antigen presenting cell, wherein the molecule comprises at least 80% sequence identity to a group consisting of CD40, CD47, TNFR2, DEC-205, PD-L1, CD36 mannose scavenger receptor 1, CLEC9A, DC-SIGN, CLEC12A, BDCA-2, OX40L, 41BBL, CD204, MARCO, CLEC5A, Dectin 1, Dectin 2, CLECIOA, CD206, CD64, CD32A, CD 16 A, HVEM, CD38, TREM2, CSF1R, or CD32B.
  • An antibody construct can further comprise an Fc domain.
  • An antibody construct can comprise, for example, a first binding domain, a second binding domain, and an Fc domain, wherein the first binding domain is attached to the Fc domain.
  • An antibody construct can comprise a first binding domain, a second binding domain, and an Fc domain, wherein the second binding domain is attached to the Fc domain.
  • a first binding domain can be attached to an Fc domain as a fusion protein.
  • a second binding domain can be attached to an Fc domain as a fusion protein.
  • a first binding domain can be attached to an Fc domain via a linker.
  • a second binding domain can be attached to an Fc domain via a linker.
  • An antibody construct can comprise a second binding domain comprising one or more CDRs.
  • a second binding domain can comprise at least 80% sequence identity to any sequence in TABLE 5.
  • a second binding domain can comprise a set of CDRs having the sequences set forth in any sete of sequence in TABLE 5.
  • An antibody construct can comprise a second binding domain that specifically binds to
  • An antibody construct can comprise a second binding domain that is a CD40 agonist.
  • An antibody construct can comprise a second binding domain that binds to CD40, wherein the second binding domain comprises at least 80% sequence identity to: a) HCDR1 comprising an amino acid sequence of SEQ ID NO: 3, HCDR2 comprising an amino acid sequence of SEQ ID NO:
  • HCDR3 comprising an amino acid sequence of SEQ ID NO: 5
  • LCDR1 comprising an amino acid sequence of SEQ ID NO: 8
  • LCDR2 comprising an amino acid sequence of SEQ ID NO:
  • HCDR1 comprising an amino acid sequence of SEQ ID NO: 582
  • HCDR2 comprising an amino acid sequence of SEQ ID NO: 583
  • HCDR3 comprising an amino acid sequence of SEQ ID NO: 584
  • LCDR1 comprising an amino acid sequence of SEQ ID NO: 587, LCDR2 comprising an amino acid sequence of SEQ ID NO: 588, and LCDR3 comprising an amino acid sequence of SEQ ID NO: 589;
  • HCDRl comprising an amino acid sequence of SEQ ID NO: 592, HCDR2 comprising an amino acid sequence of SEQ ID NO: 593, HCDR3 comprising an amino acid sequence of SEQ ID NO: 594, LCDR1 comprising an amino acid sequence of SEQ ID NO: 597, LCDR2 comprising an amino acid sequence of SEQ ID NO: 598, and LCDR3 comprising an amino acid sequence of SEQ ID NO: 599;
  • HCDRl comprising an amino acid sequence of SEQ ID NO: 602, HCDR2 comprising an amino acid sequence of SEQ ID NO: 603, HCDR3 comprising an amino acid sequence of SEQ ID NO: 604, LCDR1 comprising an amino acid sequence of SEQ ID NO: 607, LCDR2 comprising an amino acid sequence of S
  • An antibody construct can comprise a second binding domain that specifically binds DC- SIGN.
  • An antibody construct can comprise a second binding domain that binds DC-SIGN, wherein the second binding domain comprises at least 80% sequence identity to: a) HCDRl comprising an amino acid sequence of SEQ ID NO: 640, HCDR2 comprising an amino acid sequence of SEQ ID NO: 641, HCDR3 comprising an amino acid sequence of SEQ ID NO: 642, LCDR1 comprising an amino acid sequence of SEQ ID NO: 643, LCDR2 comprising an amino acid sequence of SEQ ID NO: 644, and LCDR3 comprising an amino acid sequence of SEQ ID NO: 645; b) HCDRl comprising an amino acid sequence of SEQ ID NO: 646, HCDR2 comprising an amino acid sequence of SEQ ID NO: 647, HCDR3 comprising an amino acid sequence of SEQ ID NO: 648, LCDR1 comprising an amino acid sequence of SEQ ID NO: 649, LCDR2 comprising an amino acid
  • An antibody construct can comprise a second binding domain that specifically binds DEC-205.
  • An antibody construct comprising a second binding domain that binds DEC-205 can comprise at least 80% sequence identity to: a) HCDR1 comprising an amino acid sequence of SEQ ID NO: 234, HCDR2 comprising an amino acid sequence of SEQ ID NO: 235, HCDR3 comprising an amino acid sequence of SEQ ID NO: 236, LCDR1 comprising an amino acid sequence of SEQ ID NO: 239, LCDR2 comprising an amino acid sequence of SEQ ID NO: 240, and LCDR3 comprising an amino acid sequence of SEQ ID NO: 241 ; or b) HCDR1 comprising an amino acid sequence of SEQ ID NO: 247, HCDR2 comprising an amino acid sequence of SEQ ID NO: 248, HCDR3 comprising an amino acid sequence of SEQ ID NO: 249, LCDR1 comprising an amino acid sequence of SEQ ID NO: 252, LCDR2 comprising an amino acid sequence of SEQ
  • An antibody construct can comprise a second binding domain comprising one or more variable domains.
  • An antibody construct can comprise a second binding domain comprising a light chain variable domain (V L domain).
  • a second binding domain can comprise at least 80% sequence identity to any V L sequence in TABLE 6.
  • An antibody construct can comprise a second binding domain comprising a heavy chain variable domain.
  • a second binding domain can comprise at least 80% sequence identity to any V H sequence in TABLE 6.
  • a second binding domain can comprise at least 80% sequence identity to any sequence in TABLE 6.
  • a second binding domain can comprise at a pair of V H and V L domains having a pair of sequences in
  • An antibody construct can comprise a second binding domain that specifically binds
  • An antibody construct can comprise a second binding domain that is a CD40 agonist.
  • An antibody construct can comprise a second binding domain that binds CD40, wherein the second binding domain comprises: a) a V H sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 2, and a V L sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 7; b) a V H sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 581, and a V L sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 586; c) a V H sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 591 , and a V L sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 596; d) a V H sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 601 , and a V L sequence having at least 80% sequence
  • An antibody construct can comprise a second binding domain that specifically binds DEC-205.
  • An antibody construct can comprise a second binding domain that binds DEC-205, wherein the second binding domain comprises: a) a V H sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 233, and a V L sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 238; or b) a V H sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 246, and a V L sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 251.
  • An antibody construct can comprise a second binding domain that specifically binds to CD36 mannose scavenger receptor 1.
  • An antibody construct can comprise a second binding domain that binds CD36 mannose scavenger receptor 1, wherein the second binding domain comprises a V H sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 658, and a V L sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 659.
  • An antibody construct can comprise a second binding domain that specifically binds to CLEC9A.
  • An antibody construct can comprise a second binding domain that binds CLEC9A, wherein the second binding domain comprises a V H sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 660, and a V L sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 661.
  • An antibody construct can comprise a second binding domain that specifically binds to PD-Ll.
  • An antibody construct can comprise a second binding domain that binds PD-Ll, wherein the second binding domain comprises a V H sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 901, and a V L sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 902; or wherein the second binding domain comprises a V H sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 890, and a V L sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 891; or wherein the second binding domain comprises a V H sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 892, and a V L sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 893.
  • An antibody construct can comprise a second binding domain and an Fc domain, wherein the second binding domain and the Fc domain comprise an antibody.
  • An antibody construct can comprise a heavy chain and a light chain that target an antigen expressed by an antigen presenting cell.
  • An antibody construct can comprise an antibody light chain.
  • An antibody construct can comprise a light chain comprising at least 80% sequence identity to any light chain sequence in TABLE 7.
  • An antibody construct can comprise an antibody heavy chain.
  • An antibody construct can comprise a heavy chain comprising at least 80% sequence identity to any heavy chain sequence in TABLE 7.
  • An antibody construct can comprise at least 80% sequence identity to any sequence in TABLE 7.
  • An antibody construct can comprise a heavy and light chains having a pair of sequences in TABLE 7.
  • An antibody construct can comprise a heavy chain and a light chain that target an antigen expressed by an antigen presenting cell.
  • An antibody construct can comprise a first binding domain and an Fc domain, wherein the first binding domain and the Fc domain comprise an antibody.
  • An antibody construct may comprise an anti-CD40 antibody, the antibody construct comprising: a) a heavy chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 1 and a light chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 6; b) a heavy chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 577 or SEQ ID NO: 578, and a light chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 579; c) a heavy chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 580, and a light chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 585;
  • An antibody construct can comprise a first binding domain and an Fc domain, wherein the first binding domain and the Fc domain comprise an antibody.
  • An antibody construct may comprise an anti-DEC-205 antibody, the construct comprising: a) a heavy chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 232, and a light chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 237; or b) a heavy chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 245, and a light chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 250.
  • An antibody construct can comprise a first binding domain and an Fc domain, wherein the first binding domain and the Fc domain comprise an antibody.
  • a composition may comprise an anti-CLEC12A antibody, the construct comprising: a) a heavy chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 662, and a light chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 665; b) a heavy chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 663, and a light chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 665; or c) a heavy chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 664, and a light chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 665.
  • An antibody construct can comprise a first binding domain and an Fc domain, wherein the first binding domain and the Fc domain comprise an antibody.
  • An antibody construct may comprise an anti-BDCA-2 antibody, the construct comprising: a) a heavy chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 666, and a light chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 669; b) a heavy chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 667, and a light chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 670; or c) a heavy chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 668, and a light chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 671.
  • An antibody construct can comprise a first binding domain, a second binding domain, and an Fc domain, wherein the first binding domain and the second binding domain are attached to the Fc domain.
  • the first binding domain and the second binding domain can be attached to the Fc domain as a fusion protein.
  • the first binding domain can be attached to the Fc domain at an N- terminal end of the Fc domain, wherein the second binding domain can be attached to the Fc domain at a C-terminal end.
  • the first binding domain can be attached to the Fc domain at an N- terminal end of the Fc domain, wherein the second binding domain can be attached to the Fc domain at a C-terminal end via a polypeptide linker ranging from 10 to 25 amino acids.
  • the first binding domain can be attached to the Fc domain at a C-terminal end of the Fc domain, wherein the second binding domain can be attached to the Fc domain at an N-terminal end.
  • a second binding domain and an Fc domain can comprise an antibody and a first binding domain can comprise a single chain variable fragment (scFv).
  • a single chain variable fragment can comprise a heavy chain variable domain and a light chain variable domain of an antibody.
  • the first binding domain of the fusion protein can be attached to the second binding domain at a heavy chain variable domain of the single chain variable fragment of the first binding domain (HL orientation).
  • the first binding domain of the fusion protein can be attached to the second binding domain at a light chain variable domain of the single chain variable fragment of the first binding domain (LH orientation).
  • LH orientation the first binding domain and the second binding domain can be attached via a polypeptide linker varying in length from 15 to 25 amino acids.
  • an antibody construct can comprise an antibody (having two antigen binding domains and an Fc domain) and the second binding domain can comprise a single chain variable fragment (scFv).
  • the second binding domain of the fusion protein can be attached to the first binding domain at a heavy chain variable domain of the single chain variable fragment of the first binding domain (HL orientation).
  • the second binding domain of the fusion protein can be attached to the first binding domain at a light chain variable domain of the single chain variable fragment of the first binding domain (LH orientation).
  • An antibody construct can comprise a first binding domain and a second binding domain, wherein the second binding domain can be attached to the first binding domain.
  • the antibody construct can comprise an antibody comprising a light chain and a heavy chain or pair of heavy and light chains.
  • the first binding domain can comprise an Fab fragment of the light and heavy chains.
  • the second binding domain can be attached to the light chain at a C-terminus or C- terminal end of the light chain as a fusion protein.
  • the second binding domain can comprise a single chain variable fragment (scFv).
  • a first binding domain can be specific for any of the tumor antigens, e.g., CEA, HER2, TROP2, Claudin-6 (CLDN6), Claudin- 16 (CLDN16), CLDN18.2, RON, LY6E, FRA, DLL3, PTK7, Uroplakin-IB (UPK1B), LIV1, ROR1, STRA6, TMPRSS3, TMPRSS4, TMEM238, Clorfl86, Fos-related antigen 1, VEGFR1, endoglin, VTCN1 (B7-H4), VISTA, or gpNMB, and an scFv can be a binding domain with a specificity selected from a group consisting of antigens on antigen presenting cells, such as CD40, DEC205, and PD-L1.
  • the tumor antigens e.g., CEA, HER2, TROP2, Claudin-6 (CLDN6), Claudin- 16 (CLDN16), CLDN18.2, RON
  • An antibody construct can comprise a first binding domain, a second binding domain, and an Fc domain, wherein the first binding domain and the second binding domain are attached to the Fc domain as a fusion protein.
  • the second binding domain of the fusion protein can specifically target an antigen with at least 80% sequence identity to CD40.
  • the second binding domain of the fusion protein can be a CD40 agonist.
  • the first binding domain of the fusion protein can target a tumor antigen.
  • the construct can comprise a fusion protein comprising a heavy chain (HC) attached to a single chain variable fragment.
  • the construct comprising the fusion protein can comprise at least 80% sequence identity to any sequence in TABLE 4.
  • the construct comprising the fusion protein can comprise at least 80% sequence identity to any sequence of a heavy chain CD40 monoclonal antibody (mAb) with tumor ScFv in TABLE 4.
  • the construct can comprise a fusion protein comprising at least 80% sequence identity to any sequence of a heavy chain CD40 mAb with tumor ScFv in TABLE 4 and a light chain comprising at least 80% sequence identity to SEQ ID NO: 6.
  • the construct comprising the fusion protein can comprise at least 80% sequence identity to any sequence of a heavy chain tumor mAb with CD40 ScFv in TABLE 4.
  • the construct comprising the fusion protein can comprise at least 80% sequence identity to any sequence of a heavy chain tumor antigen mAb with CD40 ScFv in TABLE 4, and at least 80% sequence identity to a light chain mAb for the tumor antigen in
  • An antibody construct can comprise a first binding domain and a second binding domain, wherein the second binding domain can be attached to the first binding domain.
  • An antibody construct can comprise a first binding domain, a second binding domain, and an Fc domain, wherein the second binding domain can be attached to the first binding domain.
  • the second binding domain can be attached at a C-terminal end of the first binding domain as a fusion protein.
  • the first binding domain can comprise a Fab fragment comprising a light chain, wherein the second binding domain can be attached at a C-terminal end of the light chain as a fusion protein.
  • the second binding domain of the fusion protein can comprise a single chain variable fragment (scFv).
  • the second binding domain of the fusion protein can be attached to the first binding domain at a heavy chain variable domain of the single chain variable fragment of the first binding domain (HL orientation).
  • the second binding domain of the fusion protein can be attached to the first binding domain at a light chain variable domain of the single chain variable fragment of the first binding domain (LH orientation).
  • HL orientation heavy chain variable domain of the single chain variable fragment of the first binding domain
  • LH orientation light chain variable domain of the single chain variable fragment of the first binding domain
  • a fusion protein comprising a light chain of an anti-CEA antibody attached to an anti-CD40 scFv in the LH orientation can be illustrated by SEQ ID NO: 842.
  • the fusion sequences comprising an scFv sequence are in the HL orientation unless indicated otherwise (e.g., sequence name recites "(LH)" indicating light heavy orientation).
  • the first binding domain of the fusion protein can target a tumor antigen.
  • the second binding domain of the fusion protein can target an APC antigen.
  • the second binding domain of the fusion protein can target CD40.
  • the first binding domain can comprise a Fab fragment comprising a light chain, wherein the second binding domain is attached at a C-terminal end of the light chain as a fusion protein.
  • the construct comprising the fusion protein can comprise at least 80% sequence identity to any sequence in TABLE 9.
  • the construct comprising the fusion protein can comprise at least 80% sequence identity to any sequence of a light chain CD40 mAb with tumor ScFv in TABLE 9.
  • the construct can comprise a fusion protein comprising at least 80% sequence identity to any sequence of a light chain CD40 mAb with tumor ScFv in TABLE 9 and a heavy chain comprising at least 80% sequence identity to SEQ ID NO: 1.
  • the construct comprising the fusion protein can comprise at least 80% sequence identity to any sequence of a light chain tumor mAb with CD40 ScFv in TABLE 9.
  • the construct can comprise a fusion protein comprising at least 80% sequence identity to any sequence of a light chain tumor antigen mAb with CD40 ScFv in TABLE 9, and at least 80% sequence identity to a heavy chain mAb for the tumor antigen in TABLE 3.
  • An antibody construct can comprise a first binding domain, a second binding domain, and an Fc domain, wherein the first binding domain and the second binding domain are attached to the Fc domain as a fusion protein.
  • the first binding domain of the fusion protein can specifically target an antigen with at least 80% sequence identity to DEC-205.
  • the second binding domain of the fusion protein can target a tumor antigen.
  • the construct can comprise a fusion protein comprising a heavy chain attached to a single chain variable fragment.
  • the construct comprising the fusion protein can comprise at least 80% sequence identity to any sequence in TABLE 8.
  • the construct comprising the fusion protein can comprise at least 80% sequence identity to any sequence of a heavy chain DEC-205 mAb with tumor ScFv in TABLE 8.
  • the construct can comprise a fusion protein comprising at least 80% sequence identity to any sequence of a heavy chain DEC-205 mAb with tumor ScFv in TABLE 8 and a peptide comprising at least 80% sequence identity to SEQ ID NO: 237.
  • the construct comprising the fusion protein can comprise at least 80% sequence identity to any sequence of a heavy chain tumor antigen mAb with CD40 ScFv in TABLE 8.
  • the construct comprising the fusion protein can comprise at least 80% sequence identity to any sequence of a heavy chain tumor antigen mAb with CD40 ScFv in TABLE 8, and at least 80% sequence identity to a heavy chain rnAb for the tumor antigen in TABLE 3.
  • the second binding domain of the fusion protein can target DEC-205.
  • the construct comprising the fusion protein can comprise at least 80% sequence identity to any sequence in
  • the construct comprising the fusion protein can comprise at least 80% sequence identity to any sequence of a light chain DEC-205 rnAb with tumor ScFv in TABLE 10.
  • the construct comprising a fusion protein can comprise at least 80% sequence identity to any sequence of a light chain DEC-205 rnAb with tumor ScFv in TABLE 10 and at least 80% sequence identity to SEQ ID NO: 237.
  • the construct comprising the fusion protein can comprise at least 80% sequence identity to any sequence of a light chain tumor mAb with DEC-205 ScFv in TABLE 10.
  • the construct comprising a fusion protein can comprise at least 80% sequence identity to any sequence of a light chain tumor antigen mAb with DEC-205 ScFv in TABLE 10, and at least 80% sequence identity to a heavy chain mAb for the tumor antigen in TABLE 3.
  • the second binding domain of the fusion protein can specifically bind to an antigen of an
  • the second binding domain of the fusion protein can specifically bind to an antigen with at least 80% sequence identity to CD40.
  • the second binding domain of the fusion protein can be a CD40 agonist.
  • the second binding domain of the fusion protein can specifically bind to an antigen with at least 80% sequence identity to CD40.
  • the second binding domain of the fusion protein can specifically bind to an antigen with at least 80% sequence identity to DC-SIGN.
  • the second binding domain of the fusion protein can specifically bind to an antigen with at least 80% sequence identity to CD36 mannose scavenger receptor.
  • the second binding domain of the fusion protein can specifically bind to an antigen with at least 80% sequence identity to CLEC12A.
  • the second binding domain of the fusion protein can specifically bind to an antigen with at least 80% sequence identity to BDCA-2.
  • the second binding domain of the fusion protein can specifically bind to an antigen with at least
  • the second binding domain of the fusion protein can specifically bind to an antigen with at least 80% sequence identity to CD40, CD47, TNFR2,
  • BDCA-2 OX40L, 41BBL, CD204, MARCO, CLEC5A, Dectin 1, Dectin 2, CLECIOA, CD206,
  • the first binding domain of the fusion protein can specifically bind to a tumor antigen.
  • the first binding domain of the fusion protein can specificall bind to an antigen with at least 80% sequence identity to CD5,
  • B7-DC (PD-L2), HLD-DR, carcinoembryonic antigen (CEA), TAG-72, EpCAM, MUC1,
  • PDGFR-B MAD-CT-2, ROR2, TRAIL 1, MAGE A4, MAGE C2, GAGE, EGFR, CMET, HER3, EPCAM, CA6, NAPI2B, TROP2, Claudin-6 (CLDN6), Claudin-16 (CLDN16),
  • the second binding domain of the fusion protein can target a tumor antigen.
  • the second binding domain of the fusion protein can specifically bind to an antigen selected from CD5, CD19, CD20, CD25, CD37, CD30, CD33, CD45, CAMPATH-1, BCMA, CS-1, PD- Ll, B7-H3, B7-DC (PD-L2), HLD-DR, carcinoembryonic antigen (CEA), TAG-72, EpCAM, MUC1, MUC15, MUC16, fo late-binding protein, A33, G250, pro state- specific membrane antigen (PSMA), ferritin, GD2, GD3, GM2, Le y , CA-125, CA19-9, epidermal growth factor, pl85HER2, IL-2 receptor, EGFRvIII (de2-7 EGFR), fibroblast activation protein (FAP), tenascin, a metalloproteinase, endosialin, vascular endothelial growth
  • FAP
  • the first binding domain of the fusion protein can specifically bind to an antigen with at least 80% sequence identity to CD40.
  • the first binding domain of the fusion protein can be a CD40 agonist.
  • the first binding domain of the fusion protein can specifically bind to an antigen with at least 80% sequence identity to DEC-205.
  • the first binding domain of the fusion protein can specifically bind to an antigen with at least 80% sequence identity to CEA, HER2, TROP2, Claudin-6 (CLDN6), Claudin-16 (CLDN16),
  • the first binding domain of the fusion protein can specifically bind to an antigen with at least 80% sequence identity to CD40, CD47, TNFR2, DEC-205, PD-L1, CD36 mannose scavenger receptor 1, CLEC9A, DC-SIGN, CLEC12A, BDCA-2, OX40L, 41BBL, CD204, MARCO, CLEC5A, Dectin 1, Dectin 2, CLECIOA, CD206, CD64, CD32A, CD 16 A, HVEM, CD38, TREM2, CSF1R, or CD32B.
  • An antibody construct can comprise a first binding domain, a second binding domain, and a third binding domain.
  • An antibody construct can comprise a first binding domain, a second binding domain, a third binding domain, and an Fc domain.
  • the first binding domain and the second binding domain can be attached to the Fc domain.
  • the first and second binding domains are described herein throughout the specification.
  • the first binding domain can be attached to the Fc domain at an N-terminal end of the Fc domain.
  • the second binding domain can be attached at a C-terminal end of the Fc domain.
  • the second binding domain can be attached at a C-terminal end of the Fc domain via a polypeptide linker having a length of 10 to 25 amino acid.
  • the third binding domain can be attached to a C-terminal end of the first binding domain.
  • the third binding domain can be attached at a C-terminal end of the Fc domain via a polypeptide linker having a length of 10 to 25 amino acid.
  • the third binding domain can be attached to a C-terminal end of a light chain of the first binding domain.
  • One or more of the first binding domain, the second binding domain, the third binding domain, and the Fc domain can be attached as a fusion protein.
  • the first binding domain can comprise a Fab fragment comprising a light chain, wherein the second binding domain is attached at a C-terminal end of the light chain as a fusion protein.
  • the second binding domain of the fusion protein can comprise a single chain variable fragment (scFv).
  • the second binding domain of the fusion protein can be attached to the Fc domain at a heavy chain variable domain of the single chain variable fragment of the second binding domain (HL orientation).
  • the second binding domain of the fusion protein can be attached to the Fc domain at a light chain variable domain of the single chain variable fragment of the second binding domain (LH orientation).
  • the third binding domain of the fusion protein can comprise a single chain variable fragment (scFv).
  • the antibody construct can comprise a fusion protein comprising the third binding domain attached to the first binding domain having at least 80% sequence identity to any sequence in TABLE 9 or TABLE 10.
  • the third binding domain of the fusion protein can be attached to the first binding domain at a heavy chain variable domain of the single chain variable fragment of the first binding domain (HL orientation).
  • the third binding domain of the fusion protein can be attached to the first binding domain at a light chain variable domain of the single chain variable fragment of the first binding domain (LH orientation).
  • the third binding domain of the fusion protein can target an antigen of an immune cell, such as an antigen presenting cell (APC).
  • the third binding domain of the fusion protein can specifically bind to an antigen with at least 80% sequence identity to CD40.
  • the third binding domain of the fusion protein can be a CD40 agonist.
  • the third binding domain of the fusion protein can specifically bind to an antigen with at least 80% sequence identity to DEC-205.
  • the third binding domain of the fusion protein can specifically bind to an antigen with at least 80% sequence identity to CD40, CD47, TNFR2, DEC-205, PD-Ll, CD36 mannose scavenger receptor 1, CLEC9A, DC-SIGN, CLEC12A, BDCA-2, OX40L, 41BBL, CD204, MARCO, CLEC5A, Dectin 1, Dectin 2, CLECIOA, CD206, CD64, CD32A, CD 16 A, HVEM, CD38, TREM2, CSF1R, or CD32B.
  • the third binding domain can target a tumor antigen.
  • the third binding domain of the fusion protein can specifically bind to an antigen with at least 80% sequence identity to CD5, CD19, CD20, CD25, CD37, CD30, CD33, CD45, CAMPATH-1, BCMA, CS-1,
  • PD-Ll B7-H3, B7-DC (PD-L2), HLD-DR, carcinoembryonic antigen (CEA), TAG-72, EpCAM,
  • PSMA pro state-specific membrane antigen
  • EGFRvIII (de2-7 EGFR), fibroblast activation protein (FAP), tenascin, a metalloproteinase, endosialin, vascular endothelial growth factor, ⁇ 3, WTl, LMP2, HPV E6, HPV E7, Her-2/neu, p53 nonmutant, NY-ESO-1, MelanA/M ART 1 , Ras mutant, gplOO, p53 mutant, PR1, bcr-abl, tyrosinase, survivin, PSA, hTERT, a Sarcoma translocation breakpoint protein, EphA2, PAP,
  • ML-IAP AFP, ERG, NA17, PAX3, ALK, androgen receptor
  • cyclin B l MYCN, RhoC, TRP-2, mesothelin (MSLN), PSCA, MAGE Al, MAGE A3, CYP1B 1, PLAV1, BORIS, GloboH, ETV6-
  • PDGFR-B MAD-CT-2, ROR2, TRAIL 1, MAGE A4, MAGE C2, GAGE, EGFR, CMET,
  • the third binding domain of the fusion protein can specifically bind to an antigen selected from CD5, CD19, CD20, CD25, CD37, CD30, CD33, CD45, CAMPATH-1, BCMA, CS-1, PD-L1, B7-H3, B7-DC (PD-L2), HLD-DR, carcinoembryonic antigen (CEA), TAG-72, EpCAM, MUC1, MUC15, MUC16, fo late-binding protein, A33, G250, pro state- specific membrane antigen (PSMA), ferritin, GD2, GD3, GM2, Le y , CA-125, CA19-9, epidermal growth factor, pl85HER2, IL-2 receptor, EGFRvIII (de2-7 EGFR), fibroblast activation protein (FAP), tenascin, a metalloproteinase, endosialin, vascular endosialin, vascular endosialin, vascular endosialin, vascular endos
  • An antibody construct can comprise a first binding domain targeting CD40 and a second binding domain targeting DEC-205.
  • an antibody construct can comprise a first binding domain targeting DEC-205 and a second binding domain targeting CD40.
  • An antibody construct can comprise a first binding domain, a second binding domain, and an Fc domain. The first binding domain and the second binding domain can be attached to the Fc domain. The first binding domain can be attached to the Fc domain at an N-terminal end of the Fc domain, wherein the second binding domain is attached to the Fc domain at a C-terminal end of the Fc domain.
  • the second binding domain can be attached to the Fc domain at an N-terminal end of the Fc domain, wherein the first binding domain is attached to the Fc domain at a C-terminal end of the Fc domain.
  • An antibody construct can comprise a fusion protein comprising a first binding domain targeting CD40 and a second binding domain targeting DEC-205.
  • the fusion protein can comprise at least 80% sequence identity to any sequence in TABLE 11.
  • antibody constucts expressing proteins having the sequences referenced in Tables 1-11 can have a dissociation constant (IQ) that is less than 10 nM for the antigen of the first binding domain.
  • the antibody constructs expressing proteins having the sequences referenced in Tables 1-11 can have a dissociation constant (IQ) that is less than 10 nM for the antigen of the second binding domain.
  • the antibody constructs expressing the sequences referenced in Tables 1-11 can have a dissociation constant (K d ) that is less than 10 nM for the antigen of the third binding domain.
  • the antibody constructs can have a dissociation constant (K d ) for the antigen of the first binding domain that is less than 1 nM, less than 100 pM, less than 10 pM, less than 1 pM, or less than 0.1 pM.
  • the antibody constructs can have a dissociation constant (K d ) for the antigen of the second binding domain that is less than 1 nM, less than 100 pM, less than 10 pM, less than 1 pM, or less than 0.1 pM.
  • the antibody constructs can have a dissociation constant (K d ) for the antigen of the third binding domain that is less than 1 nM, less than 100 pM, less than 10 pM, less than 1 pM, or less than 0.1 pM.
  • K d dissociation constant
  • An anti-CD40 light chain can be expressed with an anti-CD40 heavy chain or fragment thereof, wherein the light and heavy chains specifically bind to CD40.
  • the anti-CD40 light chain can also be expressed with an anti-CD40 heavy chain or fragment thereof to form an anti-CD40 antibody or fragment thereof, wherein the antibody or antibody fragment specifically binds to CD40.
  • the anti-CD40 antibody or fragment thereof can be purified, and can be combined with a pharmaceutically acceptable carrier.
  • An anti-DEC-205 light chain can be expressed with any anti-DEC-205 heavy chain or fragment thereof, wherein the light and heavy chains specifically bind to DEC-205.
  • the anti- DEC-205 light chain can also be expressed with an anti-DEC-205 heavy chain or fragment thereof to form an anti-DEC-205 antibody or fragment thereof, wherein the antibody or antibody fragment specifically binds to DEC-205.
  • the anti-DEC-205 antibody or fragment thereof can be purified, and can be combined with a pharmaceutically acceptable carrier.
  • An anti-PD-Ll light chain can be expressed with any anti-PD-Ll heavy chain or fragment thereof, wherein the light and heavy chains specifically bind to PD-L1.
  • the anti-PD-Ll light chain can also be expressed with an anti-PD-Ll heavy chain or fragment thereof to form an anti- PD-Ll antibody or fragment thereof, wherein the antibody or antibody fragment specifically binds to PD-L1.
  • the anti-PD-Ll antibody or fragment thereof can be purified, and can be combined with a pharmaceutically acceptable carrier.
  • An anti-tumor antigen light chain can be expressed with any anti-tumor antigen heavy chain or fragment thereof, wherein the antibody or antibody fragment specifically binds to tumor antigen.
  • the anti-tumor antigen light chain can also expressed with any anti-tumor antigen heavy chain or fragment thereof to form an anti-tumor antigen antibody or fragment thereof.
  • the antitumor antibody or fragment thereof can be purified, and can be combined with a
  • An antibody construct can comprise an antibody heavy chain.
  • a heavy chain can be a heavy chain of an anti-CD40 antibody which can bind a CD40 antigen.
  • a heavy chain of an anti- CD40 antibody can be an IgGl isotype.
  • a heavy chain of an anti-CD40 antibody can be dacetuzumab.
  • An antibody construct can comprise an antibody light chain.
  • a light chain can be a light chain of an anti-CD40 antibody which can bind a CD40 antigen.
  • a light chain of an anti-CD40 antibody can be dacetuzumab.
  • An antibody construct can comprise an antibody heavy chain.
  • a heavy chain can be a heavy chain of an anti-CD40 antibody which can bind a CD40 antigen.
  • a heavy chain of an anti- CD40 antibody can be an IgG4 isotype.
  • a heavy chain of an anti-CD40 antibody can be bleselumab.
  • An antibody construct can comprise an antibody light chain.
  • a light chain can be a light chain of an anti-CD40 antibody which can bind a CD40 antigen.
  • a light chain of an anti-CD40 antibody can be bleselumab.
  • An antibody construct can comprise an antibody heavy chain.
  • a heavy chain can be a heavy chain of an anti-CD40 antibody which can bind a CD40 antigen.
  • a heavy chain of an anti- CD40 antibody can be an IgGl isotype.
  • a heavy chain of an anti-CD40 antibody can be lucatumumab.
  • An antibody construct can comprise an antibody light chain.
  • a light chain can be a light chain of an anti-CD40 antibody which can bind a CD40 antigen.
  • a light chain of an anti-CD40 antibody can be lucatumumab.
  • An antibody construct can comprise an antibody heavy chain.
  • a heavy chain can be a heavy chain of an anti-CD40 antibody which can bind a CD40 antigen.
  • a heavy chain of an anti- CD40 antibody can be an IgGl isotype.
  • a heavy chain of an anti-CD40 antibody can be ADC- 1013.
  • An antibody construct can comprise an antibody light chain.
  • a light chain can be a light chain of an anti-CD40 antibody which can bind a CD40 antigen.
  • a light chain of an anti-CD40 antibody can be ADC- 1013.
  • An antibody construct can comprise an antibody heavy chain.
  • a heavy chain can be a heavy chain of an anti-CD40 antibody which can bind a CD40 antigen.
  • a heavy chain of an anti- CD40 antibody can be the humanized rabbit antibody APX005.
  • An antibody construct can comprise an antibody light chain.
  • a light chain can be a light chain of an anti-CD40 antibody which can bind a CD40 antigen.
  • a light chain of an anti-CD40 antibody can be the humanized rabbit antibody APX005.
  • An antibody construct can comprise an antibody heavy chain.
  • a heavy chain can be a heavy chain of an anti-CD40 antibody which can bind a CD40 antigen.
  • a heavy chain of an anti- CD40 antibody can be Chi Lob 7/4.
  • An antibody construct can comprise an antibody light chain.
  • a light chain can be a light chain of an anti-CD40 antibody which can bind a CD40 antigen.
  • a light chain of an anti-CD40 antibody can be Chi Lob 7/4.
  • An antibody construct can comprise an antibody heavy chain.
  • a heavy chain can be a heavy chain of an anti-CD40 antibody which can bind a CD40 antigen.
  • a heavy chain of an anti- CD40 antibody can be an IgGl isotype.
  • a heavy chain of an anti-CD40 antibody can be SBT- 040-G1WT.
  • An antibody construct can comprise an antibody heavy chain.
  • a heavy chain can be a heavy chain of an anti-CD40 antibody which can bind a CD40 antigen.
  • a heavy chain of an anti- CD40 antibody can be an IgGl isotype.
  • a heavy chain of an anti-CD40 antibody can be SBT- 040 VH-hlgGl wt.
  • a heavy chain of an anti-CD40 antibody can be an IgG2 isotype.
  • a heavy chain of an anti-CD40 antibody can be SBT-040-G2.
  • An antibody construct can comprise an antibody with modifications occurring at least at one amino acid residue. Modifications can be substitutions, additions, mutations, deletions, or the like. An antibody modification can be an insertion of an unnatural amino acid.
  • An antibody construct can comprise a light chain of an amino acid sequence having at least one, two, three, four, five, six, seven, eight, nine, or ten modifications but not more than 40, 35, 30, 25, 20, 15, or 10 modifications of the amino acid sequence relative to the natural or original amino acid sequence.
  • An antibody construct can comprise a heavy chain with an amino acid sequence having at least one, two, three, four, five, six, seven, eight, nine or ten
  • An antibody construct can have an Fc domain of an IgGl isotype.
  • An antibody construct can have an Fc domain of an IgG2 isotype.
  • An antibody construct can have an Fc domain of an IgG3 isotype.
  • An antibody construct can have an Fc domainof an IgG4 isotype.
  • An antibody construct can have an Fc domain of a hybrid isotype comprising constant regions from two or more isotypes.
  • An antibody construct can be an anti-CD40 antibody, in which the anti-CD40 antibody can be a monoclonal human antibody comprising a wild-type sequence of an IgGl isoform, in particular, at an Fc region of the antibody.
  • An antibody constructs disclosed herein can be non-natural, designed, and/or engineered.
  • Antibody constructs disclosed herein can be non-natural, designed, and/or engineered scaffolds comprising an antigen binding domain.
  • Antibody constructs disclosed herein can be no n- natural, designed, and/or engineered antibodies.
  • Antibody constructs can be monoclonal antibodies.
  • Antibody constructs can be human antibodies.
  • Antibody constructs can be humanized antibodies.
  • Antibody constructs can be monoclonal humanized antibodies.
  • Antibody construct can be recombinant antibodies.
  • the K d for binding of the Fc domain to an Fc receptor of an antibody construct as described herein can increase when the tumor antigen binding domain is bound to its tumor antigen as compared to the K d for binding of the Fc domain to an Fc receptor of a construct as described herein when the tumor antigen binding domain is not bound to its tumor antigen.
  • an antibody construct as described herein can have a K d for binding of the Fc domain to an Fc receptor in the presence of the binding domain that binds to an antigen on an immune cell, such as an antigen presenting cell, and the tumor target binding domain when the tumor target binding domain is bound to its tumor antigen that can be greater than or greater than about 100 nM.
  • the K d for binding of the Fc domain to an Fc receptor in the presence of the binding domain that binds to an antigen on an immune cell, such as an antigen presenting cell, and the tumor target binding domain when the tumor target binding domain is bound to its tumor antigen can be or can be about 100 nM, 200 nM, 300 nM, 400 nM, 500 nM, or 1000 nM.
  • the 3 ⁇ 4 for binding of the Fc domain to an Fc receptor in the presence of the binding domain that binds to an antigen on an immune cell, such as an antigen presenting cell, and the tumor target binding domain when the tumor target binding domain is bound to its tumor antigen can be from 100 nM to 200 nM, 100 nM to 300 nM, 100 nM to 400 nM, 100 nM to 500 nM, or 100 nM to 1000 nM.
  • the antibody construct as described herein can have a K d for binding of the Fc domain to an Fc receptor in the presence of the binding domain that binds to an antigen on an immune cell, such as an antigen presenting cell, and a tumor antigen binding domain when the tumor antigen binding domain is not bound to the tumor antigen is no greater than about 100 nM and is no greater than about 100 times a K d for binding of the Fc domain to the Fc receptor in an absence of the binding domain that binds to an antigen on an immune cell and a tumor antigen binding domain.
  • the K d for binding of the binding domain of an antibody construct as described herein that binds to an antigen on an immune cell can increase when the tumor antigen binding domain is bound to its tumor antigen as compared to the K d for binding of the binding domain that binds to an antigen on an immune cell when the tumor antigen binding domain is not bound to its tumor antigen.
  • an antibody construct can comprise a K d for binding of the binding domain that binds to an antigen on an antigen presenting cell when the tumor antigen binding domain is bound to its tumor antigen can be greater than or greater than about ⁇ .
  • the K d for binding of the binding domain that binds to an antigen on an immune cell, such as an antigen presenting cell, when the tumor antigen binding domain is bound to its tumor antigen can be or can be about 100 nM, 200 nM, 300 nM, 400 nM, 500 nM, or 1000 nM.
  • IQ for binding of the binding domain that binds to an antigen on an immune cell when the tumor antigen binding domain is bound to its tumor antigen can be from 100 nM to 200 nM, 100 nM to 300 nM, 100 nM to 400 nM, 100 nM to 500 nM, or 100 nM to 1000 nM.
  • the effect of the tumor antigen binding domain and the binding domain that binds to an antigen on the immune cell together can be to cluster the antibody constructs or conjugates on cells expressing tumor antigen, and thus clustering immune cells such as antigen presenting cells around cancerous cells and at tumor sites resulting in activation of the antigen presenting cell effector functions.
  • this activation of the antigen on the antigen presenting cell only occurs when the tumor targeting antibody construct is bound to its tumor antigen.
  • An antigen presenting cell effector function can include antibody dependent cellular cytotoxicity (ADCC) of the tumor antigen expressing cell, which can occur when a bispecific tumor targeting antibody construct is bound to its tumor antigen.
  • ADCC of the tumor antigen expressing cell only occurs with a bispecific tumor targeting antibody construct is bound to its tumor antigen.
  • a bispecific tumor targeting antibody construct density of greater than 1000, 2000, 3000, 4000, 5000, 6000, 7000, 8000, 9000, 10,000 or more per cell induces signaling in the antigen presenting cell.
  • Signaling can suitably be measured in vitro using a cell line expressing the tumor antigen bound by the target antigen binding domain, and primary antigen presenting cells isolated from a human subject. Signaling can be assessed as cytokine release, chemokine release, or increased expression of cell surface markers.
  • a bispecific tumor targeting antibody construct density of greater than 1000, 2000, 3000, 4000, 5000, 6000, 7000, 8000, 9000, 10,000 or more per cell induces ADCC of the cells expressing tumor antigen.
  • ADCC can suitably be measured in vitro using a cell line expressing the tumor antigen bound by the target antigen binding domain, and cells such as NK cells and/or macrophages isolated from a human subject. ADCC can be determined by the frequency of remaining tumor antigen expressing cells in the co-culture.
  • the antibody constructs as described herein can specifically bind to a tumor antigen in a cluster of constructs or conjugates, and this clustering can induce a signal in an antigen presenting cell.
  • the constructs as described herein can specifically bind to a tumor antigen in a cluster of constructs, and this clustering can induce antibody dependent cellular cytotoxicity.
  • the constructs as described herein can specifically bind to a tumor antigen in a cluster of constructs and this clustering can result in an increased avidity for an antigen on an antigen presenting cell.
  • the antibody constructs as described herein can specifically bind to a tumor antigen in a cluster of constructs and this clustering can result in an increased avidity of the Fc domain for an Fc receptor.
  • Sequences that can be used to produce antibodies for antibody constructs can include leader sequences.
  • Leader sequences can include signal sequences.
  • Leader sequences useful with the antibody construct and methods described herein can include, but are not limited to, an amino acid sequence comprising MRLP AQLLGLLLLWFPGS RC (SEQ ID NO: 847),
  • MDWTWRILFLVAAATGAHS SEQ ID NO: 848
  • MRAWIFFLLCLAGRALA SEQ ID NO: 849
  • An antibody construct can comprise an Fc region with an Fc domain.
  • An Fc domain is a structure that can bind to Fc receptors.
  • An antibody construct can comprise an Fc domain.
  • Fc domains can be bound by Fc receptors (FcRs).
  • Fc domains can be from antibodies.
  • An Fc domain can be at least 80% homologous to an Fc domain from an antibody.
  • An Fc region can be in a scaffold.
  • An Fc region with an Fc domain can be in an antibody scaffold.
  • An Fc region with an Fc domain can be in a non-antibody scaffold.
  • An antibody construct can comprise an Fc region with an Fc domain in an antibody scaffold.
  • An antibody construct can comprise an Fc region with an Fc domain in a non-antibody scaffold.
  • An Fc domain can be in a scaffold.
  • An Fc domain can be in an antibody scaffold.
  • An Fc domain can be in a non-antibody scaffold.
  • An antibody construct can comprise an Fc domain in an antibody scaffold.
  • An antibody construct can comprise an Fc domain in a non-antibody scaffold.
  • Fc domains of antibodies can be bound by Fc receptors (FcRs).
  • An Fc domain can be a portion of the Fc region of an antibody.
  • FcRs can bind to an Fc domain of an antibody.
  • FcRs can bind to an Fc domain of an antibody bound to an antigen.
  • FcRs are organized into classes (e.g.
  • the FcaR class can bind to IgA and includes several isoforms, FcaRI (CD89) and Fco jR.
  • the FcyR class can bind to IgG and includes several isoforms, FcyRI (CD64), FcyRIIA (CD32a), FcyRIIB (CD32b), FcyRIIIA (CD 16a), and FcyRIIIB (CD 16b).
  • An FcyRIIIA (CD 16a) can be an FcyRIIIA (CD 16a) F158 variant.
  • An FcyRIIIA (CD 16a) can be an FcyRIIIA (CD 16a) V158 variant.
  • Each FcyR isoform can differ in affinity to the Fc region of the IgG antibody.
  • FcyRI can bind to IgG with greater affinity than FcyRII or FcyRIII.
  • the affinity of a particular FcyR isoform to IgG can be controlled, in part, by a glycan (e.g., oligosaccharide) at position CH 2 84.4 of the IgG antibody.
  • a glycan e.g., oligosaccharide
  • fucose containing CH 2 84.4 glycans can reduce IgG affinity for FcyRIIIA.
  • GO glucans can have increased affinity for FcyRIIIA due to the lack of galactose and terminal GlcNAc moiety.
  • Binding of an Fc domain to an FcR can enhance an immune response.
  • FcR-mediated signaling that can result from an Fc region binding to an FcR can lead to the maturation of immune cells.
  • FcR-mediated signaling that can result from an Fc domain binding to an FcR can lead to the maturation of dendritic cells.
  • FcR-mediated signaling that can result from an Fc domain binding to an FcR can lead to antibody dependent cellular cytotoxicity.
  • FcR-mediated signaling that can result from an Fc domain binding to an FcR can lead to more efficient immune cell antigen uptake and processing.
  • FcR-mediated signaling that can result from an Fc region binding to an FcR can lead to more efficient dendritic cell antigen uptake and processing.
  • FcR- mediated signaling that can result from an Fc region binding to an FcR can increase antigen presentation.
  • FcR-mediated signaling that can result from an Fc region binding to an FcR can increase antigen presentation by immune cells.
  • FcR-mediated signaling that can result from an Fc region binding to an FcR can increase antigen presentation by antigen presenting cells.
  • FcR- mediated signaling that can result from an Fc domain binding to an FcR can increase antigen presentation by dendritic cells.
  • FcR-mediated signaling that can result from an Fc domain binding to an FcR can promote the expansion and activation of T cells.
  • FcR-mediated signaling that can result from an Fc domain binding to an FcR can promote the expansion and activation of CD8 + T cells.
  • FcR-mediated signaling that can result from an Fc domain binding to an FcR can influence immune cell regulation of T cell responses.
  • FcR-mediated signaling that can result from an Fc domain binding to an FcR can influence immune cell regulation of T cell responses.
  • FcR-mediated signaling that can result from an Fc domain binding to an FcR can influence dendritic cell regulation of T cell responses.
  • FcR-mediated signaling that can result from an Fc domain binding to an FcR can influence functional polarization of T cells (e.g., polarization can be toward a 3 ⁇ 41 cell response).
  • the profile of FcRs on a dendritic cell (DC) can impact the ability of the DC to respond upon stimulation.
  • DC dendritic cell
  • most DC can express both CD32A and CD32B, which can have opposing effects on IgG-mediated maturation and function of DCs: binding of IgG to CD32A can mature and activate DCs in contrast with CD32B, which can mediate inhibition due to phosphorylation of immunoreceptor tyro sine-based inhibition motif (ITIM), after CD32B binding of IgG. Therefore, the activity of these two receptors can establish a threshold of DC activation. Furthermore, the difference in functional avidity of these receptors for IgG can shift their functional balance. Hence, altering the Fc domain binding to FcRs can also shift their functional balance, allowing for manipulation (either enhanced activity or enhanced inhibition) of the DC immune response.
  • ITIM immunoreceptor tyro sine-based inhibition motif
  • a modification in the amino acid sequence of the Fc domain of an antibody construct can alter the recognition of an FcR for the Fc domain. However, such modifications can still allow for FcR-mediated signaling.
  • a modification can be a substitution of an amino acid at a residue (e.g., wildtype) for a different amino acid at that residue.
  • a modification can permit binding of an FcR to a site on the Fc domain or region that the FcR may not otherwise bind to.
  • a modification can increase the binding affinity of an FcR to the Fc domain that the FcR may have reduced binding affinity for.
  • a modification can decrease binding affinity of an FcR to a site on the Fc domain that the FcR may have increased binding affinity for.
  • a modification can increase the subsequent FcR-mediated signaling after Fc binding to an FcR.
  • An antibody construct can comprise an Fc region with at least one amino acid change as compared to the sequence of the wild-type Fc region.
  • An antibody construct can comprise an Fc domain with at least one amino acid change as compared to the sequence of the wild-type Fc domain.
  • An amino acid change in an Fc region can allow the construct to bind to at least one Fc receptor with greater affinity compared to a wild-type Fc region.
  • An amino acid change in an Fc domain can allow the antibody construct to bind to at least one Fc receptor with greater affinity compared to a wild-type Fc domain.
  • An Fc region can comprise an amino acid sequence having at least one, two, three, four, five, six, seven, eight, nine or ten modifications but not more than 40, 35, 30, 25, 20, 15, or 10 modifications of the amino acid sequence relative to the natural or original amino acid sequence.
  • An Fc domain can comprise an amino acid sequence having at least one, two, three, four, five, six, seven, eight, nine or ten modifications but not more than 40, 35, 30, 25, 20, 15, or 10 modifications of the amino acid sequence relative to the natural or original amino acid sequence.
  • An Fc region can be an Fc region of an IgGl antibody.
  • An Fc region can contain an Fc domain.
  • An Fc region can be an Fc domain.
  • An antibody construct can be an antibody comprising a sequence of the IgGl isoform that has been modified from the wildtype IgGl sequence.
  • a modification can comprise a substitution at more than one amino acid residue such as at 5 different amino acid residues including
  • the numbering of amino acids residues is according to the EU index of Kabat.
  • the 5 amino acid residues can be located in a portion of an antibody sequence which can encode an Fc region of the antibody and in particular, can be located in portions of the Fc region that can bind to Fc receptors (i.e., the Fc domain).
  • a modification can comprise a substitution at more than one amino acid residue such as at 3 different amino acid residues including S298A/E333A/K334A (IgGl AAA), according to the EU index of Kabat.
  • the 3 amino acid residues can be located in a portion of an antibody sequence which can encode an Fc region of the antibody and in particular, can be located in portions of the Fc region that can bind Fc receptors (i.e., the Fc domain).
  • the Fc domain or region can comprise a sequence of an IgG isoform that has been modified from the wild-type IgG sequence. In some embodiments, the Fc domain or region can comprise a sequence of the IgGl isoform that has been modified from the wild-type IgGl sequence. In some embodiments, the modification comprises substitution of one or more amino acids that reduce binding affinity of an IgG Fc domain or region to all Fey receptors.
  • a modification can be substitution of E233, L234 and L235, such as
  • a modification can be substitution of L235, F243, R292, Y300 and P396, such as
  • a modification can be a substitution of P238, such as P238A, according to the EU index of Kabat.
  • a modification can be a substitution of D265, such as D265A, according to the EU index of Kabat.
  • a modification can be a substitution of N297, such as N297A, according to the EU index of Kabat.
  • a modification can be a substitution of A327, such as A327Q, according to the EU index of Kabat.
  • a modification can be a substitution of P329, such as P239A, according to the EU index of Kabat.
  • an IgG Fc domain or region comprises at least one amino acid substitution that reduces its binding affinity to FcyRl, as compared to a wild-type or reference IgG Fc domain.
  • a modification can comprise a substitution at F241, such as F241A, according to the EU index of Kabat.
  • a modification can comprise a substitution at F243, such as F243A, according to the EU index of Kabat.
  • a modification can comprise a substitution at V264, such as V264A, according to the EU index of Kabat.
  • a modification can comprise a substitution at D265, such as D265A according to the EU index of Kabat.
  • an IgG Fc domain or region comprises at least one amino acid substitution that increases its binding affinity to FcyRl, as compared to a wild-type or reference IgG Fc domain.
  • a modification can comprise a substitution at A327 and P329, such as
  • the modification comprises substitution of one or more amino acids that reduce binding affinity of an IgG Fc domain or region to FcyRII and FcyRIIIA receptors.
  • a modification can be a substitution of D270, such as D270A, according to the EU index of Kabat.
  • a modification can be a substitution of Q295, such as Q295A, according to the EU index of Kabat.
  • a modification can be a substitution of A327, such as A237S, according to the EU index of Kabat.
  • the modification comprises substitution of one or more amino acids that increases binding affinity of an IgG Fc domain or region to FcyRII and FcyRIIIA receptors.
  • a modification can be a substitution of T256, such as T256A, according to the EU index of Kabat.
  • a modification can be a substitution of K290, such as K290A, according to the EU index of Kabat.
  • the modification comprises substitution of one or more amino acids that increases binding affinity of an IgG Fc domain or region to FcyRII receptor.
  • a modification can be a substitution of R255, such as R255A, according to the EU index of Kabat.
  • a modification can be a substitution of E258, such as E258A, according to the EU index of Kabat.
  • a modification can be a substitution of S267, such as S267A, according to the EU index of Kabat.
  • a modification can be a substitution of E272, such as E272A, according to the EU index of Kabat.
  • a modification can be a substitution of N276, such as N276A, according to the EU index of Kabat.
  • a modification can be a substitution of D280, such as D280A, according to the EU index of Kabat.
  • a modification can be a substitution of H285, such as H285A, according to the EU index of Kabat.
  • a modification can be a substitution of N286, such as N286A, according to the EU index of Kabat.
  • a modification can be a substitution of T307, such as T307A, according to the EU index of Kabat.
  • a modification can be a substitution of L309, such as L309A, according to the EU index of Kabat.
  • a modification can be a substitution of N315, such as N315A, according to the EU index of Kabat.
  • a modification can be a substitution of K326, such as K326A, according to the EU index of Kabat.
  • a modification can be a substitution of P331, such as P331A, according to the EU index of Kabat.
  • a modification can be a substitution of S337, such as S337A, according to the EU index of Kabat.
  • a modification can be a substitution of A378, such as A378A, according to the EU index of Kabat.
  • a modification can be a substitution of E430, such as E430, according to the EU index of Kabat.
  • the modification comprises substitution of one or more amino acids that increases binding affinity of an IgG Fc domain or region to FcyRII receptor and reduces the binding affinity to FcyRIIIA receptor.
  • a modification can be a substitution of H268, such as H268A, according to the EU index of Kabat.
  • a modification can be a substitution of R301, such as R301A, according to the EU index of Kabat.
  • a modification can be a substitution of K322, such as K322A, according to the EU index of Kabat.
  • the modification comprises substitution of one or more amino acids that decreases binding affinity of an IgG Fc domain or region to FcyRII receptor but does not affect the binding affinity to FcyRIIIA receptor.
  • a modification can be a substitution of R292, such as R292A, according to the EU index of Kabat.
  • a modification can be a substitution of K414, such as K414A, according to the EU index of Kabat.
  • the modification comprises substitution of one or more amino acids that decreases binding affinity of an IgG Fc domain or region to FcyRII receptor and increases the binding affinity to FcyRIIIA receptor.
  • a modification can be a substitution of S298, such as S298A, according to the EU index of Kabat.
  • a modification can be substitution of S239, 1332 and A330, such as S239D/I332E/A330L.
  • a modification can be substitution of S239 and 1332, such as S239D/I332E.
  • the modification comprises substitution of one or more amino acids that decreases binding affinity of an IgG Fc domain or region to FcyRIIIA receptor and does not affect the binding affinity to FcyRII receptor.
  • a modification can be a substitution of S239, such as S239A, according to the EU index of Kabat.
  • a modification can be a substitution of E269, such as E269A, according to the EU index of Kabat.
  • a modification can be a substitution of E293, such as E293A, according to the EU index of Kabat.
  • a modification can be a substitution of Y296, such as Y296F, according to the EU index of Kabat.
  • a modification can be a substitution of V303, such as V303A, according to the EU index of Kabat.
  • a modification can be a substitution of A327, such as A327G, according to the EU index of Kabat.
  • a modification can be a substitution of A327, such as A327G, according to the EU index of Kab
  • modification can be a substitution of K338, such as K338A, according to the EU index of Kabat.
  • a modification can be a substitution of D376, such as D376A, according to the EU index of Kabat.
  • the modification comprises substitution of one or more amino acids that increases binding affinity of an IgG Fc domain or region to FcyRIIIA receptor and does not affect the binding affinity to FcyRII receptor.
  • a modification can be a substitution of E333, such as E333A, according to the EU index of Kabat.
  • a modification can be a substitution of K334, such as K334A, according to the EU index of Kabat.
  • a modification can be a substitution of A339, such as A339T, according to the EU index of Kabat.
  • a modification can be substitution of S239 and 1332, such as S239D/I332E.
  • an IgG Fc domain or region comprises at least one amino acid substitution that reduces the binding affinity to FcRn, as compared to a wild-type or reference IgG Fc domain.
  • a modification can comprise a substitution at H435, such as H435A according to the EU index of Kabat.
  • a modification can comprise a substitution at 1253, such as 1253 A according to the EU index of Kabat.
  • a modification can comprise a substitution at H310, such as H310A according to the EU index of Kabat.
  • a modification can comprise substitutions at 1253, H310 and H435, such as I253A/H310A/H435A according to the EU index of Kabat.
  • a modification can comprise a substitution of one amino acid residue that increases the binding affinity of an IgG Fc domain for FcRn, relative to a wildtype or reference IgG Fc domain.
  • a modification can comprise a substitution at V308, such as V308P according to the EU index of Kabat.
  • a modification can comprise a substitution at M428, such as M428L according to the EU index of Kabat.
  • a modification can comprise a substitution at N434, such as N434A according to the EU index of Kabat or N434H according to the EU index of Kabat.
  • a modification can comprise substitutions at T250 and M428, such as T250Q and M428L according to the EU index of Kabat.
  • a modification can comprise substitutions at M428 and N434, such as M428L and N434S, N434A or N434H according to the EU index of Kabat.
  • a modification can comprise substitutions at M252, S254 and T256, such as M252Y/S254T/T256E according to the EU index of Kabat.
  • a modification can be a substitution of one or more amino acids selected from P257L, P257N, P257I, V279E, V279Q, V279Y, A281S, E283F, V284E, L306Y, T307V, V308F, Q311V, D376V, and N434H.
  • Other substitutions in an IgG Fc domain that affect its interaction with FcRn are disclosed in U.S. Patent No. 9,803,023 (the disclosure of which is incorporated by reference herein).
  • An antibody construct can be a monoclonal anti-CD40 human antibody comprising a sequence of the IgGl isoform that has been modified from the wildtype IgGl sequence.
  • a modification can comprise a substitution at more than one amino acid residue such as at 5 different amino acid residues including L235V/F243L/R292P/Y300L/P396L (SBT-040- G1VLPLL).
  • the numbering of amino acids residues is according to the EU index.
  • the 5 amino acid residues can be located in a portion of an antibody sequence which can encode an Fc region of the antibody and in particular, can be located in portions of the Fc region that can bind to Fc receptors (i.e., the Fc domain).
  • a modification can comprise a substitution at more than one amino acid residue such as at 3 different amino acid residues including S298A/E333A/K334A (SBT-040-G1AAA).
  • the 3 amino acid residues can be located in a portion of an antibody sequence which can encode an Fc region of the antibody and in particular, can be located in portions of the Fc region that can bind Fc receptors (i.e., the Fc domain).
  • binding of Fc receptors to an Fc region can be affected by amino acid substitutions.
  • SBT-040-VLPLL is an antibody with an amino acid sequence of a heavy chain of human anti-CD40 monoclonal antibody with modifications to a wild-type IgGl Fc domain (L235V/F243L/R292P/Y300L/P396L). Binding of some Fc receptors to the Fc region of SBT- 040-VLPLL can be enhanced compared to wild-type by as result of the
  • binding of other Fc receptors to the Fc region of SBT-040-VLPLL can be reduced compared to wild-type by the L235V/F243L/R292P/Y300L/P396L amino acid modifications.
  • the binding affinities of SBT-040-VLPLLto FcyRIIIA and to FcyRIIA can be enhanced compared to wild- type whereas the binding affinity of SBT-040-VLPLL to FcyRIIB can be reduced compared to wild-type. Binding of Fc receptors to an Fc region of are affected by amino acid substitutions.
  • SBT-040-G1AAA antibody is an antibody with an amino acid sequence of a heavy chain of a human anti-CD40 monoclonal antibody with modifications to a wild-type IgGl Fc domain (S298A/E333A/K334A). Binding of Fc receptors to an Fc region of SBT-040-G1AAA can be enhanced compared to wild-type as a result of the S298A/E333A/K334A amino acid
  • binding of some Fc receptors to the Fc region of SBT-040-G1AAA can be reduced compared to wild-type by S298A/E333A/K334A amino acid modification. Binding affinities of SBT-040-G1AAA to FcyRIIIA can be enhanced compared to wild-type whereas the binding affinity of SBT-040-G1AAA to FcyRIIB can be reduced compared to wildtype.
  • the heavy chain of a human IgG2 antibody can be mutated at cysteines as positions 127, 232, or 233.
  • the light chain of a human IgG2 antibody can be mutated at a cysteine at position 214.
  • the mutations in the heavy and light chains of the human IgG2 antibody can be from a cysteine residue to a serine residue.
  • an antibody construct of the present disclosure can comprise a first binding domain and a second binding domain (or, in some cases, a third binding domain) with wild-type or modified amino acid sequences encoding the Fc region or Fc domain
  • the modifications of the Fc region or the Fc domain from the wild-type sequence may not significantly alter binding and/or affinity of the binding domains.
  • constructcomprising a first binding domain and a second binding domain (or, in some cases, a third binding domain) and having the Fc domain modifications of SBT-040-G1WT, SBT-040- G1VLPLL, or SBT-040-G1AAA may not be significantly altered by modification of an Fc region or Fc domain amino acid sequence compared to a wild-type sequence. Modifications of an Fc region or Fc domain from a wild-type sequence may not alter binding and/or affinity of a first binding domain that binds, for example, to CD40 or DEC-205.
  • binding and/or affinity of the binding domains described herein may be comparable to the binding and/or affinity of wild-type antibodies.
  • the binding profile of the Fc domain for Fey receptors can be retained in the antibody construct or conjugate, which can allow for delivery of the antibody construct or conjugate into immune cell types comprising the Fey receptors and can further immune activation by Fey receptor signaling.
  • APCs can be activated by an antibody construct or conjugate as described herein when the antibody construct or conjugate is bound to a tumor cell, undergoes Fey receptor mediated uptake, or undergoes Fey receptor mediated Antibody Dependent Cellular Phagocytosis (ADCP).
  • ADCP Antibody Dependent Cellular Phagocytosis
  • the antibody construct or conjugate can retain weak or no Fey receptor binding to allow for maximal immune activation or decreased toxicity of the immune- stimulatory compound due to the primary delivery of the antibody construct or conjugate into tumor cells by antibody antigen mediated endocytosis.
  • the antibody construct or conjugate can comprise an IgGl Fc domain variant comprising N297A, N297G, K322A/L234A/L235A, or L234F/L235E/P331S, and lacks binding to an Fey receptor but can retain binding to FcRN in the presence of the immune-modulatory compound to allow for lower delivery of the conjugate into tumor cells or immune cells.
  • an antibody construct or conjugate can comprise an Fc domain with higher affinity to one or more Fey receptors, which can result in greater immune activation than for an antibody construct or conjugate with an Fc domain that can bind to one or more Fey receptors with lower affinity.
  • Pertuzumab V H 12 EVQLVESGGGLVQPGGSLRLSCAASGFTFTDYTMDWVRQAPGKGLEWVA
  • Adecatumum V H 142 EVQLLESGGGVVQPGRSLRLSCAASGFTFSSYGMHWVRQAPGKGLEWVA ab VISYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKD
  • Lifastuzumab V H 181 EVQLVESGGGLVQPGGSLRLSCAASGFSFSDFAMSWVRQAPGKGLEWVAT
  • V L 316 DVLMTQSPLSLPVTPGEPASISCRSSQSIVHSSGITYLEWYLQKPGQSPQLLI
  • Narnatumab V H 337 EVQLVESGGGLVQPGGSLRLSCAASGFTFSSYLMTWVRQAPGKGLEWVA
  • J591 variant 1 V H 389 EVQLQQSGPELKKPGTSVRISCKTSGYTFTEYTIHWVKQSHGKSLEWIGNIN
  • J591 variant 2 V H 402 EVQLQQSGPELVKPGTSVRISCKTSGYTFTEYTIHWVKQSHGKSLEWIGNIN
  • Atezolizumab V H 890 EVQLVESGGGLVQPGGSLRLSCAASGFTFSDSWIHWVRQAPGKGLEWVA

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Abstract

Various compositions are disclosed. The compositions of composition-immune stimulatory compound conjugates are also provided. Additionally provided are the methods of preparation and uses of the composition-immune stimulatory compound conjugates. This includes methods for treating disorders, such as cancer.

Description

ANTIBODY CONSTRUCT CONJUGATES
PRIORITY
[0001] This application claims the benefit of priority under 35 U.S.C. § 119(e) to U.S.
Provisional Application No. 62/516,667, filed June 7, 2017, the disclosure of which is incorporated herein by reference in its entirety.
BACKGROUND
[0002] One of the leading causes of death in the United States is cancer. The conventional methods of cancer treatment, like chemotherapy, surgery, or radiation therapy, tend to be either highly toxic or nonspecific to a cancer, or both, resulting in limited efficacy and harmful side effects. However, the immune system has the potential to be a powerful, specific tool in fighting cancers. In many cases tumors can specifically express genes whose products are required for inducing or maintaining the malignant state. These proteins may serve as antigen markers for the development and establishment of more specific anti-cancer immune response. The immune response may include the recruitment of immune cells that target tumors expressing these antigen markers. Additionally, the immune cells may express genes whose products are important to proper immune function and may serve as markers for specific types of immune cells. The boosting of this specific immune response has the potential to be a powerful anti-cancer treatment that can be more effective than conventional methods of cancer treatment and can have fewer side effects.
INCORPORATION BY REFERENCE
[0003] All publications, patents, and patent applications mentioned in this specification are herein incorporated by reference to the same extent as if each individual publication, patent, or patent application was specifically and individually indicated to be incorporated by reference.
SUMMARY
[0004] Provided herein are immune-modulatory and immune- stimulatory conjugates. In certain embodiments, an immune-modulatory conjugate is provided comprising: (a) an antibody construct comprising an antigen binding domain and an Fc domain, wherein the antigen binding domain binds to a first antigen; (b) a proteolysis targeting module, comprising: (i) a protein targeting moiety that binds to a target protein; (ii) an E3 ubiquitin ligase binding moiety; and (iii) a spacer S that is covalently bound to the protein targeting moiety and to the E3 ubiquitin ligase binding moiety, wherein the spacer is optionally has 1-25 consecutive non-hydrogen atoms; and (c) a linker L that is covalently attached to the antibody construct and to the proteolysis targeting module. The conjugate can be represented by one of the following formulae:
(IX),
wherein: Ab is the antibody construct; L is the linker; ULM is the E3 ubiquitin ligase binding moiety; IMC is the protein targeting moiety and comprises an immune-modulatory compound; S is the spacer; n is selected from 1 to 20; and z is selected from 1 to 20. In some embodiments, the Fc domain of the conjugate is an Fc null.
[0005] In some embodiments, the E3 ubiquitin ligase is selected from the group consisting of
Von Hippel-Lindaue E3 ubiquitin ligase (VHL), cereblon, mouse double minute 2 homolog
(MDM2), AMFR, APC/Cdc20, APC/Cdhl, C6orfl57, Cbl, CBLL1, CHFR, CHIP, DTL (Cdt2),
E6-AP, HACE1, HECTD1, HECTD2, HECTD3, HECW1, HECW2, HERC2, HERC3, HERC4,
HERC5, HUWE1, HYD, ITCH, LNX1, mahogunin, MARCH-I, MARCH-II, MARCH-III,
MARCH-IV, MARCH- VI, MARCH- VII, MARCH- VIII, MARCH-X, MEKK1, MIB 1, MIB2,
MycBP2, NEDD4, NEDD4L, Parkin, PELIl, Pirh2, PJAl, PJA2, RFFL, RFWD2, Rictor, RNF5,
RNF8, RNF19, RNF190, RNF20, RNF34, RNF40, RNF125, RNF128, RNF138, RNF168,
SCF/?-TrCP, SCF/FBW7, SCF/Skp2, SHPRH, SIAH1, SIAH2, SMURF1, SMURF2, TOPORS, TRAF6, TRAF7, TRIM63, UBE3B, UBE3C, UBR1, UBR2, UHRF2, WWP1, WWP2, and ZNRF1. In some further embodiments, the E3 ubiquitin ligase binding moiety binds to VHL, cereblon or MDM2.
[0006] In some embodiments, the protein targeting moiety binds to a protein selected from one of the groups consisting of: a) an aryl hydrocarbon receptor, androgen receptor, estrogen receptor, FK506-binding protein 12, fibroblast growth factor receptor substrate 2,
phosphatidylinositol -4,5-biphosphate 3-kinase, SMAD family member 3, bromodomain and extra-territorial family of proteins (BET), bromodomain-containing protein 4 member of the BET family, Abelson tyrosine kinase, receptor- interacting serine/threonine-protein kinase 1, estrogen- related receptor, and transforming growth factor beta (TGFP); or b) ALK, Bcr-Abl, BRAF, BTK, c-KIT, EGFR, ErbB2, JAK, MEK, MET, PDGFRB, RET, ROS 1, Syk, SRC, TNIK, TNKS, TNKS2, TrkA, TrkB, TrkC, VEGFR1, VEGFR2, VEGFR3, FGFR1, FGFR2, FGFR3, FGFR4, CSF1R, RON/MS T1R, TYR03, MERTK, AXL, PI3K, PI3K, MAP4K1, PERK, and KIT; or c) TGFpR2, TGFpRl, SMAD2, SMAD3, SMAD4, beta-catenin, CREBB2, Beta catenin/TCF4, beta catenin/LEF, beta catenin/CREBBP, YAP, TAZ, YAP/TAZ, TNKS 1, TNKS2, MST1, MST2, NRAS, HRAS, KRAS, RASmutl2, RASmutl3, PERK (EIF2AK3), RON/MS T1R, STAT3, MCT1, MCT2 and MCT4; or d) CSFR1, RON/MST1, PI3Kd, PI3Kg, PARP1, PD-L1, PP2A, A2ar, TYR03, AXL and MER.
[0007] In some embodiments, the first antigen is a tumor antigen. The tumor antigen can be MUC16, CD5, CD19, CD20, CD25, CD37, CD30, CD33, CD45, CAMPATH-1, BCMA, CS-1, PD-L1, B7-H3, B7-DC, HLD-DR, carcinoembryonic antigen (CEA), TAG-72, EpCAM, MUC1, fo late-binding protein, A33, G250, pro state- specific membrane antigen (PSMA), ferritin, GD2, GD3, GM2, Ley, CA-125, CA19-9, epidermal growth factor, pl85HER2, IL-2 receptor, fibroblast activation protein (FAP), tenascin, a metalloproteinase, endosialin, vascular endothelial growth factor, v 3, WT1, LMP2, HPV E6, HPV E7, EGFRvIII, Her-2/neu, MAGE A3, p53 nonmutant, NY-ESO-1, MelanA/M ART 1 , Ras mutant, gplOO, p53 mutant, PR1, bcr-abl, tyronsinase, survivin, PSA, hTERT, a Sarcoma translocation breakpoint fusion protein, EphA2, PAP, ML-IAP, AFP, ERG, NA17, PAX3, ALK, androgen receptor, cyclin B l, polysialic acid, MYCN, RhoC, TRP-2, fucosyl GM1, mesothelin (MSLN), PSCA, MAGE Al, sLe(animal), CYP1B 1, PLAV1, GM3, BORIS, Tn, GloboH, ETV6-AML, NY-BR-1, RGS5, SART3, STn, Carbonic anhydrase IX, PAX5, OY-TES 1, Sperm protein 17, LCK, HMWMAA, AKAP-4, SSX2, XAGE 1, Legumain, Tie 3, Page4, VEGFR2, MAD-CT-1, PDGFR-B, MAD-CT-2, ROR2, TRAIL 1, MAGE A4, MAGE C2, GAGE, EGFR, CMET, HER3, MUC15, CA6,
NAPI2B, TROP2, CLDN18.2, RON, LY6E, FRA, DLL3, PTK7, LIV1, ROR1, or Fos-related antigen 1. [0008] In some embodiments, the first antigen is an immune cell antigen, and can be an antigen present on the surface of an antigen presenting cell such as a dendritic cell or a macrophage. In some embodiments, the immune cell antigen is selected from the group consisting of CD40, DEC-205, PD-Ll, PD-1, CD36 mannose scavenger receptor 1, CLEC9A, DC-SIGN, CLEC12A, BDCA-2, OX40L, 41BBL, CD204, MARCO, CLEC5A, Dectin 1, Dectin 2, CLECIOA, CD206, CD64, CD32A, CD 16 A, HVEM, and CD32B.
[0009] In some embodiments, the protein targeting moiety can be a peptide or a non- proteinaceous molecule. In some embodiments, the proteolysis targeting module is selected from compounds 1-1, 1-2, 1-3, 1-4, 1-5, 1-6 and 2-1, as disclosed herein. In some embodiments, the linker L is a cleavable linker. In some embodiments, the linker L is a non-cleavable linker. In some embodiments, the linker L is covalently bound to the antibody construct at a cysteine residue, an engineered cysteine residue, a lysine residue, a glutamate residue, or a glutamine residue of the antibody construct or is covalently bound to the antibody construct using a Sortase linker.
[0010] In some embodiments, the spacer S is an optionally substituted Ci-25 alkylene or optionally substituted Ci-25 heteroalkylene, wherein the hetero alkylene is a Ci-24 alkylene chain interspersed with one or more groups independently selected from: -0-, -S-, -NH2-, and - C(0)NH-; and optionally substituted with a reactive group, R , that can form a functional group selected from an amide bond, an ester bond, an ether bond, a carbonate bond, a carbamate bond, or a thioether bond. The spacer can be optionally substituted with Ci-C8 alkyl, C2-C8 alkenyl, C2- C8 alkynyl, -(CH20)niH, -(CH2CH20)niH, -(CH20)niCH3, -C(0)OH or -NH2, wherein nl is from 1 to 8. In some embodiments the spacer is substituted with Rx. In some embodiments, the linker is unsubstituted.
[0011] In some embodiments, the antibody construct further comprises a second binding domain. The second binding domain can specifically bind to an antigen on an immune cell. The immune cell antigen is selected from the group consisting of CD40, PD-1, PD-Ll, DEC-205, CD36 mannose scavenger receptor 1, CLEC9A, DC-SIGN, CLEC12A, BDCA-2, OX40L, 41BBL, CD204, MARCO, CLEC5A, Dectin 1, Dectin 2, CLECIOA, CD206, CD64, CD32A, CD 16 A, HVEM, and CD32B. In some embodiments, the second binding domain is attached to the antibody construct at a C-terminal end of the Fc domain.
[0012] In some embodiments, the Fc domain is an Fc domain variant comprising at least one amino acid residue change as compared to a wild type sequence of the Fc domain. In some embodiments, the Fc domain is an Fc domain variant that decreases binding of the Fc domain to an Fc receptor. In some embodiments, the at least one amino acid residue change in the Fc domain is: a) F243L, R292P, Y300L, L235V, and P396L, wherein numbering of amino acid residues in the Fc domain is relative to SEQ ID NO: 898; b) S239D and I332E, wherein numbering of amino acid residues in the Fc domain is relative to SEQ ID NO: 898; or c) S298A, E333A, and K334A, wherein numbering of amino acid residues in the Fc domain is relative to SEQ ID NO: 898. In some embodiments, the at least one amino acid residue change in the Fc domain is: a) N297A, N297G, N297Q, N297D as in Eu index of Kabat numbering and relative to SEQ ID NO: 898; or b) K322A/L234A/L235A N296A as in the EU index of Kabat numbering and relative to SEQ ID NO: 898; or c) L234F/L235E/P331S N296A as in the EU index of Kabat numbering and relative to SEQ ID NO: 898; or d) P329G/L234A/L235A as in the EU index of Kabat numbering and relative to SEQ ID NO: 898.
[0013] Also provided is a pharmaceutical composition comprising a conjugate as described herein along with a pharmaceutically acceptable excipient. In the pharmaceutical composition, the average ratio of proteolysis targeting modules to antibody construct in the conjugate can be from 2 to 6, from 3 to 5 or 1 to 3. The pharmaceutical composition can be lyophilized.
[0014] Also provided are methods of treating cancer comprising administering a conjugate as described herein, or a pharmaceutical composition of a conjugate as described herein, to a subject in need thereof. In some embodiments, the cancer is a solid tumor, such as breast cancer, pancreatic cancer, colorectal cancer, renal cell cancer, gastric cancer, or lung cancer. In some embodiments, the pharmaceutical composition is administered parenterally. In some
embodiments, the pharmaceutical composition is administered intravenously.
[0015] In certain aspects, the disclosure also provides immune- stimulatory conjugates comprising: (a) an immune- stimulatory compound that binds to a target protein to stimulate an immune response by stimulating the activity of the target protein, reducing immune inhibition by the target protein or increasing degradation of the target protein; (b) an antibody construct comprising an antigen binding domain and an Fc domain, wherein the antigen binding domain binds to at least a first antigen and wherein the Fc domain binds to an Fc receptor; and (c) a linker, wherein the linker is covalently bound to the antibody construct and the linker is covalently bound to the immune- stimulatory compound. In some embodiments, the dissociation constant (Kd) for binding of the Fc domain of the conjugate to an Fc receptor is equal to, or up to no greater than about 100 times the Kd for binding of a control antibody construct to the Fc receptor, wherein the control antibody construct is the unconjugated antibody construct; and wherein the Kd for binding of the immune- stimulatory compound of the conjugate (i.e, attached to the conjugate) to the target protein is equal to, or up to no greater than 100 times the Kd for binding of a control compound to the target protein active site or wherein the EC50 or IC50 of the immune- stimulatory compound of the conjugate is no greater than 300-fold the EC50 or IC50 of a control compound, wherein the control compound is the unbound immune- stimulatory compound. In some embodiments, the dissociation constant (Kd) for binding of the Fc domain of the conjugate to an Fc receptor is greater than about 100 times the Kd for binding of a control antibody construct to the Fc receptor, wherein the control antibody construct is the unconjugated antibody construct; and wherein the Kd for binding of the immune- stimulatory compound of the conjugate (i.e, attached to the conjugate) to the target protein is equal to, or up to no greater than 100 times the Kd for binding of a control compound to the target protein active site or wherein the EC50 or IC50 of the immune- stimulatory compound of the conjugate is no greater than 300- fold the EC50 or IC50 of a control compound, wherein the control compound is the unbound immune- stimulatory compound. In some embodiments, the Fc domain is an Fc null.
[0016] In certain embodiments, the disclosure provides an immune- stimulatory conjugate comprising: (a) an immune- stimulatory compound that binds to a protein active site to stimulate an immune response; (b) an antibody construct comprising an antigen binding domain and an Fc domain, wherein the antigen binding domain binds to at least a first antigen and wherein the Fc domain binds to an Fc receptor; and (c) a linker, wherein the linker is covalently bound to the antibody construct and the linker is covalently bound to the immune- stimulatory compound. In some embodiments, the dissociation constant (Kd) for binding of the Fc domain of the conjugate to an Fc receptor is equal to, or up to no greater than about 100 times the Kd for binding of a control antibody construct to the Fc receptor, wherein the control antibody construct is the unconjugated antibody construct; and wherein the Kd for binding of immune- stimulatory compound of the conjugate to the protein active site is equal to, or up to no greater than 100 times the Kd for binding of a control compound to the protein active site or wherein the EC50 or IC50 of the immune- stimulatory compound of the conjugate (attached to the conjugate )is no greater than 300-fold the EC50 or IC50 of a control compound, wherein the control compound is the unbound immune- stimulatory compound. In some embodiments, the dissociation constant (Kd) for binding of the Fc domain of the conjugate to an Fc receptor is greater than about 100 times the Kd for binding of a control antibody construct to the Fc receptor, wherein the control antibody construct is the unconjugated antibody construct; and wherein the Kd for binding of immune- stimulatory compound of the conjugate to the protein active site is equal to, or up to no greater than 100 times the Kd for binding of a control compound to the protein active site or wherein the EC50 or IC50 of the immune- stimulatory compound of the conjugate (attached to the conjugate) is no greater than 300-fold the EC50 or IC50 of a control compound, wherein the control compound is the unbound immune- stimulatory compound.
[0017] In certain embodiments, the disclosure provides an immune- stimulatory conjugate comprising: (a) an immune- stimulatory compound that binds to a protein active site of a binding protein to stimulate an immune response by inhibition of the activity of the binding protein; (b) an antibody construct comprising an antigen binding domain and an Fc domain, wherein the antigen binding domain binds to at least a first antigen and wherein the Fc domain binds to an Fc receptor; and (c) a linker, wherein the linker is covalently bound to the antibody construct and the linker is covalently bound to the immune- stimulatory compound. In some embodiments, the dissociation constant (Kd) for binding of the Fc domain of the conjugate to an Fc receptor is equal to, or up to no greater than about 100 times the Kd for binding of a control antibody construct to the Fc receptor, wherein the control antibody construct is the unconjugated antibody construct; and wherein the Kd for binding of the immune- stimulatory compound of the conjugate (attached to the conjugate) to the protein active site is equal to, or up to no greater than 100 times the Kd for binding of a control compound to the protein active site or wherein the IC50 of the immune- stimulatory compound of the conjugate (attached to the conjugate) is no greater than 300-fold the IC50 of a control compound, wherein the control compound is the unbound immune- stimulatory compound. In some embodiments, the dissociation constant (Kd) for binding of the Fc domain of the conjugate to an Fc receptor is greater than about 100 times the Kd for binding of a control antibody construct to the Fc receptor, wherein the control antibody construct is the unconjugated antibody construct; and wherein the Kd for binding of the immune- stimulatory compound of the conjugate (attached to the conjugate) to the protein active site is equal to, or up to no greater than 100 times the Kd for binding of a control compound to the protein active site or wherein the IC50 of the immune- stimulatory compound of the conjugate (attached to the conjugate) is no greater than 300-fold the IC50 of a control compound, wherein the control compound is the unbound immune- stimulatory compound. In some embodiments, the Fc domain is an Fc null.
[0018] In certain embodiments, the disclosure provides an immune- stimulatory conjugate comprising: (a) an immune- stimulatory compound that binds to a protein active site of a binding protein to stimulate an immune response by increasing degradation of the binding protein; (b) an antibody construct comprising an antigen binding domain and an Fc domain, wherein the antigen binding domain binds to at least a first antigen and wherein the Fc domain binds to an Fc receptor; and (c) a linker, wherein the linker is covalently bound to the antibody construct and the linker is covalently bound to the immune- stimulatory compound. In some embodiments, the dissociation constant (Kd) for binding of the Fc domain of the conjugate to an Fc receptor is equal to, or up to no greater than about 100 times the Kd for binding of a control antibody construct to the Fc receptor, wherein the control antibody construct is the unconjugated antibody construct; and wherein the Kd for binding of the immune- stimulatory compound of the conjugate
(attached to the conjugate) to the protein active site is equal to, or up to no greater than 100 times the Kd for binding of a control compound to the protein active site or wherein the IC50 of the immune- stimulatory compound of the conjugate is no greater than 300-fold the IC50 of a control compound, wherein the control compound is the unbound immune- stimulatory compound. In some embodiments, the dissociation constant (Kd) for binding of the Fc domain of the conjugate to an Fc receptor is greater than about 100 times the Kd for binding of a control antibody construct to the Fc receptor, wherein the control antibody construct is the unconjugated antibody construct; and wherein the Kd for binding of the immune- stimulatory compound of the conjugate (attached to the conjugate) to the protein active site is equal to, or up to no greater than 100 times the Kd for binding of a control compound to the protein active site or wherein the IC50 of the immune- stimulatory compound of the conjugate is no greater than 300-fold the IC50 of a control compound, wherein the control compound is the unbound immune- stimulatory compound. In some embodiments, the Fc domain is an Fc null.
[0019] In certain embodiments, the EC50 or IC50 of the immune- stimulatory compound of the conjugate is no greater than 100-fold the EC50 or IC50 of a control compound, wherein the control compound is the unbound immune- stimulatory compound. The EC50 or IC50 of the immune- stimulatory compound of the conjugate may be no greater than 10-fold the EC50 or IC50 of a control compound, wherein the control compound is the unbound immune- stimulatory compound. The EC50 or IC50 of the immune- stimulatory compound of the conjugate may be equivalent to or less than the EC50 or IC50 of a control compound, wherein the control compound is the unbound immune- stimulatory compound.
[0020] In certain embodiments, the EC50 or IC50 of the immune stimulatory compound on an antigen bearing cell is equivalent to or less than the EC50 or IC50 of a control compound on the antigen bearing cell and the EC50 or IC50 of the immune- stimulatory conjugate is 5-fold greater or more than the EC50 or IC50 of the control compound for a non-antigen bearing cell.
[0021] The immune- stimulatory compound of the conjugate may have a Kd for binding to the protein active site of no greater than 50 times the Kd for binding of a control compound to the protein active site, wherein the control compound is the unbound immune- stimulatory
compound. The immune- stimulatory compound of the conjugate may have a Kd for binding to the protein active site of no greater than 10 times the Kd for binding of a control compound to the protein active site, wherein the control compound is the unbound immune- stimulatory
compound. The immune- stimulatory compound of the conjugate may have a Kd for binding to the protein active site of equivalent to or less than the Kd for binding of a control compound to the protein active site, wherein the control compound is the unbound immune- stimulatory compound.
[0022] In certain aspects, the disclosure provides an immune- stimulatory conjugate comprising: (a) an immune- stimulatory compound that binds to a protein active site to stimulate an immune response; (b) an antibody construct comprising an antigen binding domain and an Fc domain, wherein the antigen binding domain binds to a first antigen and wherein the Fc domain binds to an Fc receptor; and (c) a linker comprising 5- 100 consecutive atoms attachment sites, wherein one attachment site of the linker is covalently bound to the antibody construct and another attachment site of the linker is covalently bound to the immune- stimulatory compound. In some embodiments, the dissociation constant (Kd) for binding of the Fc domain of the conjugate to an Fc receptor is equal to, or up to no greater than about 100 times the Kd for binding of a control antibody construct to the Fc receptor, wherein the control antibody construct is the unconjugated antibody construct; and wherein the Kd for binding of the immune- stimulatory compound when bound to a the linker to the protein active site is no greater than 100 times the Kd for binding of a control compound to the protein active site or wherein the EC50 or IC50 of the immune- stimulatory compound of the conjugate is no greater than 300-fold the EC50 or IC50 of a control compound, wherein the control compound is the unbound immune- stimulatory compound. In some embodiments, the dissociation constant (Kd) for binding of the Fc domain of the conjugate to an Fc receptor is greater than about 100 times the Kd for binding of a control antibody construct to the Fc receptor, wherein the control antibody construct is the unconjugated antibody construct; and wherein the Kd for binding of the immune- stimulatory compound when bound to a the linker to the protein active site is no greater than 100 times the Kd for binding of a control compound to the protein active site or wherein the EC50 or IC50 of the immune- stimulatory compound of the conjugate is no greater than 300-fold the EC50 or IC50 of a control compound, wherein the control compound is the unbound immune- stimulatory compound. In some embodiments, the Fc domain is an Fc null.
[0023] In certain embodiments, the EC50 or IC50 of the immune- stimulatory compound when bound to the 5- 100 atom linker is no greater than 100-fold the EC50 or IC50 of a control compound, wherein the control compound is the unbound immune- stimulatory compound. In certain embodiments, the EC50 or IC50 of the immune- stimulatory compound when bound to the 5- 100 atom linker is no greater than 10-fold the EC50 or IC50 of a control compound, wherein the control compound is the unbound immune- stimulatory compound. The EC50 or IC50 of the immune- stimulatory compound when bound to the 5- 100 atom linker may be equivalent to or less than the EC50 or IC50 of a control compound, wherein the control compound is the unbound immune- stimulatory compound.
[0024] In certain embodiments, the Kd for binding of the immune- stimulatory compound when bound to the 5- 100 atom linker to the protein active site may be no greater than 50 times the Kd for binding of a control compound to the protein active site, wherein the control compound is the unbound immune- stimulatory compound. The Kd for binding of the of the immune- stimulatory compound when bound to a 5-100 atom linker to the protein active site may be no greater than 10 times the Kd for binding of a control compound to the protein active site, wherein the control compound is the unbound immune- stimulatory compound. The Kd for binding of the immune- stimulatory compound when bound to a 5-100 atom linker to the protein active site may be equivalent to or less than the Kd for binding of a control compound to the protein active site wherein the control compound is the unbound immune- stimulatory compound.
[0025] In certain embodiments, the immune- stimulatory conjugate described herein comprises moiety that binds to the binding protein and an E3 ubiquitin ligase binding moiety. The E3 ubiquitin ligase binding moiety may bind to, for example, VHL, cereblon or MDM2. The E3 ubiquitin ligase binding moiety may be selected from among E3 targeting compounds. The E3 ubiquitin ligase binding moiety may be attached to the linker or may be part of the linker. In certain embodiments, the E3 ubiquitin ligase binding moiety attached to the linker, wherein the E3 ubiquitin ligase binding moiety is bound through a first linker having 5-100 consecutive atoms between attachment sites to the immune- stimulatory compound and the E3 ubiquitin ligase binding moiety is bound through a second linker having 5-100 consecutive atoms between attachment points to the antibody construct.
[0026] In certain embodiments, the immune- stimulatory compound is a kinase inhibitor and the protein active site is a kinase active site. In certain embodiments, the linker is covalently bound to the kinase inhibitor at a position on the kinase inhibitor that is at or near the solvent interface of the kinase active site as determined by modeling of the kinase inhibitor in the kinase active site. In certain embodiments, the linker is covalently bound to the kinase inhibitor at a position on the kinase inhibitor such that when the kinase inhibitor is bound to the active site, the linker extends out from the kinase active site into the solvent, as determined by modeling of the kinase inhibitor in the kinase active site. In certain embodiments, the kinase inhibitor may be selected from an inhibitor of ALK, Bcr-Abl, BRAF, BTK, c-KIT, EGFR, ErbB2, JAK, MEK, MET, PDGFRB, RET, ROS 1, Syk, SRC, TGFpR2, TNIK, TNKS, TNKS2, TrkA, TrkB, TrkC, VEGF, VEGFR1, VEGFR2, VEGFR3, FGFR1, FGFR2, FGFR3, FGFR4, CSF1R, RON/MS T1R, TYR03, MERTK, AXL, PDKdelta PDKgamma, MAP4K1, PERK, and combinations thereof.
[0027] In certain embodiments, the immune- stimulatory compound is selected from a to 11- like receptor agonist, STING agonist, or RIG-I agonist. The immune- stimulatory compound may be a toll-like receptor (TLR) agonist selected from a TLR1 agonist, a TLR2 agonist, a TLR3 agonist, a TLR4 agonist, a TLR5 agonist, a TLR6 agonist, a TLR7 agonist, a TLR8 agonist, a TLR9 agonist, or a TLR10 agonist. The immune- stimulatory compound may be selected from a pyrimidine, a purine, a guanine nucleoside, an 8-oxoadenine, an imidazoquinoline, a
thiazoquinoline, a 2-aminoimidazole, a furo[2,3-c]pyridine, a furo[2,3-c]quinoline, a 2- aminobenzimidazole, a 2-aminoquinoline, and a 2-aminobenzazepine. The immune- stimulatory compound may bind to a G protein-coupled receptor (GCPR), an ion channel, a membrane transporter, a phosphatase or an endoplasmic reticulum (ER) protein.
[0028] In certain embodiments, the immune- stimulatory compound is an antagonist of the GPCR A2aR, the sphingosine 1-phosphate receptor 1, prostaglandin receptor EP3, prostanglandin receptor E2, Frizzled, CXCR4 or an LPA receptor. The immune- stimulatory compound may be an ion channel agonist for CRAC, Kvl.3 or KCa3.1. The immune- stimulatory compound may be an inhibitor of HSP90 or AAA-ATPase p97.
[0029] In certain embodiments, the immune- stimulatory conjugate has immune- stimulatory activity while attached to the conjugate or the linker and without undergoing cell processing such as by endosomal or lysosomal degradation, e.g., to release the immune- stimulatory compound or a modified form of the compound from the conjugate or linker. In certain embodiments, the linker is a non-cleavable linker. The linker may be selected from the group consisting of
Fleximer linkers. The linker may comprise one or more carbamate or amide linkages when attached to an immune- stimulatory compound. The linker ma be selected from a linker is
represented by one of the following formula:
, wherein Rx is a reactive moiety and wherein R is selected from the group consisting of hydrogen, alkyl (e.g., d- C8 alkyl), sulfonate and methyl sulfonate. The linker may be attached to the antibody construct at a cysteine or lysine residue of the antibody construct.
[0030] In certain embodiments, the first antigen is a tumor antigen. The first antigen may be at least 80% or 100% homologous to CD5, CD19, CD20, CD25, CD37, CD30, CD33, CD45, CAMPATH-1, BCMA, CS-1, PD-L1, B7-H3, B7-DC (PD-L2), HLD-DR, carcinoembryonic antigen (CEA), TAG-72, EpCAM, MUC1, MUC15, MUC16, folate-binding protein, A33, G250, pro state- specific membrane antigen (PSMA), ferritin, GD2, GD3, GM2, Ley, CA-125, CA19-9, epidermal growth factor, pl85HER2, IL-2 receptor, EGFRvIII (de2-7 EGFR), fibroblast activation protein (FAP), tenascin, a metalloproteinase, endosialin, vascular endothelial growth factor, ανβ3, WT1, LMP2, HPV E6 E7, Her-2/neu, p53 nonmutant, NY-ESO-1,
MelanA/MARTl, Ras mutant, gplOO, p53 mutant, PR1, bcr-abl, tyrosinase, survivin, PSA, hTERT, a Sarcoma translocation breakpoint protein, EphA2, PAP, ML-IAP, AFP, ERG, NA17, PAX3, ALK, androgen receptor, cyclin B l, polysialic acid, MYCN, RhoC, TRP-2, fucosyl GMl, mesothelin (MSLN), PSCA, MAGE Al, MAGE A3, sLe(animal), CYP1B 1, PLAV1, GM3, BORIS, Tn, GloboH, ETV6-AML, NY-BR-1, RGS5, SART3, STn, Carbonic anhydrase IX, PAX5, OY-TES 1, Sperm protein 17, LCK, HMWMAA, AKAP-4, SSX2, XAGE 1, Legumain, Tie 3, VEGFR2, MAD-CT-1, PDGFR-B, MAD-CT-2, ROR2, , TRAIL 1, MAGE A4, MAGE C2, GAGE, EGFR, CMET, HER3, EPCAM, CA6, NAPI2B, TROP2, Claudin-6 (CLDN6), Claudin-16 (CLDN16), CLDN18.2, RON, LY6E, FRA, DLL3, PTK7, Uroplakin-IB (UPK1B), LIV1, ROR1, STRA6, TMPRSS3, TMPRSS4, TMEM238, Clorfl86, Fos-related antigen 1, VEGFR1, endoglin, PD-L1, VTCN1 (B7-H4), VISTA, gpNMB, or any fragment thereof.
[0031] In certain embodiments, the antibody construct further comprises a second target binding domain. In certain embodiments, the target binding domain specifically binds an antigen on an immune cell. The target binding domain may be attached (e.g., conjugated or linked) to the antibody construct at a C-terminal end of the Fc domain. The antigen binding domain may be from an antibody or non-antibody scaffold. The antigen binding domain may be at least 97% homologous to an antigen binding domain from an antibody or non-antibody scaffold. The antibody construct may be a human antibody or a humanized antibody. The Fc domain may be an Fc domain variant comprising at least one amino acid residue change as compared to a wild type sequence of the Fc domain. The Fc domain may be at least about 80% homologous to an Fc domain from an antibody, wherein the Fc domain from an antibody comprises amino acid residues 216 to 447 of an IgGl (within SEQ ID NO: 898), amino acid residues 216 to 443 of an IgG2 (within SEQ ID NO: 899), or amino acid residues 216 to 444 of an IgG4 (within SEQ ID NO: 900).
[0032] In certain embodiments, the Fc domain may have at least one amino acid residue change as compared to a wildtype Fc domain. In certain embodiments, the Fc domain may have at least one amino acid residue change as compared to a wildtype Fc domain, wherein the Fc domain is at least 80% homologous to SEQ ID NO: 898. The at least one amino acid residue change may be in an IgG Fc domain: a) F243L, R292P, Y300L, L235V, and P396L, wherein numbering of amino acid residues in the Fc domain is relative to SEQ ID NO:898; b) S239D and I332E, wherein numbering of amino acid residues in the Fc domain is relative to SEQ ID NO: 898; or c)
S298A, E333A, and K334A, wherein numbering of amino acid residues in the Fc domain is relative to SEQ ID NO: 296. The Fc domain may have at least one amino acid residue change as compared to wildtype, wherein the Fc domain is at comprises at least 80% homologous to SEQ ID NO: 898, and wherein the at least one amino acid residue change is: (a) N296A wherein numbering of amino acid residues in the Fc domain is relative to SEQ ID NO: 898; or (b) N296G N296A wherein numbering of amino acid residues in the Fc domain is relative to SEQ ID NO: 898; or (c) K322A/L234A/L235A N296A wherein numbering of amino acid residues in the Fc domain is relative to SEQ ID NO: 898; or (d) L234F/L235E/P331S N296A wherein numbering of amino acid residues in the Fc domain is relative to SEQ ID NO: 898.
[0033] In certain embodiments, the IQ for binding of the antigen binding domain to the first antigen in the presence of the immune- stimulatory compound is less than about 100 nM and is equal to, or up to no greater than about 10 times the IQ of the binding of the antigen binding domain to the first antigen in the absence of the immune- stimulatory compound; and the IQ for binding of the Fc domain to the Fc receptor in the presence of the immune- stimulatory compound is equal to, or up to no greater than about 10 times the IQ for the binding of the Fc domain to the Fc receptor in the absence of the immune- stimulatory compound.
[0034] In certain embodiments, the molar ratio of immune- stimulatory compound to antibody in a conjugate is less than 5. In certain embodiments, the average molar ratio of immune- stimulatory compound to antibody construct in a composition of conjugates is less than 5. In certain embodiments, the average molar ratio of immune- stimulatory compound to antibody construct in a composition of conjugates from 1 to 3, 3 to 5, or about 2.
[0035] The linker may be bound to the antibody construct at an amino acid residue that does not interfere with the Fc domain binding to the Fc receptor. The linker may not be attached to an amino acid residue of an IgG Fc domain selected from a group consisting of: 221, 224, 227, 228, 230, 231, 223, 233, 234, 235, 236, 237, 238, 239, 240, 241, 243, 244, 245, 247, 249, 250, 258, 262, 263, 264, 265, 266, 267, 268, 269, 270, 271, 272, 273, 275, 276, 278, 280, 281, 283, 285, 286, 291, 292, 293, 294, 295, 296, 297, 298, 299, 300, 302, 305, 313, 318, 323, 324, 325, 327, 328, 329, 330, 331, 332, 333, 335, 336, 396, or 428, wherein numbering of amino acid residues in the Fc domain is according to the EU index as in Kabat.
[0036] The linker is bound to the antibody construct at an amino acid of an IgG Fc domain selected from a group consisting of: 221, 224, 227, 228, 230, 231, 223, 233, 234, 235, 236, 237, 238, 239, 240, 241, 243, 244, 245, 247, 249, 250, 258, 262, 263, 264, 265, 266, 267, 268, 269, 270, 271, 272, 273, 275, 276, 278, 280, 281, 283, 285, 286, 291, 292, 293, 294, 295, 296, 297, 298, 299, 300, 302, 305, 313, 318, 323, 324, 325, 327, 328, 329, 330, 331, 332, 333, 335, 336, 396, or 428, wherein numbering of amino acid residues in the Fc domain is according to the EU index as in Kabat. [0037] In certain embodiments, the Fc domain is an IgG Fc domain selected from a group consisting of a human IgGl Fc domain, a human IgG2 Fc domain, a human IgG3 Fc domain, and a human IgG4 Fc domain. In certain aspects, the immune- stimulatory conjugate induces the secretion of cytokines by an antigen presenting cell.
[0038] In certain aspects, the disclosure provides a pharmaceutical composition comprising immune- stimulatory conjugates as described herein and a pharmaceutically acceptable excipient. In certain aspects, the disclosure provides a method of treating cancer, comprising administering to a subject in need thereof an immune- stimulatory conjugate described herein. In certain aspects, the disclosure provides a method of treating cancer, comprising administering to a subject in need thereof a pharmaceutical composition comprising immune- stimulatory conjugates as described herein.
BRIEF DESCRIPTION OF THE DRAWINGS
[0039] The novel features of the disclosure are set forth with particularity in the appended claims. A better understanding of the features and advantages of the present disclosure will be obtained by reference to the following detailed description that sets forth illustrative aspects, in which the principles of the disclosure are utilized, and the accompanying drawings of which:
[0040] FIGURE 1 illustrates a schematic of a conjugate comprising an antibody and a second binding domain. The antibody contains two heavy chains as shown in gray and two light chains as shown in light gray. A portion of the heavy chains contain Fc domains (705 and 720). The antibody comprises a binding domain comprising two antigen binding sites (710 and 715). The second binding domain is attached to the antibody (780 and 785), for example, at the C-terminus of the heavy chains.
[0041] FIGURE 2 illustrates a schematic of an exemplary conjugate. The conjugate comprises an antibody, which contains two heavy chains as shown in gray and two light chains as shown in light gray. The antibody comprises a binding domain comprising two antigen binding sites (910 and 915), and a portion of the heavy chains contain Fc domains (905 and 920). The immune- stimulatory compounds (930 and 940) are attached to the antibody by linkers (960 and 970). A second binding domain is attached to the antibody (980 and 985), for example, at the C-terminus of the heavy chains.
[0042] FIGURE 3 illustrates a schematic of an exemplary conjugate. The conjugate comprises the Fc region of an antibody with the heavy chains shown in gray, and two scaffolds as shown in light gray. The conjugate comprises a first binding domain comprising two antigen binding sites (1110 and 1115) in the scaffolds, and a portion of the heavy chains contain Fc domains (1105 and 1120). The immune- stimulatory compounds (1130 and 1140) are conjugated to the Fc domains by linkers (1160 and 1170). A second binding domain is attached to the conjugate (1180 and 1185), for example, at the C-terminus of the heavy chains.
[0043] FIGURE 4 illustrates a schematic of an exemplary conjugate. The conjugate comprises the F(ab')2 region of an antibody with the Fab portions of heavy chains shown in gray and light chains shown in light gray, and two scaffolds as shown in dark gray. The conjugate comprises a first binding domain comprising two antigen binding sites (1310 and 1315), and a portion of two scaffolds contain Fc domains (1340 and 1345). The immune- stimulatory compounds (1330 and 1340) are attached to the conjugate by linkers (1360 and 1370). A second binding domain is attached to the Fc domains (1380 and 1385).
[0044] FIGURE 5 illustrates a schematic of an exemplary conjugate. The conjugate contains two scaffolds as shown in light gray and two scaffolds as shown in dark gray. The conjugate comprises a first binding domain comprising two antigen binding sites (1510 and 1515), and a portion of the two dark gray scaffolds contain Fc domains (1540 and 1545). The immune- stimulatory compounds (1530 and 1540) are conjugated to the conjugate by linkers (1560 and 1570). A second binding domain is attached to the conjugate (1580 and 1585).
[0045] FIGURE 6 illustrates a schematic of a conjugate comprising an antibody and a second binding domain. The antibody contains two heavy chains as shown in gray and two light chains as shown in light gray. A portion of the heavy chains contain Fc domains (1705 and 1720). The antibody comprises a binding domain comprising two antigen binding sites (1710 and 1715). The second binding domain is attached to the antibody (1780 and 1785), for example, at the C- terminus of the light chains.
[0046] FIGURE 7 illustrates a schematic of an exemplary conjugate. The conjugate comprises an antibody, which contains two heavy chains as shown in gray and two light chains as shown in light gray. The antibody comprises a binding domain comprising two antigen binding sites (1910 and 1915), and a portion of the heavy chains contain Fc domains (1905 and 1920). The immune- stimulatory compounds (1930 and 1940) are conjugated to the antibody by linkers (1960 and 1970). A second binding domain is attached to the antibody (1980 and 1985), for example, at the C-terminus of the light chains.
[0047] FIGURE 8 illustrates a schematic of an exemplary conjugate. The conjugate comprises an Fc region of an antibody shown in gray, and two scaffolds as shown in light gray. The conjugate comprises a first binding domain comprising two antigen binding sites (2110 and 2115) in the scaffolds, and a portion containing Fc domains (2105 and 2120). The immune- stimulatory compounds (2130 and 2140) are conjugated to the antibody construct by linkers (2160 and 2170). A second binding domain is attached to the antibody (2180 and 2185). [0048] FIGURE 9 illustrates a schematic of an exemplary conjugate. The conjugate comprises the F(ab')2 region of an antibody with heavy chains shown in gray and light chains shown in light gray, and two scaffolds as shown in dark gray. The conjugate comprises a first binding domain comprising two antigen binding sites (2310 and 2315), and a portion of two scaffolds contain Fc domains (2340 and 2345). The immune- stimulatory compounds (2330 and 2340) are conjugated to the antibody by linkers (2360 and 2370). A second binding domain is attached to the antibody (2380 and 2385), for example, at the C-terminus of the light chains.
[0049] FIGURE 10 illustrates a schematic of an exemplary conjugate. The conjugate contains two scaffolds as shown in light gray and two scaffolds as shown in dark gray. The conjugate comprises a first binding domain comprising two antigen binding sites (2510 and 2515), and a portion of the two dark gray scaffolds contain Fc domains (2540 and 2545). The immune- stimulatory compounds (2530 and 2540) are conjugated to the antibody construct by linkers (2560 and 2570). A second binding domain is attached to the conjugate (2580 and 2585).
[0050] FIGURE 11 illustrates a schematic of an antibody construct comprising an antibody. The antibody contains two heavy chains and two light chains. A portion of the heavy chains contain Fc domains (2705 and 2720). The antibody comprises a binding domain comprising two antigen binding sites shown in black (2710 and 2715).
[0051] FIGURE 12 illustrates a schematic of an antibody construct comprising an antibody. The antibody contains two heavy chains and two light chains. A portion of the heavy chains contain Fc domains (2925 and 2930). The antibody comprises a first binding domain comprising two antigen binding sites shown in black (2910 and 2915). The antibody comprises a second binding domain comprising two single chain variable fragments (2905 and 2920) attached to a C- terminus of the light chains. A single chain variable fragment can be attached to a light chain chain at a heavy chain variable domain of the single chain variable fragment. A single chain variable fragment can be attached to a light chain at a light chain variable domain of the single chain variable fragment.
[0052] FIGURE 13 illustrates a schematic of an antibody construct comprising an antibody. The antibody contains two heavy chains and two light chains. A portion of the heavy chains contain Fc domains (3120 and 3125). The antibody comprises a first binding domain comprising two antigen binding sites shown in black (3110 and 3115). The antibody comprises a second binding domain comprising two single chain variable fragments (3130 and 3135) attached to a C- terminus of the heavy chains. A single chain variable fragment can be attached to a heavy chain chain at a heavy chain variable domain of the single chain variable fragment. A single chain variable fragment can be attached to a heavy chain at a light chain variable domain of the single chain variable fragment. [0053] FIGURE 14 illustrates a schematic of an antibody construct comprising an antibody. The antibody contains two heavy chains and two light chains. A portion of the heavy chains contain Fc domains (3330 and 3335). The antibody comprises a first binding domain comprising two antigen binding sites shown in black (3310 and 3315). The antibody comprises a second binding domain comprising two single chain variable fragments (3320 and 3325) attached to a C- terminus of the light chains. A single chain variable fragment can be attached to a light chain chain at a heavy chain variable domain of the single chain variable fragment. A single chain variable fragment can be attached to a light chain at a light chain variable domain of the single chain variable fragment. The antibody comprises a third binding domain comprising two single chain variable fragments (3340 and 3345) attached to a C-terminus of the heavy chains. A single chain variable fragment can be attached to a heavy chain chain at a heavy chain variable domain of the single chain variable fragment. A single chain variable fragment can be attached to a heavy chain at a light chain variable domain of the single chain variable fragment.
[0054] FIGURE 15A sets forth a structure of an immune- stimulatory compound (left) and an immune- stimulatory compound with an amine handle (right top).
[0055] FIGURE 15B shows the x-ray crystal structure and binding orientation (pdb code 5D7A) of the immune- stimulatory compound as described in FIGURE 15A in a TNIK (TRAF2 and NCK- interacting protein kinase) active site. Specific interactions between the inhibitor and the TNIK active site are described in Masuda et al., TNIK inhibition abrogates colorectal cancer sternness, (2016) Nat Commun 7: 12586-12586.
[0056] FIGURE 15C shows a close up of the binding orientation of the immune- stimulatory compound as described in FIGURE 15A in a TNIK active site, where the linker and antibody portions are pointing away and sitting outside of the active site.
[0057] FIGURE 16 sets forth a structure of an immune- stimulatory compound (left) and an immune- stimulatory compound with an amine handle and linker surrogate (right). The structure on the right illustrates that the immune- stimulatory compound is predicted to sit in the enzyme active site, whereas the amine handle and linker surrogate are predicted to sit outside of the enzyme active site, in the solvent.
[0058] FIGURE 17A sets forth a structure of an immune- stimulatory compound (left) and an immune- stimulatory compound with a linker surrogate (left side of molecule on right).
[0059] FIGURE 17B shows the x-ray crystal structure and binding orientation (pdb code 5E91) of the immune- stimulatory compound as described in FIGURE 17 A in a TGFPR2 (transforming growth factor, beta receptor II) active site. Specific interactions between the inhibitor and the TGFpRII active site are described in Tebben et al., Crystal structures of apo and inhibitor-bound TGFbetaR2 kinase domain: insights into TGFbetaR isoform selectivity, (2016) Acta Crystallogr., Sect.D 72: 658-674.
[0060] FIGURE 17C shows a close up of the binding orientation of the immune- stimulatory compound as described in FIGURE 17 A in a TGFPR2 active site, where the linker and antibody construct portions are pointing away and sitting outside of the active site.
[0061] FIGURE 18A sets forth a structure of an immune- stimulatory compound (left) and an immune- stimulatory compound with an amine handle (upper left in molecule on right).
[0062] FIGURE 18B shows the x-ray crystal structure and binding orientation (pdb code 3KR8) of the immune- stimulatory compound as described in FIGURE 18A in a TNKS (tankyrase) active site.
[0063] FIGURE 18C shows a close up of the binding orientation of the immune- stimulatory compound as described in FIGURE 18A in a TNKS active site, where the linker and antibody construct portions are pointing away and sitting outside of the active site. Specific interactions between the inhibitor and the human tankyrase 2 - catalytic PARP domain active site are described in Karlberg et al., Structural basis for the interaction between tankyrase-2 and a potent Wnt-signaling inhibitor, (2010) J.Med.Chem. 53: 5352-5355.
[0064] FIGURE 19A shows a structure of an immune- stimulatory compound (left) and an immune- stimulatory compound with two amine handles (on right side of molecule on the right).
[0065] FIGURE 19B shows the x-ray crystal structure and binding orientation (pdb code 4191) of the immune- stimulatory compound as described in FIGURE 19A with dual site binding in the TNKS (tankyrase) active site. Specific interactions between the inhibitor and human tankyrase 1 are described in Bregman et al, Discovery of a class of novel tankyrase inhibitors that bind to both the nicotinamide pocket and the induced pocket, (2013) J.Med.Chem. 56: 1341-1345.
[0066] FIGURE 19C shows a close up of the binding orientation of the immune- stimulatory compound as described in FIGURE 19A in a TNKS active site, where the linker and antibody portions are pointing away and sitting outside of the active site.
[0067] FIGURE 20A illustrates a schematic of an exemplary conjugate and its molecular target. The conjugate comprises an antibody (3405) attached to a linker (3410) that is attached to a drug (3415) at the opposite end of the antibody (3405). The molecular target (3420) has an active site (3425) that is complementary to the drug (3415).
[0068] FIGURE 20B illustrates a schematic of an active exemplary conjugate that is bound to the the molecular target's active site. The drug (3415) sits within the active site of the molecular target (3420). The linker (3410) and antibody (3405) sit outside of the active site (3430).
[0069] FIGURE 20C illustrates a schematic of an active drug (3415) and linker (3410) that is bound to the molecular target (3420). [0070] FIGURE 20D illustrates a schematic of an active drug (3415) that is bound to the molecular target (3420).
[0071] FIGURE 21A, FIGURE 21B, and FIGURE 21C show the results of an assay for degradation of TFGpR2 by a TGFpR2-VHL PROTAC anti-HER2 antibody conjugate.
[0072] FIGURE 22A and FIGURE 22B show the results of an assay for antigen targeted degradation of TGFPR2 by an antibody conjugate with a PROTAC having VHL or Cereblon E3 binding moieties.
[0073] FIGURE 23A and FIGURE 23B show the results of an assay for cellular levels of TGFpR2 and TGFpRl in the presence of a TGFpR2/TGFpRl-VHL PROTAC with or without the addition of a proteasome inhibitor.
DETAILED DESCRIPTION
[0074] Additional aspects and advantages of the present disclosure will become apparent to those skilled in this art from the following detailed description, wherein illustrative aspects of the present disclosure are shown and described. As will be realized, the present disclosure is capable of other and different aspects, and its several details are capable of modifications in various respects, all without departing from the disclosure. Accordingly, the drawings and description are to be regarded as illustrative in nature, and not as restrictive.
[0075] As used herein, "potency" generally refers to measured bioactivity and may be quantified as an EC50 or IC50. Potency refers to the amount of a compound or conjugate needed to give an effect. For example, the potency of an immune- stimulatory compound which requires a lower amount of the immune- stimulatory compound compared with a different immune- stimulatory compound can be considered to have greater potency. Furthermore, the different immune- stimulatory compound requires a greater amount of the different immune- stimulatory compound to generate a response, and can therefore be considered lower potency. Potencies of bioactive compounds or conjugates may be measured over a concentration range and can be reported as those molar concentrations required to elicit or inhibit a percentage of the measured bioresponse. For example, a concentration required to stimulate 50% of observed maximal activity in the assay may be reported as an effective concentration 50 (EC50), to stimulate 90% activity as an EC90, or to stimulate 10% activity as an EC 10. For example, a concentration of an antagonist required to give 50% maximal inhibition of a biological activity may be reported as an inhibitory concentration 50 (IC50), to inhibit 90% as an IC90, or to inhibit 10% as an ICIO. This may allow for a comparison of the potencies of bioactive compounds on a molar basis by comparison of their EC or IC values for a given bioassay. For example, an immune- stimulatory conjugate or an immune- stimulatory compound with an EC50 or IC50 that is greater than 300 times the EC50 or IC50 of a control requires 300-fold higher, or more than 300-fold higher, concentration compared to the control to achieve a 50% bioresponse and has a potency weaker than the control by at least 300-fold. Therefore, an immune- stimulatory compound or conjugate that has an EC50 or IC50 not greater than about 300 times the EC50 or IC50 of a control compound may require no more than a 300-fold higher concentration than the control compound to achieve a 50% maximal bioresponse, no greater than 100 times the EC50 or IC50 requires no more than 100-fold higher concentration and no greater than 10 times the EC50 or IC50 requires no more than 10 times the concentration of the control. The potency of the immune- stimulatory compound or conjugate may be within 300-fold or better, 100-fold or better, or 10-fold or better the potency of the control.
[0076] As used herein, "control compound" refers to, in a case of an immune- stimulatory compound, the compound before attachment to a linker and antibody construct; in the case of an antibody construct, the construct before attachment of the linker and immune- stimulatory compound; or, in the case of a conjugate including an E3 ubiquitin ligase binding moiety, control compound refers to the immune- stimulatory compound attached to a linker that is attached to an E3 ubiquitin ligase binding moiety.
[0077] An "immune- stimulatory compound" as described herein, also referred to herein as an immune- stimulatory agent or molecule, refers to a molecule that stimulates the immune system by inducing activation or increasing activity of any of its components. For example, an immune stimulatory compound may bind directly to a component of the immune system and activate or increase an activity of the immune system or a component thereof. An immune stimulatory compound may also bind to an inhibitory component of the immune system and inactivate or decrease the activity of that component, thereby increasing an activity of the immune system. Immune- stimulatory compounds may have or cause antigenic specificity. In certain embodiments, an immune- stimulatory compound activates macrophages. In certain embodiments, an immune- stimulatory compound induces the secretion of cytokines by an antigen presenting cell. The increased activity or activation of the immune system may be evaluated using methods known in the field for evaluating an immune response (see, for example, J. Gratama, Cytometry A. 2008 Nov: 73(11): 971-4, the contents of which are incorporated herein in its entirety).
[0078] A "protein active site" as described herein, refers to a region of a protein target where a molecule (such as a substrate molecule or allosteric effector) binds to the target protein. The molecule may trigger a response upon binding to the the binding protein, e.g., inhibition of the activity of the binding protein or degradation of the binding protein. The protein active site includes the residues that interact with, e.g., bind covalently or non-covalently, a molecule such as an immune- stimulatory compound or a portion thereof. The protein active site may define a groove or pocket in the target protein wherein the interface between the groove or pocket and surrounding solvent is the solvent/active site interface of the protein active site.
[0079] The phrase "stimulate an immune response" refers to stimulating a response from immune cells that increases the activity or responsiveness of the immune system or a part thereof, such as, for example, inducing secretion of cytokines and/or chemokines, activating immune cells, e.g., T cells, B cells, macrophages, dendritic cells, etc., enhancing antigen presenting cell presentation of antigen, inducing antibody-dependent cell-mediated phagocytosis (ADCP), inducing antibody-dependent cell-mediated cytotoxicity (ADCC), NK cell or CD8+ T cell cytolytic activity, and/or blocking immune suppression.
[0080] As used herein, "homologous" or "homology" refer to the similarity or identity between a DNA, RNA, nucleotide, amino acid, or protein sequence to another DNA, RNA, nucleotide, amino acid, or protein sequence, respectively. Homology can be expressed in terms of a percentage of sequence identity of a first sequence to a second sequence. Percent (%) sequence identity with respect to a reference DNA sequence is the percentage of DNA nucleotides in a candidate sequence that are identical with the DNA nucleotides in the reference DNA sequence after aligning the sequences and introducing gaps, as necessary. Percent (%) sequence identity with respect to a reference amino acid sequence is the percentage of amino acid residues in a candidate sequence that are identical with the amino acid residues in the reference amino acid sequence after aligning the sequences and introducing gaps, if necessary, to achieve the maximum percent sequence identity, and not considering any conservative substitutions as part of the sequence identity.
[0081] As used herein, the abbreviations for the natural 1- enantiomeric amino acids are conventional and can be as follows: alanine (A, Ala); arginine (R, Arg); asparagine (N, Asn); aspartic acid (D, Asp); cysteine (C, Cys); glutamic acid (E, Glu); glutamine (Q, Gin); glycine (G, Gly); histidine (H, His); isoleucine (I, He); leucine (L, Leu); lysine (K, Lys); methionine (M, Met); phenylalanine (F, Phe); proline (P, Pro); serine (S, Ser); threonine (T, Thr); tryptophan (W, Trp); tyrosine (Y, Tyr); valine (V, Val). Unless otherwise specified, X can indicate any amino acid. In some aspects, X can be asparagine (N), glutamine (Q), histidine (H), lysine (K), or arginine (R).
[0082] As used herein, the term "antibody" refers to an immunoglobulin molecule that specifically binds to, or is immunologically reactive toward, a specific antigen. The term antibody can include, for example, polyclonal, monoclonal, genetically engineered, and antigen binding fragments thereof. An antibody can be, for example, murine, chimeric, humanized, bispecific, a heteroconjugate, a diabody, a triabody, a tetrabody or a hcAb (heavy chain antibody). An antigen binding fragment can include, for example, a Fab', F(ab')2, Fab, Fv, rlgG, scFv, a single domain antibody, VHH, VNAR, sdAb, or nanobody. [0083] As used herein, the term "antigen" refers to a molecule that can be bound by an antibody or a antibody construct. An antigen may elicit an immune response. An antigen can be a protein, polysaccharide, lipid, glycolipid or the like, which can be recognized by an antibody or an immune cell, such as a T cell or a B cell. Exposure of immune cells to one or more of these antigens can elicit a rapid cell division and differentiation response resulting in the formation of clones of the exposed T cells and B cells. B cells can differentiate into plasma cells which in turn can produce antibodies which selectively bind to the antigens.
[0084] As used herein, the terms "recognize," "bind," and "specifically bind" refer to the specific association or specific binding between an antigen binding domain and a corresponding antigen, or an immune stimulatory compound and a protein, as compared to the non-specific association or non-specific binding of the antigen binding domain or immune- stimulatory compound with a non-target antigen or protein.
[0085] As used herein, a "tumor antigen" is an antigen that can be expressed by or is present on, a cancer cell, a neoplastic tumor cell and/or within a tumor microenvironment.
[0086] As used herein, an "antibody construct" refers to a construct that contains an antigen binding domain and an Fc domain.
[0087] As used herein, an "antigen binding domain" refers to an antigen binding domain from an antibody or from a non-antibody that can specifically bind to the antigen. Antigen binding domains can be numbered when there is more than one antigen binding domain in a given conjugate or construct (e.g., first binding domain, second antigen binding domain, third antigen binding domain, etc.). Different antigen binding domains in the same conjugate or construct can target the same antigen or different antigens (e.g., first binding domain that can bind a tumor antigen, second antigen binding domain that can bind to a tumor antigen or an antigen presenting cell (APC) antigen, and third antigen binding domain that can bind an APC antigen).
[0088] As used herein, a "Fc domain" is an Fc domain from an antibody or from a non-antibody that can bind to an Fc receptor.
[0089] As used herein, an "Fc null" refers to an Fc domain that exhibits weak to no binding to any of the Fcgamma receptors. In some embodiments, an Fc null domain or region exhibits a reduction in binding affinity (e.g., increase in Kd) to Fc gamma receptors of at least 1000-fold.
[0090] As used herein, a "target binding domain" refers to a construct that contains an antigen binding domain from an antibody or from a non-antibody that can bind to the antigen.
[0091] The term "salt" or "pharmaceutically acceptable salt" refers to salts derived from a variety of organic and inorganic counter ions well known in the art. Pharmaceutically acceptable acid addition salts can be formed with inorganic acids and organic acids. Inorganic acids from which salts can be derived include, for example, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like. Organic acids from which salts can be derived include, for example, acetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, /?-toluenesulfonic acid, salicylic acid, and the like. Pharmaceutically acceptable base addition salts can be formed with inorganic and organic bases. Inorganic bases from which salts can be derived include, for example, sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, aluminum, and the like. Organic bases from which salts can be derived include, for example, primary, secondary, and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines, basic ion exchange resins, and the like, specifically such as
isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, and ethanolamine. In some embodiments, the pharmaceutically acceptable base addition salt is chosen from ammonium, potassium, sodium, calcium, and magnesium salts.
[0092] As used herein, "agonism" is the binding of a chemical to a receptor to induce a biological response. A chemical can be, for example, a small molecule, a compound, or a protein. An agonist causes a response, an antagonist can block the action of an agonist, and an inverse agonist can cause a response that is opposite to that of the agonist. A receptor can be activated by either endogenous or exogenous agonists.
[0093] The term "Cx-y" when used in conjunction with a chemical moiety, such as alkyl, alkenyl, or alkynyl is meant to include groups that contain from x to y carbons in the chain. For example, the term "Cx-yalkyl" refers to substituted or unsubstituted saturated hydrocarbon groups, including straight-chain alkyl and branched-chain alkyl groups that contain from x to y carbons in the chain, including haloalkyl groups such as trifluoromethyl and 2,2,2-trifluoroethyl, etc.
[0094] The terms "Cx-yalkenyl" and "Cx-yalkynyl" refer to substituted or unsubstituted
unsaturated aliphatic groups analogous in length and possible substitution to the alkyls described above, but that contain at least one double or triple bond, respectively.
[0095] The term "carbocycle" as used herein refers to a saturated, unsaturated or aromatic ring in which each atom of the ring is carbon. Carbocycle includes 3- to 10-membered monocyclic rings, 6- to 12-membered bicyclic rings, and 6- to 12-membered bridged rings. Each ring of a bicyclic carbocycle may be selected from saturated, unsaturated, and aromatic rings. In an exemplary embodiment, an aromatic ring, e.g., phenyl, may be fused to a saturated or unsaturated ring, e.g., cyclohexane, cyclopentane, or cyclohexene. Any combination of saturated, unsaturated and aromatic bicyclic rings, as valence permits, is included in the definition of carbocyclic.
Exemplary carbocycles include cyclopentyl, cyclohexyl, cyclohexenyl, adamantyl, phenyl, indanyl, and naphthyl. [0096] The term "heterocycle" as used herein refers to a saturated, unsaturated or aromatic ring comprising one or more heteroatoms. Exemplary heteroatoms include N, O, Si, P, B, and S atoms. Heterocycles include 3- to 10-membered monocyclic rings, 6- to 12-membered bicyclic rings, and 6- to 12-membered bridged rings. Each ring of a bicyclic heterocycle may be selected from saturated, unsaturated, and aromatic rings wherein at least one of the rings includes a heteroatom. In an exemplary embodiment, an aromatic ring, e.g., pyridyl, may be fused to a saturated or unsaturated ring, e.g., cyclohexane, cyclopentane, morpholine, piperidine or cyclohexene. The term "heteroaryl" includes aromatic single ring structures, preferably 5- to 7- membered rings, more preferably 5- to 6-membered rings, whose ring structures include at least one heteroatom, preferably one to four heteroatoms, more preferably one or two heteroatoms. The term "heteroaryl" also include polycyclic ring systems having two or more cyclic rings in which two or more carbons are common to two adjoining rings wherein at least one of the rings is heteroaromatic, e.g., the other cyclic rings can be aromatic or non-aromatic carbocyclic, or heterocyclic. Heteroaryl groups include, for example, pyrrole, furan, thiophene, imidazole, oxazole, thiazole, pyrazole, pyridine, pyrazine, pyridazine, and pyrimidine, and the like.
[0097] The term "substituted" refers to moieties having substituents replacing a hydrogen on one or more carbons or substitutable heteroatoms, e.g., NH, of the structure. It will be understood that "substitution" or "substituted with" includes the implicit proviso that such substitution is in accordance with permitted valence of the substituted atom and the substituent, and that the substitution results in a stable compound, i.e., a compound which does not spontaneously undergo transformation such as by rearrangement, cyclization, elimination, etc. In certain embodiments, substituted refers to moieties having substituents replacing two hydrogen atoms on the same carbon atom, such as substituting the two hydrogen atoms on a single carbon with an oxo, imino or thioxo group. As used herein, the term "substituted" is contemplated to include all permissible substituents of organic compounds. In a broad aspect, the permissible substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and non-aromatic substituents of organic compounds. The permissible substituents can be one or more and the same or different for appropriate organic compounds. For purposes of this disclosure, the heteroatoms such as nitrogen may have hydrogen substituents and/or any permissible substituents of organic compounds described herein which satisfy the valences of the heteroatoms.
[0098] In some embodiments, substituents may include any substituents described herein, for example: halogen, hydroxy, oxo (=0), thioxo (=S), cyano (-CN), nitro (-N02), imino (=N-H), oximo (=N-OH), hydrazino (=N-
NH2), -Rb-ORa, -Rb-OC(0)-Ra, -Rb-OC(0)-ORa, -Rb-OC(0)-N(Ra)2, -Rb-N(Ra)2, -Rb-C(0)Ra, -Rb -C(0)ORa, -Rb-C(0)N(Ra)2, -Rb-0-Rc-C(0)N(Ra)2, -Rb-N(Ra)C(0)ORa, -Rb-N(Ra)C(0)Ra, -Rb-N (Ra)S(0)tRa (where t is 1 or 2), -Rb-S(0)tRa (where t is 1 or 2), -Rb-S(0)tORa (where t is 1 or 2), and -Rb-S(0)tN(R )2 (where t is 1 or 2); and alkyl, alkenyl, alkynyl, aryl, aralkyl, aralkenyl, aralkynyl, cycloalkyl, cycloalkylalkyl, heterocyclo alkyl, heterocycloalkylalkyl, heteroaryl, and heteroarylalkyl any of which may be optionally substituted by alkyl, alkenyl, alkynyl, halogen, haloalkyl, haloalkenyl, haloalkynyl, oxo (=0), thioxo (=S), cyano (-CN), nitro (-N02), imino (=N-H), oximo (=N-OH), hydrazine (=N-
NH2), -Rb-ORa, -Rb-OC(0)-Ra, -Rb-OC(0)-ORa, -Rb-OC(0)-N(Ra)2, -Rb-N(Ra)2, -Rb-C(0)Ra, -Rb -C(0)ORa, -Rb-C(0)N(Ra)2, -Rb-0-Rc-C(0)N(Ra)2, -Rb-N(Ra)C(0)ORa, -Rb-N(Ra)C(0)Ra, -Rb-N (Ra)S(0)tRa (where t is 1 or 2), -Rb-S(0)tRa (where t is 1 or 2), -Rb-S(0)tORa (where t is 1 or 2) and -Rb-S(0)tN(R )2 (where t is 1 or 2); wherein each R is independently selected from hydrogen, alkyl, cycloalkyl, cycloalkylalkyl, aryl, aralkyl, heterocycloalkyl,
heterocycloalkylalkyl, heteroaryl, or heteroarylalkyl, wherein each R , valence permitting, may be optionally substituted with alkyl, alkenyl, alkynyl, halogen, haloalkyl, haloalkenyl, haloalkynyl, oxo (=0), thioxo (=S), cyano (-CN), nitro (-N02), imino (=N-H), oximo (=N-OH), hydrazine (=N-
NH2), -Rb-ORa, -Rb-OC(0)-Ra, -Rb-OC(0)-ORa, -Rb-OC(0)-N(Ra)2, -Rb-N(Ra)2, -Rb-C(0)Ra, -Rb -C(0)ORa, -Rb-C(0)N(Ra)2, -Rb-0-Rc-C(0)N(Ra)2, -Rb-N(Ra)C(0)ORa, -Rb-N(Ra)C(0)Ra, -Rb-N (Ra)S(0)tRa (where t is 1 or 2), -Rb-S(0)tRa (where t is 1 or 2), -Rb-S(0)tORa (where t is 1 or 2) and -Rb-S(0)tN(R )2 (where t is 1 or 2); and wherein each Rb is independently selected from a direct bond or a straight or branched alkylene, alkenylene, or alkynylene chain, and each Rc is a straight or branched alkylene, alkenylene or alkynylene chain.
[0099] It will be understood by those skilled in the art that substituents can themselves be substituted, if appropriate. Unless specifically stated as "unsubstituted," references to chemical moieties herein are understood to include substituted variants. For example, reference to a "heteroaryl" group or moiety implicitly includes both substituted and unsubstituted variants.
[0100] Chemical entities having carbon-carbon double bonds or carbon-nitrogen double bonds may exist in Z- or E- form (or cis- or trans- form). Furthermore, some chemical entities may exist in various tautomeric forms. Unless otherwise specified, chemical entities described herein are intended to include all Z-, E- and tautomeric forms as well.
[0101] A "tautomer" refers to a molecule wherein a proton shift from one atom of a molecule to another atom of the same molecule is possible. The compounds presented herein, in certain embodiments, exist as tautomers. In circumstances where tautomerization is possible, a chemical equilibrium of the tautomers will exist. The exact ratio of the tautomers depends on several factors, including physical state temperature, solvent, and pH. Some examples of tautomeric
equilibrium include:
[0102] The compounds, constructs and conjugates disclosed herein, in some embodiments, are used in different enriched isotopic forms, e.g., enriched in the content of 2 H, 3 H, 11 C, 13 C and/or 14C. In one particular embodiment, the compound is deuterated in at least one position. Such deuterated forms can be made by the procedure described in U.S. Patent Nos. 5,846,514 and 6,334,997, or as otherwise known in the art. As described in U.S. Patent Nos. 5,846,514 and 6,334,997, deuteration can improve the metabolic stability and or efficacy, thus increasing the duration of action of drugs.
[0103] Unless otherwise stated, structures depicted herein are intended to include compounds which differ only in the presence of one or more isotopically enriched atoms. For example, compounds having the present structures except for the replacement of a hydrogen by a deuterium or tritium, or the replacement of a carbon by 13C- or 14C-enriched carbon are within the scope of the present disclosure.
[0104] The compounds of the present disclosure optionally contain unnatural proportions of atomic isotopes at one or more atoms that constitute such compounds. For example, the compounds may be labeled with isotopes, such as for example, deuterium ( 2 H), tritium ( 3 H), iodine- 125 (125I) or carbon- 14 (14C). Isotopic substitution with 2H, UC, 13C, 14C, 15C, 12N, 13N, 15N, 16N, 160, 170, 14F, 15F, 16F, 17F, 18F, 33S, 34S, 35S, 36S, 35C1, 37C1, 79Br, 81Br, and 125I are all contemplated. All isotopic variations of the compounds of the present invention, whether radioactive or not, are encompassed within the scope of the present invention.
[0105] In certain embodiments, the compounds disclosed herein have some or all of the 1H atoms replaced with H atoms. The methods of synthesis for deuterium-containing compounds are known in the art and include, by way of non-limiting example only, the following synthetic methods.
[0106] Deuterium substituted compounds are synthesized using various methods such as described in: Dean, Dennis C; Editor. Recent Advances in the Synthesis and Applications of Radiolabeled Compounds for Drug Discovery and Development. [In: Curr., Pharm. Des., 2000; 6(10)] 2000, 110 pp; George W.; Varma, Rajender S. The Synthesis of Radiolabeled Compounds via Organometallic Intermediates, Tetrahedron, 1989, 45(21), 6601-21; and Evans, E. Anthony. Synthesis of radiolabeled compounds, J. Radioanal. Chem., 1981, 64(1-2), 9-32.
[0107] Deuterated starting materials are readily available and are subjected to the synthetic methods described herein or otherwise known to provide for the synthesis of deuterium- containing compounds. Large numbers of deuterium-containing reagents and building blocks are available commercially from chemical vendors, such as Aldrich Chemical Co.
[0108] Compounds also include crystalline and amorphous forms of those compounds, pharmaceutically acceptable salts, and active metabolites of these compounds having the same type of activity, including, for example, polymorphs, pseudopolymorphs, solvates, hydrates, unsolvated polymorphs (including anhydrates), conformational polymorphs, and amorphous forms of the compounds, as well as mixtures thereof.
[0109] The phrases "parenteral administration" and "administered parenterally" as used herein means modes of administration other than enteral and topical administration, usually by injection, and includes, without limitation, intravenous, intramuscular, intraarterial, intrathecal,
intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intraarticular, subcapsular, subarachnoid, intraspinal and intrasternal injection and infusion.
[0110] The phrase "pharmaceutically acceptable" is employed herein to refer to those
compounds, materials, compositions, and/or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit/risk ratio.
[0111] The phrase "pharmaceutically acceptable excipient" or "pharmaceutically acceptable carrier" as used herein means a pharmaceutically acceptable material, composition or vehicle, such as a liquid or solid filler, diluent, excipient, solvent or encapsulating material. Each carrier must be "acceptable" in the sense of being compatible with the other ingredients of the formulation and not injurious to the patient. Some examples of materials which can serve as pharmaceutically acceptable carriers include: (1) sugars, such as lactose, glucose and sucrose; (2) starches, such as corn starch and potato starch; (3) cellulose, and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; (4) powdered tragacanth; (5) malt; (6) gelatin; (7) talc; (8) excipients, such as cocoa butter and suppository waxes; (9) oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; (10) glycols, such as propylene glycol; (11) polyols, such as glycerin, sorbitol, mannitol and polyethylene glycol; (12) esters, such as ethyl oleate and ethyl laurate; (13) agar; (14) buffering agents, such as magnesium hydroxide and aluminum hydroxide; (15) alginic acid; (16) pyrogen- free water; (17) isotonic saline; (18) Ringer's solution; (19) ethyl alcohol; (20) phosphate buffer solutions; and (21) other non-toxic compatible substances employed in pharmaceutical formulations.
[0112] As used herein, "attached" refers to a covalent bond, between two or more groups.
Alternatively, attached may refer to the connection of two or more groups via a linker, e.g., a linker connecting a second binding domain to an antibody construct. A fusion may refer to a nucleic acid sequence of two separate domains being expressed in frame. For example, a binding domain can be attached as a fusion or by attachment (e.g., conjugation) via a linker to an antibody construct or other portion of a conjugate. For example, an antibody can be fused with an additional binding domain to create an antibody construct containing a fusion of the antibody and the additional binding domain. The antibody construct can be the result of the nucleic acid sequence of the binding domain being expressed in frame with the nucleic acid sequence of the antibody construct. The fusion can be the result of an in- frame nucleotide sequence encoding the antibody construct with the binding domain. As another example, an additional binding domain can be attached to an antibody construct via a linker, wherein the linker is attached (i.e., conjugated) to the binding domain and the linker is attached to the antibody construct. The binding domain can be linked to the linker by a chemical conjugation and the antibody construct can be linked to the linker by a chemical conjugation. The additional binding domain can be a second binding domain and/or a third binding domain as described herein. Furthermore, a binding domain can be a first binding domain attached to an Fc domain to produce the antibody construct as described herein, which may produce the first binding domain as a fusion with the Fc domain or a conjugate wherein the first binding domain can be linked to a linker and the linker can be linked to the Fc domain.
Immune-Stimulatory Conjugates
[0113] Disclosed herein are conjugates of antibody constructs, immune- stimulatory compounds and linkers, referred to as antibody construct- immune- stimulatory compound conjugates (also referred to as immune- stimulatory compound-conjugates, antibody conjugates or conjugates). The conjugates may be used in the treatment of various diseases and disorders, including cancers. Also disclosed pharmaceutical compositions of the conjugates. In certain embodiments, immune- stimulatory compounds are attached either directly or through a linker to an antibody construct to form antibody construct-immune- stimulatory compound conjugates.
[0114] In some embodiments, the immune- stimulatory compound stimulates the immune system, or a component thereof. In some embodiments, the immune- stimulatory compound has an immune-modulatory activity. In some embodiments, the immune- stimulatory compound has an inhibitory effect on a component of the immune system, thereby stimulating an immune- modulatory activity.
[0115] In certain embodiments, conjugates are represented by the following formula:
wherein A is an antibody construct, L is a linker, D is an immune- stimulatory compound, x may be from 1 to 20 (wherein each x denotes a separate compound), n may be from 1 to 20, and z may be from 1 to 20. In some embodiments, L is a cleavable linker. In some embodiments, L is a non-cleavable linker.
[0116] In some embodiments, x is 1, n is 1 and z may be from 1 to 10, such as from 1 to 9, such as from 1 to 8, such as from 2 to 8, such as from 1 to 6, such as from 3 to 5 or such as about 2.
[0117] In some embodiments, x is 1, n is 2, and z may be from 1 to 10, such as from 1 to 9, such as from 1 to 8, such as from 2 to 8, such as from 1 to 6 or such as from 3 to 5. In certain embodiments, z is 4.
[0118] In some embodiments, x may be from 1-20, n may be from 1-20, and z may be from 1 to
20.
[0119] In certain embodiments, conjugates are represented by the following formula:
wherein A is an antibody construct, L is a linker having the structure -Aa-Ww-Yy-, where A is a spacer, a is 0 or 1, W is a cleavable unit, w may be from 0 to 10, Y is a stretcher, y may be from 0 to 3, D is an immunomodulatory compound, x may be from 1 to 20 (wherein each x denotes a distinct compound), n may be from 1 to 20, and z may be from 1 to 20. [0120] In some embodiments, a is 1, w is 0, y is 0, x is 1, n is 1, and z may be from 1 to 20, 1 to 10, 1 to 9, 1 to 8, 2 to 8, 1 to 6 or 3 to 5. In certain embodiments, z is 4.
[0121] In some embodiments, a isl, w is 1, y is from 1, x is 1, n is 1 and z may be from 1 to 10, 1 to 9, 1 to 8, 2 to 8, 1 to 6, 3 to 5, 2 or 4.
[0122] In some embodiments, a is 0 or 1, w is from 0 to 10, y is from 0-3, where at least one of a, w or y is present, x may be from 1 to 20, n may be from 1-20, and z may be from 1 to 20.
[0123] In some embodiments, the immune- stimulatory conjugates include a linker (L) that may comprise from 5 to 100 linear non-hydrogen atoms that is covalently attached to an antibody construct (A) and may be:
i) covalently attached to an immune- stimulatory compound (CO as in the following formula:
ii) covalently attached to an immune- stimulatory compound (CO which itself may be covalently attached to a spacer (S) comprising from 5 to 100 linear no n- hydrogen atoms covalently attached to a second immune- stimulatory compound (C2) as in the following formula:
iii) covalently attached an immune- stimulatory compound (C2) that may be covalently attached to a spacer (S) comprising from 5 to 100 linear non-hydrogen atoms covalently attached to a formula: , or
iv) covalently attached to a spacer (S) comprising from 5 to 100 linear non- hydrogen atoms that is covalently attached immune- stimulatory compounds (Ci and C2) as in the following formula:
[0124] In some embodiments, an immune- stimulatory conjugate is provided that includes a proteolysis targeting module (PTM; also referred to as a proteolysis-targetting chimera or PROTAC) that includes an immune-modulatory compound (IMC) that is covalently attached to an E3 ubiquitin ligase binding moiety (ULM) through a spacer (S) and wherein a linker (L) is covalently attached to the protein targeting molecule and to the antibody construct as
represented by the formula < A>-(L-PTMn)z, where n is from 1-20 and z is from 1 to 20. In some embodiments, L is a cleavable linker. In some embodiments, S is a non-cleavable spacer.
[0125] Referring to the previous formula, in some embodiments, Ci is a target protein targeting moiety, such as an immodulatory compound, C2 is an E3 ubiquitin ligase binding moiety such that together C1-S-C2 may form a PTM.
[0126] In some embodiments, an immune-modulatory compound or other protein targeting moiety (IMC) is covalently attached to an E3 ubiquitin ligase binding moiety (ULM) through a spacer (S) and a linker (L) is covalently attached to the spacer, n is from 1-20 and z is from 1 to 20 as represented by the for
[0127] In some embodiments, L is a cleavable linker. In some embodiments, L is a non- cleavable linker. In some embodiments, S is non-cleavable.
[0128] In some embodiments, an immune-modulatory compound or other protein targeting moiety (IMC) is covalently attached to an E3 ubiquitin ligase binding moiety (ULM) through a spacer (S) and a linker (L) is covalently attached to the IMC, n is from 1-20 and z is from 1 to 20 as represented by the formula:
[0129] In some embodiments, L is a cleavable linker. In some embodiments, L is a non- cleavable linker.
[0130] In some embodiments, an immune-modulatory compound or other protein targeting moiety (IMC) is covalently attached to an E3 ubiquitin ligase binding moiety (ULM) through a spacer (S) and linker L is covalently attached to the ubiquitin E3 ligase moiety (ULM), n is from 1-20 and z is from 1 to 20 as represented by the formula:
[0131] In some embodiments, L is a cleavable linker. In some embodiments, L is a non- cleavable linker. In some embodiments, S is non-cleavable.
Tumor and Immune Cell Antigens
[0132] In cancer, there are several general groups of tumor antigens, including but not limited to: (i) viral tumor antigens which can be identical for any viral tumor of this type, (ii) carcinogenic tumor antigens which can be specific for patients and for the tumors, (iii) isoantigens of the transplantation type or tumor- specific transplantation antigens which can be different in all individual types of tumor but can be the same in different tumors caused by the same inciting biological event; and (iv) embryonic antigens.
[0133] As a result of the discovery of tumor antigens, tumor antigens have become important in the development of new cancer treatments that can specifically target the cancer. This has led to the development of antibodies directed against these tumor antigens. Such tumor antigens include the following: CD5, CD19, CD20, CD25, CD37, CD30, CD33, CD45, CAMPATH-1, BCMA, CS-1, PD-L1, B7-H3, B7-DC (PD-L2), HLD-DR, carcinoembryonic antigen (CEA), TAG-72, EpCAM, MUC1, MUC15, MUC16, fo late-binding protein, A33, G250, prostate- specific membrane antigen (PSMA), ferritin, GD2, GD3, GM2, Ley, CA-125, CA19-9, epidermal growth factor, pl85HER2, IL-2 receptor, EGFRvIII (de2-7 EGFR), fibroblast activation protein (FAP), tenascin, a metalloproteinase, endosialin, vascular endothelial growth factor, ανβ3, WT1, LMP2, HPV E6, HPV E7, Her-2/neu, p53 nonmutant, NY-ESO-1, MelanA/MARTl, Ras mutant, gplOO, p53 mutant, PR1, bcr-abl, tyrosinase, survivin, PSA, hTERT, a Sarcoma translocation breakpoint protein, EphA2, PAP, ML-IAP, AFP, ERG, NA17, PAX3, ALK, androgen receptor, cyclin B l, polysialic acid, MYCN, RhoC, TRP-2, fucosyl GM1, mesothelin (MSLN), PSCA, MAGE Al, MAGE A3, sLe(animal), CYP1B 1, PLAV1, GM3, BORIS, Tn, GloboH, ETV6- AML, NY-BR-1, RGS5, SART3, STn, Carbonic anhydrase IX, PAX5, OY-TES l, Sperm protein 17, LCK, HMWMAA, AKAP-4, SSX2, XAGE 1, Legumain, Tie 3, Page 4, VEGFR2, MAD- CT-1, PDGFR-B, MAD-CT-2, ROR2, TRAILl, MAGE A4, MAGE C2, GAGE, EGFR, CMET, HER3, EPCAM, CA6, NAPI2B, TROP2, Claudin-6 (CLDN6), Claudin-16 (CLDN16),
CLDN18.2, RON, LY6E, FRA, DLL3, PTK7, Uroplakin-IB (UPK1B), LIV1, ROR1, STRA6, TMPRSS3, TMPRSS4, TMEM238, Clorfl86, Fos-related antigen 1, VEGFR1, endoglin, VTCN1 (B7-H4), VISTA, and gpNMB.
[0134] In addition to the development of antibodies against tumor antigens for cancer treatment, antibodies that target immune cells to boost the immune response have also been developed, including the following: an anti-CD40 antibody, an anti-CD47 antibody, anti-TNFR2 antibody, an anti-DEC205 antibody, an anti-CD36 mannose scavenger receptor 1 antibody, an anti- CLEC9A antibody, an anti-CLEC12A antibody, an anti-DC-SIGN antibody, an anti-BDCA-2 antibody, an anti-OX40L antibody, an anti-41BBL antibody, an anti-CD204 antibody, an anti- MARCO antibody, an anti-CLEC5A antibody, an anti-Dectin 1 antibody, and anti-Dectin 2 antibody, an anti-CLEClOA antibody, an anti-CD206 antibody, an anti-CD64 antibody, an anti- CD32A antibody, an anti-CD 16A antibody, an anti-HVEM antibody, an anti-CD38 antibody, an anti-PD-Ll antibody, an anti-TREM2 antibody, an anti-CSFIR antibody, or an anti-CD32B antibody can be used to target, respectively, cell surface CD40, CD47, TNFR2, DEC-205, CD36 mannose scavenger receptor 1, CLEC9A, CLEC12A, DC-SIGN, BDCA-2, OX40L, 41BBL, CD204, MARCO, CLEC5A, Dectin 1, Dectin 2, CLECIOA, CD206, CD64, CD32A, CD 16 A, HVEM, CD38, PD-L1, TREM2, CSF1R, or CD32B molecules expressed by antigen presenting cells.
[0135] Cluster of Differentiation 40 (CD40) is a member of the Tumor Necrosis Factor Receptor (TNF-R) family. CD40 can be a 50 kDa cell surface glycoprotein that can be constitutively expressed in normal cells, such as monocytes, macrophages, B lymphocytes, dendritic cells, endothelial cells, smooth muscle cells, fibroblasts and epithelium, and in tumor cells, including B-cell lymphomas and many types of solid tumors. Expression of CD40 can be increased in antigen presenting cells in response to IL-Ιβρ, IFN-γ, GM-CSF, and LPS induced signaling events.
[0136] Examples of agonistic CD40 monoclonal antibodies include CP-870,893, dacetuzumab, Chi Lob 7/4, SEA-CD40, ADC- 1013, 3C3, or 3G5.
[0137] Cluster of Differentiation 205 (CD205 or DEC-205) is a member of the C-type multilectin family of endocytic receptors, which can include the macrophage mannose receptor (MMR) and the phospholipase A2 receptor (PLA2R). DEC-205 can be a 205 kDa endocytic receptor highly expressed in cortical thymic epithelial cells, thymic medullary dendritic cells (CDl lc+ CD8+), subpopulations of peripheral dendritic cells (CDl lc+ CD8+). The DEC-205+ CDl lc+ CD8+ dendritic cells (DCs) can function in cross-presentation of antigens derived from apoptotic cells. Additionally, DEC-205 can be significantly upregulated during DC maturation. DEC-205 can also be expressed at moderate levels in B cells and low levels in macrophages and T cells.
[0138] After antigen binding to DEC-205, the receptor- antigen complex can be internalized whereupon the antigen can be processed and be presented on the DC surface by a major histocompatibility complex class II (MHC II) or MHC class I. DEC-205 can deliver antigen to DCs for antigen presentation on MHC class II and cross-presentation on MHC class I. DEC-205 mediated antigen delivery for antigen presentation in DCs without an inflammatory stimulus can result in tolerance. Conversely, DEC-205 mediated antigen delivery in DCs in the presence of a maturational stimulus (e.g., a CD40 agonist) can result in long-term immunity via activation of antigen- specific CD4+ and CD8+ T cells.
[0139] CD36 mannose scavenger receptor 1 is an oxidized LDL receptor with two
transmembrane domains located in the caveolae of the plasma membrane. It can be classified as a Class B scavenger receptor, which can be characterized by involvement in the removal of foreign substances and waste materials. This receptor can also be involved in cell adhesion, phagocytosis of apoptotic cells, and metabolism of long-chain fatty acids.
[0140] TNFR2 (tumor necrosis factor receptor 2), also known as TNFRSF1B (tumor necrosis factor receptor super family IB) and CD 120b, is a single-pass type I membrane protein and the member of TNFR superfamily containing 4 cysteine-rich domains (CRD) repeats. In addition to the full length membrane- anchored form, soluble TNFR2 can be generated via two distinct mechanisms: (1) shedding via proteolytic processing of the full membrane anchored from, and (2) translation from an alternatively spliced message encoding the extracellular domains of TNFR2. TNFR2 is the receptor with high affinity for TNF-alpha and approximately 5-fold lower affinity for homotrimeric lymphotoxin-alpha. TNFR2 (Tumor Necrosis Factor Receptor Type II) and TNF-receptor 1 form a heterocomplex that mediates the recruitment of two anti- apoptotic proteins, c-IAPl and C-IAP2, which possess E3 ubiquitin ligase activity. c-IAPl can potentiate TNF- induced apoptosis by the ubiquitination and degradation of TNF-receptor-associated factor 2, which mediates anti- apoptotic signals. Knockout studies in mice suggest a role of TNFR2 in protecting neurons from apoptosis by stimulating antioxidative pathways.
[0141] CLEC9A is a group V C-type lectin receptor. This receptor can be expressed as on myeloid lineage cells, and can be characterized as an activation receptor.
[0142] CLEC12A is a member of the C-type lectin/C-type lectin like domain super family that can be a negative regulator of granulocyte and monocyte function. It can also be involved in cell adhesion, cell-cell signaling, and glycoprotein turnover, and can play a role in the inflammatory response. [0143] Dendritic cell-specific inter cellular adhesion molecule-3-grabbing non-integrin (DC- SIGN) or CD209, is a C-type lectin receptor that can be expressed on the surface of macrophages and dendritic cells. This receptor can recognize and bind to mannose type carbohydrates and be involved in activating phagocytosis, can mediate dendritic cell rolling, and can be involved in CD4+ T cell activation.
[0144] BDCA-2 is a C-type lectin that is a membrane protein of plasmacytoid dendritic cells. It can be involved in plasmacytoid dendritic cell function, such as ligand internalization and presentation.
[0145] OX40L, which is also referred to as CD252, is the ligand for CD 134 that can be expressed on dendritic cells. It can be involved in T cell activation.
[0146] 41BBL, which is also referred to as CD137L, is a member of the TNF superfamily, and can be expressed on B cells, dendritic cells, activated T cells, and macrophages. It can provide co-stimulatory signal for T cell activation and expansion.
[0147] CD204, which is also referred to as macrophage scavenger receptor 1, is a macrophage scavenger receptor receptor. The gene for CD204 can encode three different class A macrophage scavenger receptor isoforms. The type 1 and type 2 isoforms can be involved in binding, internalizing, and processing negatively charged macromolecules, such as low density lipoproteins. The type 3 isoform can undergo altered intracellular processing in which it can be retained within the endoplasmic reticulum, and has been shown to have a dominant negative effect on the type 1 and type 2 isoforms.
[0148] Macrophage receptor with collagenous structure (MARCO), which is also referred to as SCARA2, is a class A scavenger receptor with collagen-like and cysteine-rich domains. It can be expressed in macrophages, and can bind to modified low density lipoproteins. It can be involved in the removal of foreign substances and waste materials.
[0149] C-type lectin domain family 5 member A (CLEC5A) is a C-type lectin. It can be involved in the myeloid lineage activating pathway.
[0150] Dendritic cell-associated c-type lectin- 1 (Dectin 1), which is also referred to as CLEC7A, is member of the C-type lectin/C-type lectin- like super family. It can be expressed by myeloid dendritic cells, monocytes, macrophages, and B cells, and can be involved in antifungal immunity.
[0151] Dendritic cell-associated c-type lectin-2 (Dectin 2), which is also referred to as CLEC6A, is member of the C-type lectin/C-type lectin- like super family. It can be expressed by dendritic cells, macrophages, monocytes and neutrophils. It can be involved in antifungal immunity. [0152] CLECIOA, which is also referred to as CD301, is member of the C-type lectin/C-type lectin-like super family. It can be expressed by dendritic cells, monocytes, and CD33+ myeloid cells, and can be involved in macrophage adhesion and migration.
[0153] CD206, which is also referred to as macrophage mannose receptor, is a C-type lectin type I membrane glycoprotein. It can be expressed on dendritic cells, macrophages and endothelial cells, and can act as a pattern recognition receptor and bind high-mannose structures of viruses, bacteria, and fungi.
[0154] CD64, which is also referred to as FcyRI, is a high affinity Fc receptor for IgG. It can be expressed by monocytes and macrophages. It can be involved in mediating phagocytosis, antigen capture, and antibody dependent cell-mediated cytoxicity.
[0155] CD32A, which is also referred to as FcyRIIa, is a low affinity Fc receptor. It can be expressed on monocytes, granulocytes, B cells, and eosinophils. It can be involved in
phagocytosis, antigen capture, and antibody dependent cell-mediated cytoxicity.
[0156] CD 16 A, which is also referred to as FcyRIIIa, is low affinity Fc receptor. It can be expressed on NK cells, and can be involved in phagocytosis and antibody dependent cell- mediated cytotoxicity.
[0157] Herpesvirus entry mediator (HVEM), which is also referred to as CD270, is a member of the TNF-receptor superfamily. It can be expressed on B cells, dendritic cells, T cells, NK cells, CD33+ myeloid cells, and monocytes. It can be involved in activating the immune response.
[0158] CD32B, which is also referred to as FcyRIIb, is a low affinity Fc receptor. It can be expressed on B cells and myeloid dendritic cells. It can be involved in inhibiting maturation and cell activation of dendritic cells.
[0159] The HER2/neu (human epidermal growth factor receptor 2/receptor tyro sine-protein kinase erbB-2) is part of the human epidermal growth factor family. Overexpression of this protein can be shown to play an important role in the progression of cancer, for example, breast cancer. The HER2/neu protein can function as a receptor tyrosine kinase and autophosphorylates upon dimerization with binding partners. HER2/neu can activate several signaling pathways including, for example, mitogen-activated protein kinase, phosphoinositide 3-kinase,
phospholipase Cy, protein kinase C, and signal transducer and activator of transcription (STAT). Examples of antibodies that can target and inhibit HER2/neu can include trastuzumab and pertuzumab.
[0160] EGFR (epidermal growth factor receptor) encodes a member of the human epidermal growth factor family. Mutations that can lead to EGFR overexpression or over activity can be associated with a number of cancers, including squamous cell carcinoma and glioblastomas. EGFR can function as a receptor tyrosine kinase and ligand binding can trigger dimerization with binding partners and autophosphorylation. The phosphorylated EGFR can then activate several downstream signaling pathways including mitogen-activated protein kinase, phosphoinositide 3- kinase, phospholipase Cy, protein kinase C, and signal transducer and activator of transcription (STAT). Examples of antibodies that can target and inhibit EGFR can include cetuximab, panutumumab, nimotuzumab, and zalutumumab. One mutant variant of EGFR is EGFRvIII (epidermal growth factor receptor variant III, also referred to as de2-7EGFR). EGFRvIII is the result of an EGFR gene rearrangement in which exons 2-7 of the extracellular domain are deleted. This mutation can result in a mutant receptor incapable of binding to any known ligand. The resulting receptor can engage in a constitutive low-level signaling and can be implicated in tumor progression. Examples of antibodies that can target EGFRvIII can include AMG595 and ABT806.
[0161] C-Met (hepatocyte growth factor receptor) encodes a member of the receptor tyrosine kinase family of proteins. C-Met overexpression and over activity can be implicated in various cancers including lung adenocarcinomas, and high c-Met levels can be associated with poor patient outcome. Binding of hepatocyte growth factor can induce dimerization and
autophosphorylation of c-Met. The c-Met receptor can activate various downstream signaling pathways including mitogen-activated protein kinase, phosphoinositide 3-kinase, and protein kinase C pathways. The antibody onartuzumab can target and inhibit c-Met.
[0162] HER3 (human epidermal growth factor receptor 3) encodes a member of the human epidermal growth factor receptor family. Ligand binding can induce dimerization and autophosphorylation of cytoplasmic tyrosine residues that then can recruit signaling proteins for downstream signaling pathway activation including mitogen-activated protein kinase and phosphoinoside 3-kinase pathways. HER3 can play an active role in cell proliferation and survival, and can be overexpressed, overactive, and/or mutated in various cancers. For example, HER3 can be overexpressed in breast, ovarian, prostate, colon, pancreas, stomach, oral, and lung cancers. The antibody patritumab can target and inhibit HER3.
[0163] MUC1 (mucin 1, cell surface associated) encodes a member of the mucin family of glycosylated proteins that can play an important role in cell adhesion and forming protective mucosal layers on epithelial surfaces. MUC1 can be proteolytically cleaved into alpha and beta subunits that form a heterodimeric complex with the N-terminal alpha subunit providing cell- adhesion functionality and the C-terminal beta subunit modulating cell signaling pathways including the mitogen activated map kinase pathway. MUC1 can play a role in cancer progression, for example, by regulating TP53-mediated transcription. MUC1 overexpression, aberrant intracellular localization, and glycosylation changes can all be associated with carcinomas including pancreatic cancer cells. The antibody clivatuzumab can target MUC1. [0164] MUC16 (mucin 16, cell surface associated) encodes the largest member of the mucin family of glycosylated proteins that can play an important role in cell adhesion and forming protective mucosal layers on epithelial surfaces. MUC16 can be a highly glycosylated 2.5MDa transmembrane protein that can provide a hydrophilic lubricating barrier on epithelial cells. The cytoplasmic tail of MUC16 can be involved with various signaling pathways including the JAK2-STAT3 and Src kinase pathways. A peptide epitope of MUC16 can be used as biomarker for detecting ovarian cancer. Elevated expression of MUC16 can be present in advanced ovarian cancers and pancreatic cancers. The antibody sofituzumab can target MUC16.
[0165] EPCAM (epithelial cell adhesion molecule) encodes a transmembrane glycoprotein that can be frequently and highly expressed in carcinomas and tumor- initiating cells. EPCAM can also be a pluripotent stem cell marker. EPCAM can modulate a variety of pathways including cell-cell adhesion, cellular proliferation, migration, invasion, maintenance of a pluripotent state, and differentiation in the context of tumor cells. The antibodies edrecolomab and adecatumumab can target EPCAM.
[0166] MSLN (mesothelin) encodes a 40 kDa cell GPI-anchored membrane surface protein believed to function in cell adhesion. MSLN is overexpressed in mesothelioma and certain types of pancreatic, lung, and ovarian cancers. MSLN-related peptides that circulate in serum of patients suffering from pleural mesothelioma are used as biomarkers for monitoring the disease. MSLN may promote metastasis by inducing matrix metalloproteinase 7 and 9 expression. The monoclonal antibody anetumab has been developed to target MSLN.
[0167] CA6 (carbonic anhydrase VI) encodes one of several isozymes of carbonic anhydrase. CA6 is found in salivary glands and may play a role in the reversible hydration of carbon dioxide. CA6 is expressed in human serous ovarian adenocarcinomas. The monoclonal antibody huDS6 has been developed to target CA6.
[0168] NAPI2B (sodium/phosphate cotransporter 2B) encodes a type II sodium-phosphate cotransporter. NAPI2B is highly expressed on the tumor surface in lung, ovarian, and thyroid cancers as well as in normal lung pneumocytes. The monoclonal antibody lifastuzumab has been developed to target NAPI2B.
[0169] TROP2 (trophoblast antigen 2) encodes a transmembrane glycoprotein that acts as an intracellular calcium signal transducer. TROP2 binds to multiple factors such as IGF-1, claudin- 1, claudin-7, cyclin Dl, and PKC. TROP2 including intracellular calcium signaling and the mitogen activated protein kinase pathway. TROP 2 plays a role in cell self-renewal, proliferation, invasion, and survival. Discovered first in trophoblast cells that have the ability to invade uterine decidua during placental implantation, TROP2 overexpression has been shown to be capable of stimulating cancer growth. TROP2 overexpression has been observed in breast, cervix, colorectal, esophagus, lung, non-Hodgkin' s lymphoma, chronic lymphocytic lymphoma, Raji Burkitt lymphoma, oral squamous cell, ovarian, pancreatic, prostate, stomach, thyroid, urinary bladder, and uterine carcinomas. The monoclonal antibody sacituzumab has been developed to target TROP2.
[0170] CEA (carcinoembryonic antigen; also referred to as CEACAM5) is a member of a family of related glycoproteins involved in cell adhesion. CEA is a biomarker for gastrointestinal cancers and may promote tumor development by means of its cell adhesion function. CEA levels have been found to be elevated in serum of individuals with colorectal carcinoma. CEA levels have also been found to be elevated in gastric carcinoma, pancreatic carcinoma, lung carcinoma, breast carcinoma, and medullary thyroid carcinoma. The monoclonal antibodies PR1A3 and Ab2-3 have been developed to target CEA.
[0171] CLDN18.2 (claudin 18) encodes a member of the claudin family of integral membrane proteins. CLDN18.2 is a component of tight junctions that create a physical barrier to prevent diffusion of solutes and water through the paracellular space between epithelial cells. CLDN18.2 is overexpressed in infiltrating ductal adenocarcinomas, but is reduced in some gastric carcinomas. The monoclonal antibody claudiximab has been developed to target CLDN18.2.
[0172] FAP (fibroblast activation protein, alpha) encodes a homodimeric integral membrane protein from a family of serine proteases. FAP is believed to play a role in many processes including tissue remodeling, fibrosis, wound healing, inflammation, and tumor growth. FAP enhances tumor growth and invasion by promoting angiogenesis, collagen fiber degradation and apoptosis, and by downregulating the immune response. FAP is selectively expressed on fibroblasts within the tumor stroma. The monoclonal antibody sibrotuzumab has been developed to target FAP.
[0173] EphA2 (EPH Receptor A2) encodes a member of the ephrin receptor subfamily of the protein- tyro sine kinase family. EphA2 binds to ephrin-A ligands. Activation of EphA2 receptor upon ligand binding can result in modulation of migration, integrin- mediated adhesion, proliferation, and differentiation. EphA2 is overexpressed in various cancers including breast, prostate, urinary bladder, skin, lung, ovarian, and brain cancers. High EphA2 expression is also correlated with poor prognosis. The monoclonal antibodies DS-8895a optl, DS-8895 opt2, and the 1C1 antibody in MEDI-547 have been developed to target EphA2.
[0174] RON (macrophage stimulating 1 receptor) encodes a cell surface receptor for macrophage stimulating protein (MSP) with tyrosine kinase activity and belongs to the MET proto-oncogene family. RON has significant structural similarity and sequence identity with the cancer-related gene C-MET. RON plays a significant role in KRAS oncogene addiction and has also been shown to be overexpressed in pancreatic cancers. Altered Ron expression and activation has been associated with decreased survival and cancer progression in various cancers including gastric, colon, breast, bladder, renal cell, ovarian, and hepatocellular cancers. The monoclonal antibody narnatumab has been developed to target RON.
[0175] LY6E (lymphocyte antigen 6 complex, locus E) encodes an interferon alpha-inducible GPI-anchored cell membrane protein. LY6E is overexpressed in numerous cancers including lung, gastric, ovarian, breast, kidney, pancreatic, and head and neck carcinomas. The monoclonal antibody in RG7841 has been developed to target LY6E.
[0176] FRA (folate receptor alpha) encodes a GPI-anchored cell surface glycoprotein. FRA binds folic acid, a molecule needed for cell growth and DNA synthesis, and mediates its internalization via receptor-mediated endocytosis. FRA is overexpressed in various cancers including prostate, breast, ovarian, pancreatic, mesothelioma, non-small cell lung carcinoma, and head and neck cancer. FRA expression has also been found to enhance tumor cell proliferation. The monoclonal antibodies farletuzumab and mirvetuximab have been developed to target FRA.
[0177] PSMA (prostate specific membrane antigen) is a type II transmembrane glycoprotein belonging to the M28 peptidase family that is expressed in all types of prostate tissues. PSMA is upregulated in cancer cells within the prostate and is used as a marker for prostate cancer. PSMA expression may also serve as a predictor of disease recurrence in prostate cancer patients. The monoclonal antibodies J591 variant 1 and J591 variant 2 have been developed to target PSMA.
[0178] DLL3 (delta-like 3) encodes a ligand in the Notch signaling pathway that is associated with neuroendocrine cancer. DLL3 is most highly expressed in the fetal brain and is involved in somitogenesis in the paraxial mesoderm. DLL3 is expressed on tumor cell surfaces but not in normal tissues. The monoclonal antibody rovalpituzumab has been developed to target DLL3.
[0179] PTK7 (tyrosine protein kinase-like 7) encodes a receptor tyrosine kinase that lacks catalytic tyrosine kinase activity but is nevertheless capable of signal transduction. PTK7 interacts with the WNT signaling pathway, which itself has important roles in epithelial mesenchymal transition and various cancers such as breast cancer. PTK7 overexpression has been associated with patient prognosis depending on the cancer type. The monoclonal antibodies in PF-06647020 and the anti-PTK7 antibody described by SEQ ID NOs: 440 and 445 have been developed to target PTK7.
[0180] LIVl (LIV-1 protein, estrogen regulated) encodes a member of the LIV-1 subfamily of ZIP (Zrt-, Irt-like proteins) zinc transporters. LIVl is an estrogen regulated protein that transports zinc and/or other ions across the cell membrane. Elevated levels of LIVl have been shown in estrogen receptor positive breast cancers, and LIVl is used as a marker of ER-positive cancers. LIVl has also been implicated as a downstream target of the STAT3 transcription factor and as playing an essential role in the nuclear localization of the Snail transcription factor that modulates epithelial-to-mesenchymal transition. The monoclonal antibody in SGN-LIV1A has been developed to target LIV1.
[0181] RORl (receptor tyrosine kinase-like orphan receptor 1) encodes a member of the ROR family of orphan receptors. RORl has been found to bind Wnt5a, a non-canonical Wnt via a Frizzled domain (FZD), and plays an important role in skeletal, cardiorespiratory, and
neurological development. RORl expression is predominantly restricted to embryonic
development and is absent in most mature tissues. In contrast, RORl expression is upregulated in B-Cell chronic lymphocytic leukemia, acute lymphocytic leukemia, non-Hodgkin lymphoma, and myeloid malignancies. The monoclonal antibody cirmtuzumab has been developed to target RORl.
[0182] MAGE-A3 (melanoma-associated antigen 3) encodes a member of the melanoma- associated antigen gene family. The function of MAGE-A3 is not known, but its elevated expression has been observed in various cancers including melanoma, non-small cell lung cancer, and in putative cancer stem cell populations in bladder cancer. The monoclonal antibody described by SEQ ID NOs: 479 and 484 has been developed to target MAGE- A3.
[0183] NY-ESO-1 (New York esophageal squamous cell carcinoma 1) encodes a member of the cancer-testis family of proteins. Cancer-testis antigen expression is normally restricted to testicular germ cells in adult tissues, but has been found to be aberrantly expressed in various tumors including soft tissue sarcomas, melanoma, epithelial cancers, and myxoid and round cell liposarcomas. The monoclonal antibody described by SEQ ID Nos: 492 and 497 has been developed to target NY-ESO-1.
Binding Domain
[0184] The binding domain of an antibody construct is selected to recognize an antigen or molecule. For example, an antigen can be a cell surface marker on target cells associated with a disease or condition. An antigen can be a peptide or fragment thereof. An antigen can be expressed on an immune cell. An antigen can be expressed on an antigen-presenting cell. An antigen can be expressed on a dendritic cell, a macrophage, or a B cell. An antigen on an antigen presenting cell can be a cell lineage marker or a cell surface protein expressed preferentially on antigen presenting cells or a subset of antigen presenting cells. An antigen can be a peptide presented in a major histocompatibility complex by cell. As another example, a cell surface marker recognized by the antigen binding domain can include macromolecules associated with viral and bacterial diseases or infections, autoimmune diseases and cancerous diseases. An antigen can be a tumor antigen or fragment thereof. A tumor antigen can be any antigen listed on tumor antigen databases, such as TANTIGEN, or peptide databases for T cell-defined tumor antigens, such as the Cancer Immunity Peptide database. A tumor antigen can also be any antigen listed in the review by Chen (Chen, Cancer Immun 2004 [updated 2004 Mar 10; cited 2004 Apr 1]). An antigen can be or can be at least 80% homologous CD5, CD19, CD20, CD25, CD37, CD30, CD33, CD45, CAMPATH-1, BCMA, CS-1, PD-L1, B7-H3, B7-DC (PD-L2), HLD-DR, carcinoembryonic antigen (CEA), TAG-72, EpCAM, MUC1, MUC15, MUC16, fo late-binding protein, A33, G250, pro state- specific membrane antigen (PSMA), ferritin, GD2, GD3, GM2, Ley, CA-125, CA19-9, epidermal growth factor, pl85HER2, IL-2 receptor, EGFRvIII (de2-7 EGFR), fibroblast activation protein (FAP), tenascin, a metalloproteinase, endosialin, vascular endothelial growth factor, ανβ3, WT1, LMP2, HPV E6, HPV E7, Her-2/neu, p53 nonmutant, NY-ESO-1, MelanA/MARTl, Ras mutant, gplOO, p53 mutant, PR1, bcr-abl, tyrosinase, survivin, PSA, hTERT, a Sarcoma translocation breakpoint protein, EphA2, PAP, ML-IAP, AFP, ERG, NA17, PAX3, ALK, androgen receptor, cyclin B l, polysialic acid, MYCN, RhoC, TRP-2, fucosyl GM1, mesothelin (MSLN), PSCA, MAGE Al, MAGE A3, sLe(animal), CYP1B 1, PLAV1, GM3, BORIS, Tn, GloboH, ETV6-AML, NY-BR-1, RGS5, SART3, STn, Carbonic anhydrase IX, PAX5, OY-TES 1, Sperm protein 17, LCK, HMWMAA, AKAP-4, SSX2, XAGE 1, Legumain, Tie 3, Page 4, VEGFR2, MAD-CT-1, PDGFR-B, MAD-CT-2, ROR2, , TRAIL 1, MAGE A4, MAGE C2, GAGE, EGFR, CMET, HER3, EPCAM, CA6, NAPI2B, TROP2, Claudin-6
(CLDN6), Claudin-16 (CLDN16), CLDN18.2, RON, LY6E, FRA, DLL3, PTK7, Uroplakin-IB (UPK1B), LIV1, ROR1, STRA6, TMPRSS3, TMPRSS4, TMEM238, Clorfl86, Fos-related antigen 1, VEGFR1, endoglin, VTCN1 (B7-H4), VISTA, or gpNMB.
[0185] In some embodiments, an antigen binding domain recognizes a single antigen. In some embodiments, an antigen binding domain recognizes two or more different antigens.
[0186] An antibody construct can include one or more binding domains. For example, an antibody construct can comprise a first binding domain. An antibody construct can also comprise a second binding domain. A binding domain can specifically bind to an antigen on a cell surface. A binding domain can specifically bind to an antigen on a cell surface, for example, of a tumor or cancer cell, an immune cell, or of an antigen presenting cell, such as a dendritic cell or macrophage. A binding domain can be a cell surface receptor agonist. A binding domain can be an antigen binding domain. An antigen binding domain can be a cell surface receptor agonist. An antigen binding domain can be a domain that can specifically bind to an antigen. An antigen binding domain can specifically bind to a tumor antigen. An antigen binding domain can be an antigen-binding portion of an antibody or an antigen binding fragment thereof. A binding domain can recognize a single antigen.
[0187] An antibody construct can include, for example, one, two, three, four, five, six, seven, eight, nine, ten, or more antigen binding domains. An antibody construct, such as an antibody, can include two antigen binding domains in which each antigen binding domain can recognize the same antigen. An antibody construct, such as a bi- specific antibody, can include two antigen binding domains in which each antigen binding domain can recognize different antigens. An antibody construct can include three antigen binding domains in which each antigen binding domain can recognize different antigens. An antibody construct can include three antigen binding domains in which two of the antigen binding domains can recognize the same antigen. An antigen binding domain can be in a scaffold, in which a scaffold is a supporting framework for the antigen binding domain. An antigen binding domain can be in a non-antibody scaffold. An antigen binding domain can be in an antibody scaffold. An antibody construct can comprise an antigen binding domain in a scaffold. The antibody construct can further comprise an Fc fusion protein product. In some embodiments, the antibody construct is an Fc fusion protein.
[0188] In some embodiments, an antibody construct can comprise an antigen binding domain that can specifically bind to a tumor antigen. A tumor antigen, also referred to as a tumor associated antigen, is an antigen that can be expressed by a cancer cell, a neoplastic tumor cell and/or within a tumor microenvironment. It is preferably not expressed or expressed at low levels on normal (non-cancerous) cells. For example, a tumor antigen can be an antigen expressed on a cell associated within a tumor, such as a neoplastic cell, or a tumor associated cell such as a stromal cell, endothelial cell, fibroblast, or tumor- infiltrating immune cell. For example, the tumor antigen Her2/Neu can be overexpressed by certain types of breast and ovarian cancer. A tumor antigen can also be ectopically expressed by a tumor and contribute to deregulation of the cell cycle, reduced apoptosis, metastasis, or escape from immune surveillance. Tumor antigens can generally be proteins or polypeptides derived therefrom, but also can be glycans, lipids, or other small organic molecules. Additionally, a tumor antigen can arise through increases or decreases in post-translational processing exhibited by a cancer cell compared to a normal cell, for example, protein glycosylation, protein lipidation, protein phosphorylation, or protein acetylation.
[0189] In certain embodiments, a binding domain specifically can bind to a tumor associated antigen, such as CD5, CD19, CD20, CD25, CD37, CD30, CD33, CD45, CAMPATH- 1 antigen, BCMA, CS-1, PD-L1, B7-H3, B7-DC (PD-L2), HLD-DR, carcinoembryonic antigen (CEA), TAG-72, EpCAM, MUC1, MUC15, MUC16, fo late-binding protein, A33, G250, prostate- specific membrane antigen (PSMA), ferritin, GD2, GD3, GM2, Ley, CA-125, CA19-9, epidermal growth factor, pl85HER2, IL-2 receptor, EGFRvIII (de2-7 EGFR), fibroblast activation protein (FAP), tenascin, a metalloproteinase, endosialin, vascular endothelial growth factor, ανβ3, WT1, LMP2, HPV E6, HPV E7, Her-2/neu, p53 nonmutant, NY-ESO-1, MelanA/MARTl, Ras mutant, gplOO, p53 mutant, PR1, bcr-abl, tyrosinase, survivin, PSA, hTERT, a Sarcoma translocation breakpoint protein, EphA2, PAP, ML-IAP, AFP, ERG, NA17, PAX3, ALK, androgen receptor, cyclin B l, polysialic acid, MYCN, RhoC, TRP-2, fucosyl GM1, mesothelin (MSLN), PSCA, MAGE Al, MAGE A3, sLe(animal), CYP1B 1, PLAV1, GM3, BORIS, Tn, GloboH, ETV6- AML, NY-BR-1, RGS5, SART3, STn, Carbonic anhydrase IX, PAX5, OY-TES 1, Sperm protein 17, LCK, HMWMAA, AKAP-4, SSX2, XAGE 1, Legumain, Tie 3, Page4, VEGFR2, MAD-CT- 1, PDGFR-B, MAD-CT-2, ROR2, TRAILl, MAGE A4, MAGE C2, GAGE, EGFR, CMET, HER3, EPCAM, CA6, NAPI2B, TROP2, Claudin-6 (CLDN6), Claudin-16 (CLDN16),
CLDN18.2, RON, LY6E, FRA, DLL3, PTK7, Uroplakin-IB (UPK1B), LIV1, ROR1, STRA6, TMPRSS3, TMPRSS4, TMEM238, Clorfl86, Fos-related antigen 1, VEGFR1, endoglin, VTCN1 (B7-H4), VISTA, or gpNMB.
[0190] In some embodiments, a binding domain specifically can bind to a tumor associated antigen having at least 80%, 90%, 95%, 97%, 98%, 99% or 100% sequence identity to CD5, CD19, CD20, CD25, CD37, CD30, CD33, CD45, CAMPATH- 1 antigen, BCMA, CS-1, PD-L1, B7-H3, B7-DC (PD-L2), HLD-DR, carcinoembryonic antigen (CEA), TAG-72, EpCAM, MUCl, MUC15, MUC16, fo late-binding protein, A33, G250, pro state- specific membrane antigen
(PSMA), ferritin, CA-125, CA19-9, epidermal growth factor, pl85HER2, IL-2 receptor,
EGFRvIII (de2-7 EGFR), fibroblast activation protein (FAP), tenascin, a metalloproteinase, endosialin, vascular endothelial growth factor, ανβ3, WTl, LMP2, HPV E6, HPV E7, Her-2/neu, p53 nonmutant, NY-ESO-1, MelanA/M ART 1 , Ras mutant, gplOO, p53 mutant, PR1, bcr-abl, tyrosinase, survivin, PSA, hTERT, a Sarcoma translocation breakpoint protein, EphA2, PAP, ML-IAP, AFP, ERG, NA17, PAX3, ALK, androgen receptor, cyclin B l, MYCN, RhoC, TRP-2, mesothelin (MSLN), PSCA, MAGE Al, MAGE A3, CYP1B 1, PLAV1, BORIS, GloboH, ETV6- AML, NY-BR-1, RGS5, SART3, Carbonic anhydrase IX, PAX5, OY-TES 1, Sperm protein 17, LCK, HMWMAA, AKAP-4, SSX2, XAGE 1, Legumain, Tie 3, Page4, VEGFR2, MAD-CT-1, PDGFR-B, MAD-CT-2, ROR2, TRAILl, MAGE A4, MAGE C2, GAGE, EGFR, CMET, HER3, EPCAM, CA6, NAPI2B, TROP2, Claudin-6 (CLDN6), Claudin-16 (CLDN16), CLDN18.2, RON, LY6E, FRA, DLL3, PTK7, Uroplakin-IB (UPK1B), LIV1, ROR1, STRA6, TMPRSS3, TMPRSS4, TMEM238, Clorfl86, Fos-related antigen 1, VEGFR1, endoglin, VTCN1 (B7-H4), VISTA, or gpNMB.
[0191] In certain embodiments, a binding domain specifically binds to an antigen selected from the group consisting of comprising Her2/Neu (CD340), EGFR, CMET, HER3, MUCl, MUCl 6,
EPCAM, MSLN, CA6, NAPI2B, TROP2, CEA, CLDN18.2, EGFRvIII, FAP, EphA2, RON,
LY6E, FRA, PSMA, DLL3, PTK7, LIV1, ROR1, MAGE- A3, and NY-ESO-1.
[0192] A binding domain of an antibody construct can be selected from any domain that specifically binds to an antigen, including but not limited to, from a monoclonal antibody, a polyclonal antibody, a recombinant antibody, or a functional fragment thereof, for example, a heavy chain variable domain (VH) and a light chain variable domain (VL), or from a non-antibody such as a DARPin, an affimer, an avimer, a knottin, a monobody, an affinity clamp, an ectodomain, a receptor ectodomain, a receptor, a cytokine, a ligand, an immunocytokine, a T-cell receptor, a VNAR, an anticalin, or a recombinant T-cell receptor. In some embodiments, a binding domain of an antibody construct is from a monoclonal antibody, a polyclonal antibody, a recombinant antibody, or a functional fragment thereof, for example, a heavy chain variable domain (VH) and a light chain variable domain (VL).
[0193] The antigen binding domain of an antibody construct can be at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% homologous to an antigen binding domain selected from, but not limited to, a monoclonal antibody, a polyclonal antibody, a recombinant antibody, or a functional fragment thereof, for example, a heavy chain variable domain (VH) and a light chain variable domain (VL), or from a non-antibody, such as a DARPin, an affimer, an avimer, a knottin, a monobody, an affinity clamp, an ectodomain, a receptor ectodomain, a receptor, a cytokine, a ligand, an immunocytokine, a T-cell receptor, a VNAR, an anticalin, or a
recombinant T-cell receptor.
[0194] A binding domain, for example an antigen binding domain from a monoclonal antibody, can comprise a light chain and a heavy chain. In one aspect, the monoclonal antibody binds to an antigen present on the surface of an antigen presenting cell (APC antigen) and comprises the light chain of an anti-APC antigen antibody and the heavy chain of an anti-APC antigen antibody, which bind an APC antigen. In another aspect, the monoclonal antibody binds to a tumor antigen comprises the light chain of a tumor antigen (anti-tumor) antibody and the heavy chain of a tumor antigen (anti-tumor) antibody, which bind to the tumor antigen.
[0195] An antibody construct can be an antibody. An antibody molecule can include of two identical light protein chains (light chains) and two identical heavy protein chains (heavy chains), all held together covalently by interchain disulfide linkages. Structurally, various functions of an antibody can be confined to discrete protein regions or domains. The sites that can recognize and can bind to antigen consist of three complementarity determining regions (CDRs; also referred to as hyper- variable regions) that lie within the variable heavy chain regions and within the variable light chain regions at the N-terminal ends of the two heavy and two light chains. The constant domains can provide the general framework of the antibody and may not be involved directly in binding the antibody to an antigen, but can be involved in various effector functions, such as participation of the antibody in antibody-dependent cellular cytotoxicity (ADCC).
[0196] The variable domains of natural light and heavy chains can have the same general structures, and each domain can comprise four framework regions, whose sequences can be somewhat conserved, connected by three CDRs. The four framework regions can largely adopt a β-sheet conformation and the CDRs can form loops connecting, and in some aspects forming part of, the β -sheet structure. The CDRs in each chain can be held in close proximity by the framework regions and, with the CDRs from the other chain, can contribute to the formation of the antigen binding site.
[0197] An antibody can include an antibody of any type, which can be assigned to different classes of immunoglobins, e.g., IgA, IgD, IgE, IgG, and IgM. Several different classes can be further divided into isotypes, e.g., IgGl, IgG2, IgG3, IgG4, IgAl, and IgA2. Exemplary heavy chain sequences of reference antibodies can be used to identify residue variants and mutants.
[0198] An exemplary heavy chain sequence for human IgGl heavy chain is that of the human IgGl antibody, and can comprise:
ASTKGPS VFPLAPS S KSTS GGT AALGCLVKD YFPEPVT VS WNS G ALTS GVHTFP AVLQS S
GLYS LS S V VT VPS S S LGTQT YICN VNHKPS NTKVD KKVEPKS CD KTHTCPPCP APELLGG
PSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQY
NSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSR
EEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDK
SRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID NO: 898).
[0199] An exemplary heavy chain reference sequence for human IgG2 heavy chain can comprise:
ASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSS
GLYS LS S V VT VPS S NFGTQT YTCN VDHKPS NTKVD KT VERKCC VECPPCP APP V AGPS VF
LFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTF
RVVSVLTVVHQDWLNGKEYKCKVSNKGLPAPIEKTISKTKGQPREPQVYTLPPSREEMT
KNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQ
QGNVFSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID NO: 899).
[0200] An exemplary heavy chain reference sequence for human IgG4 heavy chain can comprise:
ASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSS GLYS LS S V VT VPS S S LGTKT YTCN VDHKPS NTKVD KRVES KYGPPCPS CP APEFLGGPS V FLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNST YRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEM TKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRW QEG NVFSCSVMHEALHNHYTQKSLSLSLGK (SEQ ID NO: 900).
[0201] The heavy-chain constant regions (Fc) that corresponds to the different classes of immunoglobulins can be α, δ, ε, γ, and μ. The light chains can be one of either kappa or κ and lambda or λ, based on the amino acid sequences of the constant domains. The Fc region can contain an Fc domain. An Fc receptor can bind an Fc domain. An Fc domain can comprise amino acid residues 216 to 447 of an IgGl, which are part of SEQ ID NO: 898. An Fc domain can comprise amino acid residues 216 to 442 of an IgG2, which are part of SEQ ID NO: 899. An Fc domain can comprise amino acid residues 216 to 444 of an IgG4, which are part of SEQ ID NO: 900. Antibody constructs can also include any fragment or recombinant forms thereof (e.g., scFVs and domain antibodies). Antibody constructs can also include any fragment or
recombinant forms thereof (e.g., scFVs and domain antibodies) of non-antibody scaffolds, including but not limited to anti-calins, affibodies, affilins, atrimers, avimers, bicyclic peptides, centyrins, Cys-knots, Darpins, fibronections, Kunitz domains, O-bodies, or peptibodies.
[0202] An antibody construct can comprise an antigen binding domain of an antibody. An antigen binding domain of an antibody can comprise one or more light chain CDRs (LCDRs) and one or more heavy chain CDRs (HCDRs), or one or more LCDRs or one or more HCDRs. For example, an antibody binding domain of an antibody construct can comprise one or more of the following: a light chain complementary determining region 1 (LCDR1), a light chain
complementary determining region 2 (LCDR2), or a light chain complementary determining region 3 (LCDR3). For another example, an antibody binding domain can comprise one or more of the following: a heavy chain complementary determining region 1 (HCDR1), a heavy chain complementary determining region 2 (HCDR2), or a heavy chain complementary determining region 3 (HCDR3). In some embodiments, an antigen binding domain comprises LCDR1, LCDR2, LCDR3, HCDR1, HCDR2 and HCDR3. Unless stated otherwise, the CDRs described herein can be defined according to the IMGT (the international ImMunoGeneTics information system).
[0203] An antigen binding domain can comprise only the heavy chain of an antibody. An antigen binding domain can comprise only the variable domain of the heavy chain of an antibody.
Alternatively, an antigen binding domain can comprise only the light chain of an antibody. An antigen binding domain can comprise only the variable light chain of an antibody.
[0204] An antibody construct can comprise an antibody fragment. An antibody fragment can include (i) a Fab fragment, a monovalent fragment consisting of the VL, VH, CL and CHI domains; (ii) a F(ab')2 fragment, a bivalent fragment comprising two Fab fragments linked by a disulfide bridge at the hinge region; and (iii) a Fv fragment consisting of the VL and VH domains of a single arm of an antibody. Although the two domains of the Fv fragment, VL and VH, can be coded for by separate genes, they can be linked by a synthetic linker to be made as a single protein chain in which the VL and VH regions pair to form monovalent molecules. F(ab')2 and Fab' moieties can be produced recombinantly. [0205] An Fv can be the minimum antibody fragment which contains a complete antigen- recognition and antigen-binding site. This region can consist of a dimer of one heavy chain and one light chain variable domain in tight, non-covalent association. In this configuration the three hypervariable regions of each variable domain can interact to define an antigen-binding site on the surface of the VH-VL dimer. A single variable domain (or half of an Fv comprising only CDRs specific for an antigen) can recognize and bind antigen, although at a lower affinity than the entire binding site.
[0206] An antibody used herein can be chimeric or "humanized." Chimeric and humanized forms of non-human (e.g., murine) antibodies can be chimeric immunoglobulins,
immunoglobulin chains or fragments thereof (such as Fv, Fab, Fab', F(ab')2 or other target- binding subdomains of antibodies), which can contain minimal sequences derived from non- human immunoglobulin. In general, the humanized antibody can comprise substantially all of at least one, and typically two, variable domains, in which all or substantially all of the CDR regions correspond to those of a non-human immunoglobulin and all or substantially all of the frame work regions (FR) are those of a human immunoglobulin sequence. The humanized antibody can also comprise at least a portion of an immunoglobulin constant region (Fc), typically that of a human immunoglobulin consensus sequence.
[0207] An antibody described herein can be a human antibody. As used herein, "human antibodies" can include antibodies having, for example, the amino acid sequence of a human immunoglobulin and include antibodies isolated from human immunoglobulin libraries or from animals transgenic for one or more human immunoglobulins that do not express endogenous immunoglobulins. Human antibodies can be produced using transgenic mice which are incapable of expressing functional endogenous immunoglobulins, but which can express human immunoglobulin genes. Completely human antibodies that recognize a selected epitope can be generated using guided selection. In this approach, a selected non-human monoclonal antibody, e.g., a mouse antibody, is used to guide the selection of a completely human antibody recognizing the same epitope
[0208] An antibody described herein can be a bispecific antibody or a dual variable domain antibody (DVD). Bispecific and DVD antibodies are monoclonal, often human or humanized, antibodies that have binding specificities for at least two different antigens.
[0209] An antibody described herein can be a derivatized antibody. For example, derivatized antibodies can be modified by glycosylation, deglycosylation, defucosylation, acetylation, pegylation, phosphorylation, amidation, derivatization by known protecting/blocking groups, proteolytic cleavage, or linkage to a cellular ligand or other protein. [0210] An antibody described herein can have a sequence that has been modified to alter at least one constant region-mediated biological effector function relative to the corresponding wild type sequence. For example, in some embodiments, the antibody can be modified to reduce at least one constant region-mediated biological effector function relative to an unmodified antibody, e.g., reduced binding or increased binding to the Fc receptor (FcR). FcR binding can be reduced or increased by, for example, mutating the immunoglobulin constant region segment of the antibody at particular regions necessary for FcR interactions.
[0211] An antibody described herein can be modified to acquire or improve at least one constant region-mediated biological effector function relative to an unmodified antibody, e.g., to enhance FcyR interactions. For example, an antibody with a constant region that binds FcyRIIA, FcyRIIB and/or FcyRIIIA with greater affinity than the corresponding wild type constant region can be produced according to the methods described herein.
[0212] An antibody described herein can bind to tumor cells, such as an antibody against a cell surface receptor or a tumor antigen.
[0213] An antibody construct can comprise a first binding domain. An antibody construct can comprise a first binding domain that specifically binds an antigen. An antibody construct can comprise a first binding domain that specifically binds a tumor antigen. A first binding domain can specifically bind a tumor antigen, wherein the tumor antigen is selected from the group consisting CD5, CD19, CD20, CD25, CD37, CD30, CD33, CD45, CAMPATH-1, BCMA, CS-1, PD-Ll, B7-H3, B7-DC (PD-L2), HLD-DR, carcinoembryonic antigen (CEA), TAG-72, EpCAM, MUC1, MUC15, MUC16, fo late-binding protein, A33, G250, pro state- specific membrane antigen (PSMA), ferritin, GD2, GD3, GM2, Ley, CA-125, CA19-9, epidermal growth factor, pl85HER2, IL-2 receptor, EGFRvIII (de2-7 EGFR), fibroblast activation protein (FAP), tenascin, a metalloproteinase, endosialin, vascular endothelial growth factor, ανβ3, WT1, LMP2, HPV E6, HPV E7, Her-2/neu, p53 nonmutant, NY-ESO-1, MelanA/MARTl, Ras mutant, gplOO, p53 mutant, PR1, bcr-abl, tyrosinase, survivin, PSA, hTERT, a Sarcoma translocation breakpoint protein, EphA2, PAP, ML-IAP, AFP, ERG, NA17, PAX3, ALK, androgen receptor, cyclin B l, polysialic acid, MYCN, RhoC, TRP-2, fucosyl GM1, mesothelin (MSLN), PSCA, MAGE Al, MAGE A3, sLe(animal), CYPIB I, PLAVl, GM3, BORIS, Tn, GloboH, ETV6-AML, NY-BR-1, RGS5, SART3, STn, Carbonic anhydrase IX, PAX5, OY-TES 1, Sperm protein 17, LCK,
HMWMAA, AKAP-4, SSX2, XAGE 1, Legumain, Tie 3, Page4, VEGFR2, MAD-CT-1,
PDGFR-B, MAD-CT-2, ROR2, TRAIL 1, MAGE A4, MAGE C2, GAGE, EGFR, CMET, HER3, EPCAM, CA6, NAPI2B, TROP2, Claudin-6 (CLDN6), Claudin-16 (CLDN16),
CLDN18.2, RON, LY6E, FRA, DLL3, PTK7, Uroplakin-IB (UPK1B), LIV1, ROR1, STRA6, TMPRSS3, TMPRSS4, TMEM238, Clorfl86, Fos-related antigen 1, VEGFR1, endoglin, VTCN1 (B7-H4), VISTA, gpNMB, or any fragment thereof. An antibody construct can comprise a first binding domain that specifically binds a tumor antigen on a tumor cell, a tumor fragment, an immune cell or an antigen presenting cell.
[0214] An antibody construct can comprise a first binding domain that specifically binds a tumor antigen. The construct can comprise a first binding domain comprising one or more CDRs, typically a set of six CDRs. A first binding domain can comprise at least 80% sequence identity to any sequence in TABLE 1. An antibody construct can comprise a first binding domain that binds a tumor antigen, wherein the first binding domain comprises at least 80% sequence identity to: a) HCDR1 comprising an amino acid sequence of SEQ ID NO: 13, HCDR2 comprising an amino acid sequence of SEQ ID NO: 14, HCDR3 comprising an amino acid sequence of SEQ ID NO: 15, LCDR1 comprising an amino acid sequence of SEQ ID NO: 18, LCDR2 comprising an amino acid sequence of SEQ ID NO: 19, and LCDR3 comprising an amino acid sequence of SEQ ID NO: 20; b) HCDR1 comprising an amino acid sequence of SEQ ID NO: 26, HCDR2 comprising an amino acid sequence of SEQ ID NO: 27, HCDR3 comprising an amino acid sequence of SEQ ID NO: 28, LCDR1 comprising an amino acid sequence of SEQ ID NO: 31, LCDR2 comprising an amino acid sequence of SEQ ID NO: 32, and LCDR3 comprising an amino acid sequence of SEQ ID NO: 33; c) HCDR1 comprising an amino acid sequence of SEQ ID NO: 39, HCDR2 comprising an amino acid sequence of SEQ ID NO: 40, HCDR3 comprising an amino acid sequence of SEQ ID NO: 41, LCDR1 comprising an amino acid sequence of SEQ ID NO: 44, LCDR2 comprising an amino acid sequence of SEQ ID NO: 45, and LCDR3 comprising an amino acid sequence of SEQ ID NO: 46; d) HCDR1 comprising an amino acid sequence of SEQ ID NO: 52, HCDR2 comprising an amino acid sequence of SEQ ID NO: 53, HCDR3 comprising an amino acid sequence of SEQ ID NO: 54, LCDR1 comprising an amino acid sequence of SEQ ID NO: 57, LCDR2 comprising an amino acid sequence of SEQ ID NO: 58, and LCDR3 comprising an amino acid sequence of SEQ ID NO: 59; e) HCDR1 comprising an amino acid sequence of SEQ ID NO: 65, HCDR2 comprising an amino acid sequence of SEQ ID NO: 66, HCDR3 comprising an amino acid sequence of SEQ ID NO: 67, LCDR1 comprising an amino acid sequence of SEQ ID NO: 70, LCDR2 comprising an amino acid sequence of SEQ ID NO: 71, and LCDR3 comprising an amino acid sequence of SEQ ID NO: 72; f) HCDR1 comprising an amino acid sequence of SEQ ID NO: 78, HCDR2 comprising an amino acid sequence of SEQ ID NO: 79, HCDR3 comprising an amino acid sequence of SEQ ID NO: 80, LCDR1 comprising an amino acid sequence of SEQ ID NO: 83, LCDR2 comprising an amino acid sequence of SEQ ID NO: 84, and LCDR3 comprising an amino acid sequence of SEQ ID NO: 85; g) HCDR1 comprising an amino acid sequence of SEQ ID NO: 91, HCDR2 comprising an amino acid sequence of SEQ ID NO: 92, HCDR3 comprising an amino acid sequence of SEQ ID NO: 93, LCDRl comprising an amino acid sequence of SEQ ID NO: 96, LCDR2 comprising an amino acid sequence of SEQ ID NO: 97, and LCDR3 comprising an amino acid sequence of SEQ ID NO: 98; h) HCDRl comprising an amino acid sequence of SEQ ID NO: 104, HCDR2 comprising an amino acid sequence of SEQ ID NO: 105, HCDR3 comprising an amino acid sequence of SEQ ID NO: 106, LCDRl comprising an amino acid sequence of SEQ ID NO: 109, LCDR2 comprising an amino acid sequence of SEQ ID NO: 110, and LCDR3 comprising an amino acid sequence of SEQ ID NO: 111; i) HCDRl comprising an amino acid sequence of SEQ ID NO: 117, HCDR2 comprising an amino acid sequence of SEQ ID NO: 118, HCDR3 comprising an amino acid sequence of SEQ ID NO: 119, LCDRl comprising an amino acid sequence of SEQ ID NO: 122, LCDR2 comprising an amino acid sequence of SEQ ID NO: 123, and LCDR3 comprising an amino acid sequence of SEQ ID NO: 124; j) HCDRl comprising an amino acid sequence of SEQ ID NO: 130, HCDR2 comprising an amino acid sequence of SEQ ID NO: 131, HCDR3 comprising an amino acid sequence of SEQ ID NO: 132, LCDRl comprising an amino acid sequence of SEQ ID NO: 135, LCDR2 comprising an amino acid sequence of SEQ ID NO: 136, and LCDR3 comprising an amino acid sequence of SEQ ID NO: 137; k) HCDRl comprising an amino acid sequence of SEQ ID NO: 143, HCDR2 comprising an amino acid sequence of SEQ ID NO: 144, HCDR3 comprising an amino acid sequence of SEQ ID NO: 145, LCDRl comprising an amino acid sequence of SEQ ID NO: 148, LCDR2 comprising an amino acid sequence of SEQ ID NO: 149, and LCDR3 comprising an amino acid sequence of SEQ ID NO: 150; 1) HCDRl comprising an amino acid sequence of SEQ ID NO: 156, HCDR2 comprising an amino acid sequence of SEQ ID NO: 157, HCDR3 comprising an amino acid sequence of SEQ ID NO: 158, LCDRl comprising an amino acid sequence of SEQID NO: 161, LCDR2 comprising an amino acid sequence of SEQ ID NO: 162, and LCDR3 comprising an amino acid sequence of SEQ ID NO: 163; m) HCDRl comprising an amino acid sequence of SEQ ID NO: 169, HCDR2 comprising an amino acid sequence of SEQ ID NO: 170, HCDR3 comprising an amino acid sequence of SEQ ID NO: 171, LCDRl comprising an amino acid sequence of SEQ ID NO: 174, LCDR2 comprising an amino acid sequence of SEQ ID NO: 175, and LCDR3 comprising an amino acid sequence of SEQ ID NO: 176; n) HCDRl comprising an amino acid sequence of SEQ ID NO: 182, HCDR2 comprising an amino acid sequence of SEQ ID NO: 183, HCDR3 comprising an amino acid sequence of SEQ ID NO: 184, LCDRl comprising an amino acid sequence of SEQ ID NO: 187, LCDR2 comprising an amino acid sequence of SEQ ID NO: 188, and LCDR3 comprising an amino acid sequence of SEQ ID NO: 189; o) HCDRl comprising an amino acid sequence of SEQ ID NO: 195, HCDR2 comprising an amino acid sequence of SEQ ID NO: 196, HCDR3 comprising an amino acid sequence of SEQ ID NO: 197, LCDRl comprising an amino acid sequence of SEQ ID NO: 200, LCDR2 comprising an amino acid sequence of SEQ ID NO: 201, and LCDR3 comprising an amino acid sequence of SEQ ID NO: 202; p) HCDRl comprising an amino acid sequence of
SEQ ID NO: 208, HCDR2 comprising an amino acid sequence of SEQ ID NO: 209, HCDR3 comprising an amino acid sequence of SEQ ID NO: 210, LCDR1 comprising an amino acid sequence of SEQ ID NO: 213, LCDR2 comprising an amino acid sequence of SEQ ID NO: 214, and LCDR3 comprising an amino acid sequence of SEQ ID NO: 215; q) HCDRl comprising an amino acid sequence of SEQ ID NO: 805, HCDR2 comprising an amino acid sequence of SEQ
ID NO: 806, HCDR3 comprising an amino acid sequence of SEQ ID NO: 807, LCDR1 comprising an amino acid sequence of SEQ ID NO: 808, LCDR2 comprising an amino acid sequence of SEQ ID NO: 809, and LCDR3 comprising an amino acid sequence of SEQ ID NO:
810; r) HCDRl comprising an amino acid sequence of SEQ ID NO: 823, HCDR2 comprising an amino acid sequence of SEQ ID NO: 824, HCDR3 comprising an amino acid sequence of SEQ
ID NO: 825, LCDR1 comprising an amino acid sequence of SEQ ID NO: 826, LCDR2 comprising an amino acid sequence of SEQ ID NO: 827, and LCDR3 comprising an amino acid sequence of SEQ ID NO: 828; s) HCDRl comprising an amino acid sequence of SEQ ID NO:
221, HCDR2 comprising an amino acid sequence of SEQ ID NO: 222, HCDR3 comprising an amino acid sequence of SEQ ID NO: 223, LCDR1 comprising an amino acid sequence of SEQ
ID NO: 226, LCDR2 comprising an amino acid sequence of SEQ ID NO: 227, and LCDR3 comprising an amino acid sequence of SEQ ID NO: 228; t) HCDRl comprising an amino acid sequence of SEQ ID NO: 260, HCDR2 comprising an amino acid sequence of SEQ ID NO: 261,
HCDR3 comprising an amino acid sequence of SEQ ID NO: 262, LCDR1 comprising an amino acid sequence of SEQ ID NO: 265, LCDR2 comprising an amino acid sequence of SEQ ID NO:
266, and LCDR3 comprising an amino acid sequence of SEQ ID NO: 267; u) HCDRl comprising an amino acid sequence of SEQ ID NO: 273, HCDR2 comprising an amino acid sequence of SEQ ID NO: 274, HCDR3 comprising an amino acid sequence of SEQ ID NO: 275,
LCDR1 comprising an amino acid sequence of SEQ ID NO: 278, LCDR2 comprising an amino acid sequence of SEQ ID NO: 279, and LCDR3 comprising an amino acid sequence of SEQ ID
NO: 280; v) HCDRl comprising an amino acid sequence of SEQ ID NO: 286, HCDR2 comprising an amino acid sequence of SEQ ID NO: 287, HCDR3 comprising an amino acid sequence of SEQ ID NO: 288, LCDR1 comprising an amino acid sequence of SEQ ID NO: 291,
LCDR2 comprising an amino acid sequence of SEQ ID NO: 292, and LCDR3 comprising an amino acid sequence of SEQ ID NO: 293; w) HCDRl comprising an amino acid sequence of
SEQ ID NO: 299, HCDR2 comprising an amino acid sequence of SEQ ID NO: 300, HCDR3 comprising an amino acid sequence of SEQ ID NO: 301, LCDR1 comprising an amino acid sequence of SEQ ID NO: 304, LCDR2 comprising an amino acid sequence of SEQ ID NO: 305, and LCDR3 comprising an amino acid sequence of SEQ ID NO: 306; x) HCDRl comprising an amino acid sequence of SEQ ID NO: 312, HCDR2 comprising an amino acid sequence of SEQ ID NO: 313, HCDR3 comprising an amino acid sequence of SEQ ID NO: 314, LCDR1 comprising an amino acid sequence of SEQ ID NO: 317, LCDR2 comprising an amino acid sequence of SEQ ID NO: 318, and LCDR3 comprising an amino acid sequence of SEQ ID NO: 319; y) HCDRl comprising an amino acid sequence of SEQ ID NO: 325, HCDR2 comprising an amino acid sequence of SEQ ID NO: 326, HCDR3 comprising an amino acid sequence of SEQ ID NO: 327, LCDR1 comprising an amino acid sequence of SEQ ID NO: 330, LCDR2 comprising an amino acid sequence of SEQ ID NO: 331, and LCDR3 comprising an amino acid sequence of SEQ ID NO: 332; z) HCDRl comprising an amino acid sequence of SEQ ID NO: 338, HCDR2 comprising an amino acid sequence of SEQ ID NO: 339, HCDR3 comprising an amino acid sequence of SEQ ID NO: 340, LCDR1 comprising an amino acid sequence of SEQ ID NO: 343, LCDR2 comprising an amino acid sequence of SEQ ID NO: 344, and LCDR3 comprising an amino acid sequence of SEQ ID NO: 345; aa) HCDRl comprising an amino acid sequence of SEQ ID NO: 351, HCDR2 comprising an amino acid sequence of SEQ ID NO: 352, HCDR3 comprising an amino acid sequence of SEQ ID NO: 353, LCDR1 comprising an amino acid sequence of SEQ ID NO: 356, LCDR2 comprising an amino acid sequence of SEQ ID NO: 357, and LCDR3 comprising an amino acid sequence of SEQ ID NO: 358; bb) HCDRl comprising an amino acid sequence of SEQ ID NO: 364, HCDR2 comprising an amino acid sequence of SEQ ID NO: 365, HCDR3 comprising an amino acid sequence of SEQ ID NO: 366, LCDR1 comprising an amino acid sequence of SEQ ID NO: 369, LCDR2 comprising an amino acid sequence of SEQ ID NO: 370, and LCDR3 comprising an amino acid sequence of SEQ ID NO: 371; cc) HCDRl comprising an amino acid sequence of SEQ ID NO: 377, HCDR2 comprising an amino acid sequence of SEQ ID NO: 378, HCDR3 comprising an amino acid sequence of SEQ ID NO: 379, LCDR1 comprising an amino acid sequence of SEQ ID NO: 382, LCDR2 comprising an amino acid sequence of SEQ ID NO: 383, and LCDR3 comprising an amino acid sequence of SEQ ID NO: 384; dd) HCDRl comprising an amino acid sequence of SEQ ID NO: 390, HCDR2 comprising an amino acid sequence of SEQ ID NO: 391, HCDR3 comprising an amino acid sequence of SEQ ID NO: 392, LCDR1 comprising an amino acid sequence of SEQ ID NO: 395, LCDR2 comprising an amino acid sequence of SEQ ID NO: 396, and LCDR3 comprising an amino acid sequence of SEQ ID NO: 397; ee) HCDRl comprising an amino acid sequence of SEQ ID NO: 403, HCDR2 comprising an amino acid sequence of SEQ ID NO: 404, HCDR3 comprising an amino acid sequence of SEQ ID NO: 405, LCDR1 comprising an amino acid sequence of SEQ ID NO: 408, LCDR2 comprising an amino acid sequence of SEQ ID NO: 409, and LCDR3 comprising an amino acid sequence of SEQ ID NO: 410; ff) HCDRl comprising an amino acid sequence of SEQ ID NO: 416, HCDR2 comprising an amino acid sequence of SEQ ID NO: 417, HCDR3 comprising an amino acid sequence of SEQ
ID NO: 418, LCDR1 comprising an amino acid sequence of SEQ ID NO: 421, LCDR2 comprising an amino acid sequence of SEQ ID NO: 422, and LCDR3 comprising an amino acid sequence of SEQ ID NO: 423; gg) HCDRl comprising an amino acid sequence of SEQ ID NO:
429, HCDR2 comprising an amino acid sequence of SEQ ID NO: 430, HCDR3 comprising an amino acid sequence of SEQ ID NO: 431, LCDR1 comprising an amino acid sequence of SEQ
ID NO: 434, LCDR2 comprising an amino acid sequence of SEQ ID NO: 435, and LCDR3 comprising an amino acid sequence of SEQ ID NO: 436; hh) HCDRl comprising an amino acid sequence of SEQ ID NO: 442, HCDR2 comprising an amino acid sequence of SEQ ID NO: 443,
HCDR3 comprising an amino acid sequence of SEQ ID NO: 444, LCDR1 comprising an amino acid sequence of SEQ ID NO: 447, LCDR2 comprising an amino acid sequence of SEQ ID NO:
448, and LCDR3 comprising an amino acid sequence of SEQ ID NO: 449; ii) HCDRl comprising an amino acid sequence of SEQ ID NO: 455, HCDR2 comprising an amino acid sequence of SEQ ID NO: 456, HCDR3 comprising an amino acid sequence of SEQ ID NO: 457,
LCDR1 comprising an amino acid sequence of SEQ ID NO: 460, LCDR2 comprising an amino acid sequence of SEQ ID NO: 461, and LCDR3 comprising an amino acid sequence of SEQ ID
NO: 462; jj) HCDRl comprising an amino acid sequence of SEQ ID NO: 468, HCDR2 comprising an amino acid sequence of SEQ ID NO: 469, HCDR3 comprising an amino acid sequence of SEQ ID NO: 470, LCDR1 comprising an amino acid sequence of SEQ ID NO: 473,
LCDR2 comprising an amino acid sequence of SEQ ID NO: 474, and LCDR3 comprising an amino acid sequence of SEQ ID NO: 475; kk) HCDRl comprising an amino acid sequence of
SEQ ID NO: 481, HCDR2 comprising an amino acid sequence of SEQ ID NO: 482, HCDR3 comprising an amino acid sequence of SEQ ID NO: 483, LCDR1 comprising an amino acid sequence of SEQ ID NO: 486, LCDR2 comprising an amino acid sequence of SEQ ID NO: 487, and LCDR3 comprising an amino acid sequence of SEQ ID NO: 488; 11) HCDRl comprising an amino acid sequence of SEQ ID NO: 494, HCDR2 comprising an amino acid sequence of SEQ
ID NO: 495, HCDR3 comprising an amino acid sequence of SEQ ID NO: 496, LCDR1 comprising an amino acid sequence of SEQ ID NO: 499, LCDR2 comprising an amino acid sequence of SEQ ID NO: 500, and LCDR3 comprising an amino acid sequence of SEQ ID NO:
501; mm) HCDRl comprising an amino acid sequence of SEQ ID NO: 673, HCDR2 comprising an amino acid sequence of SEQ ID NO: 674, HCDR3 comprising an amino acid sequence of
SEQ ID NO: 675, LCDR1 comprising an amino acid sequence of SEQ ID NO: 676, LCDR2 comprising an amino acid sequence of SEQ ID NO: 677, and LCDR3 comprising an amino acid sequence of SEQ ID NO: 678; nn) HCDRl comprising an amino acid sequence of SEQ ID NO: 850, HCDR2 comprising an amino acid sequence of SEQ ID NO: 851, HCDR3 comprising an amino acid sequence of SEQ ID NO: 852, LCDR1 comprising an amino acid sequence of SEQ ID NO: 853, LCDR2 comprising an amino acid sequence of SEQ ID NO: 854, and LCDR3 comprising an amino acid sequence of SEQ ID NO: 855; oo) HCDR1 comprising an amino acid sequence of SEQ ID NO: 856, HCDR2 comprising an amino acid sequence of SEQ ID NO: 857, HCDR3 comprising an amino acid sequence of SEQ ID NO: 858, LCDR1 comprising an amino acid sequence of SEQ ID NO: 859, LCDR2 comprising an amino acid sequence of SEQ ID NO: 860, and LCDR3 comprising an amino acid sequence of SEQ ID NO: 861 ; pp) HCDR1 comprising an amino acid sequence of SEQ ID NO: 862, HCDR2 comprising an amino acid sequence of SEQ ID NO: 863, HCDR3 comprising an amino acid sequence of SEQ ID NO: 864, LCDR1 comprising an amino acid sequence of SEQ ID NO: 865, LCDR2 comprising an amino acid sequence of SEQ ID NO: 866, and LCDR3 comprising an amino acid sequence of SEQ ID NO: 867; qq) HCDR1 comprising an amino acid sequence of SEQ ID NO: 868, HCDR2 comprising an amino acid sequence of SEQ ID NO: 869, HCDR3 comprising an amino acid sequence of SEQ ID NO: 870, LCDR1 comprising an amino acid sequence of SEQ ID NO: 871 , LCDR2 comprising an amino acid sequence of SEQ ID NO: 872, and LCDR3 comprising an amino acid sequence of SEQ ID NO: 873 ; rr) HCDR1 comprising an amino acid sequence of SEQ ID NO: 874, HCDR2 comprising an amino acid sequence of SEQ ID NO: 875, HCDR3 comprising an amino acid sequence of SEQ ID NO: 876, LCDR1 comprising an amino acid sequence of SEQ ID NO: 877, LCDR2 comprising an amino acid sequence of SEQ ID NO: 878, and LCDR3 comprising an amino acid sequence of SEQ ID NO: 879; or ss) HCDR1 comprising an amino acid sequence of SEQ ID NO: 880, HCDR2 comprising an amino acid sequence of SEQ ID NO: 881, HCDR3 comprising an amino acid sequence of SEQ ID NO: 882, LCDR1 comprising an amino acid sequence of SEQ ID NO: 883, LCDR2 comprising an amino acid sequence of SEQ ID NO: 884, and LCDR3 comprising an amino acid sequence of SEQ ID NO: 885.
[0215] An antibody construct can comprise a first binding domain that specifically binds a tumor antigen. An antibody construct can comprise a first binding domain comprising one or more variable domains. An antibody construct can comprise a first binding domain comprising a light chain variable domain (VL domain). A first binding domain can have at least 80% or 100% sequence identity to any VL sequence in TABLE 2. An antibody construct can comprise a first binding domain comprising a heavy chain variable domain (VH domain). A first binding domain can comprise at least 80% or 100% sequence identity to any VH sequence in TABLE 2. A first binding domain can comprise at least 80% sequence identity to any sequence in TABLE 2. A first binding domain can have a pair of VH and VL regions, having sequence selected from the pairs in TABLE 2.
[0216] An antibody construct can comprise a first binding domain that specifically binds a tumor antigen, wherein the first binding domain comprises: a) a VH sequence having at least 80% or 100% sequence identity to an amino acid sequence of SEQ ID NO: 12, and a VL sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 17; b) a VH sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 25, and a VL sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 30; c) a VH sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 38, and a VL sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 43; d) a VH sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 51, and a VL sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 56; e) a VH sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 64, and a VL sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 69; f) a VH sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 77, and a VL sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 82; g) a VH sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 90, and a VL sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 95; h) a VH sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 103, and a VL sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 108; i) a VH sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 116, and a VL sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 121; j) a VH sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 129, and a VL sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 134; k) a VH sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 142, and a VL sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 147; 1) a VH sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 155, and a VL sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 160; m) a VH sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 168, and a VL sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 173; n) a VH sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 181, and a VL sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 186; o) a VH sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 194, and a VL sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 199; p) a VH sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 207, and a VL sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 212; q) a VH sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 811, and a VL sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 812; r) a VH sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 829, and a VL sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 830; s) a VH sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO:
220, and a VL sequence having at least 80% sequence identity to an amino acid sequence of SEQ
ID NO: 225; t) a VH sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 259, and a VL sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 264; u) a VH sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 272, and a VL sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 277; v) a VH sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 285, and a VL sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 290; w) a VH sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 298, and a VL sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 303; x) a VH sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO:
311, and a VL sequence having at least 80% sequence identity to an amino acid sequence of SEQ
ID NO: 316; y) a VH sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 324, and a VL sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 328; z) a VH sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 337, and a VL sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 342; aa) a VH sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 350, and a VL sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 355; bb) a VH sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 363, and a VL sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 368; cc) a VH sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID
NO: 376, and a VL sequence having at least 80% sequence identity to an amino acid sequence of
SEQ ID NO: 381; dd) a VH sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 389, and a VL sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 394; ee) a VH sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 402, and a VL sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 407; ff) a VH sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 415, and a VL sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 420; gg) a VH sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 428, and a VL sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 433; hh) a VH sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 441, and a VL sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 446; ii) a VH sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 454, and a VL sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 459; jj) a VH sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 467, and a VL sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 472; kk) a VH sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 480, and a VL sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 485; 11) a VH sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 493, and a VL sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 498; mm) a VH sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 679, and a VL sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 680; nn) a VH sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 886, and a VL sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 887; oo) a VH sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 888, and a VL sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 889; pp) a VH sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 890, and a VL sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 891; qq) a VH sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 892, and a VL sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 893; rr) a VH sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 894, and a VL sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO:895; ss) a VH sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 896, and a VL sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 897.
[0217] An antibody construct can comprise a first binding domain and an Fc domain, wherein the first binding domain and the Fc domain comprise an antibody. A first binding domain can specifically bind a tumor antigen. An antibody construct can comprise an antibody light chain wherein the antibody construct specifically binds to an antigen. An antibody construct can comprise a light chain comprising at least 80% or 100% sequence identity to a light chain sequence in TABLE 3, wherein the antibody construct specifically binds to an antigen. An antibody construct can comprise an antibody heavy chain, wherein the antibody construct specifically binds to an antigen. An antibody construct can comprise a heavy chain comprising at least 80% or 100% sequence identity to a heavy chain sequence in TABLE 3, wherein the antibody construct specifically binds to an antigen. An antibody construct can comprise at least 80% sequence identity to any sequence in TABLE 3, wherein the antibody construct specifically binds to an antigen. An antibody construct can have a pair of heavy and light chains having sequences selected from the pairs of sequences in TABLE 3, wherein the antibody construct specifically binds to an antigen.
[0218] An antibody construct can comprise an anti-tumor antibody, wherein the antibody comprises: a) a heavy chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 11, and a light chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 16; b) a heavy chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 24, and a light chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 29; c) a heavy chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 37, and a light chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 42; d) a heavy chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 50, and a light chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 55; e) a heavy chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 63, and a light chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 68; f) a heavy chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 76, and a light chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 81; g) a heavy chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 89, and a light chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 94; h) a heavy chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 102, and a light chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 107; i) a heavy chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 115, and a light chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 120; j) a heavy chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 128, and a light chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 133; k) a heavy chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 141, and a light chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 146; 1) a heavy chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 154, and a light chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 159; m) a heavy chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 167, and a light chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 172; n) a heavy chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 180, and a light chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 185; o) a heavy chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 193, and a light chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 198; p) a heavy chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 206, and a light chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 211; q) a heavy chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 813, and a light chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 814; r) a heavy chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 831, and a light chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 832; s) a heavy chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 219, and a light chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 224; t) a heavy chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 258, and a light chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 263; u) a heavy chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 271, and a light chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 276; v) a heavy chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 284, and a light chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 289; w) a heavy chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 297, and a light chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 302; x) a heavy chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 310, and a light chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 315; y) a heavy chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 323, and a light chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 328; z) a heavy chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 336, and a light chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 341; aa) a heavy chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 349, and a light chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 354; bb) a heavy chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 362, and a light chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 367; cc) a heavy chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 375, and a light chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 380; dd) a heavy chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 388, and a light chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 393; ee) a heavy chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 401, and a light chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 406; ff) a heavy chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 414, and a light chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 419; gg) a heavy chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 427, and a light chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 432; hh) a heavy chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 440, and a light chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 445; ii) a heavy chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 453, and a light chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 458; jj) a heavy chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 466, and a light chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 471; kk) a heavy chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 479, and a light chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 484; 11) a heavy chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 492, and a light chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 497; or mm) a heavy chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 681, and a light chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 682.
[0219] An antibody construct can comprise a second binding domain that specifically binds to an antigen. An antibody construct can comprise a second binding domain that specifically binds to an antigen on an antigen presenting cell. An antigen presenting cell can be a dendritic cell or a macrophage. A second binding domain can specifically bind to an antigen on an immune cell such as an antigen presenting cell, wherein the molecule comprises at least 80% sequence identity to a group consisting of CD40, CD47, TNFR2, DEC-205, PD-L1, CD36 mannose scavenger receptor 1, CLEC9A, DC-SIGN, CLEC12A, BDCA-2, OX40L, 41BBL, CD204, MARCO, CLEC5A, Dectin 1, Dectin 2, CLECIOA, CD206, CD64, CD32A, CD 16 A, HVEM, CD38, TREM2, CSF1R, or CD32B.
[0220] An antibody construct can further comprise an Fc domain. An antibody construct can comprise, for example, a first binding domain, a second binding domain, and an Fc domain, wherein the first binding domain is attached to the Fc domain. An antibody construct can comprise a first binding domain, a second binding domain, and an Fc domain, wherein the second binding domain is attached to the Fc domain. A first binding domain can be attached to an Fc domain as a fusion protein. A second binding domain can be attached to an Fc domain as a fusion protein. A first binding domain can be attached to an Fc domain via a linker. A second binding domain can be attached to an Fc domain via a linker.
[0221] An antibody construct can comprise a second binding domain comprising one or more CDRs. A second binding domain can comprise at least 80% sequence identity to any sequence in TABLE 5. A second binding domain can comprise a set of CDRs having the sequences set forth in any sete of sequence in TABLE 5.
[0222] An antibody construct can comprise a second binding domain that specifically binds to
CD40. An antibody construct can comprise a second binding domain that is a CD40 agonist. An antibody construct can comprise a second binding domain that binds to CD40, wherein the second binding domain comprises at least 80% sequence identity to: a) HCDR1 comprising an amino acid sequence of SEQ ID NO: 3, HCDR2 comprising an amino acid sequence of SEQ ID
NO: 4, HCDR3 comprising an amino acid sequence of SEQ ID NO: 5, LCDR1 comprising an amino acid sequence of SEQ ID NO: 8, LCDR2 comprising an amino acid sequence of SEQ ID
NO: 9, and LCDR3 comprising an amino acid sequence of SEQ ID NO: 10; b) HCDR1 comprising an amino acid sequence of SEQ ID NO: 582, HCDR2 comprising an amino acid sequence of SEQ ID NO: 583, HCDR3 comprising an amino acid sequence of SEQ ID NO: 584,
LCDR1 comprising an amino acid sequence of SEQ ID NO: 587, LCDR2 comprising an amino acid sequence of SEQ ID NO: 588, and LCDR3 comprising an amino acid sequence of SEQ ID NO: 589; c) HCDRl comprising an amino acid sequence of SEQ ID NO: 592, HCDR2 comprising an amino acid sequence of SEQ ID NO: 593, HCDR3 comprising an amino acid sequence of SEQ ID NO: 594, LCDR1 comprising an amino acid sequence of SEQ ID NO: 597, LCDR2 comprising an amino acid sequence of SEQ ID NO: 598, and LCDR3 comprising an amino acid sequence of SEQ ID NO: 599; d) HCDRl comprising an amino acid sequence of SEQ ID NO: 602, HCDR2 comprising an amino acid sequence of SEQ ID NO: 603, HCDR3 comprising an amino acid sequence of SEQ ID NO: 604, LCDR1 comprising an amino acid sequence of SEQ ID NO: 607, LCDR2 comprising an amino acid sequence of SEQ ID NO: 608, and LCDR3 comprising an amino acid sequence of SEQ ID NO: 609; e) HCDRl comprising an amino acid sequence of SEQ ID NO: 612, HCDR2 comprising an amino acid sequence of SEQ ID NO: 613, HCDR3 comprising an amino acid sequence of SEQ ID NO: 614, LCDR1 comprising an amino acid sequence of SEQ ID NO: 617, LCDR2 comprising an amino acid sequence of SEQ ID NO: 618, and LCDR3 comprising an amino acid sequence of SEQ ID NO: 619; f) HCDRl comprising an amino acid sequence of SEQ ID NO: 622, HCDR2 comprising an amino acid sequence of SEQ ID NO: 623, HCDR3 comprising an amino acid sequence of SEQ ID NO: 624, LCDR1 comprising an amino acid sequence of SEQ ID NO: 627, LCDR2 comprising an amino acid sequence of SEQ ID NO: 628, and LCDR3 comprising an amino acid sequence of SEQ ID NO: 629; or g) HCDRl comprising an amino acid sequence of SEQ ID NO: 632, HCDR2 comprising an amino acid sequence of SEQ ID NO: 633, HCDR3 comprising an amino acid sequence of SEQ ID NO: 634, LCDR1 comprising an amino acid sequence of SEQ ID NO: 637, LCDR2 comprising an amino acid sequence of SEQ ID NO: 638, and LCDR3 comprising an amino acid sequence of SEQ ID NO: 639.
[0223] An antibody construct can comprise a second binding domain that specifically binds DC- SIGN. An antibody construct can comprise a second binding domain that binds DC-SIGN, wherein the second binding domain comprises at least 80% sequence identity to: a) HCDRl comprising an amino acid sequence of SEQ ID NO: 640, HCDR2 comprising an amino acid sequence of SEQ ID NO: 641, HCDR3 comprising an amino acid sequence of SEQ ID NO: 642, LCDR1 comprising an amino acid sequence of SEQ ID NO: 643, LCDR2 comprising an amino acid sequence of SEQ ID NO: 644, and LCDR3 comprising an amino acid sequence of SEQ ID NO: 645; b) HCDRl comprising an amino acid sequence of SEQ ID NO: 646, HCDR2 comprising an amino acid sequence of SEQ ID NO: 647, HCDR3 comprising an amino acid sequence of SEQ ID NO: 648, LCDR1 comprising an amino acid sequence of SEQ ID NO: 649, LCDR2 comprising an amino acid sequence of SEQ ID NO: 650, and LCDR3 comprising an amino acid sequence of SEQ ID NO: 651; or c) HCDRl comprising an amino acid sequence of SEQ ID NO: 652, HCDR2 comprising an amino acid sequence of SEQ ID NO: 653, HCDR3 comprising an amino acid sequence of SEQ ID NO: 654, LCDR1 comprising an amino acid sequence of SEQ ID NO: 655, LCDR2 comprising an amino acid sequence of SEQ ID NO: 656, and LCDR3 comprising an amino acid sequence of SEQ ID NO: 657.
[0224] An antibody construct can comprise a second binding domain that specifically binds DEC-205. An antibody construct comprising a second binding domain that binds DEC-205 can comprise at least 80% sequence identity to: a) HCDR1 comprising an amino acid sequence of SEQ ID NO: 234, HCDR2 comprising an amino acid sequence of SEQ ID NO: 235, HCDR3 comprising an amino acid sequence of SEQ ID NO: 236, LCDR1 comprising an amino acid sequence of SEQ ID NO: 239, LCDR2 comprising an amino acid sequence of SEQ ID NO: 240, and LCDR3 comprising an amino acid sequence of SEQ ID NO: 241 ; or b) HCDR1 comprising an amino acid sequence of SEQ ID NO: 247, HCDR2 comprising an amino acid sequence of SEQ ID NO: 248, HCDR3 comprising an amino acid sequence of SEQ ID NO: 249, LCDR1 comprising an amino acid sequence of SEQ ID NO: 252, LCDR2 comprising an amino acid sequence of SEQ ID NO: 253, and LCDR3 comprising an amino acid sequence of SEQ ID NO: 254.
[0225] An antibody construct can comprise a second binding domain comprising one or more variable domains. An antibody construct can comprise a second binding domain comprising a light chain variable domain (VL domain). A second binding domain can comprise at least 80% sequence identity to any VL sequence in TABLE 6. An antibody construct can comprise a second binding domain comprising a heavy chain variable domain. A second binding domain can comprise at least 80% sequence identity to any VH sequence in TABLE 6. A second binding domain can comprise at least 80% sequence identity to any sequence in TABLE 6. A second binding domain can comprise at a pair of VH and VL domains having a pair of sequences in
TABLE 6.
[0226] An antibody construct can comprise a second binding domain that specifically binds
CD40. An antibody construct can comprise a second binding domain that is a CD40 agonist. An antibody construct can comprise a second binding domain that binds CD40, wherein the second binding domain comprises: a) a VH sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 2, and a VL sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 7; b) a VH sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 581, and a VL sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 586; c) a VH sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 591 , and a VL sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 596; d) a VH sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 601 , and a VL sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 606; e) a VH sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 611, and a VL sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 616; f) a VH sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 621, and a VL sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 626; g) a VH sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 631, and a VL sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 636.
[0227] An antibody construct can comprise a second binding domain that specifically binds DEC-205. An antibody construct can comprise a second binding domain that binds DEC-205, wherein the second binding domain comprises: a) a VH sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 233, and a VL sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 238; or b) a VH sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 246, and a VL sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 251.
[0228] An antibody construct can comprise a second binding domain that specifically binds to CD36 mannose scavenger receptor 1. An antibody construct can comprise a second binding domain that binds CD36 mannose scavenger receptor 1, wherein the second binding domain comprises a VH sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 658, and a VL sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 659.
[0229] An antibody construct can comprise a second binding domain that specifically binds to CLEC9A. An antibody construct can comprise a second binding domain that binds CLEC9A, wherein the second binding domain comprises a VH sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 660, and a VL sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 661.
[0230] An antibody construct can comprise a second binding domain that specifically binds to PD-Ll. An antibody construct can comprise a second binding domain that binds PD-Ll, wherein the second binding domain comprises a VH sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 901, and a VL sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 902; or wherein the second binding domain comprises a VH sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 890, and a VL sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 891; or wherein the second binding domain comprises a VH sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 892, and a VL sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 893.
[0231] An antibody construct can comprise a second binding domain and an Fc domain, wherein the second binding domain and the Fc domain comprise an antibody. An antibody construct can comprise a heavy chain and a light chain that target an antigen expressed by an antigen presenting cell. An antibody construct can comprise an antibody light chain. An antibody construct can comprise a light chain comprising at least 80% sequence identity to any light chain sequence in TABLE 7. An antibody construct can comprise an antibody heavy chain. An antibody construct can comprise a heavy chain comprising at least 80% sequence identity to any heavy chain sequence in TABLE 7. An antibody construct can comprise at least 80% sequence identity to any sequence in TABLE 7. An antibody construct can comprise a heavy and light chains having a pair of sequences in TABLE 7.
[0232] An antibody construct can comprise a heavy chain and a light chain that target an antigen expressed by an antigen presenting cell. An antibody construct can comprise a first binding domain and an Fc domain, wherein the first binding domain and the Fc domain comprise an antibody. An antibody construct may comprise an anti-CD40 antibody, the antibody construct comprising: a) a heavy chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 1 and a light chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 6; b) a heavy chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 577 or SEQ ID NO: 578, and a light chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 579; c) a heavy chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 580, and a light chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 585; d) a heavy chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 590, and a light chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 595; e) a heavy chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 600, and a light chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 605; f) a heavy chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 610, and a light chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 615; g) a heavy chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 620, and a light chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 625; or h) a heavy chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 630, and a light chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 635.
[0233] An antibody construct can comprise a first binding domain and an Fc domain, wherein the first binding domain and the Fc domain comprise an antibody. An antibody construct may comprise an anti-DEC-205 antibody, the construct comprising: a) a heavy chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 232, and a light chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 237; or b) a heavy chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 245, and a light chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 250.
[0234] An antibody construct can comprise a first binding domain and an Fc domain, wherein the first binding domain and the Fc domain comprise an antibody. A composition may comprise an anti-CLEC12A antibody, the construct comprising: a) a heavy chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 662, and a light chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 665; b) a heavy chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 663, and a light chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 665; or c) a heavy chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 664, and a light chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 665.
[0235] An antibody construct can comprise a first binding domain and an Fc domain, wherein the first binding domain and the Fc domain comprise an antibody. An antibody construct may comprise an anti-BDCA-2 antibody, the construct comprising: a) a heavy chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 666, and a light chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 669; b) a heavy chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 667, and a light chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 670; or c) a heavy chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 668, and a light chain sequence having at least 80% sequence identity to an amino acid sequence of SEQ ID NO: 671.
Antibody-ScFv Fusion Protein Products
[0236] An antibody construct can comprise a first binding domain, a second binding domain, and an Fc domain, wherein the first binding domain and the second binding domain are attached to the Fc domain. The first binding domain and the second binding domain can be attached to the Fc domain as a fusion protein. The first binding domain can be attached to the Fc domain at an N- terminal end of the Fc domain, wherein the second binding domain can be attached to the Fc domain at a C-terminal end. The first binding domain can be attached to the Fc domain at an N- terminal end of the Fc domain, wherein the second binding domain can be attached to the Fc domain at a C-terminal end via a polypeptide linker ranging from 10 to 25 amino acids. In some embodiments, the polypeptide linker has the sequence [G4S]n where n = 2 to 5. Alternatively, the first binding domain can be attached to the Fc domain at a C-terminal end of the Fc domain, wherein the second binding domain can be attached to the Fc domain at an N-terminal end. A second binding domain and an Fc domain can comprise an antibody and a first binding domain can comprise a single chain variable fragment (scFv). A single chain variable fragment can comprise a heavy chain variable domain and a light chain variable domain of an antibody. The first binding domain of the fusion protein can be attached to the second binding domain at a heavy chain variable domain of the single chain variable fragment of the first binding domain (HL orientation). Alternatively, the first binding domain of the fusion protein can be attached to the second binding domain at a light chain variable domain of the single chain variable fragment of the first binding domain (LH orientation). In either orientation, the first binding domain and the second binding domain can be attached via a polypeptide linker varying in length from 15 to 25 amino acids. In some embodiments, the polypeptide linker has the sequence [G4S]n where n = 3 to 5.
[0237] Alternatively, an antibody construct can comprise an antibody (having two antigen binding domains and an Fc domain) and the second binding domain can comprise a single chain variable fragment (scFv). The second binding domain of the fusion protein can be attached to the first binding domain at a heavy chain variable domain of the single chain variable fragment of the first binding domain (HL orientation). Alternatively, the second binding domain of the fusion protein can be attached to the first binding domain at a light chain variable domain of the single chain variable fragment of the first binding domain (LH orientation).
[0238] An antibody construct can comprise a first binding domain and a second binding domain, wherein the second binding domain can be attached to the first binding domain. The antibody construct can comprise an antibody comprising a light chain and a heavy chain or pair of heavy and light chains. The first binding domain can comprise an Fab fragment of the light and heavy chains. The second binding domain can be attached to the light chain at a C-terminus or C- terminal end of the light chain as a fusion protein. The second binding domain can comprise a single chain variable fragment (scFv). In some embodiments, a first binding domain can be specific for any of the tumor antigens, e.g., CEA, HER2, TROP2, Claudin-6 (CLDN6), Claudin- 16 (CLDN16), CLDN18.2, RON, LY6E, FRA, DLL3, PTK7, Uroplakin-IB (UPK1B), LIV1, ROR1, STRA6, TMPRSS3, TMPRSS4, TMEM238, Clorfl86, Fos-related antigen 1, VEGFR1, endoglin, VTCN1 (B7-H4), VISTA, or gpNMB, and an scFv can be a binding domain with a specificity selected from a group consisting of antigens on antigen presenting cells, such as CD40, DEC205, and PD-L1.
[0239] An antibody construct can comprise a first binding domain, a second binding domain, and an Fc domain, wherein the first binding domain and the second binding domain are attached to the Fc domain as a fusion protein. The second binding domain of the fusion protein can specifically target an antigen with at least 80% sequence identity to CD40. The second binding domain of the fusion protein can be a CD40 agonist. The first binding domain of the fusion protein can target a tumor antigen. The construct can comprise a fusion protein comprising a heavy chain (HC) attached to a single chain variable fragment. The construct comprising the fusion protein can comprise at least 80% sequence identity to any sequence in TABLE 4. The construct comprising the fusion protein can comprise at least 80% sequence identity to any sequence of a heavy chain CD40 monoclonal antibody (mAb) with tumor ScFv in TABLE 4. The construct can comprise a fusion protein comprising at least 80% sequence identity to any sequence of a heavy chain CD40 mAb with tumor ScFv in TABLE 4 and a light chain comprising at least 80% sequence identity to SEQ ID NO: 6. The construct comprising the fusion protein can comprise at least 80% sequence identity to any sequence of a heavy chain tumor mAb with CD40 ScFv in TABLE 4. The construct comprising the fusion protein can comprise at least 80% sequence identity to any sequence of a heavy chain tumor antigen mAb with CD40 ScFv in TABLE 4, and at least 80% sequence identity to a light chain mAb for the tumor antigen in
TABLE 3.
[0240] An antibody construct can comprise a first binding domain and a second binding domain, wherein the second binding domain can be attached to the first binding domain. An antibody construct can comprise a first binding domain, a second binding domain, and an Fc domain, wherein the second binding domain can be attached to the first binding domain. The second binding domain can be attached at a C-terminal end of the first binding domain as a fusion protein. The first binding domain can comprise a Fab fragment comprising a light chain, wherein the second binding domain can be attached at a C-terminal end of the light chain as a fusion protein. The second binding domain of the fusion protein can comprise a single chain variable fragment (scFv). The second binding domain of the fusion protein can be attached to the first binding domain at a heavy chain variable domain of the single chain variable fragment of the first binding domain (HL orientation). Alternatively, the second binding domain of the fusion protein can be attached to the first binding domain at a light chain variable domain of the single chain variable fragment of the first binding domain (LH orientation). For example, a fusion protein comprising a light chain of an anti-CEA antibody attached to an anti-CD40 scFv in the LH orientation can be illustrated by SEQ ID NO: 842. The fusion sequences comprising an scFv sequence are in the HL orientation unless indicated otherwise (e.g., sequence name recites "(LH)" indicating light heavy orientation).
[0241] The first binding domain of the fusion protein can target a tumor antigen. The second binding domain of the fusion protein can target an APC antigen. The second binding domain of the fusion protein can target CD40. The first binding domain can comprise a Fab fragment comprising a light chain, wherein the second binding domain is attached at a C-terminal end of the light chain as a fusion protein. The construct comprising the fusion protein can comprise at least 80% sequence identity to any sequence in TABLE 9. The construct comprising the fusion protein can comprise at least 80% sequence identity to any sequence of a light chain CD40 mAb with tumor ScFv in TABLE 9. The construct can comprise a fusion protein comprising at least 80% sequence identity to any sequence of a light chain CD40 mAb with tumor ScFv in TABLE 9 and a heavy chain comprising at least 80% sequence identity to SEQ ID NO: 1. The construct comprising the fusion protein can comprise at least 80% sequence identity to any sequence of a light chain tumor mAb with CD40 ScFv in TABLE 9. The construct can comprise a fusion protein comprising at least 80% sequence identity to any sequence of a light chain tumor antigen mAb with CD40 ScFv in TABLE 9, and at least 80% sequence identity to a heavy chain mAb for the tumor antigen in TABLE 3.
[0242] An antibody construct can comprise a first binding domain, a second binding domain, and an Fc domain, wherein the first binding domain and the second binding domain are attached to the Fc domain as a fusion protein. The first binding domain of the fusion protein can specifically target an antigen with at least 80% sequence identity to DEC-205. The second binding domain of the fusion protein can target a tumor antigen. The construct can comprise a fusion protein comprising a heavy chain attached to a single chain variable fragment. The construct comprising the fusion protein can comprise at least 80% sequence identity to any sequence in TABLE 8. The construct comprising the fusion protein can comprise at least 80% sequence identity to any sequence of a heavy chain DEC-205 mAb with tumor ScFv in TABLE 8. The construct can comprise a fusion protein comprising at least 80% sequence identity to any sequence of a heavy chain DEC-205 mAb with tumor ScFv in TABLE 8 and a peptide comprising at least 80% sequence identity to SEQ ID NO: 237. The construct comprising the fusion protein can comprise at least 80% sequence identity to any sequence of a heavy chain tumor antigen mAb with CD40 ScFv in TABLE 8. The construct comprising the fusion protein can comprise at least 80% sequence identity to any sequence of a heavy chain tumor antigen mAb with CD40 ScFv in TABLE 8, and at least 80% sequence identity to a heavy chain rnAb for the tumor antigen in TABLE 3.
[0243] The second binding domain of the fusion protein can target DEC-205. The construct comprising the fusion protein can comprise at least 80% sequence identity to any sequence in
TABLE 10. The construct comprising the fusion protein can comprise at least 80% sequence identity to any sequence of a light chain DEC-205 rnAb with tumor ScFv in TABLE 10. The construct comprising a fusion protein can comprise at least 80% sequence identity to any sequence of a light chain DEC-205 rnAb with tumor ScFv in TABLE 10 and at least 80% sequence identity to SEQ ID NO: 237. The construct comprising the fusion protein can comprise at least 80% sequence identity to any sequence of a light chain tumor mAb with DEC-205 ScFv in TABLE 10. The construct comprising a fusion protein can comprise at least 80% sequence identity to any sequence of a light chain tumor antigen mAb with DEC-205 ScFv in TABLE 10, and at least 80% sequence identity to a heavy chain mAb for the tumor antigen in TABLE 3.
[0244] The second binding domain of the fusion protein can specifically bind to an antigen of an
Antigen Presenting Cell (APC) or other immune cell. The second binding domain of the fusion protein can specifically bind to an antigen with at least 80% sequence identity to CD40. The second binding domain of the fusion protein can be a CD40 agonist. The second binding domain of the fusion protein can specifically bind to an antigen with at least 80% sequence identity to
DEC-205. The second binding domain of the fusion protein can specifically bind to an antigen with at least 80% sequence identity to DC-SIGN. The second binding domain of the fusion protein can specifically bind to an antigen with at least 80% sequence identity to CD36 mannose scavenger receptor. The second binding domain of the fusion protein can specifically bind to an antigen with at least 80% sequence identity to CLEC12A. The second binding domain of the fusion protein can specifically bind to an antigen with at least 80% sequence identity to BDCA-2.
The second binding domain of the fusion protein can specifically bind to an antigen with at least
80% sequence identity to PD-L1. The second binding domain of the fusion protein can specifically bind to an antigen with at least 80% sequence identity to CD40, CD47, TNFR2,
DEC-205, PD-L1, CD36 mannose scavenger receptor 1, CLEC9A, DC-SIGN, CLEC12A,
BDCA-2, OX40L, 41BBL, CD204, MARCO, CLEC5A, Dectin 1, Dectin 2, CLECIOA, CD206,
CD64, CD32A, CD 16 A, HVEM, CD38, TREM2, CSF1R, or CD32B. The first binding domain of the fusion protein can specifically bind to a tumor antigen. The first binding domain of the fusion protein can specificall bind to an antigen with at least 80% sequence identity to CD5,
CD19, CD20, CD25, CD37, CD30, CD33, CD45, CAMPATH-1, BCMA, CS-1, PD-L1, B7-H3,
B7-DC (PD-L2), HLD-DR, carcinoembryonic antigen (CEA), TAG-72, EpCAM, MUC1,
MUC15, MUC16, fo late-binding protein, A33, G250, pro state- specific membrane antigen (PSMA), ferritin, CA-125, CA19-9, epidermal growth factor, pl85HER2, IL-2 receptor, EGFRvIII (de2-7 EGFR), fibroblast activation protein (FAP), tenascin, a metalloproteinase, endosialin, vascular endothelial growth factor, ανβ3, WTl, LMP2, HPV E6, HPV E7, Her-2/neu, p53 nonmutant, NY-ESO-1, MelanA/M ART 1 , Ras mutant, gplOO, p53 mutant, PR1, bcr-abl, tyrosinase, survivin, PSA, hTERT, a Sarcoma translocation breakpoint protein, EphA2, PAP, ML-IAP, AFP, ERG, NA17, PAX3, ALK, androgen receptor, cyclin B l, MYCN, RhoC, TRP-2, mesothelin (MSLN), PSCA, MAGE Al, MAGE A3, CYP1B 1, PLAV1, BORIS, ETV6-AML, NY-BR-1, RGS5, SART3, Carbonic anhydrase IX, PAX5, OY-TES 1, Sperm protein 17, LCK, HMWMAA, AKAP-4, SSX2, XAGE 1, Legumain, Tie 3, Page4, VEGFR2, MAD-CT-1,
PDGFR-B, MAD-CT-2, ROR2, TRAIL 1, MAGE A4, MAGE C2, GAGE, EGFR, CMET, HER3, EPCAM, CA6, NAPI2B, TROP2, Claudin-6 (CLDN6), Claudin-16 (CLDN16),
CLDN18.2, RON, LY6E, FRA, DLL3, PTK7, Uroplakin-IB (UPK1B), LIV1, ROR1, STRA6, TMPRSS3, TMPRSS4, TMEM238, Clorfl86, Fos-related antigen 1, VEGFR1, endoglin, PD- Ll, VTCN1 (B7-H4), VISTA, gpNMB, or any fragment thereof.
[0245] Alternatively, the second binding domain of the fusion protein can target a tumor antigen. The second binding domain of the fusion protein can specifically bind to an antigen selected from CD5, CD19, CD20, CD25, CD37, CD30, CD33, CD45, CAMPATH-1, BCMA, CS-1, PD- Ll, B7-H3, B7-DC (PD-L2), HLD-DR, carcinoembryonic antigen (CEA), TAG-72, EpCAM, MUC1, MUC15, MUC16, fo late-binding protein, A33, G250, pro state- specific membrane antigen (PSMA), ferritin, GD2, GD3, GM2, Ley, CA-125, CA19-9, epidermal growth factor, pl85HER2, IL-2 receptor, EGFRvIII (de2-7 EGFR), fibroblast activation protein (FAP), tenascin, a metalloproteinase, endosialin, vascular endothelial growth factor, ανβ3, WTl, LMP2, HPV E6 E7, Her-2/neu, p53 nonmutant, NY-ESO-1, MelanA/M ART 1 , Ras mutant, gplOO, p53 mutant, PR1, bcr-abl, tyrosinase, survivin, PSA, hTERT, a Sarcoma translocation breakpoint protein, EphA2, PAP, ML-IAP, AFP, ERG, NA17, PAX3, ALK, androgen receptor, cyclin B l, polysialic acid, MYCN, RhoC, TRP-2, fucosyl GM1, mesothelin (MSLN), PSCA, MAGE Al, MAGE A3, sLe(animal), CYP1B 1, PLAV1, GM3, BORIS, Tn, GloboH, ETV6-AML, NY-BR-1, RGS5, SART3, STn, Carbonic anhydrase IX, PAX5, OY-TES 1, Sperm protein 17, LCK, HMWMAA, AKAP-4, SSX2, XAGE 1, Legumain, Tie 3, Page4, VEGFR2, MAD-CT-1, PDGFR-B, MAD- CT-2, ROR2, TRAIL 1, MAGE A4, MAGE C2, GAGE, EGFR, CMET, HER3, EPCAM, CA6, NAPI2B, TROP2, Claudin-6 (CLDN6), Claudin-16 (CLDN16), CLDN18.2, RON, LY6E, FRA, DLL3, PTK7, Uroplakin-IB (UPK1B), LIV1, ROR1, STRA6, TMPRSS3, TMPRSS4,
TMEM238, Clorfl86, Fos-related antigen 1, VEGFR1, endoglin, PD-L1, VTCN1 (B7-H4), VISTA, gpNMB, or any fragment thereof. The first binding domain of the fusion protein can specifically bind to an antigen with at least 80% sequence identity to CD40. The first binding domain of the fusion protein can be a CD40 agonist. The first binding domain of the fusion protein can specifically bind to an antigen with at least 80% sequence identity to DEC-205. The first binding domain of the fusion protein can specifically bind to an antigen with at least 80% sequence identity to CEA, HER2, TROP2, Claudin-6 (CLDN6), Claudin-16 (CLDN16),
CLDN18.2, RON, LY6E, FRA, DLL3, PTK7, Uroplakin-IB (UPK1B), LIV1, ROR1, STRA6, TMPRSS3, TMPRSS4, TMEM238, Clorfl86, Fos-related antigen 1, VEGFR1, endoglin, VTCN1 (B7-H4), VISTA or gpNMB. The first binding domain of the fusion protein can specifically bind to an antigen with at least 80% sequence identity to CD40, CD47, TNFR2, DEC-205, PD-L1, CD36 mannose scavenger receptor 1, CLEC9A, DC-SIGN, CLEC12A, BDCA-2, OX40L, 41BBL, CD204, MARCO, CLEC5A, Dectin 1, Dectin 2, CLECIOA, CD206, CD64, CD32A, CD 16 A, HVEM, CD38, TREM2, CSF1R, or CD32B.
[0246] An antibody construct can comprise a first binding domain, a second binding domain, and a third binding domain. An antibody construct can comprise a first binding domain, a second binding domain, a third binding domain, and an Fc domain. The first binding domain and the second binding domain can be attached to the Fc domain. The first and second binding domains are described herein throughout the specification. The first binding domain can be attached to the Fc domain at an N-terminal end of the Fc domain. The second binding domain can be attached at a C-terminal end of the Fc domain. The second binding domain can be attached at a C-terminal end of the Fc domain via a polypeptide linker having a length of 10 to 25 amino acid. In some embodiments, the polypeptide linker has the sequence [G4S]n where n = 2 to 5. The third binding domain can be attached to a C-terminal end of the first binding domain. The third binding domain can be attached at a C-terminal end of the Fc domain via a polypeptide linker having a length of 10 to 25 amino acid. In some embodiments, the polypeptide linker has the sequence [G4S]n where n = 2 to 5. The third binding domain can be attached to a C-terminal end of a light chain of the first binding domain. One or more of the first binding domain, the second binding domain, the third binding domain, and the Fc domain can be attached as a fusion protein. The first binding domain can comprise a Fab fragment comprising a light chain, wherein the second binding domain is attached at a C-terminal end of the light chain as a fusion protein. The second binding domain of the fusion protein can comprise a single chain variable fragment (scFv). The second binding domain of the fusion protein can be attached to the Fc domain at a heavy chain variable domain of the single chain variable fragment of the second binding domain (HL orientation). The second binding domain of the fusion protein can be attached to the Fc domain at a light chain variable domain of the single chain variable fragment of the second binding domain (LH orientation). The third binding domain of the fusion protein can comprise a single chain variable fragment (scFv). The antibody construct can comprise a fusion protein comprising the third binding domain attached to the first binding domain having at least 80% sequence identity to any sequence in TABLE 9 or TABLE 10. The third binding domain of the fusion protein can be attached to the first binding domain at a heavy chain variable domain of the single chain variable fragment of the first binding domain (HL orientation). Alternatively, the third binding domain of the fusion protein can be attached to the first binding domain at a light chain variable domain of the single chain variable fragment of the first binding domain (LH orientation). The third binding domain of the fusion protein can target an antigen of an immune cell, such as an antigen presenting cell (APC). The third binding domain of the fusion protein can specifically bind to an antigen with at least 80% sequence identity to CD40. The third binding domain of the fusion protein can be a CD40 agonist. The third binding domain of the fusion protein can specifically bind to an antigen with at least 80% sequence identity to DEC-205. The third binding domain of the fusion protein can specifically bind to an antigen with at least 80% sequence identity to CD40, CD47, TNFR2, DEC-205, PD-Ll, CD36 mannose scavenger receptor 1, CLEC9A, DC-SIGN, CLEC12A, BDCA-2, OX40L, 41BBL, CD204, MARCO, CLEC5A, Dectin 1, Dectin 2, CLECIOA, CD206, CD64, CD32A, CD 16 A, HVEM, CD38, TREM2, CSF1R, or CD32B.
[0247] Alternatively, the third binding domain can target a tumor antigen. The third binding domain of the fusion protein can specifically bind to an antigen with at least 80% sequence identity to CD5, CD19, CD20, CD25, CD37, CD30, CD33, CD45, CAMPATH-1, BCMA, CS-1,
PD-Ll, B7-H3, B7-DC (PD-L2), HLD-DR, carcinoembryonic antigen (CEA), TAG-72, EpCAM,
MUC1, MUC15, MUC16, fo late-binding protein, A33, G250, pro state- specific membrane antigen (PSMA), ferritin, CA-125, CA19-9, epidermal growth factor, pl85HER2, IL-2 receptor,
EGFRvIII (de2-7 EGFR), fibroblast activation protein (FAP), tenascin, a metalloproteinase, endosialin, vascular endothelial growth factor, ανβ3, WTl, LMP2, HPV E6, HPV E7, Her-2/neu, p53 nonmutant, NY-ESO-1, MelanA/M ART 1 , Ras mutant, gplOO, p53 mutant, PR1, bcr-abl, tyrosinase, survivin, PSA, hTERT, a Sarcoma translocation breakpoint protein, EphA2, PAP,
ML-IAP, AFP, ERG, NA17, PAX3, ALK, androgen receptor, cyclin B l, MYCN, RhoC, TRP-2, mesothelin (MSLN), PSCA, MAGE Al, MAGE A3, CYP1B 1, PLAV1, BORIS, GloboH, ETV6-
AML, NY-BR-1, RGS5, SART3, Carbonic anhydrase IX, PAX5, OY-TES 1, Sperm protein 17,
LCK, HMWMAA, AKAP-4, SSX2, XAGE 1, Legumain, Tie 3, PAGE4, VEGFR2, MAD-CT-1,
PDGFR-B, MAD-CT-2, ROR2, TRAIL 1, MAGE A4, MAGE C2, GAGE, EGFR, CMET,
HER3, EPCAM, CA6, NAPI2B, TROP2, Claudin-6 (CLDN6), Claudin-16 (CLDN16),
CLDN18.2, RON, LY6E, FRA, DLL3, PTK7, Uroplakin-IB (UPK1B), LIV1, ROR1, STRA6,
TMPRSS3, TMPRSS4, TMEM238, Clorfl86, Fos-related antigen 1, VEGFR1, endoglin,
VTCN1 (B7-H4), VISTA, gpNMB, or any fragment thereof. In additional embodiments, the third binding domain of the fusion protein can specifically bind to an antigen selected from CD5, CD19, CD20, CD25, CD37, CD30, CD33, CD45, CAMPATH-1, BCMA, CS-1, PD-L1, B7-H3, B7-DC (PD-L2), HLD-DR, carcinoembryonic antigen (CEA), TAG-72, EpCAM, MUC1, MUC15, MUC16, fo late-binding protein, A33, G250, pro state- specific membrane antigen (PSMA), ferritin, GD2, GD3, GM2, Ley, CA-125, CA19-9, epidermal growth factor, pl85HER2, IL-2 receptor, EGFRvIII (de2-7 EGFR), fibroblast activation protein (FAP), tenascin, a metalloproteinase, endosialin, vascular endothelial growth factor, ανβ3, WT1, LMP2, HPV E6 E7, Her-2/neu, p53 nonmutant, NY-ESO-1, MelanA/M ART 1 , Ras mutant, gplOO, p53 mutant, PR1, bcr-abl, tyrosinase, survivin, PSA, hTERT, a Sarcoma translocation breakpoint protein, EphA2, PAP, ML-IAP, AFP, ERG, NA17, PAX3, ALK, androgen receptor, cyclin B l, polysialic acid, MYCN, RhoC, TRP-2, fucosyl GM1, mesothelin (MSLN), PSCA, MAGE Al, MAGE A3, sLe(animal), CYP1B 1, PLAV1, GM3, BORIS, Tn, GloboH, ETV6-AML, NY-BR-1, RGS5, SART3, STn, Carbonic anhydrase IX, PAX5, OY-TES 1, Sperm protein 17, LCK, HMWMAA, AKAP-4, SSX2, XAGE 1, Legumain, Tie 3, PAGE4, VEGFR2, MAD-CT-1, PDGFR-B, MAD- CT-2, ROR2, TRAIL 1, MAGE A4, MAGE C2, GAGE, EGFR, CMET, HER3, EPCAM, CA6, NAPI2B, TROP2, Claudin-6 (CLDN6), Claudin-16 (CLDN16), CLDN18.2, RON, LY6E, FRA, DLL3, PTK7, Uroplakin-IB (UPK1B), LIV1, ROR1, STRA6, TMPRSS3, TMPRSS4,
TMEM238, Clorfl86, Fos-related antigen 1, VEGFR1, endoglin, VTCN1 (B7-H4), VISTA, gpNMB, or any fragment thereof.
[0248] An antibody construct can comprise a first binding domain targeting CD40 and a second binding domain targeting DEC-205. Alternatively, an antibody construct can comprise a first binding domain targeting DEC-205 and a second binding domain targeting CD40. An antibody construct can comprise a first binding domain, a second binding domain, and an Fc domain. The first binding domain and the second binding domain can be attached to the Fc domain. The first binding domain can be attached to the Fc domain at an N-terminal end of the Fc domain, wherein the second binding domain is attached to the Fc domain at a C-terminal end of the Fc domain. Alternatively, the second binding domain can be attached to the Fc domain at an N-terminal end of the Fc domain, wherein the first binding domain is attached to the Fc domain at a C-terminal end of the Fc domain. An antibody construct can comprise a fusion protein comprising a first binding domain targeting CD40 and a second binding domain targeting DEC-205. The fusion protein can comprise at least 80% sequence identity to any sequence in TABLE 11.
[0249] Additionally, antibody constucts expressing proteins having the sequences referenced in Tables 1-11 can have a dissociation constant (IQ) that is less than 10 nM for the antigen of the first binding domain. The antibody constructs expressing proteins having the sequences referenced in Tables 1-11 can have a dissociation constant (IQ) that is less than 10 nM for the antigen of the second binding domain. The antibody constructs expressing the sequences referenced in Tables 1-11 can have a dissociation constant (Kd) that is less than 10 nM for the antigen of the third binding domain. The antibody constructs can have a dissociation constant (Kd) for the antigen of the first binding domain that is less than 1 nM, less than 100 pM, less than 10 pM, less than 1 pM, or less than 0.1 pM. The antibody constructs can have a dissociation constant (Kd) for the antigen of the second binding domain that is less than 1 nM, less than 100 pM, less than 10 pM, less than 1 pM, or less than 0.1 pM. The antibody constructs can have a dissociation constant (Kd) for the antigen of the third binding domain that is less than 1 nM, less than 100 pM, less than 10 pM, less than 1 pM, or less than 0.1 pM.
[0250] An anti-CD40 light chain can be expressed with an anti-CD40 heavy chain or fragment thereof, wherein the light and heavy chains specifically bind to CD40. The anti-CD40 light chain can also be expressed with an anti-CD40 heavy chain or fragment thereof to form an anti-CD40 antibody or fragment thereof, wherein the antibody or antibody fragment specifically binds to CD40. The anti-CD40 antibody or fragment thereof can be purified, and can be combined with a pharmaceutically acceptable carrier.
[0251] An anti-DEC-205 light chain can be expressed with any anti-DEC-205 heavy chain or fragment thereof, wherein the light and heavy chains specifically bind to DEC-205. The anti- DEC-205 light chain can also be expressed with an anti-DEC-205 heavy chain or fragment thereof to form an anti-DEC-205 antibody or fragment thereof, wherein the antibody or antibody fragment specifically binds to DEC-205. The anti-DEC-205 antibody or fragment thereof can be purified, and can be combined with a pharmaceutically acceptable carrier.
[0252] An anti-PD-Ll light chain can be expressed with any anti-PD-Ll heavy chain or fragment thereof, wherein the light and heavy chains specifically bind to PD-L1. The anti-PD-Ll light chain can also be expressed with an anti-PD-Ll heavy chain or fragment thereof to form an anti- PD-Ll antibody or fragment thereof, wherein the antibody or antibody fragment specifically binds to PD-L1. The anti-PD-Ll antibody or fragment thereof can be purified, and can be combined with a pharmaceutically acceptable carrier.
[0253] An anti-tumor antigen light chain can be expressed with any anti-tumor antigen heavy chain or fragment thereof, wherein the antibody or antibody fragment specifically binds to tumor antigen. The anti-tumor antigen light chain can also expressed with any anti-tumor antigen heavy chain or fragment thereof to form an anti-tumor antigen antibody or fragment thereof. The antitumor antibody or fragment thereof can be purified, and can be combined with a
pharmaceutically acceptable carrier.
[0254] An antibody construct can comprise an antibody heavy chain. A heavy chain can be a heavy chain of an anti-CD40 antibody which can bind a CD40 antigen. A heavy chain of an anti- CD40 antibody can be an IgGl isotype. A heavy chain of an anti-CD40 antibody can be dacetuzumab.
[0255] An antibody construct can comprise an antibody light chain. A light chain can be a light chain of an anti-CD40 antibody which can bind a CD40 antigen. A light chain of an anti-CD40 antibody can be dacetuzumab.
[0256] An antibody construct can comprise an antibody heavy chain. A heavy chain can be a heavy chain of an anti-CD40 antibody which can bind a CD40 antigen. A heavy chain of an anti- CD40 antibody can be an IgG4 isotype. A heavy chain of an anti-CD40 antibody can be bleselumab.
[0257] An antibody construct can comprise an antibody light chain. A light chain can be a light chain of an anti-CD40 antibody which can bind a CD40 antigen. A light chain of an anti-CD40 antibody can be bleselumab.
[0258] An antibody constructcan comprise an antibody heavy chain. A heavy chain can be a heavy chain of an anti-CD40 antibody which can bind a CD40 antigen. A heavy chain of an anti- CD40 antibody can be an IgGl isotype. A heavy chain of an anti-CD40 antibody can be lucatumumab.
[0259] An antibody construct can comprise an antibody light chain. A light chain can be a light chain of an anti-CD40 antibody which can bind a CD40 antigen. A light chain of an anti-CD40 antibody can be lucatumumab.
[0260] An antibody construct can comprise an antibody heavy chain. A heavy chain can be a heavy chain of an anti-CD40 antibody which can bind a CD40 antigen. A heavy chain of an anti- CD40 antibody can be an IgGl isotype. A heavy chain of an anti-CD40 antibody can be ADC- 1013.
[0261] An antibody construct can comprise an antibody light chain. A light chain can be a light chain of an anti-CD40 antibody which can bind a CD40 antigen. A light chain of an anti-CD40 antibody can be ADC- 1013.
[0262] An antibody construct can comprise an antibody heavy chain. A heavy chain can be a heavy chain of an anti-CD40 antibody which can bind a CD40 antigen. A heavy chain of an anti- CD40 antibody can be the humanized rabbit antibody APX005.
[0263] An antibody construct can comprise an antibody light chain. A light chain can be a light chain of an anti-CD40 antibody which can bind a CD40 antigen. A light chain of an anti-CD40 antibody can be the humanized rabbit antibody APX005.
[0264] An antibody construct can comprise an antibody heavy chain. A heavy chain can be a heavy chain of an anti-CD40 antibody which can bind a CD40 antigen. A heavy chain of an anti- CD40 antibody can be Chi Lob 7/4. [0265] An antibody construct can comprise an antibody light chain. A light chain can be a light chain of an anti-CD40 antibody which can bind a CD40 antigen. A light chain of an anti-CD40 antibody can be Chi Lob 7/4.
[0266] An antibody construct can comprise an antibody heavy chain. A heavy chain can be a heavy chain of an anti-CD40 antibody which can bind a CD40 antigen. A heavy chain of an anti- CD40 antibody can be an IgGl isotype. A heavy chain of an anti-CD40 antibody can be SBT- 040-G1WT.
[0267] An antibody construct can comprise an antibody heavy chain. A heavy chain can be a heavy chain of an anti-CD40 antibody which can bind a CD40 antigen. A heavy chain of an anti- CD40 antibody can be an IgGl isotype. A heavy chain of an anti-CD40 antibody can be SBT- 040 VH-hlgGl wt.
[0268] A heavy chain of an anti-CD40 antibody can be an IgG2 isotype. A heavy chain of an anti-CD40 antibody can be SBT-040-G2.
[0269] An antibody construct can comprise an antibody with modifications occurring at least at one amino acid residue. Modifications can be substitutions, additions, mutations, deletions, or the like. An antibody modification can be an insertion of an unnatural amino acid.
[0270] An antibody construct can comprise a light chain of an amino acid sequence having at least one, two, three, four, five, six, seven, eight, nine, or ten modifications but not more than 40, 35, 30, 25, 20, 15, or 10 modifications of the amino acid sequence relative to the natural or original amino acid sequence. An antibody construct can comprise a heavy chain with an amino acid sequence having at least one, two, three, four, five, six, seven, eight, nine or ten
modifications but not more than 40, 35, 30, 25, 20, 15, or 10 modifications of the amino acid sequence relative to the natural or original amino acid sequence.
[0271] An antibody construct can have an Fc domain of an IgGl isotype. An antibody construct can have an Fc domain of an IgG2 isotype. An antibody construct can have an Fc domain of an IgG3 isotype. An antibody construct can have an Fc domainof an IgG4 isotype. An antibody construct can have an Fc domain of a hybrid isotype comprising constant regions from two or more isotypes. An antibody construct can be an anti-CD40 antibody, in which the anti-CD40 antibody can be a monoclonal human antibody comprising a wild-type sequence of an IgGl isoform, in particular, at an Fc region of the antibody.
[0272] An antibody constructs disclosed herein can be non-natural, designed, and/or engineered. Antibody constructs disclosed herein can be non-natural, designed, and/or engineered scaffolds comprising an antigen binding domain. Antibody constructs disclosed herein can be no n- natural, designed, and/or engineered antibodies. Antibody constructs can be monoclonal antibodies. Antibody constructs can be human antibodies. Antibody constructs can be humanized antibodies. Antibody constructs can be monoclonal humanized antibodies. Antibody construct can be recombinant antibodies.
[0273] The Kd for binding of the Fc domain to an Fc receptor of an antibody construct as described herein can increase when the tumor antigen binding domain is bound to its tumor antigen as compared to the Kd for binding of the Fc domain to an Fc receptor of a construct as described herein when the tumor antigen binding domain is not bound to its tumor antigen. For example, an antibody construct as described herein can have a Kd for binding of the Fc domain to an Fc receptor in the presence of the binding domain that binds to an antigen on an immune cell, such as an antigen presenting cell, and the tumor target binding domain when the tumor target binding domain is bound to its tumor antigen that can be greater than or greater than about 100 nM. The Kd for binding of the Fc domain to an Fc receptor in the presence of the binding domain that binds to an antigen on an immune cell, such as an antigen presenting cell, and the tumor target binding domain when the tumor target binding domain is bound to its tumor antigen can be or can be about 100 nM, 200 nM, 300 nM, 400 nM, 500 nM, or 1000 nM. The ¾ for binding of the Fc domain to an Fc receptor in the presence of the binding domain that binds to an antigen on an immune cell, such as an antigen presenting cell, and the tumor target binding domain when the tumor target binding domain is bound to its tumor antigen can be from 100 nM to 200 nM, 100 nM to 300 nM, 100 nM to 400 nM, 100 nM to 500 nM, or 100 nM to 1000 nM. Additionally, the antibody construct as described herein can have a Kd for binding of the Fc domain to an Fc receptor in the presence of the binding domain that binds to an antigen on an immune cell, such as an antigen presenting cell, and a tumor antigen binding domain when the tumor antigen binding domain is not bound to the tumor antigen is no greater than about 100 nM and is no greater than about 100 times a Kd for binding of the Fc domain to the Fc receptor in an absence of the binding domain that binds to an antigen on an immune cell and a tumor antigen binding domain.
[0274] The Kd for binding of the binding domain of an antibody construct as described herein that binds to an antigen on an immune cell, such as an antigen presenting cell, can increase when the tumor antigen binding domain is bound to its tumor antigen as compared to the Kd for binding of the binding domain that binds to an antigen on an immune cell when the tumor antigen binding domain is not bound to its tumor antigen. For example, an antibody construct can comprise a Kd for binding of the binding domain that binds to an antigen on an antigen presenting cell when the tumor antigen binding domain is bound to its tumor antigen can be greater than or greater than about ΙΟΟηΜ. The Kd for binding of the binding domain that binds to an antigen on an immune cell, such as an antigen presenting cell, when the tumor antigen binding domain is bound to its tumor antigen can be or can be about 100 nM, 200 nM, 300 nM, 400 nM, 500 nM, or 1000 nM. IQ for binding of the binding domain that binds to an antigen on an immune cell when the tumor antigen binding domain is bound to its tumor antigen can be from 100 nM to 200 nM, 100 nM to 300 nM, 100 nM to 400 nM, 100 nM to 500 nM, or 100 nM to 1000 nM.
[0275] The effect of the tumor antigen binding domain and the binding domain that binds to an antigen on the immune cell together can be to cluster the antibody constructs or conjugates on cells expressing tumor antigen, and thus clustering immune cells such as antigen presenting cells around cancerous cells and at tumor sites resulting in activation of the antigen presenting cell effector functions. This can include the activation of the antigen on the antigen presenting cell when a an antibody construct is bound to its antigen, such as activation of CD40, CD47, TNFR2, DEC-205, PD-L1, CD36 mannose scavenger receptor 1, DC-SIGN, CLEC9A, CLEC12A, BDCA-2, OX40L, 41BBL, CD204, MARCO, CLEC5A, Dectin 1, Dectin 2, CLECIOA, CD206, CD64, CD32A, CD 16 A, HVEM, CD38, TREM2, CSFIR, or CD32B. In some embodiments, this activation of the antigen on the antigen presenting cell only occurs when the tumor targeting antibody construct is bound to its tumor antigen. An antigen presenting cell effector function can include antibody dependent cellular cytotoxicity (ADCC) of the tumor antigen expressing cell, which can occur when a bispecific tumor targeting antibody construct is bound to its tumor antigen. In some embodiments, ADCC of the tumor antigen expressing cell only occurs with a bispecific tumor targeting antibody construct is bound to its tumor antigen. In certain
embodiments, a bispecific tumor targeting antibody construct density of greater than 1000, 2000, 3000, 4000, 5000, 6000, 7000, 8000, 9000, 10,000 or more per cell, resulting from the bispecific tumor targeting antibody construct binding to the tumor antigen, induces signaling in the antigen presenting cell. Signaling can suitably be measured in vitro using a cell line expressing the tumor antigen bound by the target antigen binding domain, and primary antigen presenting cells isolated from a human subject. Signaling can be assessed as cytokine release, chemokine release, or increased expression of cell surface markers. In certain embodiments, a bispecific tumor targeting antibody construct density of greater than 1000, 2000, 3000, 4000, 5000, 6000, 7000, 8000, 9000, 10,000 or more per cell, resulting from the bispecific tumor targeting antibody construct binding to the tumor antigen, induces ADCC of the cells expressing tumor antigen. ADCC can suitably be measured in vitro using a cell line expressing the tumor antigen bound by the target antigen binding domain, and cells such as NK cells and/or macrophages isolated from a human subject. ADCC can be determined by the frequency of remaining tumor antigen expressing cells in the co-culture.
[0276] In some embodiments, the antibody constructs as described herein can specifically bind to a tumor antigen in a cluster of constructs or conjugates, and this clustering can induce a signal in an antigen presenting cell. The constructs as described herein can specifically bind to a tumor antigen in a cluster of constructs, and this clustering can induce antibody dependent cellular cytotoxicity. The constructs as described herein can specifically bind to a tumor antigen in a cluster of constructs and this clustering can result in an increased avidity for an antigen on an antigen presenting cell. The antibody constructs as described herein can specifically bind to a tumor antigen in a cluster of constructs and this clustering can result in an increased avidity of the Fc domain for an Fc receptor.
[0277] Sequences that can be used to produce antibodies for antibody constructs can include leader sequences. Leader sequences can include signal sequences. Leader sequences useful with the antibody construct and methods described herein can include, but are not limited to, an amino acid sequence comprising MRLP AQLLGLLLLWFPGS RC (SEQ ID NO: 847),
MDWTWRILFLVAAATGAHS (SEQ ID NO: 848), and MRAWIFFLLCLAGRALA (SEQ ID NO: 849).
[0278] An antibody construct can comprise an Fc region with an Fc domain. An Fc domain is a structure that can bind to Fc receptors. An antibody construct can comprise an Fc domain. Fc domains can be bound by Fc receptors (FcRs). Fc domains can be from antibodies. An Fc domain can be at least 80% homologous to an Fc domain from an antibody. An Fc region can be in a scaffold. An Fc region with an Fc domain can be in an antibody scaffold. An Fc region with an Fc domain can be in a non-antibody scaffold. An antibody construct can comprise an Fc region with an Fc domain in an antibody scaffold. An antibody construct can comprise an Fc region with an Fc domain in a non-antibody scaffold. An Fc domain can be in a scaffold. An Fc domain can be in an antibody scaffold. An Fc domain can be in a non-antibody scaffold. An antibody construct can comprise an Fc domain in an antibody scaffold. An antibody construct can comprise an Fc domain in a non-antibody scaffold. Fc domains of antibodies, including those of the present disclosure, can be bound by Fc receptors (FcRs). An Fc domain can be a portion of the Fc region of an antibody. FcRs can bind to an Fc domain of an antibody. FcRs can bind to an Fc domain of an antibody bound to an antigen. FcRs are organized into classes (e.g. , gamma (γ), alpha (a) and epsilon (ε)) based on the class of antibody that the FcR recognizes. The FcaR class can bind to IgA and includes several isoforms, FcaRI (CD89) and Fco jR. The FcyR class can bind to IgG and includes several isoforms, FcyRI (CD64), FcyRIIA (CD32a), FcyRIIB (CD32b), FcyRIIIA (CD 16a), and FcyRIIIB (CD 16b). An FcyRIIIA (CD 16a) can be an FcyRIIIA (CD 16a) F158 variant. An FcyRIIIA (CD 16a) can be an FcyRIIIA (CD 16a) V158 variant. Each FcyR isoform can differ in affinity to the Fc region of the IgG antibody. For example, FcyRI can bind to IgG with greater affinity than FcyRII or FcyRIII. The affinity of a particular FcyR isoform to IgG can be controlled, in part, by a glycan (e.g., oligosaccharide) at position CH2 84.4 of the IgG antibody. For example, fucose containing CH2 84.4 glycans can reduce IgG affinity for FcyRIIIA. In addition, GO glucans can have increased affinity for FcyRIIIA due to the lack of galactose and terminal GlcNAc moiety.
[0279] Binding of an Fc domain to an FcR can enhance an immune response. FcR-mediated signaling that can result from an Fc region binding to an FcR can lead to the maturation of immune cells. FcR-mediated signaling that can result from an Fc domain binding to an FcR can lead to the maturation of dendritic cells. FcR-mediated signaling that can result from an Fc domain binding to an FcR can lead to antibody dependent cellular cytotoxicity. FcR-mediated signaling that can result from an Fc domain binding to an FcR can lead to more efficient immune cell antigen uptake and processing. FcR-mediated signaling that can result from an Fc region binding to an FcR can lead to more efficient dendritic cell antigen uptake and processing. FcR- mediated signaling that can result from an Fc region binding to an FcR can increase antigen presentation. FcR-mediated signaling that can result from an Fc region binding to an FcR can increase antigen presentation by immune cells. FcR-mediated signaling that can result from an Fc region binding to an FcR can increase antigen presentation by antigen presenting cells. FcR- mediated signaling that can result from an Fc domain binding to an FcR can increase antigen presentation by dendritic cells. FcR-mediated signaling that can result from an Fc domain binding to an FcR can promote the expansion and activation of T cells. FcR-mediated signaling that can result from an Fc domain binding to an FcR can promote the expansion and activation of CD8+ T cells. FcR-mediated signaling that can result from an Fc domain binding to an FcR can influence immune cell regulation of T cell responses. FcR-mediated signaling that can result from an Fc domain binding to an FcR can influence immune cell regulation of T cell responses. FcR-mediated signaling that can result from an Fc domain binding to an FcR can influence dendritic cell regulation of T cell responses. FcR-mediated signaling that can result from an Fc domain binding to an FcR can influence functional polarization of T cells (e.g., polarization can be toward a ¾1 cell response).
[0280] The profile of FcRs on a dendritic cell (DC) can impact the ability of the DC to respond upon stimulation. For example, most DC can express both CD32A and CD32B, which can have opposing effects on IgG-mediated maturation and function of DCs: binding of IgG to CD32A can mature and activate DCs in contrast with CD32B, which can mediate inhibition due to phosphorylation of immunoreceptor tyro sine-based inhibition motif (ITIM), after CD32B binding of IgG. Therefore, the activity of these two receptors can establish a threshold of DC activation. Furthermore, the difference in functional avidity of these receptors for IgG can shift their functional balance. Hence, altering the Fc domain binding to FcRs can also shift their functional balance, allowing for manipulation (either enhanced activity or enhanced inhibition) of the DC immune response.
[0281] A modification in the amino acid sequence of the Fc domain of an antibody construct can alter the recognition of an FcR for the Fc domain. However, such modifications can still allow for FcR-mediated signaling. A modification can be a substitution of an amino acid at a residue (e.g., wildtype) for a different amino acid at that residue. A modification can permit binding of an FcR to a site on the Fc domain or region that the FcR may not otherwise bind to. A modification can increase the binding affinity of an FcR to the Fc domain that the FcR may have reduced binding affinity for. A modification can decrease binding affinity of an FcR to a site on the Fc domain that the FcR may have increased binding affinity for. A modification can increase the subsequent FcR-mediated signaling after Fc binding to an FcR.
[0282] An antibody construct can comprise an Fc region with at least one amino acid change as compared to the sequence of the wild-type Fc region. An antibody construct can comprise an Fc domain with at least one amino acid change as compared to the sequence of the wild-type Fc domain. An amino acid change in an Fc region can allow the construct to bind to at least one Fc receptor with greater affinity compared to a wild-type Fc region. An amino acid change in an Fc domain can allow the antibody construct to bind to at least one Fc receptor with greater affinity compared to a wild-type Fc domain. An Fc region can comprise an amino acid sequence having at least one, two, three, four, five, six, seven, eight, nine or ten modifications but not more than 40, 35, 30, 25, 20, 15, or 10 modifications of the amino acid sequence relative to the natural or original amino acid sequence. An Fc domain can comprise an amino acid sequence having at least one, two, three, four, five, six, seven, eight, nine or ten modifications but not more than 40, 35, 30, 25, 20, 15, or 10 modifications of the amino acid sequence relative to the natural or original amino acid sequence. An Fc region can be an Fc region of an IgGl antibody. An Fc region can contain an Fc domain. An Fc region can be an Fc domain.
[0283] An antibody construct can be an antibody comprising a sequence of the IgGl isoform that has been modified from the wildtype IgGl sequence. A modification can comprise a substitution at more than one amino acid residue such as at 5 different amino acid residues including
L235V/F243L/R292P/Y300L/P396L (IgGlVLPLL). The numbering of amino acids residues is according to the EU index of Kabat. The 5 amino acid residues can be located in a portion of an antibody sequence which can encode an Fc region of the antibody and in particular, can be located in portions of the Fc region that can bind to Fc receptors (i.e., the Fc domain). A modification can comprise a substitution at more than one amino acid residue such as at 3 different amino acid residues including S298A/E333A/K334A (IgGl AAA), according to the EU index of Kabat. The 3 amino acid residues can be located in a portion of an antibody sequence which can encode an Fc region of the antibody and in particular, can be located in portions of the Fc region that can bind Fc receptors (i.e., the Fc domain).
[0284] In some embodiments, the Fc domain or region can comprise a sequence of an IgG isoform that has been modified from the wild-type IgG sequence. In some embodiments, the Fc domain or region can comprise a sequence of the IgGl isoform that has been modified from the wild-type IgGl sequence. In some embodiments, the modification comprises substitution of one or more amino acids that reduce binding affinity of an IgG Fc domain or region to all Fey receptors. A modification can be substitution of E233, L234 and L235, such as
E233P/L234V/L235A or E233P/L234V/L235A/AG236, according to the EU index of Kabat. A modification can be substitution of L235, F243, R292, Y300 and P396, such as
L235V/F243L/R292P/Y300L/P396L (IgGlVLPLL) according to the EU index of Kabat. A modification can be a substitution of P238, such as P238A, according to the EU index of Kabat. A modification can be a substitution of D265, such as D265A, according to the EU index of Kabat. A modification can be a substitution of N297, such as N297A, according to the EU index of Kabat. A modification can be a substitution of A327, such as A327Q, according to the EU index of Kabat. A modification can be a substitution of P329, such as P239A, according to the EU index of Kabat.
[0285] In some embodiments, an IgG Fc domain or region comprises at least one amino acid substitution that reduces its binding affinity to FcyRl, as compared to a wild-type or reference IgG Fc domain. A modification can comprise a substitution at F241, such as F241A, according to the EU index of Kabat. A modification can comprise a substitution at F243, such as F243A, according to the EU index of Kabat. A modification can comprise a substitution at V264, such as V264A, according to the EU index of Kabat. A modification can comprise a substitution at D265, such as D265A according to the EU index of Kabat.
[0286] In some embodiments, an IgG Fc domain or region comprises at least one amino acid substitution that increases its binding affinity to FcyRl, as compared to a wild-type or reference IgG Fc domain. A modification can comprise a substitution at A327 and P329, such as
A327Q/P329A, according to the EU index of Kabat.
[0287] In some embodiments, the modification comprises substitution of one or more amino acids that reduce binding affinity of an IgG Fc domain or region to FcyRII and FcyRIIIA receptors. A modification can be a substitution of D270, such as D270A, according to the EU index of Kabat. A modification can be a substitution of Q295, such as Q295A, according to the EU index of Kabat. A modification can be a substitution of A327, such as A237S, according to the EU index of Kabat. [0288] In some embodiments, the modification comprises substitution of one or more amino acids that increases binding affinity of an IgG Fc domain or region to FcyRII and FcyRIIIA receptors. A modification can be a substitution of T256, such as T256A, according to the EU index of Kabat. A modification can be a substitution of K290, such as K290A, according to the EU index of Kabat.
[0289] In some embodiments, the modification comprises substitution of one or more amino acids that increases binding affinity of an IgG Fc domain or region to FcyRII receptor. A modification can be a substitution of R255, such as R255A, according to the EU index of Kabat. A modification can be a substitution of E258, such as E258A, according to the EU index of Kabat. A modification can be a substitution of S267, such as S267A, according to the EU index of Kabat. A modification can be a substitution of E272, such as E272A, according to the EU index of Kabat. A modification can be a substitution of N276, such as N276A, according to the EU index of Kabat. A modification can be a substitution of D280, such as D280A, according to the EU index of Kabat. A modification can be a substitution of H285, such as H285A, according to the EU index of Kabat. A modification can be a substitution of N286, such as N286A, according to the EU index of Kabat. A modification can be a substitution of T307, such as T307A, according to the EU index of Kabat. A modification can be a substitution of L309, such as L309A, according to the EU index of Kabat. A modification can be a substitution of N315, such as N315A, according to the EU index of Kabat. A modification can be a substitution of K326, such as K326A, according to the EU index of Kabat. A modification can be a substitution of P331, such as P331A, according to the EU index of Kabat. A modification can be a substitution of S337, such as S337A, according to the EU index of Kabat. A modification can be a substitution of A378, such as A378A, according to the EU index of Kabat. A modification can be a substitution of E430, such as E430, according to the EU index of Kabat.
[0290] In some embodiments, the modification comprises substitution of one or more amino acids that increases binding affinity of an IgG Fc domain or region to FcyRII receptor and reduces the binding affinity to FcyRIIIA receptor. A modification can be a substitution of H268, such as H268A, according to the EU index of Kabat. A modification can be a substitution of R301, such as R301A, according to the EU index of Kabat. A modification can be a substitution of K322, such as K322A, according to the EU index of Kabat.
[0291] In some embodiments, the modification comprises substitution of one or more amino acids that decreases binding affinity of an IgG Fc domain or region to FcyRII receptor but does not affect the binding affinity to FcyRIIIA receptor. A modification can be a substitution of R292, such as R292A, according to the EU index of Kabat. A modification can be a substitution of K414, such as K414A, according to the EU index of Kabat. [0292] In some embodiments, the modification comprises substitution of one or more amino acids that decreases binding affinity of an IgG Fc domain or region to FcyRII receptor and increases the binding affinity to FcyRIIIA receptor. A modification can be a substitution of S298, such as S298A, according to the EU index of Kabat. A modification can be substitution of S239, 1332 and A330, such as S239D/I332E/A330L. A modification can be substitution of S239 and 1332, such as S239D/I332E.
[0293] In some embodiments, the modification comprises substitution of one or more amino acids that decreases binding affinity of an IgG Fc domain or region to FcyRIIIA receptor and does not affect the binding affinity to FcyRII receptor. A modification can be a substitution of S239, such as S239A, according to the EU index of Kabat. A modification can be a substitution of E269, such as E269A, according to the EU index of Kabat. A modification can be a substitution of E293, such as E293A, according to the EU index of Kabat. A modification can be a substitution of Y296, such as Y296F, according to the EU index of Kabat. A modification can be a substitution of V303, such as V303A, according to the EU index of Kabat. A modification can be a substitution of A327, such as A327G, according to the EU index of Kabat. A
modification can be a substitution of K338, such as K338A, according to the EU index of Kabat. A modification can be a substitution of D376, such as D376A, according to the EU index of Kabat.
[0294] In some embodiments, the modification comprises substitution of one or more amino acids that increases binding affinity of an IgG Fc domain or region to FcyRIIIA receptor and does not affect the binding affinity to FcyRII receptor. A modification can be a substitution of E333, such as E333A, according to the EU index of Kabat. A modification can be a substitution of K334, such as K334A, according to the EU index of Kabat. A modification can be a substitution of A339, such as A339T, according to the EU index of Kabat. A modification can be substitution of S239 and 1332, such as S239D/I332E.
[0295] In some embodiments, an IgG Fc domain or region comprises at least one amino acid substitution that reduces the binding affinity to FcRn, as compared to a wild-type or reference IgG Fc domain. A modification can comprise a substitution at H435, such as H435A according to the EU index of Kabat. A modification can comprise a substitution at 1253, such as 1253 A according to the EU index of Kabat. A modification can comprise a substitution at H310, such as H310A according to the EU index of Kabat. A modification can comprise substitutions at 1253, H310 and H435, such as I253A/H310A/H435A according to the EU index of Kabat.
[0296] A modification can comprise a substitution of one amino acid residue that increases the binding affinity of an IgG Fc domain for FcRn, relative to a wildtype or reference IgG Fc domain. A modification can comprise a substitution at V308, such as V308P according to the EU index of Kabat. A modification can comprise a substitution at M428, such as M428L according to the EU index of Kabat. A modification can comprise a substitution at N434, such as N434A according to the EU index of Kabat or N434H according to the EU index of Kabat. A modification can comprise substitutions at T250 and M428, such as T250Q and M428L according to the EU index of Kabat. A modification can comprise substitutions at M428 and N434, such as M428L and N434S, N434A or N434H according to the EU index of Kabat. A modification can comprise substitutions at M252, S254 and T256, such as M252Y/S254T/T256E according to the EU index of Kabat. A modification can be a substitution of one or more amino acids selected from P257L, P257N, P257I, V279E, V279Q, V279Y, A281S, E283F, V284E, L306Y, T307V, V308F, Q311V, D376V, and N434H. Other substitutions in an IgG Fc domain that affect its interaction with FcRn are disclosed in U.S. Patent No. 9,803,023 (the disclosure of which is incorporated by reference herein).
[0297] An antibody construct can be a monoclonal anti-CD40 human antibody comprising a sequence of the IgGl isoform that has been modified from the wildtype IgGl sequence. A modification can comprise a substitution at more than one amino acid residue such as at 5 different amino acid residues including L235V/F243L/R292P/Y300L/P396L (SBT-040- G1VLPLL). The numbering of amino acids residues is according to the EU index. The 5 amino acid residues can be located in a portion of an antibody sequence which can encode an Fc region of the antibody and in particular, can be located in portions of the Fc region that can bind to Fc receptors (i.e., the Fc domain). A modification can comprise a substitution at more than one amino acid residue such as at 3 different amino acid residues including S298A/E333A/K334A (SBT-040-G1AAA). The 3 amino acid residues can be located in a portion of an antibody sequence which can encode an Fc region of the antibody and in particular, can be located in portions of the Fc region that can bind Fc receptors (i.e., the Fc domain).
[0298] Binding of Fc receptors to an Fc region can be affected by amino acid substitutions. For example, SBT-040-VLPLL is an antibody with an amino acid sequence of a heavy chain of human anti-CD40 monoclonal antibody with modifications to a wild-type IgGl Fc domain (L235V/F243L/R292P/Y300L/P396L). Binding of some Fc receptors to the Fc region of SBT- 040-VLPLL can be enhanced compared to wild-type by as result of the
L235V/F243L/R292P/Y300L/P396L amino acid modifications. However, binding of other Fc receptors to the Fc region of SBT-040-VLPLL can be reduced compared to wild-type by the L235V/F243L/R292P/Y300L/P396L amino acid modifications. For example, the binding affinities of SBT-040-VLPLLto FcyRIIIA and to FcyRIIA can be enhanced compared to wild- type whereas the binding affinity of SBT-040-VLPLL to FcyRIIB can be reduced compared to wild-type. Binding of Fc receptors to an Fc region of are affected by amino acid substitutions. SBT-040-G1AAA antibody is an antibody with an amino acid sequence of a heavy chain of a human anti-CD40 monoclonal antibody with modifications to a wild-type IgGl Fc domain (S298A/E333A/K334A). Binding of Fc receptors to an Fc region of SBT-040-G1AAA can be enhanced compared to wild-type as a result of the S298A/E333A/K334A amino acid
modification. However, binding of some Fc receptors to the Fc region of SBT-040-G1AAA can be reduced compared to wild-type by S298A/E333A/K334A amino acid modification. Binding affinities of SBT-040-G1AAA to FcyRIIIA can be enhanced compared to wild-type whereas the binding affinity of SBT-040-G1AAA to FcyRIIB can be reduced compared to wildtype.
[0299] In some embodiments, the heavy chain of a human IgG2 antibody can be mutated at cysteines as positions 127, 232, or 233. In some embodiments, the light chain of a human IgG2 antibody can be mutated at a cysteine at position 214. The mutations in the heavy and light chains of the human IgG2 antibody can be from a cysteine residue to a serine residue.
[0300] While an antibody construct of the present disclosure can comprise a first binding domain and a second binding domain (or, in some cases, a third binding domain) with wild-type or modified amino acid sequences encoding the Fc region or Fc domain, the modifications of the Fc region or the Fc domain from the wild-type sequence may not significantly alter binding and/or affinity of the binding domains. For example, binding and/or affinity of an antibody
constructcomprising a first binding domain and a second binding domain (or, in some cases, a third binding domain) and having the Fc domain modifications of SBT-040-G1WT, SBT-040- G1VLPLL, or SBT-040-G1AAA may not be significantly altered by modification of an Fc region or Fc domain amino acid sequence compared to a wild-type sequence. Modifications of an Fc region or Fc domain from a wild-type sequence may not alter binding and/or affinity of a first binding domain that binds, for example, to CD40 or DEC-205. Additionally, the binding and/or affinity of the binding domains described herein, for example, a first binding domain, a second binding domain (or, in some cases, a third binding domain), and an Fc domain modification selected from SBT-040-G1WT, SBT-040-G1VLPLL, and SBT-040-G1AAA, may be comparable to the binding and/or affinity of wild-type antibodies.
[0301] In some embodiments, the binding profile of the Fc domain for Fey receptors can be retained in the antibody construct or conjugate, which can allow for delivery of the antibody construct or conjugate into immune cell types comprising the Fey receptors and can further immune activation by Fey receptor signaling. In some embodiments, APCs can be activated by an antibody construct or conjugate as described herein when the antibody construct or conjugate is bound to a tumor cell, undergoes Fey receptor mediated uptake, or undergoes Fey receptor mediated Antibody Dependent Cellular Phagocytosis (ADCP). In some embodiments, the antibody construct or conjugate can retain weak or no Fey receptor binding to allow for maximal immune activation or decreased toxicity of the immune- stimulatory compound due to the primary delivery of the antibody construct or conjugate into tumor cells by antibody antigen mediated endocytosis.
[0302] In some embodiments, the antibody construct or conjugate can comprise an IgGl Fc domain variant comprising N297A, N297G, K322A/L234A/L235A, or L234F/L235E/P331S, and lacks binding to an Fey receptor but can retain binding to FcRN in the presence of the immune-modulatory compound to allow for lower delivery of the conjugate into tumor cells or immune cells.
[0303] In some embodiments, an antibody construct or conjugate can comprise an Fc domain with higher affinity to one or more Fey receptors, which can result in greater immune activation than for an antibody construct or conjugate with an Fc domain that can bind to one or more Fey receptors with lower affinity.
TABLE 1. Tumor Antibody CDRs
Antibody Region SEQ ID Sequence
NO:
Pertuzumab HCDR1 13 GFTFTDYT
HCDR2 14 VNPNSGGS
HCDR3 15 ARNLGPSFYFDY
LCDR1 18 QDVSIG
LCDR2 19 SAS
LCDR3 20 QQYYIYPYT
Cetuximab HCDR1 26 GFSLTNYG
HCDR2 27 IWSGGNT
HCDR3 28 ARALTYYDYEFAY
LCDR1 31 QSIGTN
LCDR2 32 YAS
LCDR3 33 QQNNNWPTT
Panitumumab HCDR1 39 GGSVSSGDYY
HCDR2 40 IYYSGNT
HCDR3 41 VRDRVTGAFDI
LCDR1 44 QDISNY
LCDR2 45 DAS
LCDR3 46 QHFDHLPLA
Nimotuzumab HCDR1 52 GYTFTNYY
HCDR2 53 INPTSGGS
HCDR3 54 ARQGLWFDSDGRGFDF
LCDR1 57 QNIVHSNGNTY
LCDR2 58 KVS
LCDR3 59 FQYSHVPWT
Zalutumumab HCDR1 65 GFTFSTYG
HCDR2 66 IWDDGSYK
HCDR3 67 ARDGITMVRGVMKDYFDY
LCDR1 70 QDISSA
LCDR2 71 DAS
LCDR3 72 QQFNSYPLT
Onartuzumab HCDR1 78 GYTFTSYW
HCDR2 79 IDPSNSDT
HCDR3 80 ATYRSYVTPLDY
LCDR1 83 QSLLYTSSQKNY
LCDR2 84 WAS Antibody Region SEQ ID Sequence
NO:
LCDR3 85 QQYYAYPWT
Patritumab HCDR1 91 GGSFSGYY
HCDR2 92 INHSGST
HCDR3 93 ARDKWTWYFDL
LCDR1 96 QSVLYSSSNRNY
LCDR2 97 WAS
LCDR3 98 QQYYSTPRT
Clivatuzumab HCDR1 104 GYTFPSYV
HCDR2 105 INPYNDGT
HCDR3 106 ARGFGGSYGFAY
LCDR1 109 SSVSSSY
LCDR2 110 STS
LCDR3 111 HQWNRYPYT
Sofituzumab HCDR1 117 GYSITNDYA
HCDR2 118 ISYSGYT
HCDR3 119 ARWT SGLDY
LCDR1 122 DLIHNW
LCDR2 123 GAT
LCDR3 124 QQYWTTPFT
Edrecolomab HCDR1 130 GYAFTNYL
HCDR2 131 INPGSGGT
HCDR3 132 ARDGPWFAY
LCDR1 135 ENVVTY
LCDR2 136 GAS
LCDR3 137 GQGYSYPYT
Adecatumumab HCDR1 143 GFTFSSYG
HCDR2 144 ISYDGSNK
HCDR3 145 AKDMGWGSGWRPYYYYGMDV
LCDR1 148 QSISSY
LCDR2 149 WAS
LCDR3 150 QQSYDIPYT
Anetumab HCDR1 156 GYSFTSYW
HCDR2 157 IDPGDSRT
HCDR3 158 ARGQLYGGTYMDG
LCDR1 161 SSDIGGYNS
LCDR2 162 GVN
LCDR3 163 SSYDIESATPV huDS6 HCDR1 169 GYTFTSYN
HCDR2 170 IYPGNGAT
HCDR3 171 ARGDSVPFAY
LCDR1 174 SSVSF
LCDR2 175 STS
LCDR3 176 QQRSSFPLT
Lifastuzumab HCDR1 182 GFSFSDFA
HCDR2 183 IGRVAFHT
HCDR3 184 ARHRGFDVGHFDF
LCDR1 187 ETLV HSSGNTY
LCDR2 188 RVS
LCDR3 189 FQGSFNPLT
Sacituzumab HCDR1 195 GYTFTNYG
HCDR2 196 INTYTGEP
HCDR3 197 ARGGFGSSYWYFDV
LCDR1 200 QDVSIA
LCDR2 201 SAS
LCDR3 202 QQHYITPLT
PR1A3 HCDR1 208 GYTFTEFG
HCDR2 209 INTKTGEA
HCDR3 210 ARWDFYDYVEAMDY Antibody Region SEQ ID Sequence
NO:
LCDR1 213 QNVGTN
LCDR2 214 SAS
LCDR3 215 HQYYTYPLFT
Humanized PR1A3 HCDR1 805 GYTFTEFG
HCDR2 806 INTKTGEA
HCDR3 807 ARWDFAYYVEAMDY
LCDR1 808 AAVGTY
LCDR2 809 SAS
LCDR3 810 HQYYTYPLFT
Humanized Ab2-3 HCDR1 823 GFVFSSYD
HCDR2 824 YISSGGGIT
HCDR3 825 AAHYFGSSGPFAY
LCDR1 826 ENIFSY
LCDR2 827 NTR
LCDR3 828 QHHYGTPFT
IMAB362, HCDR1 221 GYTFTSYW
CLAUDIXIMAB HCDR2 222 IYPSDSYT
HCDR3 223 TRSWRGNSFDY
LCDR1 226 QSLLNSGNQKNY
LCDR2 227 WAS
LCDR3 228 QNDYSYPFT
AMG595 HCDR1 260 GFTFRNYG
HCDR2 261 IWYDGSDK
HCDR3 262 ARDGYDILTGNPRDFDY
LCDR1 265 QSLVHSDGNTY
LCDR2 266 RIS
LCDR3 267 MQSTHVPRT
ABT806 HCDR1 273 GYSISRDFA
HCDR2 274 ISYNGNT
HCDR3 275 VTASRGFPY
LCDR1 278 QDINSN
LCDR2 279 HGT
LCDR3 280 VQYAQFPWT
Sibrotuzumab HCDR1 286 RYTFTEYT
HCDR2 287 INPNNGIP
HCDR3 288 ARRRIAYGYDEGHAMDY
LCDR1 291 QSLLYSRNQKNY
LCDR2 292 WAS
LCDR3 293 QQYFSYPLT
DS-8895a variant 1 HCDR1 299 GYTFIDYS
HCDR2 300 INTYTGEP
HCDR3 301 ATYYRYERDFDY
LCDR1 304 QSIVHSSGITY
LCDR2 305 KVS
LCDR3 306 FQGSHVPYT
DS-8895a variant 2 HCDR1 312 GYTFIDYS
HCDR2 313 INTYTGEP
HCDR3 314 ATYYRYERDFDY
LCDR1 317 QSIVHSSGITY
LCDR2 318 KVS
LCDR3 319 FQGSHVPYT
MEDI-547 HCDR1 325 GFTFSHYM
HCDR2 326 IGPSGGPT
HCDR3 327 AGYDSGYDYVAVAGPAEYFQH
LCDR1 330 QSISTW
LCDR2 331 KAS
LCDR3 332 QQYNSYSRT
Narnatumab HCDR1 338 GFTFSSYL Antibody Region SEQ ID Sequence
NO:
HCDR2 339 IKQDGSEK
HCDR3 340 TRDGYSSGRHYGMDV
LCDR1 343 QSVSRY
LCDR2 344 DAS
LCDR3 345 QQRSNWPRT
RG7841 HCDR1 351 GFSLTGYS
HCDR2 352 IWGDGST
HCDR3 353 ARDYYFNYASWFAY
LCDR1 356 QGISNYL
LCDR2 357 YTS
LCDR3 358 QQYSELPWT
Farletuzumab HCDR1 364 GFTFSGYG
HCDR2 365 ISSGGSYT
HCDR3 366 ARHGDDPAWFAY
LCDR1 369 SSISSNN
LCDR2 370 GTS
LCDR3 371 QQWSSYPYMYT
Mirvetuximab HCDR1 377 GYTFTGYF
HCDR2 378 IHPYDGDT
HCDR3 379 TRYDGSRAMDY
LCDR1 382 QSVSFAGTSL
LCDR2 383 RAS
LCDR3 384 QQSREYPYT
J591 variant 1 HCDR1 390 GYTFTEYT
HCDR2 391 INPNNGGT
HCDR3 392 AAGWNFDY
LCDR1 395 QDVGTA
LCDR2 396 WAS
LCDR3 397 QQYNSYPLT
J591 variant 2 HCDR1 403 GYTFTEYT
HCDR2 404 INPNNGGT
HCDR3 405 AAGWNFDY
LCDR1 408 ENVVTY
LCDR2 409 GAS
LCDR3 410 GQGYSYPYT
Rovalpituzumab HCDR1 416 GYTFTNYG
HCDR2 417 INTYTGEP
HCDR3 418 ARIGDSSPSDY
LCDR1 421 QSVSND
LCDR2 422 YAS
LCDR3 423 QQDYTSPWT
PF-06647020 HCDR1 429 GYTFTDYA
HCDR2 430 ISTYNDYT
HCDR3 431 ARGNSYFYALDY
LCDR1 434 ESVDSYGKSF
LCDR2 435 RAS
LCDR3 436 QQSNEDPWT
Antibody to PTK7 HCDR1 442 GFTFSSYA
HCDR2 443 ISYDGSIK
HCDR3 444 ARTYYFDY
LCDR1 447 QSIGSS
LCDR2 448 YAS
LCDR3 449 HQSSSLPIT
Ladiratuzumab HCDR1 455 GLTIEDYY
HCDR2 456 IDPENGDT
HCDR3 457 AVHNAHYGTWFAY
LCDR1 460 QSLLHS SGNT Y
LCDR2 461 KIS Antibody Region SEQ ID Sequence
NO:
LCDR3 462 FQGSHVPYT
Cirmtuzumab HCDR1 468 GYAFTAYN
HCDR2 469 FDPYDGGS
HCDR3 470 ARGWYYFDY
LCDR1 473 KSISKY
LCDR2 474 SGS
LCDR3 475 QQHDESPYT
Antibody to HCDR1 481 GFRFRSHG
MAGE-A3 HCDR2 482 SYDGNNK
HCDR3 483 ASPYTSDWQYFQY
LCDR1 486 QNISTT
LCDR2 487 DTS
LCDR3 488 QQSNSWPLT
Antibody to NY- HCDR1 494 GFSFIDYG
ESO-1 HCDR2 495 MNWSGDKK
HCDR3 496 ARGEYSNRFDP
LCDR1 499 QSLVFTDGNTY
LCDR2 500 KVS
LCDR3 501 MQGTHWPPI
Trastuzumab HCDR1 673 GFNIKDTY
HCDR2 674 IYPTNGYT
HCDR3 675 SRWGGDGFYAMDY
LCDR1 676 QDVNTA
LCDR2 677 SAS
LCDR3 678 QQHYTTPPT
Ipilumumab HCDR1 850 GFTFSSYT
HCDR2 851 ISYDGNNK
HCDR3 852 ARTGWLGPFDY
LCDR1 853 QSVGSSY
LCDR2 854 SSY
LCDR3 855 QQYGSSPWT
Vonlerolizumab HCDR1 856 GYTFTDSY
HCDR2 857 MYPDNGDS
HCDR3 858 VLAPRWYFSV
LCDR1 859 QDISNY
LCDR2 860 YTS
LCDR3 861 QQGHTLPPT
Anti-CD27 HCDR1 862 GFTFSSYD
Antibody HCDR2 863 IWYDGSNK
HCDR3 864 ARGSGNWGFFDY
LCDR1 865 QGISRW
LCDR2 866 AAS
LCDR3 867 QQYNTYPRT
Atezolizumab HCDR1 868 GFTFSDSW
HCDR2 869 ISPYGGST
HCDR3 870 ARRHWPGGFDY
LCDR1 871 QDVSTA
LCDR2 872 SAS
LCDR3 873 QQYLYHPAT
Durvalumab HCDR1 874 GFTFSRYW
HCDR2 875 IKQDGSEK
HCDR3 876 AREGGWFGELAFDY
LCDR1 877 QRVSSSY
LCDR2 878 DAS
LCDR3 879 QQYGSLPWT
MDX-1106 HCDR1 880 GDTFSTYA
HCDR2 881 IIPIFGKA
HCDR3 882 ARKFHFVSGSPFGMDV Antibody Region SEQ ID Sequence
NO:
LCDR1 883 QSVSSY
LCDR2 884 DAS
LCDR3 885 QQRSNWPT
TABLE 2. Tumor Antibody VH sequences and VL sequences
Antibody Region SEQ Sequence
ID
NO:
Pertuzumab VH 12 EVQLVESGGGLVQPGGSLRLSCAASGFTFTDYTMDWVRQAPGKGLEWVA
DVNPNSGGSIYNQRFKGRFTLSVDRSKNTLYLQMNSLRAEDTAVYYCARN LGPSFYFDYWGQGTLVTVSS
VL 17 DIQMTQSPSSLSASVGDRVTITCKASQDVSIGVAWYQQKPGKAPKLLIYSAS
YRYTGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYYIYPYTFGQGTKV EIK
Cetuximab VH 25 QVQLKQSGPGLVQPSQSLSITCTVSGFSLTNYGVHWVRQSPGKGLEWLGVI
WSGGNTDYNTPFTSRLSINKDNSKSQVFFKMNSLQSNDTAIYYCARALTYY DYEFAYWGQGTLVTVSA
VL 30 DILLTQSPVILSVSPGERVSFSCRASQSIGTNIHWYQQRTNGSPRLLIKYASES
ISGIPSRFSGSGSGTDFTLSINSVESEDIADYYCQQNNNWPTTFGAGTKLELK
Panitumumab VH 38 QVQLQESGPGLVKPSETLSLTCTVSGGSVSSGDYYWTWIRQSPGKGLEWIG fflYYSGNTNYNPSLKSRLTISIDTSKTQFSLKLSSVTAADTAIYYCVRDRVTG AFDIWGQGTMVTVSS
VL 43 DIQMTQSPSSLSASVGDRVTITCQASQDISNYLNWYQQKPGKAPKLLIYDA
SNLETGVPSRFSGSGSGTDFTFTISSLQPEDIATYFCQHFDHLPLAFGGGTKV EIK
Nimotuzumab VH 51 QVQLQQSGAEVKKPGSSVKVSCKASGYTFTNYYIYWVRQAPGQGLEWIG
GINPTSGGSNFNEKFKTRVTITVDESTNTAYMELSSLRSEDTAFYFCARQGL
WFDSDGRGFDFWGQGSTVTVSS
VL 56 DIQMTQSPSSLSASVGDRVTITCRSSQNIVHSNGNTYLDWYQQTPGKAPKL
LIYKVSNRFSGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCFQYSHVPWTFG QGTKLEIK
Zalutumumab VH 64 QVQLVESGGGVVQPGRSLRLSCAASGFTFSTYGMHWVRQAPGKGLEWVA
VIWDDGSYKYYGDSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAR
DGITMVRGVMKDYFDYWGQGTLVTVSS
VL 69 AIQLTQSPSSLSASVGDRVTITCRASQDISSALVWYQQKPGKAPKLLIYDAS
SLESGVPSRFSGSESGTDFTLTISSLQPEDFATYYCQQFNSYPLTFGGGTKVE IK
Onartuzumab VH 77 EVQLVESGGGLVQPGGSLRLSCAASGYTFTSYWLHWVRQAPGKGLEWVG
MIDPSNSDTRFNPNFKDRFTISADTSKNTAYLQMNSLRAEDTAVYYCATYR SYVTPLDYWGQGTLVTVSS
VL 82 DIQMTQSPSSLSASVGDRVTITCKSSQSLLYTSSQKNYLAWYQQKPGKAPK
LLIYWASTRESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYYAYPWT FGQGTKVEIK
Patritumab VH 90 QVQLQQWGAGLLKPSETLSLTCAVYGGSFSGYYWSWIRQPPGKGLEWIGE
INHSGSTNYNPSLKSRVTISVETSKNQFSLKLSSVTAADTAVYYCARDKWT WYFDLWGRGTLVTVSS Antibody Region SEQ Sequence
ID
NO:
vL 95 DIEMTQSPDSLAVSLGERATINCRSSQSVLYSSSNRNYLAWYQQNPGQPPK
LLIYWASTRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQYYSTPRT FGQGTKVEIK
Clivatuzumab VH 103 QVQLQQSGAEVKKFGASVKVSCEASGYTFPSYVLHWVKQAPGQGLEWIG
YINPYNDGTQTNKKFKGKATLTRDTSINTAYMELSRLRSDDTAVYYCARG FGGSYGFAYNGQGTLVTVSS
VL 108 DIQLTQSPSSLSASVGDRVTMTCSASSSVSSSYLYWYQQKPGKAPKLWIYS
TSNLASGVPARFSGSGSGTDFTLTISSLQPEDSASYFCHQWNRYPYTFGGGT
RLEIK
Sofituzumab VH 116 EVQLVESGGGLVQPGGSLRLSCAASGYSITNDYAWNWVRQAPGKGLEWV
GYISYSGYTTYNPSLKSRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARWT SGLDYWGQGTLVTVSS
VL 121 DIQMTQSPSSLSASVGDRVTITCKASDLIHNWLAWYQQKPGKAPKLLIYGA
TSLETGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYWTTPFTFGQGTK VEIK
Edrecolomab VH 129 QVQLQQSGAELVRPGTSVKVSCKASGYAFTNYLIEWVKQRPGQGLEWIGV
INPGSGGTNYNEKFKGKATLTADKSSSTAYMQLSSLTSDDSAVYFCARDGP WFAYWGQGTLVTVSA
VL 134 NIVMTQSPKSMSMSVGERVTLTCKASENVVTYVSWYQQKPEQSPKLLIYG
ASNRYTGVPDRFTGSGSATDFTLTISSVQAEDLADYHCGQGYSYPYTFGGG
TKLEIK
Adecatumum VH 142 EVQLLESGGGVVQPGRSLRLSCAASGFTFSSYGMHWVRQAPGKGLEWVA ab VISYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKD
MGWGSGWRPYYYYGMDVWGQGTTVTVSS
VL 147 ELQMTQSPSSLSASVGDRVTITCRTSQSISSYLNWYQQKPGQPPKLLIYWAS
TRESGVPDRFSGSGSGTDFTLTISSLQPEDSATYYCQQSYDIPYTFGQGTKLE IK
Anetumab VH 155 QVELVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWVRQAPGKGLEWMGII
DPGDSRTRYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCARGQLY GGTYMDGWGQGTLVTVSS
VL 160 DIALTQPASVSGSPGQSITISCTGTSSDIGGYNSVSWYQQHPGKAPKLMIYG
VNNRPSGVSNRFSGSKSGNTASLTISGLQAEDEADYYCSSYDIESATPVFGG
GTKLTVL
huDS6 VH 168 QAQLVQSGAEVVKPGASVKMSCKASGYTFTSYNMHWVKQTPGQGLEWIG
YIYPGNGATNYNQKFQGKATLTADPSSSTAYMQISSLTSEDSAVYFCARGD SVPFAYWGQGTLVTVSA
VL 173 EIVLTQSPATMSASPGERVTITCSAHSSVSFMHWFQQKPGTSPKLWIYSTSS
LASGVPARFGGSGSGTSYSLTISSMEAEDAATYYCQQRSSFPLTFGAGTKLE
LK
Lifastuzumab VH 181 EVQLVESGGGLVQPGGSLRLSCAASGFSFSDFAMSWVRQAPGKGLEWVAT
IGRVAFHTYYPDSMKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARHR
GFDVGHFDFWGQGTLVTVSS
VL 186 DIQMTQSPSSLSASVGDRVTITCRSSETLVHSSGNTYLEWYQQKPGKAPKL Antibody Region SEQ Sequence
ID
NO:
LIYRVSNRFSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCFQGSFNPLTFG QGTKVEIK
Sacituzumab VH 194 QVQLQQSGSELKKPGASVKVSCKASGYTFTNYGMNWVKQAPGQGLKWM
GWINTYTGEPTYTDDFKGRFAFSLDTSVSTAYLQISSLKADDTAVYFCARG GFGSSYWYFDVWGQGSLVTVSS
VL 199 DIQLTQSPSSLSASVGDRVSITCKASQDVSIAVAWYQQKPGKAPKLLIYSAS
YRYTGVPDRFSGSGSGTDFTLTISSLQPEDFAVYYCQQHYITPLTFGAGTKV EIK
PR1A3 VH 207 QVKLQQSGPELKKPGETVKISCKASGYTFTEFGMNWVKQAPGKGLKWMG
WINTKTGEATYVEEFKGRFAFSLETSATTAYLQINNLKNEDTAKYFCARW
DFYDYVEAMDYWGQGTTVTVSS
VL 212 DIVMTQSQRFMSTSVGDRVSVTCKASQNVGTNVAWYQQKPGQSPKALIYS
ASYRYSGVPDRFTGSGSGTDFTLTISNVQSEDLAEYFCHQYYTYPLFTFGSG TKLEMK
Humanized VH 811 QVQLVQSGAEVKKPGASVKVSCKASGYTFTEFGMNWVRQAPGQGLEWM PR1A3 GWINTKTGEATYVEEFKGRVTFTTDTSTSTAYMELRSLRSDDTAVYYCAR
WDFAYYVEAMDYWGQGTTVTVSS
VL 812 DIQMTQSPSSLSASVGDRVTITCKASAAVGTYVAWYQQKPGKAPKLLIYSA
SYRKRGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCHQYYTYPLFTFGQGT
KLEIK
Humanized VH 829 EVQLQESGPGLVKPGGSLSLSCAASGFVFSSYDMSWVRQTPERGLEWVAYI Ab2-3 SSGGGITYAPSTVKGRFTVSRDNAKNTLYLQMNSLTSEDTAVYYCAAHYF
GSSGPFAYWGQGTLVTVSS
VL 830 DIQMTQSPASLSASVGDRVTITCRASENIFSYLAWYQQKPGKSPKLLVYNT
RTLAEGVPSRFSGSGSGTDFSLTISSLQPEDFATYYCQHHYGTPFTFGSGTKL
EIK
IMAB362, VH 220 QVQLQQPGAELVRPGASVKLSCKASGYTFTSYWINWVKQRPGQGLEWIGN
CLAUDIXIM IYPSDSYTNYNQKFKDKATLTVDKSSSTAYMQLSSPTSEDSAVYYCTRSWR
AB GNSFDYWGQGTTLTVSS
VL 225 DIVMTQSPSSLTVTAGEKVTMSCKSSQSLLNSGNQKNYLTWYQQKPGQPP
KLLIYWASTRESGVPDRFTGSGSGTDFTLTISSVQAEDLAVYYCQNDYSYPF
TFGSGTKLEIK
AMG595 VH 259 QVQLVESGGGVVQSGRSLRLSCAASGFTFRNYGMHWVRQAPGKGLEWVA
VIWYDGSDKYYADSVRGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARD GYDILTGNPRDFDYWGQGTLVTVSS
VL 264 DTVMTQTPLSSHVTLGQPASISCRSSQSLVHSDGNTYLSWLQQRPGQPPRL
LIYRISRRFSGVPDRFSGSGAGTDFTLEISRVEAEDVGVYYCMQSTHVPRTF GQGTKVEIK
ABT806 VH 272 EVQLQESGPGLVKPSQTLSLTCTVSGYSISRDFAWNWIRQPPGKGLEWMG
YISYNGNTRYQPSLKSRITISRDTSKNQFFLKLNSVTAADTATYYCVTASRG
FPYWGQGTLVTVSS
VL 277 DIQMTQSPSSMSVSVGDRVTITCHSSQDINSNIGWLQQKPGKSFKGLIYHGT
NLDDGVPSRFSGSGSGTDYTLTISSLQPEDFATYYCVQYAQFPWTFGGGTK LEIK
Sibrotuzumab VH 285 QVQLVQSGAEVKKPGASVKVSCKTSRYTFTEYTIHWVRQAPGQRLEWIGG
INPNNGIPNYNQKFKGRVTITVDTSASTAYMELSSLRSEDTAVYYCARRRIA YGYDEGHAMDYWGQGTLVTVSS
VL 290 DIVMTQSPDSLAVSLGERATINCKSSQSLLYSRNQKNYLAWYQQKPGQPPK
LLIFWASTRESGVPDRFSGSGFGTDFTLTISSLQAEDVAVYYCQQYFSYPLT Antibody Region SEQ Sequence
ID
NO:
FGQGTKVEIK
DS-8895a VH 298 QVQLVQSGAEVKKPGASVKVSCKASGYTFIDYSMHWVRQAPGQGLEWM variant 1 GWINTYTGEPTYSDDFKGRVTITADTSTSTAYLELSSLRSEDTAVYYCATY
YRYERDFDYWGQGTLVTVSS
VL 303 DIVMTQSPLSLPVTPGEPASISCRSSQSIVHSSGITYLEWYLQKPGQSPQLLIY
KVSNRFSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCFQGSHVPYTFGQ GTKVEIK
DS-8895a VH 311 QIQLVQSGAEVKKPGASVKVSCKASGYTFIDYSMHWVRQAPGQGLKWMG variant 2 WINTYTGEPTYSDDFKGRFAFSLDTSTSTAYLELSSLRSEDTAVYYCATYY
RYERDFDYWGQGTLVTVSS
VL 316 DVLMTQSPLSLPVTPGEPASISCRSSQSIVHSSGITYLEWYLQKPGQSPQLLI
YKVSNRFSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCFQGSHVPYTFG QGTKVEIK
MEDI-547 VH 324 EVQLLESGGGLVQPGGSLRLSCAASGFTFSHYMMAWVRQAPGKGLEWVS
RIGPSGGPTHYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAGY DSGYDYVAVAGPAEYFQHWGQGTLVTVSS
VL 329 DIQMTQSPSSLSASVGDRVTITCRASQSISTWLAWYQQKPGKAPKLLIYKAS
NLHTGVPSRFSGSGSGTEFSLTISGLQPDDFATYYCQQYNSYSRTFGQGTKV EIK
Narnatumab VH 337 EVQLVESGGGLVQPGGSLRLSCAASGFTFSSYLMTWVRQAPGKGLEWVA
NIKQDGSEKYYVDSVKGRFTISRDNAKNSLNLQMNSLRAEDTAVYYCTRD
GYSSGRHYGMDVWGQGTTVIVSS
VL 342 EIVLTQSPATLSLSPGERATLSCRASQSVSRYLAWYQQKPGQAPRLLIYDAS
NRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRSNWPRTFGQGTKV EIK
RG7841 VH 350 EVQLVESGPALVKPTQTLTLTCTVSGFSLTGYSVNWIRQPPGKALEWLGMI
WGDGSTDYNSALKSRLTISKDTSKNQVVLTMTNMDPVDTATYYCARDYY FNYASWFAYWGQGTLVTVSS
VL 355 DIQMTQSPSSLSASVGDRVTITCSASQGISNYLNWYQQKPGKTVKLLIYYTS
NLHSGVPSRFSGSGSGTDYTLTISSLQPEDFATYYCQQYSELPWTFGQGTKV EIK
Farletuzumab VH 363 EVQLVESGGGVVQPGRSLRLSCSASGFTFSGYGLSWVRQAPGKGLEWVA
MISSGGSYTYYADSVKGRFAISRDNAKNTLFLQMDSLRPEDTGVYFCARH GDDPAWFAYWGQGTPVTVSS
VL 368 DIQLTQSPSSLSASVGDRVTITCSVSSSISSNNLHWYQQKPGKAPKPWIYGTS
NLASGVPSRFSGSGSGTDYTFTISSLQPEDIATYYCQQWSSYPYMYTFGQGT KVEIK
Mirvetuximab VH 376 QVQLVQSGAEVVKPGASVKISCKASGYTFTGYFMNWVKQSPGQSLEWIGR
IHPYDGDTFYNQKFQGKATLTVDKSSNTAHMELLSLTSEDFAVYYCTRYD
GSRAMDYWGQGTTVTVSS
VL 381 DIVLTQSPLSLAVSLGQPAIISCKASQSVSFAGTSLMHWYHQKPGQQPRLLI
YRASNLEAGVPDRFSGSGSKTDFTLTISPVEAEDAATYYCQQSREYPYTFG GGTKLEIK Antibody Region SEQ Sequence
ID
NO:
J591 variant 1 VH 389 EVQLQQSGPELKKPGTSVRISCKTSGYTFTEYTIHWVKQSHGKSLEWIGNIN
PNNGGTTYNQKFEDKATLTVDKSSSTAYMELRSLTSEDSAVYYCAAGWNF DYWGQGTTLTVSS
VL 394 DIVMTQSHKFMSTSVGDRVSIICKASQDVGTAVDWYQQKPGQSPKLLIYW
ASTRHTGVPDRFTGSGSGTDFTLTITNVQSEDLADYFCQQYNSYPLTFGAG TMLDLK
J591 variant 2 VH 402 EVQLQQSGPELVKPGTSVRISCKTSGYTFTEYTIHWVKQSHGKSLEWIGNIN
PNNGGTTYNQKFEDKATLTVDKSSSTAYMELRSLTSEDSAVYYCAAGWNF DYWGQGTTLTVSS
VL 407 NIVMTQSPKSMSMSVGERVTLTCKASENVVTYVSWYQQKPEQSPKLLIYG
ASNRYTGVPDRFTGSGSATDFTLTISSVQAEDLADYHCGQGYSYPYTFGGG
TKLEIK
Rovalpituzum VH 415 QVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGMNWVRQAPGQGLEWM ab GWINTYTGEPTYADDFKGRVTMTTDTSTSTAYMELRSLRSDDTAVYYCAR
IGDSSPSDYWGQGTLVTVSS
VL 420 EIVMTQSPATLSVSPGERATLSCKASQSVSNDVVWYQQKPGQAPRLLIYYA
SNRYTGIPARFSGSGSGTEFTLTISSLQSEDFAVYYCQQDYTSPWTFGQGTK LEIK
PF-06647020 VH 428 QVQLVQSGPEVKKPGASVKVSCKASGYTFTDYAVHWVRQAPGKRLEWIG
VISTYNDYTYNNQDFKGRVTMTRDTSASTAYMELSRLRSEDTAVYYCARG NSYFYALDYWGQGTSVTVSS
VL 433 EIVLTQSPATLSLSPGERATLSCRASESVDSYGKSFMHWYQQKPGQAPRLLI
YRASNLESGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQSNEDPWTFGG
GTKLEIK
Antibody to VH 441 QVQLVESGGGVVQPGRSLRLSCAASGFTFSSYAFHWVRQAPGKGLEWVA PTK7 VISYDGSIKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARTY
YFDYWGQGTLVTVSS
VL 446 EIVLTQSPDFQSVTPKEKVTITCRASQSIGSSLHWYQQKPDQSPKLLIKYASQ
SFSGVPSRFSGSGSGTDFTLTINSLEAEDAAAYYCHQSSSLPITFGQGTRLEI
K
Ladiratuxuma VH 454 QVQLVQSGAEVKKPGASVKVSCKASGLTIEDYYMHWVRQAPGQGLEWM b GWIDPENGDTEYGPKFQGRVTMTRDTSINTAYMELSRLRSDDTAVYYCAV
HNAHYGTWFAYWGQGTLVTVSS
VL 459 DVVMTQSPLSLPVTLGQPASISCRSSQSLLHSSGNTYLEYFQQRPGQSPRPLI
YKISTRFSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCFQGSHVPYTFGG GTKVEIK
Cirmtuzumab VH 467 QVQLQESGPGLVKPSQTLSLTCTVSGYAFTAYNIHWVRQAPGQGLEWMGS
FDPYDGGSSYNQKFKDRLTISKDTSKNQVVLTMTNMDPVDTATYYCARG
WYYFDYWGHGTLVTVSS
VL 472 DIVMTQTPLSLPVTPGEPASISCRASKSISKYLAWYQQKPGQAPRLLIYSGST
LQSGIPPRFSGSGYGTDFTLTINNIESEDAAYYFCQQHDESPYTFGEGTKVEI K
Antibody to VH 480 QVQLVESGGGVVQPGRSLRLSCTASGFRFRSHGMHWVRQAPGKGLEWVA MAGE-A3 VISYDGNNKLYADSVKGRITISRDNSKNTLFLQMNNVRAEDTAVYYCASP
YTSDWQYFQYWGQGTLVIVSS Antibody Region SEQ Sequence
ID
NO:
vL 485 EIVMTQSPATLSVSPGERATFSCRASQNISTTLAWYQQKPGQAPRLLIYDTS
TRATGIPARFSGSGSGTEFTLTISSLQSEDLAVYYCQQSNSWPLTFGGGTKV EIK
Antibody to VH 493 QVQLVQSGGGVVRPGGSLRLSCAASGFSFIDYGMSWVRQVPGKGLEWVA NY-ESO-1 GMNWSGDKKGHAESVKGRFIISRDNAKNTLYLEMSSLRVEDTALYFCARG
EYSNRFDPRGRGTLVTVSS
VL 498 DIVMTQTPLSLPVTLGQPASLSCRSSQSLVFTDGNTYLNWFQQRPGQSPRRL
IYKVSSRDPGVPDRFSGTGSGTDFTLEISRVEAEDIGVYYCMQGTHWPPIFG
QGTKVEIK
Trastuzumab VH 679 EVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVAR
IYPTNGYTRYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRWG GDGFYAMDYWGQGTLVTVSS
VL 680 DIQMTQSPSSLSASVGDRVTITCRASQDVNTAVAWYQQKPGKAPKLLIYSA
SFLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQHYTTPPTFGQGTKV EIK
Vonlerolizum VH 886 EVQLVQSGAEVKKPGASVKVSCKASGYTFTDSYMSWVRQAPGQGLEWIG ab DMYPDNGDSSYNQKFRERVTITRDTSTSTAYLELSSLRSEDTAVYYCVLAP
RWYFSVWGQGTLVTVSS
VL 887 DIQMTQSPSSLSASVGDRVTITCRASQDISNYLNWYQQKPGKAPKLLIYYTS
RLRSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQGHTLPPTFGQGTKV
EIK
Varlilumab VH 888 QVQLVESGGGVVQPGRSLRLSCAASGFTFSSYDMHWVRQAPGKGLEWVA
VIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAR
GSGNWGFFDYWGQGTLVTVSS
VL 889 DIQMTQSPSSLSASVGDRVTITCRASQGISRWLAWYQQKPEKAPKSLIYAAS
SLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYNTYPRTFGQGTKV
EIK
Atezolizumab VH 890 EVQLVESGGGLVQPGGSLRLSCAASGFTFSDSWIHWVRQAPGKGLEWVA
WISPYGGSTYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARR
HWPGGFDYWGQGTLVTVSS
VL 891 DIQMTQSPSSLSASVGDRVTITCRASQDVSTAVAWYQQKPGKAPKLLIYSA
SFLYSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYLYHPATFGQGTK VEIK
Durvalumab VH 892 EVQLVESGGGLVQPGGSLRLSCAASGFTFSRYWMSWVRQAPGKGLEWVA
NIKQDGSEKYYVDSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCARE GGWFGELAFDYWGQGTLVTVSS
VL 893 EIVLTQSPGTLSLSPGERATLSCRASQRVSSSYLAWYQQKPGQAPRLLIYDA
SSRATGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQYGSLPWTFGQGTK VEIK
MDX-1106 VH 894 QVQLVQSGAEVKKPGSSVKVSCKTSGDTFSTYAISWVRQAPGQGLEWMG
GIIPIFGKAHYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYFCARKFH FVSGSPFGMDVWGQGTTVTVSS
VL 895 EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLIYDAS
NRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRSNWPTFGQGTKVE IK
Ipilumumab VH 896 QVQLVESGGGVVQPGRSLRLSCAASGFTFSSYTMHWVRQAPGKGLEWVT
FISYDGNNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAIYYCARTG
WLGPFDYWGQGTLVTVSS
VL 897 EIVLTQSPGTLSLSPGERATLSCRASQSVGSSYLAWYQQKPGQAPRLLIYGA
FSRATGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQYGSSPWTFGQGTK VEIK
MDX-1105 VH 901 QVQLVQSGAEVKKPGSSVKVSCKTSGDTFSTYAISWVRQAPGQGLEWMG
GIIPIFGKAHYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYFCARKFH Antibody Region SEQ Sequence
ID
NO:
FVSGSPFGMDVWGQGTTVTVSS
VL 902 EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLIYDAS
NRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRSNWPTFGQGTKVE IK
TABLE 3. Tumor Antibody Heavy Chain and Light Chain sequences
Antibody Region SEQ Sequence
ID
NO:
Pertuzumab Heavy 11 EVQLVESGGGLVQPGGSLRLSCAASGFTFTDYTMDWVRQAPGKGLEWVA
Chain DVNPNSGGSIYNQRFKGRFTLSVDRSKNTLYLQMNSLRAEDTAVYYCARN
LGPSFYFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKD
YFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYIC
NVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTL
MISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYR
VVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLP
PSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGS
FFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG
Light 16 DIQMTQSPSSLSASVGDRVTITCKASQDVSIGVAWYQQKPGKAPKLLIYSAS Chain YRYTGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYYIYPYTFGQGTKV
EIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQ
SGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVT
KSFNRGEC
Cetuximab Heavy 24 QVQLKQSGPGLVQPSQSLSITCTVSGFSLTNYGVHWVRQSPGKGLEWLGVI
Chain WSGGNTDYNTPFTSRLSINKDNSKSQVFFKMNSLQSNDTAIYYCARALTYY
DYEFAYWGQGTLVTVSAASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFP
EPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVN
HKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISR
TPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVS
VLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSR
DELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFL
YSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK
Light 29 DILLTQSPVILSVSPGERVSFSCRASQSIGTNIHWYQQRTNGSPRLLIKYASES Chain ISGIPSRFSGSGSGTDFTLSINSVESEDIADYYCQQNNNWPTTFGAGTKLELK
RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGN
SQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSF
NRGEC
Panitumumab Heavy 37 QVQLQESGPGLVKPSETLSLTCTVSGGSVSSGDYYWTWIRQSPGKGLEWIG
Chain HIYYSGNTNYNPSLKSRLTISIDTSKTQFSLKLSSVTAADTAIYYCVRDRVTG
AFDIWGQGTMVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPV
TVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSNFGTQTYTCNVDHK
PSNTKVDKTVERKCCVECPPCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTC
VVVDVSHEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVH
QDWLNGKEYKCKVSNKGLPAPIEKTISKTKGQPREPQVYTLPPSREEMTKN
QVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTV
DKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK
Light 42 DIQMTQSPSSLSASVGDRVTITCQASQDISNYLNWYQQKPGKAPKLLIYDA Chain SNLETGVPSRFSGSGSGTDFTFTISSLQPEDIATYFCQHFDHLPLAFGGGTKV
EIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQ SGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVT KSFNRGEC Antibody Region SEQ Sequence
ID
NO:
Nimotuzumab Heavy 50 QVQLQQSGAEVKKPGSSVKVSCKASGYTFTNYYIYWVRQAPGQGLEWIG
Chain GINPTSGGSNFNEKFKTRVTITVDESTNTAYMELSSLRSEDTAFYFCARQGL
WFDSDGRGFDFWGQGSTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLV
KDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTY
ICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDT
LMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTY
RVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYT
LPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSD
GSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK
Light 55 DIQMTQSPSSLSASVGDRVTITCRSSQNIVHSNGNTYLDWYQQTPGKAPKL Chain LIYKVSNRFSGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCFQYSHVPWTFG
QGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKV DNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQG LSSPVTKSFNRGEC
Zalutumumab Heavy 63 QVQLVESGGGVVQPGRSLRLSCAASGFTFSTYGMHWVRQAPGKGLEWVA
Chain VIWDDGSYKYYGDSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAR
DGITMVRGVMKDYFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTA
ALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSS
SLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLF
PPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPRE
EQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQP
REPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTT
PPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSP
GK
Light 68 AIQLTQSPSSLSASVGDRVTITCRASQDISSALVWYQQKPGKAPKLLIYDAS Chain SLESGVPSRFSGSESGTDFTLTISSLQPEDFATYYCQQFNSYPLTFGGGTKVE
IKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQS GNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTK SFNRGEC
Onartuzumab Heavy 76 EVQLVESGGGLVQPGGSLRLSCAASGYTFTSYWLHWVRQAPGKGLEWVG
Chain MIDPSNSDTRFNPNFKDRFTISADTSKNTAYLQMNSLRAEDTAVYYCATYR
SYVTPLDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDY
FPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICN
VNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMI
SRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRV
VSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPP
SREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSF
FLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK
Light 81 DIQMTQSPSSLSASVGDRVTITCKSSQSLLYTSSQKNYLAWYQQKPGKAPK Chain LLIYWASTRESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYYAYPWT
FGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQW KVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTH QGLSSPVTKSFNRGEC
Patritumab Heavy 89 QVQLQQWGAGLLKPSETLSLTCAVYGGSFSGYYWSWIRQPPGKGLEWIGE
Chain INHSGSTNYNPSLKSRVTISVETSKNQFSLKLSSVTAADTAVYYCARDKWT
WYFDLWGRGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPE
PVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNH
KPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRT
PEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSV
LTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRE
EMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLY
SKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK
Light 94 DIEMTQSPDSLAVSLGERATINCRSSQSVLYSSSNRNYLAWYQQNPGQPPK Chain LLIYWASTRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQYYSTPRT
FGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQW
KVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTH
QGLSSPVTKSFNRGEC
Clivatuzumab Heavy 102 QVQLQQSGAEVKKPGASVKVSCEASGYTFPSYVLHWVKQAPGQGLEWIG Antibody Region SEQ Sequence
ID
NO:
Chain YINPYNDGTQYNEKFKGKATLTRDTSINTAYMELSRLRSDDTAVYYCARG
FGGSYGFAYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKD
YFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYIC
NVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTL
MISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYR
VVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLP
PSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGS
FFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK
Light 107 DIQLTQSPSSLSASVGDRVTMTCSASSSVSSSYLYWYQQKPGKAPKLWIYS Chain TSNLASGVPARFSGSGSGTDFTLTISSLQPEDSASYFCHQWNRYPYTFGGGT
RLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNA
LQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSP
VTKSFNRGEC
Sofituzumab Heavy 115 EVQLVESGGGLVQPGGSLRLSCAASGYSITNDYAWNWVRQAPGKGLEWV
Chain GYISYSGYTTYNPSLKSRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARWT
SGLDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPE
PVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNH
KPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRT
PEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSV
LTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRE
EMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLY
SKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK
Light 120 DIQMTQSPSSLSASVGDRVTITCKASDLIHNWLAWYQQKPGKAPKLLIYGA Chain TSLETGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYWTTPFTFGQGTK
VEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNAL QSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPV TKSFNRGEC
Edrecolomab Heavy 128 QVQLQQSGAELVRPGTSVKVSCKASGYAFTNYLIEWVKQRPGQGLEWIGV
Chain INPGSGGTNYNEKFKGKATLTADKSSSTAYMQLSSLTSDDSAVYFCARDGP
WFAYWGQGTLVTVSAASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEP
VTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNH
KPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRT
PEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSV
LTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRE
EMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLY
SKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK
Light 133 NIVMTQSPKSMSMSVGERVTLTCKASENVVTYVSWYQQKPEQSPKLLIYG Chain ASNRYTGVPDRFTGSGSATDFTLTISSVQAEDLADYHCGQGYSYPYTFGGG
TKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDN
ALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSS
PVTKSFNRGEC
Adecatumum Heavy 141 EVQLLESGGGVVQPGRSLRLSCAASGFTFSSYGMHWVRQAPGKGLEWVA ab Chain VISYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKD
MGWGSGWRPYYYYGMDVWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGG
TAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVP
SSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVF
LFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKP
REEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKG
QPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYK
TTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLS
LSPGK
Light 146 ELQMTQSPSS LSASVGDRVT ITCRTSQSIS SYLNWYQQKP GQPPKLLIYW Chain ASTRESGVPD RFSGSGSGTD FTLTISSLQP EDSATYYCQQ SYDIPYTFGQ
GTKLEIKRTV AAPSVFIFPP SDEQLKSGTA SVVCLLNNFY PREAKVQWKV DNALQSGNSQ ESVTEQDSKD STYSLSSTLT LSKADYEKHK VYACEVTHQG LSSPVTKSFN RGEC
Anetumab Heavy 154 QVELVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWVRQAPGKGLEWMGII Antibody Region SEQ Sequence
ID
NO:
Chain DPGDSRTRYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCARGQLY
GGTYMDGWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDY
FPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICN
VNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMI
SRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRV
VSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPP
SRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSF
FLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK
Light 159 DIALTQPASVSGSPGQSITISCTGTSSDIGGYNSVSWYQQHPGKAPKLMIYG Chain VNNRPSGVSNRFSGSKSGNTASLTISGLQAEDEADYYCSSYDIESATPVFGG
GTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKG DSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGST VEKT VAPTECS
huDS6 Heavy 167 QAQLVQSGAEVVKPGASVKMSCKASGYTFTSYNMHWVKQTPGQGLEWIG
Chain YIYPGNGATNYNQKFQGKATLTADPSSSTAYMQISSLTSEDSAVYFCARGD
SVPFAYWGQGTLVTVSAASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFP
EPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVN
HKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISR
TPEVTCWVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSV
LTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRD
ELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLY
SKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK
Light 172 EIVLTQSPATMSASPGERVTITCSAHSSVSFMHWFQQKPGTSPKLWIYSTSS Chain LASGVPARFGGSGSGTSYSLTISSMEAEDAATYYCQQRSSFPLTFGAGTKLE
LKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQS
GNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTK
SFNRGEC
Lifastuzumab Heavy 180 EVQLVESGGGLVQPGGSLRLSCAASGFSFSDFAMSWVRQAPGKGLEWVAT
Chain IGRVAFHTYYPDSMKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARHR
GFDVGHFDFWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKD
YFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYIC
NVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTL
MISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYR
VVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLP
PSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGS
FFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK
Light 185 DIQMTQSPSSLSASVGDRVTITCRSSETLVHSSGNTYLEWYQQKPGKAPKL Chain LIYRVSNRFSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCFQGSFNPLTFG
QGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKV DNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQG LSSPVTKSFNRGEC
Sacituzumab Heavy 193 QVQLQQSGSELKKPGASVKVSCKASGYTFTNYGMNWVKQAPGQGLKWM
Chain GWINTYTGEPTYTDDFKGRFAFSLDTSVSTAYLQISSLKADDTAVYFCARG
GFGSSYWYFDVWGQGSLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLV
KDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTY
ICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDT
LMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTY
RVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYT
LPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSD
GSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK
Light 198 DIQLTQSPSS LSASVGDRVS ITCKASQDVS IAVAWYQQKP GKAPKLLIYS Chain ASYRYTGVPD RFSGSGSGTD FTLTISSLQP EDFAVYYCQQ HYITPLTFGA
GTKVEIKRTV AAPSVFIFPP SDEQLKSGTA SVVCLLNNFY PREAKVQWKV DNALQSGNSQ ESVTEQDSKD STYSLSSTLT LSKADYEKHK VYACEVTHQG LSSPVTKSFN RGEC
PR1A3 Heavy 206 QVKLQQSGPELKKPGETVKISCKASGYTFTEFGMNWVKQAPGKGLKWMG
Chain WINTKTGEATYVEEFKGRFAFSLETSATTAYLQINNLKNEDTAKYFCARW
DFYDYVEAMDYWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCL Antibody Region SEQ Sequence
ID
NO:
VKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQT
YICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKD
TLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNST
YRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVY
TLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDS
DGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK
Light 211 DIVMTQSQRFMSTSVGDRVSVTCKASQNVGTNVAWYQQKPGQSPKALIYS Chain ASYRYSGVPDRFTGSGSGTDFTLTISNVQSEDLAEYFCHQYYTYPLFTFGSG
TKLEMKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLS
SPVTKSFNRGEC
Humanized Heavy 813 QVQLVQSGAEVKKPGASVKVSCKASGYTFTEFGMNWVRQAPGQGLEWM PR1A3 Chain GWINTKTGEATYVEEFKGRVTFTTDTSTSTAYMELRSLRSDDTAVYYCAR
WDFAYYVEAMDYWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGC
LVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQ
TYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPK
DTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNS
TYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQV
YTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLD
SDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK
Light 814 DIQMTQSPSSLSASVGDRVTITCKASAAVGTYVAWYQQKPGKAPKLLIYSA Chain SYRKRGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCHQYYTYPLFTFGQGT
KLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNA
LQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSP
VTKSFNRGEC
Humanized Heavy 831 EVQLQESGPGLVKPGGSLSLSCAASGFVFSSYDMSWVRQTPERGLEWVAYI Ab2-3 Chain SSGGGITYAPSTVKGRFTVSRDNAKNTLYLQMNSLTSEDTAVYYCAAHYF
GSSGPFAYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDY
FPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICN
VNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMI
SRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRV
VSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPP
SREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSF
FLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK
Light 832 DIQMTQSPASLSASVGDRVTITCRASENIFSYLAWYQQKPGKSPKLLVYNT Chain RTLAEGVPSRFSGSGSGTDFSLTISSLQPEDFATYYCQHHYGTPFTFGSGTKL
EIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQ
SGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVT
KSFNRGEC
IMAB362, Heavy 219 QVQLQQPGAELVRPGASVKLSCKASGYTFTSYWINWVKQRPGQGLEWIGN
CLAUDIXIM Chain IYPSDSYTNYNQKFKDKATLTVDKSSSTAYMQLSSPTSEDSAVYYCTRSWR
AB GNSFDYWGQGTTLTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFP
EPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVN
HKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISR
TPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVS
VLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSR
EEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFL
YSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK
Light 224 DIVMTQSPSSLTVTAGEKVTMSCKSSQSLLNSGNQKNYLTWYQQKPGQPP Chain KLLIYWASTRESGVPDRFTGSGSGTDFTLTISSVQAEDLAVYYCQNDYSYPF
TFGSGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQW
KVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTH
QGLSSPVTKSFNRGEC
AMG595 Heavy 258 QVQLVESGGGVVQSGRSLRLSCAASGFTFRNYGMHWVRQAPGKGLEWVA
Chain VIWYDGSDKYYADSVRGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARD
GYDILTGNPRDFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALG
CLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGT
QTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKP
KDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQY Antibody Region SEQ Sequence
ID
NO:
NSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREP
QVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPV
LDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK
Light 263 DTVMTQTPLSSHVTLGQPASISCRSSQSLVHSDGNTYLSWLQQRPGQPPRL Chain LIYRISRRFSGVPDRFSGSGAGTDFTLEISRVEAEDVGVYYCMQSTHVPRTF
GQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWK VDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQ GLSSPVTKSFNRGEC
ABT806 Heavy 271 EVQLQESGPGLVKPSQTLSLTCTVSGYSISRDFAWNWIRQPPGKGLEWMG
Chain YISYNGNTRYQPSLKSRITISRDTSKNQFFLKLNSVTAADTATYYCVTASRG
FPYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPV
TVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKP
SNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPE
VTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLT
VLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEM
TKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSK
LTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK
Light 276 DIQMTQSPSSMSVSVGDRVTITCHSSQDINSNIGWLQQKPGKSFKGLIYHGT Chain NLDDGVPSRFSGSGSGTDYTLTISSLQPEDFATYYCVQYAQFPWTFGGGTK
LEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNAL QSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPV TKSFNRGEC
Sibrotuzumab Heavy 284 QVQLVQSGAEVKKPGASVKVSCKTSRYTFTEYTIHWVRQAPGQRLEWIGG
Chain INPNNGIPNYNQKFKGRVTITVDTSASTAYMELSSLRSEDTAVYYCARRRIA
YGYDEGHAMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCL
VKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQT
YICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKD
TLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNST
YRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVY
TLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDS
DGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK
Light 289 DIVMTQSPDSLAVSLGERATINCKSSQSLLYSRNQKNYLAWYQQKPGQPPK Chain LLIFWASTRESGVPDRFSGSGFGTDFTLTISSLQAEDVAVYYCQQYFSYPLT
FGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQW
KVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTH
QGLSSPVTKSFNRGEC
DS-8895a Heavy 297 QVQLVQSGAEVKKPGASVKVSCKASGYTFIDYSMHWVRQAPGQGLEWM variant 1 Chain GWINTYTGEPTYSDDFKGRVTITADTSTSTAYLELSSLRSEDTAVYYCATY
YRYERDFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKD
YFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYIC
NVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTL
MISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYR
VVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLP
PSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGS
FFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK
Light 302 DIVMTQSPLSLPVTPGEPASISCRSSQSIVHSSGITYLEWYLQKPGQSPQLLIY Chain KVSNRFSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCFQGSHVPYTFGQ
GTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLS
SPVTKSFNRGEC
DS-8895a Heavy 310 QIQLVQSGAEVKKPGASVKVSCKASGYTFIDYSMHWVRQAPGQGLKWMG variant 2 Chain WINTYTGEPTYSDDFKGRFAFSLDTSTSTAYLELSSLRSEDTAVYYCATYY
RYERDFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDY
FPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICN
VNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMI
SRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRV
VSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPP
SREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSF
FLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK Antibody Region SEQ Sequence
ID
NO:
Light 315 DVLMTQSPLSLPVTPGEPASISCRSSQSIVHSSGITYLEWYLQKPGQSPQLLI Chain YKVSNRFSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCFQGSHVPYTFG
QGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKV
DNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQG
LSSPVTKSFNRGEC
MEDI-547 Heavy 323 EVQLLESGGGLVQPGGSLRLSCAASGFTFSHYMMAWVRQAPGKGLEWVS
Chain RIGPSGGPTHYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAGY
DSGYDYVAVAGPAEYFQHWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGG
TAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVP
SSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVF
LFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKP
REEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKG
QPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYK
TTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLS
LSPGK
Light 328 DIQMTQSPSSLSASVGDRVTITCRASQSISTWLAWYQQKPGKAPKLLIYKAS Chain NLHTGVPSRFSGSGSGTEFSLTISGLQPDDFATYYCQQYNSYSRTFGQGTKV
EIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQ
SGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVT
KSFNRGEC
Narnatumab Heavy 336 EVQLVESGGGLVQPGGSLRLSCAASGFTFSSYLMTWVRQAPGKGLEWVA
Chain NIKQDGSEKYYVDSVKGRFTISRDNAKNSLNLQMNSLRAEDTAVYYCTRD
GYSSGRHYGMDVWGQGTTVIVSSASTKGPSVFPLAPSSKSTSGGTAALGCL
VKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQT
YICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKD
TLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNST
YRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVY
TLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDS
DGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK
Light 341 EIVLTQSPATLSLSPGERATLSCRASQSVSRYLAWYQQKPGQAPRLLIYDAS Chain NRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRSNWPRTFGQGTKV
EIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQ SGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVT KSFNRGEC
RG7841 Heavy 349 EVQLVESGPALVKPTQTLTLTCTVSGFSLTGYSVNWIRQPPGKALEWLGMI
Chain WGDGSTDYNSALKSRLTISKDTSKNQVVLTMTNMDPVDTATYYCARDYY
FNYASWFAYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVK
DYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYI
CNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDT
LMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTY
RVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYT
LPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSD
GSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK
Light 354 DIQMTQSPSSLSASVGDRVTITCSASQGISNYLNWYQQKPGKTVKLLIYYTS Chain NLHSGVPSRFSGSGSGTDYTLTISSLQPEDFATYYCQQYSELPWTFGQGTKV
EIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQ
SGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVT
KSFNRGEC
Farletuzumab Heavy 362 EVQLVESGGGVVQPGRSLRLSCSASGFTFSGYGLSWVRQAPGKGLEWVA
Chain MISSGGSYTYYADSVKGRFAISRDNAKNTLFLQMDSLRPEDTGVYFCARH
GDDPAWFAYWGQGTPVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVK
DYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYI
CNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDT
LMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTY
RVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYT
LPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSD
GSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK
Light 367 DIQLTQSPSSLSASVGDRVTITCSVSSSISSNNLHWYQQKPGKAPKPWIYGTS Antibody Region SEQ Sequence
ID
NO:
Chain NLASGVPSRFSGSGSGTDYTFTISSLQPEDIATYYCQQWSSYPYMYTFGQGT
KVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNA LQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSP VTKSFNRGEC
Mirvetuximab Heavy 375 QVQLVQSGAEVVKPGASVKISCKASGYTFTGYFMNWVKQSPGQSLEWIGR
Chain IHPYDGDTFY
NQKFQGKATLTVDKSSNTAHMELLSLTSEDFAVYYCTRYDGSRAMDYWG QGTTVTVSSAS
TKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTF PAVLQSSGL
YSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPA PELLGGPS
VFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKT KPREEQYNST
YRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVY TLPPSRDELT
KNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKL TVDKSRWQQ
GNVFSCSVMHEALHNHYTQKSLSLSPG
Light 380 DIVLTQSPLSLAVSLGQPAIISCKASQSVSFAGTSLMHWYHQKPGQQPRLLI Chain YRASNLEAGVPDRFSGSGSKTDFTLTISPVEAEDAATYYCQQSREYPYTFG
GGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKV DNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQG LSSPVTKSFNRGEC
J591 variant 1 Heavy 388 EVQLQQSGPELKKPGTSVRISCKTSGYTFTEYTIHWVKQSHGKSLEWIGNIN
Chain PNNGGTTYNQKFEDKATLTVDKSSSTAYMELRSLTSEDSAVYYCAAGWNF
DYWGQGTTLTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVT
VSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPS
NTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEV
TCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTV
LHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMT
KNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKL
TVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK
Light 393 DIVMTQSHKFMSTSVGDRVSIICKASQDVGTAVDWYQQKPGQSPKLLIYW Chain ASTRHTGVPDRFTGSGSGTDFTLTITNVQSEDLADYFCQQYNSYPLTFGAG
TMLDLKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLS
SPVTKSFNRGEC
J591 variant 2 Heavy 401 EVQLQQSGPELVKPGTSVRISCKTSGYTFTEYTIHWVKQSHGKSLEWIGNIN
Chain PNNGGTTYNQKFEDKATLTVDKSSSTAYMELRSLTSEDSAVYYCAAGWNF
DYWGQGTTLTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVT
VSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPS
NTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEV
TCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTV
LHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMT
KNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKL
TVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK
Light 406 NIVMTQSPKSMSMSVGERVTLTCKASENVVTYVSWYQQKPEQSPKLLIYG Chain ASNRYTGVPDRFTGSGSATDFTLTISSVQAEDLADYHCGQGYSYPYTFGGG
TKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDN
ALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSS
PVTKSFNRGEC
Rovalpituzum Heavy 414 QVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGMNWVRQAPGQGLEWM ab Chain GWINTYTGEPTYADDFKGRVTMTTDTSTSTAYMELRSLRSDDTAVYYCAR
IGDSSPSDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKD
YFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYIC
NVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTL Antibody Region SEQ Sequence
ID
NO:
MISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYR VVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLP PSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGS FFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG
Light 419 EIVMTQSPATLSVSPGERATLSCKASQSVSNDVVWYQQKPGQAPRLLIYYA Chain SNRYTGIPARFSGSGSGTEFTLTISSLQSEDFAVYYCQQDYTSPWTFGQGTK
LEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNAL
QSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPV
TKSFNRGEC
PF-06647020 Heavy 427 QVQLVQSGPEVKKPGASVKVSCKASGYTFTDYAVHWVRQAPGKRLEWIG
Chain VISTYNDYTYNNQDFKGRVTMTRDTSASTAYMELSRLRSEDTAVYYCARG
NSYFYALDYWGQGTSVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKD
YFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYIC
NVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTL
MISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYR
VVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLP
PSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGS
FFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK
Light 432 EIVLTQSPATLSLSPGERATLSCRASESVDSYGKSFMHWYQQKPGQAPRLLI Chain YRASNLESGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQSNEDPWTFGG
GTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLS
SPVTKSFNRGEC
Antibody to Heavy 440 QVQLVESGGGVVQPGRSLRLSCAASGFTFSSYAFHWVRQAPGKGLEWVA PTK7 Chain VISYDGSIKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARTY
YFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEP
VTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNH
KPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRT
PEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSV
LTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRE
EMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLY
SKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK
Light 445 EIVLTQSPDFQSVTPKEKVTITCRASQSIGSSLHWYQQKPDQSPKLLIKYASQ Chain SFSGVPSRFSGSGSGTDFTLTINSLEAEDAAAYYCHQSSSLPITFGQGTRLEI
KRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSG
NSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKS
FNRGEC
Ladiratuzuma Heavy 453 QVQLVQSGAEVKKPGASVKVSCKASGLTIEDYYMHWVRQAPGQGLEWM b Chain GWIDPENGDTEYGPKFQGRVTMTRDTSINTAYMELSRLRSDDTAVYYCAV
HNAHYGTWFAYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCL
VKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQT
YICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKD
TLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNST
YRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVY
TLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDS
DGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK
Light 458 DVVMTQSPLSLPVTLGQPASISCRSSQSLLHSSGNTYLEYFQQRPGQSPRPLI Chain YKISTRFSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCFQGSHVPYTFGG
GTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLS SPVTKSFNRGEC
Cirmtuzumab Heavy 466 QVQLQESGPGLVKPSQTLSLTCTVSGYAFTAYNIHWVRQAPGQGLEWMGS
Chain FDPYDGGSSYNQKFKDRLTISKDTSKNQVVLTMTNMDPVDTATYYCARG
WYYFDYWGHGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYF
PEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNV
NHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMIS
RTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVV
SVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPS
REEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFF Antibody Region SEQ Sequence
ID
NO:
LYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK
Light 471 DIVMTQTPLSLPVTPGEPASISCRASKSISKYLAWYQQKPGQAPRLLIYSGST Chain LQSGIPPRFSGSGYGTDFTLTINNIESEDAAYYFCQQHDESPYTFGEGTKVEI
KRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSG
NSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKS
FNRGEC
Antibody to Heavy 479 QVQLVESGGGVVQPGRSLRLSCTASGFRFRSHGMHWVRQAPGKGLEWVA MAGE-A3 Chain VISYDGNNKLYADSVKGRITISRDNSKNTLFLQMNNVRAEDTAVYYCASP
YTSDWQYFQYWGQGTLVIVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVK
DYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYI
CNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDT
LMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTY
RVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYT
LPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSD
GSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK
Light 484 EIVMTQSPATLSVSPGERATFSCRASQNISTTLAWYQQKPGQAPRLLIYDTS Chain TRATGIPARFSGSGSGTEFTLTISSLQSEDLAVYYCQQSNSWPLTFGGGTKV
EIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQ SGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVT KSFNRGEC
Antibody to Heavy 492 QVQLVQSGGGVVRPGGSLRLSCAASGFSFIDYGMSWVRQVPGKGLEWVA NY-ESO-1 Chain GMNWSGDKKGHAESVKGRFIISRDNAKNTLYLEMSSLRVEDTALYFCARG
EYSNRFDPRGRGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYF
PEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNV
NHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMIS
RTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVV
SVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPS
REEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFF
LYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK
Light 497 DIVMTQTPLSLPVTLGQPASLSCRSSQSLVFTDGNTYLNWFQQRPGQSPRRL Chain IYKVSSRDPGVPDRFSGTGSGTDFTLEISRVEAEDIGVYYCMQGTHWPPIFG
QGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKV
DNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQG
LSSPVTKSFNRGEC
Trastuzumab Heavy 681 EVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVAR
Chain IYPTNGYTRYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRWG
GDGFYAMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVK
DYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYI
CNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDT
LMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTY
RVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYT
LPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSD
GSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK
Light 682 DIQMTQSPSSLSASVGDRVTITCRASQDVNTAVAWYQQKPGKAPKLLIYSA Chain SFLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQHYTTPPTFGQGTKV
EIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQ
SGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVT
KSFNRGEC
TABLE 4. Fusion Sequences - CD40 fusions via the heavy chain
Antibody Region SEQ Sequence
ID
NO:
Pertuzumab HC CD40 21 QVQLVQSGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLE mAb with WMGWINPDSGGTNYAQKFQGRVTMTRDTSISTAYMELNRLRSDDTAV tumor mAb YYCARDQPLGYCTNGVCSYFDYWGQGTLVTVSSASTKGPSVFPLAPSS ScFv KSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS Antibody Region SEQ Sequence
ID
NO:
LSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCP
APELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYV
DGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK
ALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSD
IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFS
CSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSEVQLVESGGGLVQPGG
SLRLSCAASGFTFTDYTMDWVRQAPGKGLEWVADVNPNSGGSIYNQRF
KGRFTLSVDRSKNTLYLQMNSLRAEDTAVYYCARNLGPSFYFDYWGQ
GTLVTVSSGGGGSGGGGSGGGGSGGGGSDIQMTQSPSSLSASVGDRVTI
TCKASQDVSIGVAWYQQKPGKAPKLLIYSASYRYTGVPSRFSGSGSGTD
FTLTISSLQPEDFATYYCQQYYIYPYTFGQGTKVEIK
HC tumor 22 EVQLVESGGGLVQPGGSLRLSCAASGFTFTDYTMDWVRQAPGKGLEW mAb with VADVNPNSGGSIYNQRFKGRFTLSVDRSKNTLYLQMNSLRAEDTAVYY CD40 mAb CARNLGPSFYFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAAL ScFv GCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSS
SLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSV
FLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAK
TKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTIS
KAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNG
QPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALH
NHYTQKSLSLSPGGGGGSGGGGSQVQLVQSGAEVKKPGASVKVSCKAS
GYTFTGYYMHWVRQAPGQGLEWMGWINPDSGGTNYAQKFQGRVTMT
RDTSISTAYMELNRLRSDDTAVYYCARDQPLGYCTNGVCSYFDYWGQ
GTLVTVSSGGGGSGGGGSGGGGSGGGGSDIQMTQSPSSVSASVGDRVTI
TCRASQGIYSWLAWYQQKPGKAPNLLIYTASTLQSGVPSRFSGSGSGTD
FTLTISSLQPEDFATYYCQQANIFPLTFGGGTKVEIK
HC tumor 845 EVQLVESGGGLVQPGGSLRLSCAASGFTFTDYTMDWVRQAPGKGLEW mAb with VADVNPNSGGSIYNQRFKGRFTLSVDRSKNTLYLQMNSLRAEDTAVYY CD40 mAb CARNLGPSFYFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAAL scFv (LH) GCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSS
SLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSV
FLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAK
TKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTIS
KAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNG
QPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALH
NHYTQKSLSLSPGGGGGSGGGGSDIQMTQSPSSVSASVGDRVTITCRAS
QGIYSWLAWYQQKPGKAPNLLIYTASTLQSGVPSRFSGSGSGTDFTLTIS
SLQPEDFATYYCQQANIFPLTFGGGTKVEIKGGGGSGGGGSGGGGSGGG
GSQVQLVQSGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQG
LEWMGWINPDSGGTNYAQKFQGRVTMTRDTSISTAYMELNRLRSDDT
AVYYCARDQPLGYCTNGVCSYFDYWGQGTLVTVSS
Cetuximab HC CD40 34 QVQLVQSGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLE mAb with WMGWINPDSGGTNYAQKFQGRVTMTRDTSISTAYMELNRLRSDDTAV tumor mAb YYCARDQPLGYCTNGVCSYFDYWGQGTLVTVSSASTKGPSVFPLAPSS ScFv KSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS
LSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCP
APELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYV
DGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK
ALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSD
IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFS
CSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSQVQLKQSGPGLVQPSQ
SLSITCTVSGFSLTNYGVHWVRQSPGKGLEWLGVIWSGGNTDYNTPFTS
RLSINKDNSKSQVFFKMNSLQSNDTAIYYCARALTYYDYEFAYWGQGT
LVTVSAGGGGSGGGGSGGGGSGGGGSDILLTQSPVILSVSPGERVSFSCR
ASQSIGTNIHWYQQRTNGSPRLLIKYASESISGIPSRFSGSGSGTDFTLSIN
SVESEDIADYYCQQNNNWPTTFGAGTKLELK
HC tumor 35 QVQLKQSGPGLVQPSQSLSITCTVSGFSLTNYGVHWVRQSPGKGLEWL mAb with GVIWSGGNTDYNTPFTSRLSINKDNSKSQVFFKMNSLQSNDTAIYYCAR CD40 mAb ALTYYDYEFAYWGQGTLVTVSAASTKGPSVFPLAPSSKSTSGGTAALG Antibody Region SEQ Sequence
ID
NO:
ScFv CLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSL
GTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFL
FPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTK
PREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKA
KGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPE
NNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHY
TQKSLSLSPGGGGGSGGGGSQVQLVQSGAEVKKPGASVKVSCKASGYT
FTGYYMHWVRQAPGQGLEWMGWINPDSGGTNYAQKFQGRVTMTRDT
SISTAYMELNRLRSDDTAVYYCARDQPLGYCTNGVCSYFDYWGQGTL
VTVSSGGGGSGGGGSGGGGSGGGGSDIQMTQSPSSVSASVGDRVTITCR
ASQGIYSWLAWYQQKPGKAPNLLIYTASTLQSGVPSRFSGSGSGTDFTL
TISSLQPEDFATYYCQQANIFPLTFGGGTKVEIK
Panitumuma HC CD40 47 QVQLVQSGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLE b mAb with WMGWINPDSGGTNYAQKFQGRVTMTRDTSISTAYMELNRLRSDDTAV tumor mAb YYCARDQPLGYCTNGVCSYFDYWGQGTLVTVSSASTKGPSVFPLAPSS ScFv KSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS
LSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCP
APELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYV
DGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK
ALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSD
IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFS
CSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSQVQLQESGPGLVKPSE
TLSLTCTVSGGSVSSGDYYWTWIRQSPGKGLEWIGHIYYSGNTNYNPSL
KSRLTISIDTSKTQFSLKLSSVTAADTAIYYCVRDRVTGAFDIWGQGTMV
TVSSGGGGSGGGGSGGGGSGGGGSDIQMTQSPSSLSASVGDRVTITCQA
SQDISNYLNWYQQKPGKAPKLLIYDASNLETGVPSRFSGSGSGTDFTFTI
SSLQPEDIATYFCQHFDHLPLAFGGGTKVEIK
HC tumor 48 QVQLQESGPGLVKPSETLSLTCTVSGGSVSSGDYYWTWIRQSPGKGLE mAb with WIGHIYYSGNTNYNPSLKSRLTISIDTSKTQFSLKLSSVTAADTAIYYCVR CD40 mAb DRVTGAFDIWGQGTMVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLV ScFv KDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQ
TYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPK
PKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREE
QYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQ
PREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNY
KTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQK
SLSLSPGGGGGSGGGGSQVQLVQSGAEVKKPGASVKVSCKASGYTFTG
YYMHWVRQAPGQGLEWMGWINPDSGGTNYAQKFQGRVTMTRDTSIS
TAYMELNRLRSDDTAVYYCARDQPLGYCTNGVCSYFDYWGQGTLVTV
SSGGGGSGGGGSGGGGSGGGGSDIQMTQSPSSVSASVGDRVTITCRASQ
GIYSWLAWYQQKPGKAPNLLIYTASTLQSGVPSRFSGSGSGTDFTLTISS
LQPEDFATYYCQQANIFPLTFGGGTKVEIK
Nimotuzuma HC CD40 60 QVQLVQSGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLE b mAb with WMGWINPDSGGTNYAQKFQGRVTMTRDTSISTAYMELNRLRSDDTAV tumor mAb YYCARDQPLGYCTNGVCSYFDYWGQGTLVTVSSASTKGPSVFPLAPSS ScFv KSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS
LSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCP
APELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYV
DGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK
ALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSD
IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFS
CSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSQVQLQQSGAEVKKPG
SSVKVSCKASGYTFTNYYIYWVRQAPGQGLEWIGGINPTSGGSNFNEKF
KTRVTITVDESTNTAYMELSSLRSEDTAFYFCARQGLWFDSDGRGFDF
WGQGSTVTVSSGGGGSGGGGSGGGGSGGGGSDIQMTQSPSSLSASVGD
RVTITCRSSQNIVHSNGNTYLDWYQQTPGKAPKLLIYKVSNRFSGVPSRF
SGSGSGTDFTFTISSLQPEDIATYYCFQYSHVPWTFGQGTKLEIK
HC tumor 61 QVQLQQSGAEVKKPGSSVKVSCKASGYTFTNYYIYWVRQAPGQGLEWI mAb with GGINPTSGGSNFNEKFKTRVTITVDESTNTAYMELSSLRSEDTAFYFCAR Antibody Region SEQ Sequence
ID
NO:
CD40 mAb QGLWFDSDGRGFDFWGQGSTVTVSSASTKGPSVFPLAPSSKSTSGGTAA ScFv LGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPS
SSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPS
VFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNA
KTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKT
ISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEAL
HNHYTQKSLSLSPGGGGGSGGGGSQVQLVQSGAEVKKPGASVKVSCK
ASGYTFTGYYMHWVRQAPGQGLEWMGWINPDSGGTNYAQKFQGRVT
MTRDTSISTAYMELNRLRSDDTAVYYCARDQPLGYCTNGVCSYFDYW
GQGTLVTVSSGGGGSGGGGSGGGGSGGGGSDIQMTQSPSSVSASVGDR
VTITCRASQGIYSWLAWYQQKPGKAPNLLIYTASTLQSGVPSRFSGSGS
GTDFTLTISSLQPEDFATYYCQQANIFPLTFGGGTKVEIK
Zalutumuma HC CD40 73 QVQLVQSGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLE b mAb with WMGWINPDSGGTNYAQKFQGRVTMTRDTSISTAYMELNRLRSDDTAV tumor mAb YYCARDQPLGYCTNGVCSYFDYWGQGTLVTVSSASTKGPSVFPLAPSS ScFv KSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS
LSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCP
APELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYV
DGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK
ALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSD
IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFS
CSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSQVQLVESGGGVVQPG
RSLRLSCAASGFTFSTYGMHWVRQAPGKGLEWVAVIWDDGSYKYYGD
SVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDGITMVRGVMK
DYFDYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSAIQLTQSPSSLS
ASVGDRVTITCRASQDISSALVWYQQKPGKAPKLLIYDASSLESGVPSRF
SGSESGTDFTLTISSLQPEDFATYYCQQFNSYPLTFGGGTKVEIK
HC tumor 74 QVQLVESGGGVVQPGRSLRLSCAASGFTFSTYGMHWVRQAPGKGLEW mAb with VAVIWDDGSYKYYGDSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVY CD40 mAb YCARDGITMVRGVMKDYFDYWGQGTLVTVSSASTKGPSVFPLAPSSKS ScFv TSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLS
SVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAP
ELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDG
VEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKAL
PAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIA
VEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCS
VMHEALHNHYTQKSLSLSPGGGGGSGGGGSQVQLVQSGAEVKKPGAS
VKVSCKASGYTFTGYYMHWVRQAPGQGLEWMGWINPDSGGTNYAQK
FQGRVTMTRDTSISTAYMELNRLRSDDTAVYYCARDQPLGYCTNGVCS
YFDYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSDIQMTQSPSSVSA
SVGDRVTITCRASQGIYSWLAWYQQKPGKAPNLLIYTASTLQSGVPSRF
SGSGSGTDFTLTISSLQPEDFATYYCQQANIFPLTFGGGTKVEIK
Onartuzuma HC CD40 86 QVQLVQSGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLE b mAb with WMGWINPDSGGTNYAQKFQGRVTMTRDTSISTAYMELNRLRSDDTAV tumor mAb YYCARDQPLGYCTNGVCSYFDYWGQGTLVTVSSASTKGPSVFPLAPSS ScFv KSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS
LSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCP
APELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYV
DGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK
ALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSD
IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFS
CSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSEVQLVESGGGLVQPGG
SLRLSCAASGYTFTSYWLHWVRQAPGKGLEWVGMIDPSNSDTRFNPNF
KDRFTISADTSKNTAYLQMNSLRAEDTAVYYCATYRSYVTPLDYWGQ
GTLVTVSSGGGGSGGGGSGGGGSGGGGSDIQMTQSPSSLSASVGDRVTI
TCKSSQSLLYTSSQKNYLAWYQQKPGKAPKLLIYWASTRESGVPSRFSG
SGSGTDFTLTISSLQPEDFATYYCQQYYAYPWTFGQGTKVEIK
HC tumor 87 EVQLVESGGGLVQPGGSLRLSCAASGYTFTSYWLHWVRQAPGKGLEW Antibody Region SEQ Sequence
ID
NO:
mAb with VGMIDPSNSDTRFNPNFKDRFTISADTSKNTAYLQMNSLRAEDTAVYYC CD40 mAb ATYRSYVTPLDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALG ScFv CLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSL
GTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFL
FPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTK
PREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKA
KGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPE
NNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHY
TQKSLSLSPGGGGGSGGGGSQVQLVQSGAEVKKPGASVKVSCKASGYT
FTGYYMHWVRQAPGQGLEWMGWINPDSGGTNYAQKFQGRVTMTRDT
SISTAYMELNRLRSDDTAVYYCARDQPLGYCTNGVCSYFDYWGQGTL
VTVSSGGGGSGGGGSGGGGSGGGGSDIQMTQSPSSVSASVGDRVTITCR
ASQGIYSWLAWYQQKPGKAPNLLIYTASTLQSGVPSRFSGSGSGTDFTL
TISSLQPEDFATYYCQQANIFPLTFGGGTKVEIK
Patritumab HC CD40 99 QVQLVQSGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLE mAb with WMGWINPDSGGTNYAQKFQGRVTMTRDTSISTAYMELNRLRSDDTAV tumor mAb YYCARDQPLGYCTNGVCSYFDYWGQGTLVTVSSASTKGPSVFPLAPSS ScFv KSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS
LSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCP
APELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYV
DGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK
ALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSD
IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFS
CSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSQVQLQQWGAGLLKPS
ETLSLTCAVYGGSFSGYYWSWIRQPPGKGLEWIGEINHSGSTNYNPSLK
SRVTISVETSKNQFSLKLSSVTAADTAVYYCARDKWTWYFDLWGRGTL
VTVSSGGGGSGGGGSGGGGSGGGGSDIEMTQSPDSLAVSLGERATINCR
SSQSVLYSSSNRNYLAWYQQNPGQPPKLLIYWASTRESGVPDRFSGSGS
GTDFTLTISSLQAEDVAVYYCQQYYSTPRTFGQGTKVEIK
HC tumor 100 QVQLQQWGAGLLKPSETLSLTCAVYGGSFSGYYWSWIRQPPGKGLEWI mAb with GEINHSGSTNYNPSLKSRVTISVETSKNQFSLKLSSVTAADTAVYYCARD CD40 mAb KWTWYFDLWGRGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLV ScFv KDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQ
TYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPK
PKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREE
QYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQ
PREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNY
KTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQK
SLSLSPGGGGGSGGGGSQVQLVQSGAEVKKPGASVKVSCKASGYTFTG
YYMHWVRQAPGQGLEWMGWINPDSGGTNYAQKFQGRVTMTRDTSIS
TAYMELNRLRSDDTAVYYCARDQPLGYCTNGVCSYFDYWGQGTLVTV
SSGGGGSGGGGSGGGGSGGGGSDIQMTQSPSSVSASVGDRVTITCRASQ
GIYSWLAWYQQKPGKAPNLLIYTASTLQSGVPSRFSGSGSGTDFTLTISS
LQPEDFATYYCQQANIFPLTFGGGTKVEIK
Clivatuzuma HC CD40 112 QVQLVQSGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLE b mAb with WMGWINPDSGGTNYAQKFQGRVTMTRDTSISTAYMELNRLRSDDTAV tumor mAb YYCARDQPLGYCTNGVCSYFDYWGQGTLVTVSSASTKGPSVFPLAPSS ScFv KSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS
LSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCP
APELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYV
DGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK
ALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSD
IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFS
CSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSQVQLQQSGAEVKKPG
ASVKVSCEASGYTFPSYVLHWVKQAPGQGLEWIGYINPYNDGTQYNEK
FKGKATLTRDTSINTAYMELSRLRSDDTAVYYCARGFGGSYGFAYWGQ
GTLVTVSSGGGGSGGGGSGGGGSGGGGSDIQLTQSPSSLSASVGDRVT
MTCSASSSVSSSYLYWYQQKPGKAPKLWIYSTSNLASGVPARFSGSGSG
TDFTLTISSLQPEDSASYFCHQWNRYPYTFGGGTRLEIK Antibody Region SEQ Sequence
ID
NO:
HC tumor 113 QVQLQQSGAEVKKPGASVKVSCEASGYTFPSYVLHWVKQAPGQGLEW mAb with IGYINPYNDGTQYNEKFKGKATLTRDTSINTAYMELSRLRSDDTAVYYC CD40 mAb ARGFGGSYGFAYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALG ScFv CLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSL
GTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFL
FPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTK
PREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKA
KGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPE
NNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHY
TQKSLSLSPGGGGGSGGGGSQVQLVQSGAEVKKPGASVKVSCKASGYT
FTGYYMHWVRQAPGQGLEWMGWINPDSGGTNYAQKFQGRVTMTRDT
SISTAYMELNRLRSDDTAVYYCARDQPLGYCTNGVCSYFDYWGQGTL
VTVSSGGGGSGGGGSGGGGSGGGGSDIQMTQSPSSVSASVGDRVTITCR
ASQGIYSWLAWYQQKPGKAPNLLIYTASTLQSGVPSRFSGSGSGTDFTL
TISSLQPEDFATYYCQQANIFPLTFGGGTKVEIK
Sofituzumab HC CD40 125 QVQLVQSGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLE mAb with WMGWINPDSGGTNYAQKFQGRVTMTRDTSISTAYMELNRLRSDDTAV tumor mAb YYCARDQPLGYCTNGVCSYFDYWGQGTLVTVSSASTKGPSVFPLAPSS ScFv KSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS
LSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCP
APELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYV
DGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK
ALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSD
IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFS
CSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSEVQLVESGGGLVQPGG
SLRLSCAASGYSITNDYAWNWVRQAPGKGLEWVGYISYSGYTTYNPSL
KSRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARWTSGLDYWGQGTLV
TVSSGGGGSGGGGSGGGGSGGGGSDIQMTQSPSSLSASVGDRVTITCKA
SDLIHNWLAWYQQKPGKAPKLLIYGATSLETGVPSRFSGSGSGTDFTLTI
SSLQPEDFATYYCQQYWTTPFTFGQGTKVEIK
HC tumor 126 EVQLVESGGGLVQPGGSLRLSCAASGYSITNDYAWNWVRQAPGKGLE mAb with WVGYISYSGYTTYNPSLKSRFTISRDTSKNTLYLQMNSLRAEDTAVYYC CD40 mAb ARWTSGLDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCL ScFv VKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGT
QTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPP
KPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPRE
EQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKG
QPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENN
YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQ
KSLSLSPGGGGGSGGGGSQVQLVQSGAEVKKPGASVKVSCKASGYTFT
GYYMHWVRQAPGQGLEWMGWINPDSGGTNYAQKFQGRVTMTRDTSI
STAYMELNRLRSDDTAVYYCARDQPLGYCTNGVCSYFDYWGQGTLVT
VSSGGGGSGGGGSGGGGSGGGGSDIQMTQSPSSVSASVGDRVTITCRAS
QGIYSWLAWYQQKPGKAPNLLIYTASTLQSGVPSRFSGSGSGTDFTLTIS
SLQPEDFATYYCQQANIFPLTFGGGTKVEIK
Edrecoloma HC CD40 138 QVQLVQSGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLE b mAb with WMGWINPDSGGTNYAQKFQGRVTMTRDTSISTAYMELNRLRSDDTAV tumor mAb YYCARDQPLGYCTNGVCSYFDYWGQGTLVTVSSASTKGPSVFPLAPSS ScFv KSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS
LSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCP
APELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYV
DGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK
ALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSD
IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFS
CSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSQVQLQQSGAELVRPGT
SVKVSCKASGYAFTNYLIEWVKQRPGQGLEWIGVINPGSGGTNYNEKF
KGKATLTADKSSSTAYMQLSSLTSDDSAVYFCARDGPWFAYWGQGTL
VTVSAGGGGSGGGGSGGGGSGGGGSNIVMTQSPKSMSMSVGERVTLT
CKASENVVTYVSWYQQKPEQSPKLLIYGASNRYTGVPDRFTGSGSATD Antibody Region SEQ Sequence
ID
NO:
FTLTISSVQAEDLADYHCGQGYSYPYTFGGGTKLEIK
HC tumor 139 QVQLQQSGAELVRPGTSVKVSCKASGYAFTNYLIEWVKQRPGQGLEWI mAb with GVINPGSGGTNYNEKFKGKATLTADKSSSTAYMQLSSLTSDDSAVYFCA CD40 mAb RDGPWFAYWGQGTLVTVSAASTKGPSVFPLAPSSKSTSGGTAALGCLV ScFv KDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQ
TYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPK
PKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREE
QYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQ
PREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNY
KTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQK
SLSLSPGGGGGSGGGGSQVQLVQSGAEVKKPGASVKVSCKASGYTFTG
YYMHWVRQAPGQGLEWMGWINPDSGGTNYAQKFQGRVTMTRDTSIS
TAYMELNRLRSDDTAVYYCARDQPLGYCTNGVCSYFDYWGQGTLVTV
SSGGGGSGGGGSGGGGSGGGGSDIQMTQSPSSVSASVGDRVTITCRASQ
GIYSWLAWYQQKPGKAPNLLIYTASTLQSGVPSRFSGSGSGTDFTLTISS
LQPEDFATYYCQQANIFPLTFGGGTKVEIK
Adecatumu HC CD40 151 QVQLVQSGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLE mab mAb with WMGWINPDSGGTNYAQKFQGRVTMTRDTSISTAYMELNRLRSDDTAV tumor mAb YYCARDQPLGYCTNGVCSYFDYWGQGTLVTVSSASTKGPSVFPLAPSS ScFv KSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS
LSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCP
APELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYV
DGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK
ALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSD
IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFS
CSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSEVQLLESGGGVVQPGR
SLRLSCAASGFTFSSYGMHWVRQAPGKGLEWVAVISYDGSNKYYADS
VKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKDMGWGSGWRPYY
YYGMDVWGQGTTVTVSSGGGGSGGGGSGGGGSGGGGSELQMTQSPSS
LSASVGDRVTITCRTSQSISSYLNWYQQKPGQPPKLLIYWASTRESGVPD
RFSGSGSGTDFTLTISSLQPEDSATYYCQQSYDIPYTFGQGTKLEIK
HC tumor 152 EVQLLESGGGVVQPGRSLRLSCAASGFTFSSYGMHWVRQAPGKGLEW mAb with VAVISYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYY CD40 mAb CAKDMGWGSGWRPYYYYGMDVWGQGTTVTVSSASTKGPSVFPLAPSS ScFv KSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS
LSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCP
APELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYV
DGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK
ALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSD
IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFS
CSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSQVQLVQSGAEVKKPG
ASVKVSCKASGYTFTGYYMHWVRQAPGQGLEWMGWINPDSGGTNYA
QKFQGRVTMTRDTSISTAYMELNRLRSDDTAVYYCARDQPLGYCTNGV
CSYFDYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSDIQMTQSPSSV
SASVGDRVTITCRASQGIYSWLAWYQQKPGKAPNLLIYTASTLQSGVPS
RFSGSGSGTDFTLTISSLQPEDFATYYCQQANIFPLTFGGGTKVEIK
Anetumab HC CD40 164 QVQLVQSGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLE mAb with WMGWINPDSGGTNYAQKFQGRVTMTRDTSISTAYMELNRLRSDDTAV tumor mAb YYCARDQPLGYCTNGVCSYFDYWGQGTLVTVSSASTKGPSVFPLAPSS ScFv KSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS
LSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCP
APELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYV
DGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK
ALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSD
IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFS
CSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSQVELVQSGAEVKKPGE
SLKISCKGSGYSFTSYWIGWVRQAPGKGLEWMGIIDPGDSRTRYSPSFQ
GQVTISADKSISTAYLQWSSLKASDTAMYYCARGQLYGGTYMDGWGQ
GTLVTVSSGGGGSGGGGSGGGGSGGGGSDIALTQPASVSGSPGQSITISC Antibody Region SEQ Sequence
ID
NO:
TGTSSDIGGYNSVSWYQQHPGKAPKLMIYGVNNRPSGVSNRFSGSKSG NTASLTISGLQAEDEADYYCSSYDIESATPVFGGGTKLTVL
HC tumor 165 QVELVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWVRQAPGKGLEWM mAb with GIIDPGDSRTRYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCAR CD40 mAb GQLYGGTYMDGWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALG ScFv CLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSL
GTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFL
FPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTK
PREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKA
KGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPE
NNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHY
TQKSLSLSPGGGGGSGGGGSQVQLVQSGAEVKKPGASVKVSCKASGYT
FTGYYMHWVRQAPGQGLEWMGWINPDSGGTNYAQKFQGRVTMTRDT
SISTAYMELNRLRSDDTAVYYCARDQPLGYCTNGVCSYFDYWGQGTL
VTVSSGGGGSGGGGSGGGGSGGGGSDIQMTQSPSSVSASVGDRVTITCR
ASQGIYSWLAWYQQKPGKAPNLLIYTASTLQSGVPSRFSGSGSGTDFTL
TISSLQPEDFATYYCQQANIFPLTFGGGTKVEIK
huDS6 HC CD40 177 QVQLVQSGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLE mAb with WMGWINPDSGGTNYAQKFQGRVTMTRDTSISTAYMELNRLRSDDTAV tumor mAb YYCARDQPLGYCTNGVCSYFDYWGQGTLVTVSSASTKGPSVFPLAPSS ScFv KSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS
LSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCP
APELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYV
DGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK
ALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSD
IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFS
CSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSQAQLVQSGAEVVKPG
ASVKMSCKASGYTFTSYNMHWVKQTPGQGLEWIGYIYPGNGATNYNQ
KFQGKATLTADPSSSTAYMQISSLTSEDSAVYFCARGDSVPFAYWGQGT
LVTVSAGGGGSGGGGSGGGGSGGGGSEIVLTQSPATMSASPGERVTITC
SAHSSVSFMHWFQQKPGTSPKLWIYSTSSLASGVPARFGGSGSGTSYSL
TISSMEAEDAATYYCQQRSSFPLTFGAGTKLELK
HC tumor 178 QAQLVQSGAEVVKPGASVKMSCKASGYTFTSYNMHWVKQTPGQGLE mAb with WIGYIYPGNGATNYNQKFQGKATLTADPSSSTAYMQISSLTSEDSAVYF CD40 mAb CARGDSVPFAYWGQGTLVTVSAASTKGPSVFPLAPSSKSTSGGTAALGC ScFv LVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLG
TQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFP
PKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPR
EEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAK
GQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPEN
NYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYT
QKSLSLSPGGGGGSGGGGSQVQLVQSGAEVKKPGASVKVSCKASGYTF
TGYYMHWVRQAPGQGLEWMGWINPDSGGTNYAQKFQGRVTMTRDTS
ISTAYMELNRLRSDDTAVYYCARDQPLGYCTNGVCSYFDYWGQGTLV
TVSSGGGGSGGGGSGGGGSGGGGSDIQMTQSPSSVSASVGDRVTITCRA
SQGIYSWLAWYQQKPGKAPNLLIYTASTLQSGVPSRFSGSGSGTDFTLTI
SSLQPEDFATYYCQQANIFPLTFGGGTKVEIK
Lifastuzuma HC CD40 190 QVQLVQSGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLE b mAb with WMGWINPDSGGTNYAQKFQGRVTMTRDTSISTAYMELNRLRSDDTAV tumor mAb YYCARDQPLGYCTNGVCSYFDYWGQGTLVTVSSASTKGPSVFPLAPSS ScFv KSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS
LSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCP
APELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYV
DGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK
ALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSD
IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFS
CSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSEVQLVESGGGLVQPGG
SLRLSCAASGFSFSDFAMSWVRQAPGKGLEWVATIGRVAFHTYYPDSM
KGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARHRGFDVGHFDFWG Antibody Region SEQ Sequence
ID
NO:
QGTLVTVSSGGGGSGGGGSGGGGSGGGGSDIQMTQSPSSLSASVGDRV TITCRSSETLVHSSGNTYLEWYQQKPGKAPKLLIYRVSNRFSGVPSRFSG SGSGTDFTLTISSLQPEDFATYYCFQGSFNPLTFGQGTKVEIK
HC tumor 191 EVQLVESGGGLVQPGGSLRLSCAASGFSFSDFAMSWVRQAPGKGLEWV mAb with ATIGRVAFHTYYPDSMKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYC CD40 mAb ARHRGFDVGHFDFWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAAL ScFv GCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSS
SLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSV
FLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAK
TKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTIS
KAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNG
QPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALH
NHYTQKSLSLSPGGGGGSGGGGSQVQLVQSGAEVKKPGASVKVSCKAS
GYTFTGYYMHWVRQAPGQGLEWMGWINPDSGGTNYAQKFQGRVTMT
RDTSISTAYMELNRLRSDDTAVYYCARDQPLGYCTNGVCSYFDYWGQ
GTLVTVSSGGGGSGGGGSGGGGSGGGGSDIQMTQSPSSVSASVGDRVTI
TCRASQGIYSWLAWYQQKPGKAPNLLIYTASTLQSGVPSRFSGSGSGTD
FTLTISSLQPEDFATYYCQQANIFPLTFGGGTKVEIK
Sacituzumab HC CD40 203 QVQLVQSGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLE mAb with WMGWINPDSGGTNYAQKFQGRVTMTRDTSISTAYMELNRLRSDDTAV tumor mAb YYCARDQPLGYCTNGVCSYFDYWGQGTLVTVSSASTKGPSVFPLAPSS ScFv KSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS
LSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCP
APELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYV
DGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK
ALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSD
IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFS
CSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSQVQLQQSGSELKKPGA
SVKVSCKASGYTFTNYGMNWVKQAPGQGLKWMGWINTYTGEPTYTD
DFKGRFAFSLDTSVSTAYLQISSLKADDTAVYFCARGGFGSSYWYFDV
WGQGSLVTVSSGGGGSGGGGSGGGGSGGGGSDIQLTQSPSSLSASVGD
RVSITCKASQDVSIAVAWYQQKPGKAPKLLIYSASYRYTGVPDRFSGSG
SGTDFTLTISSLQPEDFAVYYCQQHYITPLTFGAGTKVEIK
HC tumor 204 QVQLQQSGSELKKPGASVKVSCKASGYTFTNYGMNWVKQAPGQGLK mAb with WMGWINTYTGEPTYTDDFKGRFAFSLDTSVSTAYLQISSLKADDTAVYF CD40 mAb CARGGFGSSYWYFDVWGQGSLVTVSSASTKGPSVFPLAPSSKSTSGGTA ScFv ALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVP
SSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPS
VFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNA
KTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKT
ISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEAL
HNHYTQKSLSLSPGGGGGSGGGGSQVQLVQSGAEVKKPGASVKVSCK
ASGYTFTGYYMHWVRQAPGQGLEWMGWINPDSGGTNYAQKFQGRVT
MTRDTSISTAYMELNRLRSDDTAVYYCARDQPLGYCTNGVCSYFDYW
GQGTLVTVSSGGGGSGGGGSGGGGSGGGGSDIQMTQSPSSVSASVGDR
VTITCRASQGIYSWLAWYQQKPGKAPNLLIYTASTLQSGVPSRFSGSGS
GTDFTLTISSLQPEDFATYYCQQANIFPLTFGGGTKVEIK
PR1A3 HC CD40 216 QVQLVQSGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLE mAb with WMGWINPDSGGTNYAQKFQGRVTMTRDTSISTAYMELNRLRSDDTAV tumor mAb YYCARDQPLGYCTNGVCSYFDYWGQGTLVTVSSASTKGPSVFPLAPSS ScFv KSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS
LSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCP
APELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYV
DGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK
ALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSD
IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFS
CSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSQVKLQQSGPELKKPGE
TVKISCKASGYTFTEFGMNWVKQAPGKGLKWMGWINTKTGEATYVEE Antibody Region SEQ Sequence
ID
NO:
FKGRFAFSLETSATTAYLQINNLKNEDTAKYFCARWDFYDYVEAMDY WGQGTTVTVSSGGGGSGGGGSGGGGSGGGGSDIVMTQSQRFMSTSVG DRVSVTCKASQNVGTNVAWYQQKPGQSPKALIYSASYRYSGVPDRFTG SGSGTDFTLTISNVQSEDLAEYFCHQYYTYPLFTFGSGTKLEMK
HC tumor 217 QVKLQQSGPELKKPGETVKISCKASGYTFTEFGMNWVKQAPGKGLKW mAb with MGWINTKTGEATYVEEFKGRFAFSLETSATTAYLQINNLKNEDTAKYFC CD40 mAb ARWDFYDYVEAMDYWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTA ScFv ALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVP
SSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPS
VFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNA
KTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKT
ISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEAL
HNHYTQKSLSLSPGGGGGSGGGGSQVQLVQSGAEVKKPGASVKVSCK
ASGYTFTGYYMHWVRQAPGQGLEWMGWINPDSGGTNYAQKFQGRVT
MTRDTSISTAYMELNRLRSDDTAVYYCARDQPLGYCTNGVCSYFDYW
GQGTLVTVSSGGGGSGGGGSGGGGSGGGGSDIQMTQSPSSVSASVGDR
VTITCRASQGIYSWLAWYQQKPGKAPNLLIYTASTLQSGVPSRFSGSGS
GTDFTLTISSLQPEDFATYYCQQANIFPLTFGGGTKVEIK
Humanized HC CD40 815 QVQLVQSGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLE PR1A3 mAb with WMGWINPDSGGTNYAQKFQGRVTMTRDTSISTAYMELNRLRSDDTAV tumor mAb YYCARDQPLGYCTNGVCSYFDYWGQGTLVTVSSASTKGPSVFPLAPSS ScFv KSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS
LSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCP
APELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYV
DGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK
ALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSD
IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFS
CSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSQVQLVQSGAEVKKPG
ASVKVSCKASGYTFTEFGMNWVRQAPGQGLEWMGWINTKTGEATYV
EEFKGRVTFTTDTSTSTAYMELRSLRSDDTAVYYCARWDFAYYVEAM
DYWGQGTTVTVSSGGGGSGGGGSGGGGSGGGGSDIQMTQSPSSLSASV
GDRVTITCKASAAVGTYVAWYQQKPGKAPKLLIYSASYRKRGVPSRFS
GSGSGTDFTLTISSLQPEDFATYYCHQYYTYPLFTFGQGTKLEIK
HC tumor 816 QVQLVQSGAEVKKPGASVKVSCKASGYTFTEFGMNWVRQAPGQGLE mAb with WMGWINTKTGEATYVEEFKGRVTFTTDTSTSTAYMELRSLRSDDTAVY CD40 mAb YCARWDFAYYVEAMDYWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGG ScFv TAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVT
VPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLG
GPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVH
NAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIE
KTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWE
SNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHE
ALHNHYTQKSLSLSPGGGGGSGGGGSQVQLVQSGAEVKKPGASVKVSC
KASGYTFTGYYMHWVRQAPGQGLEWMGWINPDSGGTNYAQKFQGRV
TMTRDTSISTAYMELNRLRSDDTAVYYCARDQPLGYCTNGVCSYFDY
WGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSDIQMTQSPSSVSASVGD
RVTITCRASQGIYSWLAWYQQKPGKAPNLLIYTASTLQSGVPSRFSGSGS
GTDFTLTISSLQPEDFATYYCQQANIFPLTFGGGTKVEIK
HC tumor 843 QVQLVQSGAEVKKPGASVKVSCKASGYTFTEFGMNWVRQAPGQGLE mAb with WMGWINTKTGEATYVEEFKGRVTFTTDTSTSTAYMELRSLRSDDTAVY CD40 mAb YCARWDFAYYVEAMDYWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGG scFv (LH) TAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVT
VPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLG
GPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVH
NAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIE
KTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWE
SNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHE
ALHNHYTQKSLSLSPGGGGGSGGGGSDIQMTQSPSSVSASVGDRVTITC Antibody Region SEQ Sequence
ID
NO:
RASQGIYSWLAWYQQKPGKAPNLLIYTASTLQSGVPSRFSGSGSGTDFT
LTISSLQPEDFATYYCQQANIFPLTFGGGTKVEIKGGGGSGGGGSGGGGS
GGGGSQVQLVQSGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAP
GQGLEWMGWINPDSGGTNYAQKFQGRVTMTRDTSISTAYMELNRLRS
DDTAVYYCARDQPLGYCTNGVCSYFDYWGQGTLVTVSS
Humanized HC CD40 833 QVQLVQSGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLE Ab2-3 mAb with WMGWINPDSGGTNYAQKFQGRVTMTRDTSISTAYMELNRLRSDDTAV tumor mAb YYCARDQPLGYCTNGVCSYFDYWGQGTLVTVSSASTKGPSVFPLAPSS ScFv KSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS
LSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCP
APELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYV
DGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK
ALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSD
IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFS
CSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSEVQLQESGPGLVKPGG
SLSLSCAASGFVFSSYDMSWVRQTPERGLEWVAYISSGGGITYAPSTVK
GRFTVSRDNAKNTLYLQMNSLTSEDTAVYYCAAHYFGSSGPFAYWGQ
GTLVTVSSGGGGSGGGGSGGGGSGGGGSDIQMTQSPASLSASVGDRVTI
TCRASENIFSYLAWYQQKPGKSPKLLVYNTRTLAEGVPSRFSGSGSGTD
FSLTISSLQPEDFATYYCQHHYGTPFTFGSGTKLEIK
HC tumor 834 EVQLQESGPGLVKPGGSLSLSCAASGFVFSSYDMSWVRQTPERGLEWV mAb with AYISSGGGITYAPSTVKGRFTVSRDNAKNTLYLQMNSLTSEDTAVYYCA CD40 mAb AHYFGSSGPFAYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALG ScFv CLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSL
GTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFL
FPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTK
PREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKA
KGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPE
NNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHY
TQKSLSLSPGGGGGSGGGGSQVQLVQSGAEVKKPGASVKVSCKASGYT
FTGYYMHWVRQAPGQGLEWMGWINPDSGGTNYAQKFQGRVTMTRDT
SISTAYMELNRLRSDDTAVYYCARDQPLGYCTNGVCSYFDYWGQGTL
VTVSSGGGGSGGGGSGGGGSGGGGSDIQMTQSPSSVSASVGDRVTITCR
ASQGIYSWLAWYQQKPGKAPNLLIYTASTLQSGVPSRFSGSGSGTDFTL
TISSLQPEDFATYYCQQANIFPLTFGGGTKVEIK
HC tumor 841 EVQLQESGPGLVKPGGSLSLSCAASGFVFSSYDMSWVRQTPERGLEWV mAb with AYISSGGGITYAPSTVKGRFTVSRDNAKNTLYLQMNSLTSEDTAVYYCA CD40 mAb AHYFGSSGPFAYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALG scFv (LH) CLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSL
GTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFL
FPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTK
PREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKA
KGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPE
NNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHY
TQKSLSLSPGGGGGSGGGGSDIQMTQSPSSVSASVGDRVTITCRASQGIY
SWLAWYQQKPGKAPNLLIYTASTLQSGVPSRFSGSGSGTDFTLTISSLQP
EDFATYYCQQANIFPLTFGGGTKVEIKGGGGSGGGGSGGGGSGGGGSQ
VQLVQSGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLEW
MGWINPDSGGTNYAQKFQGRVTMTRDTSISTAYMELNRLRSDDTAVY
YCARDQPLGYCTNGVCSYFDYWGQGTLVTVSS
IMAB362, HC CD40 229 QVQLVQSGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLE
CLAUDIXI mAb with WMGWINPDSGGTNYAQKFQGRVTMTRDTSISTAYMELNRLRSDDTAV
MAB tumor mAb YYCARDQPLGYCTNGVCSYFDYWGQGTLVTVSSASTKGPSVFPLAPSS
ScFv KSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS
LSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCP
APELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYV
DGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK
ALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSD
IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFS Antibody Region SEQ Sequence
ID
NO:
CSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSQVQLQQPGAELVRPG
ASVKLSCKASGYTFTSYWINWVKQRPGQGLEWIGNIYPSDSYTNYNQK
FKDKATLTVDKSSSTAYMQLSSPTSEDSAVYYCTRSWRGNSFDYWGQG
TTLTVSSGGGGSGGGGSGGGGSGGGGSDIVMTQSPSSLTVTAGEKVTM
SCKSSQSLLNSGNQKNYLTWYQQKPGQPPKLLIYWASTRESGVPDRFTG
SGSGTDFTLTISSVQAEDLAVYYCQNDYSYPFTFGSGTKLEIK
HC tumor 230 QVQLQQPGAELVRPGASVKLSCKASGYTFTSYWINWVKQRPGQGLEWI mAb with GNIYPSDSYTNYNQKFKDKATLTVDKSSSTAYM
CD40 mAb QLSSPTSEDSAVYYCTRSWRGNSFDYWGQGTTLTVSSASTKGPSVFPLA ScFv PSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSG
LYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPP
CPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNW
YVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVS
NKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYP
SDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNV
FSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSQVQLVQSGAEVKKP
GASVKVSCKASGYTFTGYYMHWVRQAPGQGLEWMGWINPDSGGTNY
AQKFQGRVTMTRDTSISTAYMELNRLRSDDTAVYYCARDQPLGYCTNG
VCSYFDYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSDIQMTQSPSS
VSASVGDRVTITCRASQGIYSWLAWYQQKPGKAPNLLIYTASTLQSGVP
SRFSGSGSGTDFTLTISSLQPEDFATYYCQQANIFPLTFGGGTKVEIK
AMG595 HC CD40 268 QVQLVQSGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLE mAb with WMGWINPDSGGTNYAQKFQGRVTMTRDTSISTAYMELNRLRSDDTAV tumor mAb YYCARDQPLGYCTNGVCSYFDYWGQGTLVTVSSASTKGPSVFPLAPSS ScFv KSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS
LSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCP
APELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYV
DGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK
ALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSD
IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFS
CSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSQVQLVESGGGVVQSG
RSLRLSCAASGFTFRNYGMHWVRQAPGKGLEWVAVIWYDGSDKYYA
DSVRGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDGYDILTGNPR
DFDYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSDTVMTQTPLSSH
VTLGQPASISCRSSQSLVHSDGNTYLSWLQQRPGQPPRLLIYRISRRFSGV
PDRFSGSGAGTDFTLEISRVEAEDVGVYYCMQSTHVPRTFGQGTKVEIK
HC tumor 269 QVQLVESGGGVVQSGRSLRLSCAASGFTFRNYGMHWVRQAPGKGLEW mAb with VAVIWYDGSDKYYADSVRGRFTISRDNSKNTLYLQMNSLRAEDTAVYY CD40 mAb CARDGYDILTGNPRDFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSG ScFv GTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVV
TVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLG
GPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVH
NAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIE
KTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWE
SNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHE
ALHNHYTQKSLSLSPGGGGGSGGGGSQVQLVQSGAEVKKPGASVKVSC
KASGYTFTGYYMHWVRQAPGQGLEWMGWINPDSGGTNYAQKFQGRV
TMTRDTSISTAYMELNRLRSDDTAVYYCARDQPLGYCTNGVCSYFDY
WGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSDIQMTQSPSSVSASVGD
RVTITCRASQGIYSWLAWYQQKPGKAPNLLIYTASTLQSGVPSRFSGSGS
GTDFTLTISSLQPEDFATYYCQQANIFPLTFGGGTKVEIK
ABT806 HC CD40 281 QVQLVQSGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLE mAb with WMGWINPDSGGTNYAQKFQGRVTMTRDTSISTAYMELNRLRSDDTAV tumor mAb YYCARDQPLGYCTNGVCSYFDYWGQGTLVTVSSASTKGPSVFPLAPSS ScFv KSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS
LSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCP
APELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYV
DGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK
ALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSD Antibody Region SEQ Sequence
ID
NO:
IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFS
CSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSEVQLQESGPGLVKPSQ
TLSLTCTVSGYSISRDFAWNWIRQPPGKGLEWMGYISYNGNTRYQPSLK
SRITISRDTSKNQFFLKLNSVTAADTATYYCVTASRGFPYWGQGTLVTV
SSGGGGSGGGGSGGGGSGGGGSDIQMTQSPSSMSVSVGDRVTITCHSSQ
DINSNIGWLQQKPGKSFKGLIYHGTNLDDGVPSRFSGSGSGTDYTLTISS
LQPEDFATYYCVQYAQFPWTFGGGTKLEIK
HC tumor 282 EVQLQESGPGLVKPSQTLSLTCTVSGYSISRDFAWNWIRQPPGKGLEWM mAb with GYISYNGNTRYQPSLKSRITISRDTSKNQFFLKLNSVTAADTATYYCVTA CD40 mAb SRGFPYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDY ScFv FPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYI
CNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPK
DTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQY
NSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPR
EPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKT
TPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSL
SLSPGGGGGSGGGGSQVQLVQSGAEVKKPGASVKVSCKASGYTFTGYY
MHWVRQAPGQGLEWMGWINPDSGGTNYAQKFQGRVTMTRDTSISTA
YMELNRLRSDDTAVYYCARDQPLGYCTNGVCSYFDYWGQGTLVTVSS
GGGGSGGGGSGGGGSGGGGSDIQMTQSPSSVSASVGDRVTITCRASQGI
YSWLAWYQQKPGKAPNLLIYTASTLQSGVPSRFSGSGSGTDFTLTISSLQ
PEDFATYYCQQANIFPLTFGGGTKVEIK
Sibrotuzuma HC CD40 294 QVQLVQSGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLE b mAb with WMGWINPDSGGTNYAQKFQGRVTMTRDTSISTAYMELNRLRSDDTAV tumor mAb YYCARDQPLGYCTNGVCSYFDYWGQGTLVTVSSASTKGPSVFPLAPSS ScFv KSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS
LSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCP
APELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYV
DGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK
ALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSD
IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFS
CSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSQVQLVQSGAEVKKPG
ASVKVSCKTSRYTFTEYTIHWVRQAPGQRLEWIGGINPNNGIPNYNQKF
KGRVTITVDTSASTAYMELSSLRSEDTAVYYCARRRIAYGYDEGHAMD
YWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSDIVMTQSPDSLAVSL
GERATINCKSSQSLLYSRNQKNYLAWYQQKPGQPPKLLIFWASTRESGV
PDRFSGSGFGTDFTLTISSLQAEDVAVYYCQQYFSYPLTFGQGTKVEIK
HC tumor 295 QVQLVQSGAEVKKPGASVKVSCKTSRYTFTEYTIHWVRQAPGQRLEWI mAb with GGINPNNGIPNYNQKFKGRVTITVDTSASTAYMELSSLRSEDTAVYYCA CD40 mAb RRRIAYGYDEGHAMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGG ScFv TAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVT
VPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLG
GPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVH
NAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIE
KTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWE
SNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHE
ALHNHYTQKSLSLSPGGGGGSGGGGSQVQLVQSGAEVKKPGASVKVSC
KASGYTFTGYYMHWVRQAPGQGLEWMGWINPDSGGTNYAQKFQGRV
TMTRDTSISTAYMELNRLRSDDTAVYYCARDQPLGYCTNGVCSYFDY
WGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSDIQMTQSPSSVSASVGD
RVTITCRASQGIYSWLAWYQQKPGKAPNLLIYTASTLQSGVPSRFSGSGS
GTDFTLTISSLQPEDFATYYCQQANIFPLTFGGGTKVEIK
DS-8895a HC CD40 307 QVQLVQSGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLE variant 1 mAb with WMGWINPDSGGTNYAQKFQGRVTMTRDTSISTAYMELNRLRSDDTAV tumor mAb YYCARDQPLGYCTNGVCSYFDYWGQGTLVTVSSASTKGPSVFPLAPSS ScFv KSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS
LSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCP
APELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYV
DGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK Antibody Region SEQ Sequence
ID
NO:
ALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSD
IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFS
CSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSQVQLVQSGAEVKKPG
ASVKVSCKASGYTFIDYSMHWVRQAPGQGLEWMGWINTYTGEPTYSD
DFKGRVTITADTSTSTAYLELSSLRSEDTAVYYCATYYRYERDFDYWG
QGTLVTVSSGGGGSGGGGSGGGGSGGGGSDIVMTQSPLSLPVTPGEPAS
ISCRSSQSIVHSSGITYLEWYLQKPGQSPQLLIYKVSNRFSGVPDRFSGSG
SGTDFTLKISRVEAEDVGVYYCFQGSHVPYTFGQGTKVEIK
HC tumor 308 QVQLVQSGAEVKKPGASVKVSCKASGYTFIDYSMHWVRQAPGQGLEW mAb with MGWINTYTGEPTYSDDFKGRVTITADTSTSTAYLELSSLRSEDTAVYYC CD40 mAb ATYYRYERDFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALG ScFv CLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSL
GTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFL
FPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTK
PREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKA
KGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPE
NNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHY
TQKSLSLSPGGGGGSGGGGSQVQLVQSGAEVKKPGASVKVSCKASGYT
FTGYYMHWVRQAPGQGLEWMGWINPDSGGTNYAQKFQGRVTMTRDT
SISTAYMELNRLRSDDTAVYYCARDQPLGYCTNGVCSYFDYWGQGTL
VTVSSGGGGSGGGGSGGGGSGGGGSDIQMTQSPSSVSASVGDRVTITCR
ASQGIYSWLAWYQQKPGKAPNLLIYTASTLQSGVPSRFSGSGSGTDFTL
TISSLQPEDFATYYCQQANIFPLTFGGGTKVEIK
DS-8895a HC CD40 320 QVQLVQSGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLE variant 2 mAb with WMGWINPDSGGTNYAQKFQGRVTMTRDTSISTAYMELNRLRSDDTAV tumor mAb YYCARDQPLGYCTNGVCSYFDYWGQGTLVTVSSASTKGPSVFPLAPSS ScFv KSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS
LSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCP
APELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYV
DGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK
ALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSD
IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFS
CSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSQIQLVQSGAEVKKPGA
SVKVSCKASGYTFIDYSMHWVRQAPGQGLKWMGWINTYTGEPTYSDD
FKGRFAFSLDTSTSTAYLELSSLRSEDTAVYYCATYYRYERDFDYWGQ
GTLVTVSSGGGGSGGGGSGGGSGGGGSDVLMTQSPLSLPVTPGEPASIS
CRSSQSIVHSSGITYLEWYLQKPGQSPQLLIYKVSNRFSGVPDRFSGSGS
GTDFTLKISRVEAEDVGVYYCFQGSHVPYTFGQGTKVEIK
HC tumor 321 QIQLVQSGAEVKKPGASVKVSCKASGYTFIDYSMHWVRQAPGQGLKW mAb with MGWINTYTGEPTYSDDFKGRFAFSLDTSTSTAYLELSSLRSEDTAVYYC CD40 mAb ATYYRYERDFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALG ScFv CLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSL
GTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFL
FPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTK
PREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKA
KGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPE
NNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHY
TQKSLSLSPGGGGGSGGGGSQVQLVQSGAEVKKPGASVKVSCKASGYT
FTGYYMHWVRQAPGQGLEWMGWINPDSGGTNYAQKFQGRVTMTRDT
SISTAYMELNRLRSDDTAVYYCARDQPLGYCTNGVCSYFDYWGQGTL
VTVSSGGGGSGGGGSGGGGSGGGGSDIQMTQSPSSVSASVGDRVTITCR
ASQGIYSWLAWYQQKPGKAPNLLIYTASTLQSGVPSRFSGSGSGTDFTL
TISSLQPEDFATYYCQQANIFPLTFGGGTKVEIK
MEDI-547 HC CD40 333 QVQLVQSGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLE mAb with WMGWINPDSGGTNYAQKFQGRVTMTRDTSISTAYMELNRLRSDDTAV tumor mAb YYCARDQPLGYCTNGVCSYFDYWGQGTLVTVSSASTKGPSVFPLAPSS ScFv KSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS
LSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCP
APELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYV Antibody Region SEQ Sequence
ID
NO:
DGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK
ALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSD
IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFS
CSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSEVQLLESGGGLVQPGG
SLRLSCAASGFTFSHYMMAWVRQAPGKGLEWVSRIGPSGGPTHYADSV
KGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAGYDSGYDYVAVAGP
AEYFQHWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSDIQMTQSPSSL
SASVGDRVTITCRASQSISTWLAWYQQKPGKAPKLLIYKASNLHTGVPS
RFSGSGSGTEFSLTISGLQPDDFATYYCQQYNSYSRTFGQGTKVEIK
HC tumor 334 EVQLLESGGGLVQPGGSLRLSCAASGFTFSHYMMAWVRQAPGKGLEW mAb with VSRIGPSGGPTHYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYC CD40 mAb AGYDSGYDYVAVAGPAEYFQHWGQGTLVTVSSASTKGPSVFPLAPSSK ScFv STSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSL
SSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAP
ELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDG
VEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKAL
PAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIA
VEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCS
VMHEALHNHYTQKSLSLSPGGGGGSGGGGSQVQLVQSGAEVKKPGAS
VKVSCKASGYTFTGYYMHWVRQAPGQGLEWMGWINPDSGGTNYAQK
FQGRVTMTRDTSISTAYMELNRLRSDDTAVYYCARDQPLGYCTNGVCS
YFDYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSDIQMTQSPSSVSA
SVGDRVTITCRASQGIYSWLAWYQQKPGKAPNLLIYTASTLQSGVPSRF
SGSGSGTDFTLTISSLQPEDFATYYCQQANIFPLTFGGGTKVEIK
Narnatumab HC CD40 346 QVQLVQSGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLE mAb with WMGWINPDSGGTNYAQKFQGRVTMTRDTSISTAYMELNRLRSDDTAV tumor mAb YYCARDQPLGYCTNGVCSYFDYWGQGTLVTVSSASTKGPSVFPLAPSS ScFv KSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS
LSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCP
APELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYV
DGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK
ALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSD
IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFS
CSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSEVQLVESGGGLVQPGG
SLRLSCAASGFTFSSYLMTWVRQAPGKGLEWVANIKQDGSEKYYVDSV
KGRFTISRDNAKNSLNLQMNSLRAEDTAVYYCTRDGYSSGRHYGMDV
WGQGTTVIVSSGGGGSGGGGSGGGGSGGGGSEIVLTQSPATLSLSPGER
ATLSCRASQSVSRYLAWYQQKPGQAPRLLIYDASNRATGIPARFSGSGS
GTDFTLTISSLEPEDFAVYYCQQRSNWPRTFGQGTKVEIK
HC tumor 347 EVQLVESGGGLVQPGGSLRLSCAASGFTFSSYLMTWVRQAPGKGLEWV mAb with ANIKQDGSEKYYVDSVKGRFTISRDNAKNSLNLQMNSLRAEDTAVYYC CD40 mAb TRDGYSSGRHYGMDVWGQGTTVIVSSASTKGPSVFPLAPSSKSTSGGTA ScFv ALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVP
SSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPS
VFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNA
KTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKT
ISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEAL
HNHYTQKSLSLSPGGGGGSGGGGSQVQLVQSGAEVKKPGASVKVSCK
ASGYTFTGYYMHWVRQAPGQGLEWMGWINPDSGGTNYAQKFQGRVT
MTRDTSISTAYMELNRLRSDDTAVYYCARDQPLGYCTNGVCSYFDYW
GQGTLVTVSSGGGGSGGGGSGGGGSGGGGSDIQMTQSPSSVSASVGDR
VTITCRASQGIYSWLAWYQQKPGKAPNLLIYTASTLQSGVPSRFSGSGS
GTDFTLTISSLQPEDFATYYCQQANIFPLTFGGGTKVEIK
RG7841 HC CD40 359 QVQLVQSGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLE mAb with WMGWINPDSGGTNYAQKFQGRVTMTRDTSISTAYMELNRLRSDDTAV tumor mAb YYCARDQPLGYCTNGVCSYFDYWGQGTLVTVSSASTKGPSVFPLAPSS ScFv KSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS
LSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCP Antibody Region SEQ Sequence
ID
NO:
APELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYV
DGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK
ALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSD
IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFS
CSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSEVQLVESGPALVKPTQ
TLTLTCTVSGFSLTGYSVNWIRQPPGKALEWLGMIWGDGSTDYNSALK
SRLTISKDTSKNQVVLTMTNMDPVDTATYYCARDYYFNYASWFAYWG
QGTLVTVSSGGGGSGGGGSGGGGSGGGGSDIQMTQSPSSLSASVGDRV
TITCSASQGISNYLNWYQQKPGKTVKLLIYYTSNLHSGVPSRFSGSGSGT
DYTLTISSLQPEDFATYYCQQYSELPWTFGQGTKVEIK
HC tumor 360 EVQLVESGPALVKPTQTLTLTCTVSGFSLTGYSVNWIRQPPGKALEWLG mAb with MIWGDGSTDYNSALKSRLTISKDTSKNQVVLTMTNMDPVDTATYYCA CD40 mAb RDYYFNYASWFAYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAA ScFv LGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPS
SSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPS
VFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNA
KTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKT
ISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEAL
HNHYTQKSLSLSPGGGGGSGGGGSQVQLVQSGAEVKKPGASVKVSCK
ASGYTFTGYYMHWVRQAPGQGLEWMGWINPDSGGTNYAQKFQGRVT
MTRDTSISTAYMELNRLRSDDTAVYYCARDQPLGYCTNGVCSYFDYW
GQGTLVTVSSGGGGSGGGGSGGGGSGGGGSDIQMTQSPSSVSASVGDR
VTITCRASQGIYSWLAWYQQKPGKAPNLLIYTASTLQSGVPSRFSGSGS
GTDFTLTISSLQPEDFATYYCQQANIFPLTFGGGTKVEIK
Farletuzuma HC CD40 372 QVQLVQSGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLE b mAb with WMGWINPDSGGTNYAQKFQGRVTMTRDTSISTAYMELNRLRSDDTAV tumor mAb YYCARDQPLGYCTNGVCSYFDYWGQGTLVTVSSASTKGPSVFPLAPSS ScFv KSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS
LSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCP
APELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYV
DGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK
ALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSD
IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFS
CSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSEVQLVESGGGVVQPG
RSLRLSCSASGFTFSGYGLSWVRQAPGKGLEWVAMISSGGSYTYYADS
VKGRFAISRDNAKNTLFLQMDSLRPEDTGVYFCARHGDDPAWFAYWG
QGTPVTVSSGGGGSGGGGSGGGGSGGGGSDIQLTQSPSSLSASVGDRVT
ITCSVSSSISSNNLHWYQQKPGKAPKPWIYGTSNLASGVPSRFSGSGSGT
DYTFTISSLQPEDIATYYCQQWSSYPYMYTFGQGTKVEIK
HC tumor 373 EVQLVESGGGVVQPGRSLRLSCSASGFTFSGYGLSWVRQAPGKGLEWV mAb with AMISSGGSYTYYADSVKGRFAISRDNAKNTLFLQMDSLRPEDTGVYFCA CD40 mAb RHGDDPAWFAYWGQGTPVTVSSASTKGPSVFPLAPSSKSTSGGTAALG ScFv CLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSL
GTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFL
FPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTK
PREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKA
KGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPE
NNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHY
TQKSLSLSPGGGGGSGGGGSQVQLVQSGAEVKKPGASVKVSCKASGYT
FTGYYMHWVRQAPGQGLEWMGWINPDSGGTNYAQKFQGRVTMTRDT
SISTAYMELNRLRSDDTAVYYCARDQPLGYCTNGVCSYFDYWGQGTL
VTVSSGGGGSGGGGSGGGGSGGGGSDIQMTQSPSSVSASVGDRVTITCR
ASQGIYSWLAWYQQKPGKAPNLLIYTASTLQSGVPSRFSGSGSGTDFTL
TISSLQPEDFATYYCQQANIFPLTFGGGTKVEIK
Mirvetuxima HC CD40 385 QVQLVQSGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLE b mAb with WMGWINPDSGGTNYAQKFQGRVTMTRDTSISTAYMELNRLRSDDTAV tumor mAb YYCARDQPLGYCTNGVCSYFDYWGQGTLVTVSSASTKGPSVFPLAPSS ScFv KSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS Antibody Region SEQ Sequence
ID
NO:
LSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCP
APELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYV
DGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK
ALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSD
IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFS
CSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSQVQLVQSGAEVVKPG
ASVKISCKASGYTFTGYFMNWVKQSPGQSLEWIGRIHPYDGDTFYNQK
FQGKATLTVDKSSNTAHMELLSLTSEDFAVYYCTRYDGSRAMDYWGQ
GTTVTVSSGGGGSGGGGSGGGGSGGGGSDIVLTQSPLSLAVSLGQPAIIS
CKASQSVSFAGTSLMHWYHQKPGQQPRLLIYRASNLEAGVPDRFSGSG
SKTDFTLTISPVEAEDAATYYCQQSREYPYTFGGGTKLEIK
HC tumor 386 QVQLVQSGAEVVKPGASVKISCKASGYTFTGYFMNWVKQSPGQSLEWI mAb with GRIHPYDGDTFYNQKFQGKATLTVDKSSNTAHMELLSLTSEDFAVYYC CD40 mAb TRYDGSRAMDYWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALG ScFv CLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSL
GTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFL
FPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTK
PREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKA
KGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPE
NNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHY
TQKSLSLSPGGGGGSGGGGSQVQLVQSGAEVKKPGASVKVSCKASGYT
FTGYYMHWVRQAPGQGLEWMGWINPDSGGTNYAQKFQGRVTMTRDT
SISTAYMELNRLRSDDTAVYYCARDQPLGYCTNGVCSYFDYWGQGTL
VTVSSGGGGSGGGGSGGGGSGGGGSDIQMTQSPSSVSASVGDRVTITCR
ASQGIYSWLAWYQQKPGKAPNLLIYTASTLQSGVPSRFSGSGSGTDFTL
TISSLQPEDFATYYCQQANIFPLTFGGGTKVEIK
J591 HC CD40 398 QVQLVQSGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLE variantl mAb with WMGWINPDSGGTNYAQKFQGRVTMTRDTSISTAYMELNRLRSDDTAV tumor mAb YYCARDQPLGYCTNGVCSYFDYWGQGTLVTVSSASTKGPSVFPLAPSS ScFv KSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS
LSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCP
APELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYV
DGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK
ALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSD
IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFS
CSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSEVQLQQSGPELKKPGT
SVRISCKTSGYTFTEYTIHWVKQSHGKSLEWIGNINPNNGGTTYNQKFE
DKATLTVDKSSSTAYMELRSLTSEDSAVYYCAAGWNFDYWGQGTTLT
VSSGGGGSGGGGSGGGGSGGGGSDIVMTQSHKFMSTSVGDRVSIICKAS
QDVGTAVDWYQQKPGQSPKLLIYWASTRHTGVPDRFTGSGSGTDFTLT
ITNVQSEDLADYFCQQYNSYPLTFGAGTMLDLK
HC tumor 399 EVQLQQSGPELKKPGTSVRISCKTSGYTFTEYTIHWVKQSHGKSLEWIG mAb with NINPNNGGTTYNQKFEDKATLTVDKSSSTAYMELRSLTSEDSAVYYCA CD40 mAb AGWNFDYWGQGTTLTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVK ScFv DYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQT
YICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKP
KDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQ
YNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQP
REPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYK
TTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKS
LSLSPGGGGGSGGGGSQVQLVQSGAEVKKPGASVKVSCKASGYTFTGY
YMHWVRQAPGQGLEWMGWINPDSGGTNYAQKFQGRVTMTRDTSIST
AYMELNRLRSDDTAVYYCARDQPLGYCTNGVCSYFDYWGQGTLVTVS
SGGGGSGGGGSGGGGSGGGGSDIQMTQSPSSVSASVGDRVTITCRASQG
IYSWLAWYQQKPGKAPNLLIYTASTLQSGVPSRFSGSGSGTDFTLTISSL
QPEDFATYYCQQANIFPLTFGGGTKVEIK
J591 variant HC CD40 411 QVQLVQSGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLE 2 mAb with WMGWINPDSGGTNYAQKFQGRVTMTRDTSISTAYMELNRLRSDDTAV tumor mAb YYCARDQPLGYCTNGVCSYFDYWGQGTLVTVSSASTKGPSVFPLAPSS Antibody Region SEQ Sequence
ID
NO:
ScFv KSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS
LSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCP
APELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYV
DGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK
ALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSD
IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFS
CSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSEVQLQQSGPELVKPGT
SVRISCKTSGYTFTEYTIHWVKQSHGKSLEWIGNINPNNGGTTYNQKFE
DKATLTVDKSSSTAYMELRSLTSEDSAVYYCAAGWNFDYWGQGTTLT
VSSGGGGSGGGGSGGGGSGGGGSNIVMTQSPKSMSMSVGERVTLTCKA
SENVVTYVSWYQQKPEQSPKLLIYGASNRYTGVPDRFTGSGSATDFTLT
ISSVQAEDLADYHCGQGYSYPYTFGGGTKLEIK
HC tumor 412 EVQLQQSGPELVKPGTSVRISCKTSGYTFTEYTIHWVKQSHGKSLEWIG mAb with NINPNNGGTTYNQKFEDKATLTVDKSSSTAYMELRSLTSEDSAVYYCA CD40 mAb AGWNFDYWGQGTTLTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVK ScFv DYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQT
YICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKP
KDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQ
YNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQP
REPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYK
TTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKS
LSLSPGGGGGSGGGGSQVQLVQSGAEVKKPGASVKVSCKASGYTFTGY
YMHWVRQAPGQGLEWMGWINPDSGGTNYAQKFQGRVTMTRDTSIST
AYMELNRLRSDDTAVYYCARDQPLGYCTNGVCSYFDYWGQGTLVTVS
SGGGGSGGGGSGGGGSGGGGSDIQMTQSPSSVSASVGDRVTITCRASQG
IYSWLAWYQQKPGKAPNLLIYTASTLQSGVPSRFSGSGSGTDFTLTISSL
QPEDFATYYCQQANIFPLTFGGGTKVEIK
Rovalpituzu HC CD40 424 QVQLVQSGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLE mab mAb with WMGWINPDSGGTNYAQKFQGRVTMTRDTSISTAYMELNRLRSDDTAV tumor mAb YYCARDQPLGYCTNGVCSYFDYWGQGTLVTVSSASTKGPSVFPLAPSS ScFv KSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS
LSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCP
APELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYV
DGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK
ALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSD
IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFS
CSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSQVQLVQSGAEVKKPG
ASVKVSCKASGYTFTNYGMNWVRQAPGQGLEWMGWINTYTGEPTYA
DDFKGRVTMTTDTSTSTAYMELRSLRSDDTAVYYCARIGDSSPSDYWG
QGTLVTVSSGGGGSGGGGSGGGGSGGGGSEIVMTQSPATLSVSPGERAT
LSCKASQSVSNDVVWYQQKPGQAPRLLIYYASNRYTGIPARFSGSGSGT
EFTLTISSLQSEDFAVYYCQQDYTSPWTFGQGTKLEIK
HC tumor 425 QVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGMNWVRQAPGQGLE mAb with WMGWINTYTGEPTYADDFKGRVTMTTDTSTSTAYMELRSLRSDDTAV CD40 mAb YYCARIGDSSPSDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAAL ScFv GCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSS
SLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSV
FLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAK
TKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTIS
KAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNG
QPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALH
NHYTQKSLSLSPGGGGGSGGGGSQVQLVQSGAEVKKPGASVKVSCKAS
GYTFTGYYMHWVRQAPGQGLEWMGWINPDSGGTNYAQKFQGRVTMT
RDTSISTAYMELNRLRSDDTAVYYCARDQPLGYCTNGVCSYFDYWGQ
GTLVTVSSGGGGSGGGGSGGGGSGGGGSDIQMTQSPSSVSASVGDRVTI
TCRASQGIYSWLAWYQQKPGKAPNLLIYTASTLQSGVPSRFSGSGSGTD
FTLTISSLQPEDFATYYCQQANIFPLTFGGGTKVEIK
PF- HC CD40 437 QVQLVQSGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLE
06647020 mAb with WMGWINPDSGGTNYAQKFQGRVTMTRDTSISTAYMELNRLRSDDTAV Antibody Region SEQ Sequence
ID
NO:
tumor mAb YYCARDQPLGYCTNGVCSYFDYWGQGTLVTVSSASTKGPSVFPLAPSS ScFv KSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS
LSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCP
APELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYV
DGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK
ALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSD
IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFS
CSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSQVQLVQSGPEVKKPG
ASVKVSCKASGYTFTDYAVHWVRQAPGKRLEWIGVISTYNDYTYNNQ
DFKGRVTMTRDTSASTAYMELSRLRSEDTAVYYCARGNSYFYALDYW
GQGTSVTVSSGGGGSGGGGSGGGGSGGGGSEIVLTQSPATLSLSPGERA
TLSCRASESVDSYGKSFMHWYQQKPGQAPRLLIYRASNLESGIPARFSG
SGSGTDFTLTISSLEPEDFAVYYCQQSNEDPWTFGGGTKLEIK
HC tumor 438 QVQLVQSGPEVKKPGASVKVSCKASGYTFTDYAVHWVRQAPGKRLEW mAb with IGVISTYNDYTYNNQDFKGRVTMTRDTSASTAYMELSRLRSEDTAVYY CD40 mAb CARGNSYFYALDYWGQGTSVTVSSASTKGPSVFPLAPSSKSTSGGTAAL ScFv GCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSS
SLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSV
FLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAK
TKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTIS
KAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNG
QPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALH
NHYTQKSLSLSPGGGGGSGGGGSQVQLVQSGAEVKKPGASVKVSCKAS
GYTFTGYYMHWVRQAPGQGLEWMGWINPDSGGTNYAQKFQGRVTMT
RDTSISTAYMELNRLRSDDTAVYYCARDQPLGYCTNGVCSYFDYWGQ
GTLVTVSSGGGGSGGGGSGGGGSGGGGSDIQMTQSPSSVSASVGDRVTI
TCRASQGIYSWLAWYQQKPGKAPNLLIYTASTLQSGVPSRFSGSGSGTD
FTLTISSLQPEDFATYYCQQANIFPLTFGGGTKVEIK
Antibody to HC CD40 450 QVQLVQSGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLE PTK7 mAb with WMGWINPDSGGTNYAQKFQGRVTMTRDTSISTAYMELNRLRSDDTAV tumor mAb YYCARDQPLGYCTNGVCSYFDYWGQGTLVTVSSASTKGPSVFPLAPSS ScFv KSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS
LSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCP
APELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYV
DGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK
ALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSD
IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFS
CSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSQVQLVESGGGVVQPG
RSLRLSCAASGFTFSSYAFHWVRQAPGKGLEWVAVISYDGSIKYYADSV
KGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARTYYFDYWGQGTLV
TVSSGGGGSGGGGSGGGGSGGGGSEIVLTQSPDFQSVTPKEKVTITCRA
SQSIGSSLHWYQQKPDQSPKLLIKYASQSFSGVPSRFSGSGSGTDFTLTIN
SLEAEDAAAYYCHQSSSLPITFGQGTRLEIK
HC tumor 451 QVQLVESGGGVVQPGRSLRLSCAASGFTFSSYAFHWVRQAPGKGLEWV mAb with AVISYDGSIKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCA CD40 mAb RTYYFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKD ScFv YFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTY
ICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPK
DTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQY
NSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPR
EPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKT
TPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSL
SLSPGGGGGSGGGGSQVQLVQSGAEVKKPGASVKVSCKASGYTFTGYY
MHWVRQAPGQGLEWMGWINPDSGGTNYAQKFQGRVTMTRDTSISTA
YMELNRLRSDDTAVYYCARDQPLGYCTNGVCSYFDYWGQGTLVTVSS
GGGGSGGGGSGGGGSGGGGSDIQMTQSPSSVSASVGDRVTITCRASQGI
YSWLAWYQQKPGKAPNLLIYTASTLQSGVPSRFSGSGSGTDFTLTISSLQ
PEDFATYYCQQANIFPLTFGGGTKVEIK
SGN-LIV1A HC CD40 463 QVQLVQSGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLE Antibody Region SEQ Sequence
ID
NO:
mAb with WMGWINPDSGGTNYAQKFQGRVTMTRDTSISTAYMELNRLRSDDTAV tumor mAb YYCARDQPLGYCTNGVCSYFDYWGQGTLVTVSSASTKGPSVFPLAPSS ScFv KSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS
LSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCP
APELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYV
DGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK
ALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSD
IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFS
CSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSQVQLVQSGAEVKKPG
ASVKVSCKASGLTIEDYYMHWVRQAPGQGLEWMGWIDPENGDTEYGP
KFQGRVTMTRDTSINTAYMELSRLRSDDTAVYYCAVHNAHYGTWFAY
WGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSDVVMTQSPLSLPVTLG
QPASISCRSSQSLLHSSGNTYLEYFQQRPGQSPRPLIYKISTRFSGVPDRFS
GSGSGTDFTLKISRVEAEDVGVYYCFQGSHVPYTFGGGTKVEIK
HC tumor 464 QVQLVQSGAEVKKPGASVKVSCKASGLTIEDYYMHWVRQAPGQGLEW mAb with MGWIDPENGDTEYGPKFQGRVTMTRDTSINTAYMELSRLRSDDTAVYY CD40 mAb CAVHNAHYGTWFAYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTA ScFv ALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVP
SSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPS
VFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNA
KTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKT
ISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEAL
HNHYTQKSLSLSPGGGGGSGGGGSQVQLVQSGAEVKKPGASVKVSCK
ASGYTFTGYYMHWVRQAPGQGLEWMGWINPDSGGTNYAQKFQGRVT
MTRDTSISTAYMELNRLRSDDTAVYYCARDQPLGYCTNGVCSYFDYW
GQGTLVTVSSGGGGSGGGGSGGGGSGGGGSDIQMTQSPSSVSASVGDR
VTITCRASQGIYSWLAWYQQKPGKAPNLLIYTASTLQSGVPSRFSGSGS
GTDFTLTISSLQPEDFATYYCQQANIFPLTFGGGTKVEIK
Cirmtuzuma HC CD40 476 QVQLVQSGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLE b mAb with WMGWINPDSGGTNYAQKFQGRVTMTRDTSISTAYMELNRLRSDDTAV tumor mAb YYCARDQPLGYCTNGVCSYFDYWGQGTLVTVSSASTKGPSVFPLAPSS ScFv KSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS
LSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCP
APELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYV
DGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK
ALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSD
IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFS
CSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSQVQLQESGPGLVKPSQ
TLSLTCTVSGYAFTAYNIHWVRQAPGQGLEWMGSFDPYDGGSSYNQKF
KDRLTISKDTSKNQVVLTMTNMDPVDTATYYCARGWYYFDYWGHGT
LVTVSSGGGGSGGGGSGGGGSGGGGSDIVMTQTPLSLPVTPGEPASISC
RASKSISKYLAWYQQKPGQAPRLLIYSGSTLQSGIPPRFSGSGYGTDFTL
TINNIESEDAAYYFCQQHDESPY TFGEGTKVEIK
HC tumor 477 QVQLQESGPGLVKPSQTLSLTCTVSGYAFTAYNIHWVRQAPGQGLEWM mAb with GSFDPYDGGSSYNQKFKDRLTISKDTSKNQVVLTMTNMDPVDTATYYC CD40 mAb ARGWYYFDYWGHGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCL ScFv VKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGT
QTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPP
KPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPRE
EQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKG
QPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENN
YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQ
KSLSLSPGGGGGSGGGGSQVQLVQSGAEVKKPGASVKVSCKASGYTFT
GYYMHWVRQAPGQGLEWMGWINPDSGGTNYAQKFQGRVTMTRDTSI
STAYMELNRLRSDDTAVYYCARDQPLGYCTNGVCSYFDYWGQGTLVT
VSSGGGGSGGGGSGGGGSGGGGSDIQMTQSPSSVSASVGDRVTITCRAS
QGIYSWLAWYQQKPGKAPNLLIYTASTLQSGVPSRFSGSGSGTDFTLTIS
SLQPEDFATYYCQQANIFPLTFGGGTKVEIK Antibody Region SEQ Sequence
ID
NO:
Antibody to HC CD40 489 QVQLVQSGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLE MAGE-A3 mAb with WMGWINPDSGGTNYAQKFQGRVTMTRDTSISTAYMELNRLRSDDTAV tumor mAb YYCARDQPLGYCTNGVCSYFDYWGQGTLVTVSSASTKGPSVFPLAPSS ScFv KSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS
LSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCP
APELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYV
DGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK
ALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSD
IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFS
CSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSQVQLVESGGGVVQPG
RSLRLSCTASGFRFRSHGMHWVRQAPGKGLEWVAVISYDGNNKLYAD
SVKGRITISRDNSKNTLFLQMNNVRAEDTAVYYCASPYTSDWQYFQYW
GQGTLVIVSSGGGGSGGGGSGGGGSGGGGSEIVMTQSPATLSVSPGERA
TFSCRASQNISTTLAWYQQKPGQAPRLLIYDTSTRATGIPARFSGSGSGT
EFTLTISSLQSEDLAVYYCQQSNSWPLTFGGGTKVEIK
HC tumor 490 QVQLVESGGGVVQPGRSLRLSCTASGFRFRSHGMHWVRQAPGKGLEW mAb with VAVISYDGNNKLYADSVKGRITISRDNSKNTLFLQMNNVRAEDTAVYY CD40 mAb CASPYTSDWQYFQYWGQGTLVIVSSASTKGPSVFPLAPSSKSTSGGTAA ScFv LGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPS
SSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPS
VFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNA
KTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKT
ISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEAL
HNHYTQKSLSLSPGGGGGSGGGGSQVQLVQSGAEVKKPGASVKVSCK
ASGYTFTGYYMHWVRQAPGQGLEWMGWINPDSGGTNYAQKFQGRVT
MTRDTSISTAYMELNRLRSDDTAVYYCARDQPLGYCTNGVCSYFDYW
GQGTLVTVSSGGGGSGGGGSGGGGSGGGGSDIQMTQSPSSVSASVGDR
VTITCRASQGIYSWLAWYQQKPGKAPNLLIYTASTLQSGVPSRFSGSGS
GTDFTLTISSLQPEDFATYYCQQANIFPLTFGGGTKVEIK
Antibody to HC CD40 502 QVQLVQSGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLE NY-ESO-1 mAb with WMGWINPDSGGTNYAQKFQGRVTMTRDTSISTAYMELNRLRSDDTAV tumor mAb YYCARDQPLGYCTNGVCSYFDYWGQGTLVTVSSASTKGPSVFPLAPSS ScFv KSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS
LSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCP
APELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYV
DGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK
ALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSD
IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFS
CSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSQVQLVQSGGGVVRPG
GSLRLSCAASGFSFIDYGMSWVRQVPGKGLEWVAGMNWSGDKKGHA
ESVKGRFIISRDNAKNTLYLEMSSLRVEDTALYFCARGEYSNRFDPRGR
GTLVTVSSGGGGSGGGGSGGGGSGGGGSDIVMTQTPLSLPVTLGQPASL
SCRSSQSLVFTDGNTYLNWFQQRPGQSPRRLIYKVSSRDPGVPDRFSGT
GSGTDFTLEISRVEAEDIGVYYCMQGTHWPPIFGQGTKVEIK
HC tumor 503 QVQLVQSGGGVVRPGGSLRLSCAASGFSFIDYGMSWVRQVPGKGLEW mAb with VAGMNWSGDKKGHAESVKGRFIISRDNAKNTLYLEMSSLRVEDTALYF CD40 mAb CARGEYSNRFDPRGRGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGC ScFv LVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLG
TQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFP
PKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPR
EEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAK
GQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPEN
NYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYT
QKSLSLSPGGGGGSGGGGSQVQLVQSGAEVKKPGASVKVSCKASGYTF
TGYYMHWVRQAPGQGLEWMGWINPDSGGTNYAQKFQGRVTMTRDTS
ISTAYMELNRLRSDDTAVYYCARDQPLGYCTNGVCSYFDYWGQGTLV
TVSSGGGGSGGGGSGGGGSGGGGSDIQMTQSPSSVSASVGDRVTITCRA
SQGIYSWLAWYQQKPGKAPNLLIYTASTLQSGVPSRFSGSGSGTDFTLTI Antibody Region SEQ Sequence
ID
NO:
SSLQPEDFATYYCQQANIFPLTFGGGTKVEIK
Trastuzumab HC CD40 683 QVQLVQSGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLE mAb with WMGWINPDSGGTNYAQKFQGRVTMTRDTSISTAYMELNRLRSDDTAV tumor mAb YYCARDQPLGYCTNGVCSYFDYWGQGTLVTVSSASTKGPSVFPLAPSS ScFv KSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS
LSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCP
APELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYV
DGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK
ALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSD
IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFS
CSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSEVQLVESGGGLVQPGG
SLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYADSV
KGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYW
GQGTLVTVSSGGGGSGGGGSGGGGSGGGGSDIQMTQSPSSLSASVGDR
VTITCRASQDVNTAVAWYQQKPGKAPKLLIYSASFLYSGVPSRFSGSRS
GTDFTLTISSLQPEDFATYYCQQHYTTPPTFGQGTKVEIK
HC tumor 684 EVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWV mAb with ARIYPTNGYTRYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYC CD40 mAb SRWGGDGFYAMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAA ScFv LGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPS
SSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPS
VFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNA
KTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKT
ISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEAL
HNHYTQKSLSLSPGGGGGSGGGGSQVQLVQSGAEVKKPGASVKVSCK
ASGYTFTGYYMHWVRQAPGQGLEWMGWINPDSGGTNYAQKFQGRVT
MTRDTSISTAYMELNRLRSDDTAVYYCARDQPLGYCTNGVCSYFDYW
GQGTLVTVSSGGGGSGGGGSGGGGSGGGGSDIQMTQSPSSVSASVGDR
VTITCRASQGIYSWLAWYQQKPGKAPNLLIYTASTLQSGVPSRFSGSGS
GTDFTLTISSLQPEDFATYYCQQANIFPLTFGGGTKVEIK
HC CD40 796 QVQLVQSGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLE mAb with WMGWINPDSGGTNYAQKFQGRVTMTRDTSISTAYMELNRLRSDDTAV tumor mAb YYCARDQPLGYCTNGVCSYFDYWGQGTLVTVSSASTKGPSVFPLAPSS ScFv KSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS
(LH,25mer) LSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCP
APELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYV
DGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK
ALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSD
IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFS
CSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSDIQMTQSPSSLSASVG
DRVTITCRASQDVNTAVAWYQQKPGKAPKLLIYSASFLYSGVPSRFSGS
RSGTDFTLTISSLQPEDFATYYCQQHYTTPPTFGQGTKVEIKGGGGSGGG
GSGGGGSGGGGSGGGGSEVQLVESGGGLVQPGGSLRLSCAASGFNIKD
TYIHWVRQAPGKGLEWVARIYPTNGYTRYADSVKGRFTISADTSKNTA
YLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSS
TABLE 5. APC Antibody CDRs
Antibody Region SEQ ID NO: Sequence
CP-8709893 HCDR1 3 GYTFTGYY
HCDR2 4 INPDSGGT
HCDR3 5 ARDQPLGYCTNGVCSYFDY
LCDR1 8 QGIYSW
LCDR2 9 TAS
LCDR3 10 QQANIFPLT
SBT-040 (G1/G2) HCDR1 3 GYTFTGYY
HCDR2 4 INPDSGGT Antibody Region SEQ ID NO: Sequence
HCDR3 5 ARDQPLGYCTNGVCSYFDY
LCDR1 8 QGIYSW
LCDR2 9 TAS
LCDR3 10 QQANIFPLT
Dacetuzumab HCDR1 582 GYSFTGYY
HCDR2 583 VIPNAGGT
HCDR3 584 AREGIYW
LCDR1 587 QSLVHSNGNTF
LCDR2 588 TVS
LCDR3 589 SQTTHVPWT
Bleselumab HCDR1 592 GGSISSPGYY
HCDR2 593 IYKSGST
HCDR3 594 TRPVVRYFGWFDP
LCDR1 597 QGISSA
LCDR2 598 DAS
LCDR3 599 QQFNSYPT
lucatumumab HCDR1 602 GFTFSSYG
HCDR2 603 ISYEESNR
HCDR3 604 ARDGGIAAPGPDY
LCDR1 607 QSLLYSNGYNY
LCDR2 608 LGS
LCDR3 609 MQARQTPFT
ADC-1013 HCDR1 612 GFTFSTYG
HCDR2 613 ISGGSSYI
HCDR3 614 ARILRGGSGMDL
LCDR1 617 SSNIGAGYN
LCDR2 618 GNI
LCDR3 619 AAWDKSISGLV
APX005 HCDR1 622 GFSFSSTY
HCDR2 623 IYTGDGTN
HCDR3 624 ARPDITYGFAINFW
LCDR1 627 QSISSR
LCDR2 628 RAS
LCDR3 629 QCTGYGISWP
Chi Lob 7/4 HCDR1 632 GYTFTEYI
HCDR2 633 IIPNNGGT
HCDR3 634 TRREVYGRNYYALDY
LCDR1 637 QGINNY
LCDR2 638 YTS
LCDR3 639 QQYSNLPYT
DEC-205 variant 1 HCDR1 234 GFTFSNYG
HCDR2 235 IWYDGSNK
HCDR3 236 ARDLWGWYFDY
LCDR1 239 QSVSSY
LCDR2 240 DAS
LCDR3 241 QQRRNWPLT
DEC-205 variant 2 HCDR1 247 GDSFTTYW
HCDR2 248 IYPGDSDT
HCDR3 249 TRGDRGVDY
LCDR1 252 QGISRW
LCDR2 253 AAS
LCDR3 254 QQYNSYPRT
DC-SIGN variant 1 HCDR1 640 QHFWNTPWT
HCDR2 641 QQGHTLPYT
HCDR3 642 SNDGYYS
LCDR1 643 RYYLGVD
LCDR2 644 DDSGRFP
LCDR3 645 YGYAVDY
DC-SIGN variant 2 HCDR1 646 YYGIYVDY Antibody Region SEQ ID NO: Sequence
HCDR2 647 FLVY
HCDR3 648 NFGILGY
LCDR1 649 YPNALDY
LCDR2 650 GLKSFYAMDH
LCDR3 651 QQGKTLPWT
DC-SIGN variant 3 HCDR1 652 QQGNTLPPT
HCDR2 653 QQHYITPLT
HCDR3 654 QQYGNLPYT
LCDR1 655 QQYYSTPRT
LCDR2 656 GQSYNYPPT
LCDR3 657 WQDTHFPHV
TABLE 6. APC Antibody VH sequences and VL sequences
Antibody Region SEQ Sequence
ID
NO:
CP-8709893 VH 2 QVQLVQSGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLEWM
GWINPDSGGTNYAQKFQGRVTMTRDTSISTAYMELNRLRSDDTAVYYCAR DQPLGYCTNGVCSYFDYWGQGTLVTVSS
VL 7 DIQMTQSPSSVSASVGDRVTITCRASQGIYSWLAWYQQKPGKAPNLLIYTA
STLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQANIFPLTFGGGTKV EIK
SBT-040 VH 2 QVQLVQSGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLEWM
GWINPDSGGTNYAQKFQGRVTMTRDTSISTAYMELNRLRSDDTAVYYCAR DQPLGYCTNGVCSYFDYWGQGTLVTVSS
VL 7 DIQMTQSPSSVSASVGDRVTITCRASQGIYSWLAWYQQKPGKAPNLLIYTA
STLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQANIFPLTFGGGTKV EIK
Dacetuzumab VH 581 EVQLVESGGGLVQPGGSLRLSCAASGYSFTGYYIHWVRQAPGKGLEWVAR
VIPNAGGTSYNQKFKGRFTLSVDNSKNTAYLQMNSLRAEDTAVYYCAREG IYWWGQGTLVTVSS
VL 586 DIQMTQSPSSLSASVGDRVTITCRSSQSLVHSNGNTFLHWYQQKPGKAPKL
LIYTVSNRFSGVPSRFSGSGSGTDFTLTISSLQPEDFATYFCSQTTHVPWTFG QGTKVEIK
Bleselumab VH 591 QLQLQESGPGLLKPSETLSLTCTVSGGSISSPGYYGGWIRQPPGKGLEWIGSI
YKSGSTYHNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCTRPVVRY FGWFDPWGQGTLVTVSS
VL 596 AIQLTQSPSSLSASVGDRVTITCRASQGISSALAWYQQKPGKAPKLLIYDAS
NLESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQFNSYPTFGQGTKVEI
K
Lucatumuma VH 601 QVQLVESGGGVVQPGRSLRLSCAASGFTFSSYGMHWVRQAPGKGLEWVA b VISYEESNRYHADSVKGRFTISRDNSKITLYLQMNSLRTEDTAVYYCARDG
GIAAPGPDYWGQGTLVTVSS
VL 606 DIVMTQSPLSLTVTPGEPASISCRSSQSLLYSNGYNYLDWYLQKPGQSPQVL
ISLGSNRASGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCMQARQTPFTFG PGTKVDIR
ADC-1013 VH 611 EVQLLESGGGLVQPGGSLRLSCAASGFTFSTYGMHWVRQAPGKGLEWLSY
ISGGSSYIFYADSVRGRFTISRDNSENALYLQMNSLRAEDTAVYYCARILRG GSGMDLWGQGTLVTVSS
VL 616 QSVLTQPPSASGTPGQRVTISCTGSSSNIGAGYNVYWYQQLPGTAPKLLIYG
NINRPSGVPDRFSGSKSGTSASLAISGLRSEDEADYYCAAWDKSISGLVFGG GTKLTVL
APX005 VH 621 QVQLVESGGGVVQPGRSLRLSCAASGFSFSSTYVCWVRQAPGKGLEWIACI
YTGDGTNYSASWAKGRFTISKDSSKNTVYLQMNSLRAEDTAVYFCARPDI
TYGFAINFWGPGTLVTVSS
VL 626 DIQMTQSPSSLSASVGDRVTIKCQASQSISSRLAWYQQKPGKPPKLLIYRAS
TLASGVPSRFSGSGSGTDFTLTISSLQPEDVATYYCQCTGYGISWPIGGGTK Antibody Region SEQ Sequence
ID
NO:
VEIK
Chi Lob 7/4 VH 631 EVQLQQSGPDLVKPGASVKISCKTSGYTFTEYIMHWVKQSHGKSLEWIGGI
IPNNGGTSYNQKFKDKATMTVDKSSSTGYMELRSLTSEDSAVYYCTRREV YGRNYYALDYWGQGTLVTVSS
VL 636 DIQMTQTTSSLSASLGDRVTITCSASQGINNYLNWYQQKPDGTVKLLIYYTS
SLHSGVPSRFSGSGSGTDYSLTISNLEPEDIATYYCQQYSNLPYTFGGGTKLE IK
VH 233 QVQLVESGGGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGLEWVA
DEC-205 VIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAR variant 1 DLWGWYFDYWGQGTLVTVSS
VL 238 EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLIYDAS
NRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRRNWPLTFGGGTKV EIK
VH 246 EVQLVQSGAEVKKPGESLRISCKGSGDSFTTYWIGWVRQMPGKGLEWMGI
DEC-205 IYPGDSDTIYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCTRGDRG variant 2 VDYWGQGTLVTVSS
VL 251 DIQMTQSPSSLSASVGDRVTITCRASQGISRWLAWYQQKPEKAPKSLIYAAS
SLQSGVPSRFSGSGSGTDFTLTISGLQPEDFATYYCQQYNSYPRTFGQGTKV EIK
VH 658 DIQMTQSPSSLSASVGDRVTITCRASQGISRWLAWYQQKPEKAPKSLIYAAS
CD36 SLQSGVPSRFSGSGSGTDFTLTISGLQPEDFATYYCQQYNSYPRTFGQGTKV mannose EIK
Scavenger VL 659 EVQLVQSGAEVKKPGESLRISCKGSGDSFTTYWIGWVRQMPGKGLEWMGI Receptor IYPGDSDTIYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCTRGDRG
VDYWGQGTLVTVSS
VH 660 QIVESGGGLVQPKESLKISCTASGFTFSNAAIYWVRQTPGKGLEWVGRIRTR
CLEC9A PSKYATDYADSVRGRFTISRDDSKSMVYLQMDNLRTEDTAMYYCTPRATE
DVPFYWGQGVMVTVSS
VL 661 DIVMTQTPSSQAVSAGEKVTMNCKSSQSVLYDENKKNYLAWYQQKSGQS
PKLLIYWASTGESGVPDRFIGSGSGTDFTLTISSVQAEDLAVYYCQQYYDFP PTFGGGTK
TABLE 7. APC Antibody Heavy Chain and Light Chain sequences
Antibody Region SEQ Sequence
ID
NO:
CP-8709893 Heavy 1 QVQLVQSGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLEWM
Chain GWINPDSGGTNYAQKFQGRVTMTRDTSISTAYMELNRLRSDDTAVYYCAR
DQPLGYCTNGVCSYFDYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTA
ALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSS
NFGTQTYTCNVDHKPSNTKVDKTVERKCCVECPPCPAPPVAGPSVFLFPPK
PKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYVDGVEVHNAKTKPREEQF
NSTFRVVSVLTVVHQDWLNGKEYKCKVSNKGLPAPIEKTISKTKGQPREPQ
VYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPML
DSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK
Light 6 DIQMTQSPSSVSASVGDRVTITCRASQGIYSWLAWYQQKPGKAPNLLIYTA Chain STLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQANIFPLTFGGGTKV
EIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQ SGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVT KSFNRGEC
SBT-040 Heavy 577 MDWTWRILFLVAAATGAHSQVQLVQSGAEVKKPGASVKVSCKASGYTFT
Chain GYYMHWVRQAPGQGLEWMGWINPDSGGTNYAQKFQGRVTMTRDTSIST (IgGl) AYMELNRLRSDDTAVYYCARDQPLGYCTNGVCSYFDYWGQGTLVTVSSA
STKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTF
PAVLQSSGLYSLSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVEPKSCD
KTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEV
KFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKC Antibody Region SEQ Sequence
ID
NO:
KVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFY PSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFS CSVMHEALHNHYTQKSLSLSPGK
Heavy 578 MDWTWRILFLVAAATGAHSQVQLVQSGAEVKKPGASVKVSCKASGYTFT Chain GYYMHWVRQAPGQGLEWMGWINPDSGGTNYAQKFQGRVTMTRDTSIST (IgG2) AYMELNRLRSDDTAVYYCARDQPLGYCTNGVCSYFDYWGQGTLVTVSSA
STKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTF
PAVLQSSGLYSLSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKCCV
ECPPCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNW
YVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNK
GLPAPIEKTISKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIA
VEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVM
HEALHNHYTQKSLSLSPGK
Light 579 DIQMTQSPSSVSASVGDRVTITCRASQGIYSWLAWYQQKPGKAPNLLIYTA Chain STLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQANIFPLTFGGGTKV
EIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQ SGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVT KSFNRGEC
Dacetuzumab Heavy 580 EVQLVESGGGLVQPGGSLRLSCAASGYSFTGYYIHWVRQAPGKGLEWVAR
Chain VIPNAGGTSYNQKFKGRFTLSVDNSKNTAYLQMNSLRAEDTAVYYCAREG
IYWWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPV
TVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKP
SNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPE
VTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLT
VLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEM
TKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSK
LTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK
Light 585 DIQMTQSPSSLSASVGDRVTITCRSSQSLVHSNGNTFLHWYQQKPGKAPKL Chain LIYTVSNRFSGVPSRFSGSGSGTDFTLTISSLQPEDFATYFCSQTTHVPWTFG
QGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKV DNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQG LSSPVTKSFNRGEC
Bleselumab Heavy 590 QLQLQESGPGLLKPSETLSLTCTVSGGSISSPGYYGGWIRQPPGKGLEWIGSI
Chain YKSGSTYHNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCTRPVVRY
FGWFDPWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFP
EPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNV
DHKPSNTKVDKRVESKYGPPCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTP
EVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVL
TVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEE
MTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYS
RLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK
Light 595 AIQLTQSPSSLSASVGDRVTITCRASQGISSALAWYQQKPGKAPKLLIYDAS Chain NLESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQFNSYPTFGQGTKVEI
KRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSG
NSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKS
FNRGEC
Lucatumuma Heavy 600 QVQLVESGGGVVQPGRSLRLSCAASGFTFSSYGMHWVRQAPGKGLEWVA b Chain VISYEESNRYHADSVKGRFTISRDNSKITLYLQMNSLRTEDTAVYYCARDG
GIAAPGPDYWGQGTLVTVSSASTKGPSVFPLAPASKSTSGGTAALGCLVKD
YFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYIC
NVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTL
MISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYR
VVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLP
PSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGS
FFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK
Light 605 DIVMTQSPLSLTVTPGEPASISCRSSQSLLYSNGYNYLDWYLQKPGQSPQVL Chain ISLGSNRASGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCMQARQTPFTFG
PGTKVDIRRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKV Antibody Region SEQ Sequence
ID
NO:
DNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQG LSSPVTKSFNRGEC
ADC-1013 Heavy 610 EVQLLESGGGLVQPGGSLRLSCAASGFTFSTYGMHWVRQAPGKGLEWLSY
Chain ISGGSSYIFYADSVRGRFTISRDNSENALYLQMNSLRAEDTAVYYCARILRG
GSGMDLWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFP
EPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTQTYICNVN
HKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISR
TPEVTCNAVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRWSV
LTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRD
ELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLY
SKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK
Light 615 QSVLTQPPSASGTPGQRVTISCTGSSSNIGAGYNVYWYQQLPGTAPKLLIYG Chain NINRPSGVPDRFSGSKSGTSASLAISGLRSEDEADYYCAAWDKSISGLVFGG
GTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKA DSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGST VEKTVAPTECS
APX005 Heavy 620 QVQLVESGGGVVQPGRSLRLSCAASGFSFSSTYVCWVRQAPGKGLEWIACI
Chain YTGDGTNYSASWAKGRFTISKDSSKNTVYLQMNSLRAEDTAVYFCARPDI
TYGFAINFWGPGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYF
PEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNV
NHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMIS
RTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVV
SVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPS
REEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFF
LYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK
Light 625 DIQMTQSPSSLSASVGDRVTIKCQASQSISSRLAWYQQKPGKPPKLLIYRAS Chain TLASGVPSRFSGSGSGTDFTLTISSLQPEDVATYYCQCTGYGISWPIGGGTK
VEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNAL QSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPV TKSFNRGEC
Chi Lob 7/4 Heavy 630 EVQLQQSGPDLVKPGASVKISCKTSGYTFTEYIMHWVKQSHGKSLEWIGGI
Chain IPNNGGTSYNQKFKDKATMTVDKSSSTGYMELRSLTSEDSAVYYCTRREV
YGRNYYALDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLV
KDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTY
ICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDT
LMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTY
RVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYT
LPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSD
GSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK
Light 635 DIQMTQTTSSLSASLGDRVTITCSASQGINNYLNWYQQKPDGTVKLLIYYTS Chain SLHSGVPSRFSGSGSGTDYSLTISNLEPEDIATYYCQQYSNLPYTFGGGTKLE
IKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQS
GNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTK
SFNRGEC
DEC-205 Heavy 232 QVQLVESGGGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGLEWVA (variant 1) Chain VIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAR
DLWGWYFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVK
DYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYI
CNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDT
LMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTY
RVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYT
LPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSD
GSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK
Light 237 EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLIYDAS Chain NRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRRNWPLTFGGGTKV
EIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQ SGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVT Antibody Region SEQ Sequence
ID
NO:
KSFNRGEC
DEC-205 Heavy 245 EVQLVQSGAEVKKPGESLRISCKGSGDSFTTYWIGWVRQMPGKGLEWMGI (variant 2) Chain IYPGDSDTIYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCTRGDRG
VDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPV
TVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKP
SNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPE
VTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLT
VLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEM
TKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSK
LTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK
Light 250 DIQMTQSPSSLSASVGDRVTITCRASQGISRWLAWYQQKPEKAPKSLIYAAS Chain SLQSGVPSRFSGSGSGTDFTLTISGLQPEDFATYYCQQYNSYPRTFGQGTKV
EIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQ
SGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVT
KSFNRGEC
CLEC12A Heavy 662 QVQLQESGPGLVKPSETLSLTCVVSGGSISSSNWWSWVRQPPGKGLEWIGE
Chain IYHSGSPDYNPSLKSRVTISVDKSRNQFSLKLSSVTAADTAVYYCAKVSTG variant GFFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPE 1 PVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNH
KPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRT
PEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSV
LTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRE
EMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLY
SKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK
Heavy 663 QVQLQESGPGLVKPSETLSLTCVVSGGSISSSNWWSWVRQPPGKGLEWIGE Chain IYHSGSPNYNPSLKSRVTISVDKSKNQFSLKLSSVTAADTAVYYCARSSSGG variant FFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEP 2 VTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNH
KPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRT
PEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSV
LTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRE
EMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLY
SKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK
Heavy 664 QVQLQESGPGLVKPSETLSLTCVVSGGSISSSNWWSWVRQPPGKGLEWIGE Chain IYHSGSPNYNPSLKSRVTISVDKSKNQFSLKLSSVTAADTAVYYCARQTTA variant GSFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPE 3 PVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNH
KPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRT
PEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSV
LTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRE
EMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLY
SKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK
Light 665 DIQMTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYAAS Chain SLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPPTFGQGTKVE
IKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQS
GNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTK
SFNRGEC
BDCA-2 Heavy 666 QVQLVESGGGVVQPGRSLRLSCAASGFTLSSYGMHWVRQAPGKGLEWVA Variant 1 Chain VIWYDGNDKYYADSVKGRFTISRDNSKNTLYLQVNSLRAEDTAVYYCAR
GTGTPYWYFDLWGRGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLV
KDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTY
ICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDT
LMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTY
RVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYT
LPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSD
GSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK
Light 669 EIVLTQSPATLSLSPGERATLSCRASQSVNNYLAWYQQKPGQAPRLLIYDAS Chain NRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRSTWPPYTFGQGTK
LEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNAL Antibody Region SEQ Sequence
ID
NO:
QSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPV TKSFNRGEC
BDCA-2 Heavy 667 EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYLMNWVRQAPGKGLEWVA Variant 2 Chain NIEQDGSEKYYVDSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYFCARD
GDTAMITFDFWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVK
DYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYI
CNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDT
LMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTY
RVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYT
LPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSD
GSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK
Light 670 DIQMTQSPSSVSASVGDRVTITCRASQGIRRWLAWYQQKPGKAPKLLIYAA Chain SSLQRGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQANSFPWTFGQGTK
VEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNAL QSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPV TKSFNRGEC
BDCA-2 Heavy 668 QVQLQESGPGLVKPSQTLSLTCTVSGGSISSGGYYWNWIRQHPGKGLEWIG Variant 3 Chain YIYYSGNTYYNPSLKSRVTISVDTSKNQFSLKLSSVTAADAAVYHCARGYG
DYGGGYFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVK
DYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYI
CNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDT
LMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTY
RVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYT
LPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSD
GSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK
Light 671 DIQMTQSPSSLSASVGDRVTITCQASQDISNYLNWYQQKPGKAPKFLIYDVS Chain NLETGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCQQYDNLPYTFGQGTKL
EIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQ
SGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVT
KSFNRGEC
TABLE 8. Fusion Sequences - DEC-205 fusions via the heavy chain
Antibody Region SEQ Sequence
ID
NO:
PCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKF
NWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEY
KCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLT
CLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTV
DKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSQ
VQLVESGGGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGLE
WVAVIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDT
AVYYCARDLWGWYFDYWGQGTLVTVSSGGGGSGGGGSGGGGSG
GGGSEIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQA
PRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQR
RNWPLTFGGGTKVEIK
LC tumor 846 DIQMTQSPSSLSASVGDRVTITCKASQDVSIGVAWYQQKPGKAPKL mAb LIYSASYRYTGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYYIY containing PYTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPR CD40 mab EAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYE scFv (LH) KHKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSDIQMTQSPS
SVSASVGDRVTITCRASQGIYSWLAWYQQKPGKAPNLLIYTASTLQ
SGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQANIFPLTFGGGTK
VEIKGGGGSGGGGSGGGGSGGGGSQVQLVQSGAEVKKPGASVKVS
CKASGYTFTGYYMHWVRQAPGQGLEWMGWINPDSGGTNYAQKF
QGRVTMTRDTSISTAYMELNRLRSDDTAVYYCARDQPLGYCTNGV
CSYFDYWGQGTLVTVSS
Cetuximab HC DEC-205 507 QVQLVESGGGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGL mAb with EWVAVIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAED tumor mAb TAVYYCARDLWGWYFDYWGQGTLVTVSSASTKGPSVFPLAPSSKS ScFv TSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLY
SLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCP
PCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKF
NWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEY
KCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLT
CLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTV
DKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSQ
VQLKQSGPGLVQPSQSLSITCTVSGFSLTNYGVHWVRQSPGKGLEW
LGVIWSGGNTDYNTPFTSRLSINKDNSKSQVFFKMNSLQSNDTAIYY
CARALTYYDYEFAYWGQGTLVTVSAGGGGSGGGGSGGGGSGGGG
SDILLTQSPVILSVSPGERVSFSCRASQSIGTNIHWYQQRTNGSPRLLI
KYASESISGIPSRFSGSGSGTDFTLSINSVESEDIADYYCQQNNNWPT
TFGAGTKLELK
HC tumor 508 QVQLKQSGPGLVQPSQSLSITCTVSGFSLTNYGVHWVRQSPGKGLE mAb with WLGVIWSGGNTDYNTPFTSRLSINKDNSKSQVFFKMNSLQSNDTAI DEC-205 mAb YYCARALTYYDYEFAYWGQGTLVTVSAASTKGPSVFPLAPSSKSTS ScFv GGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSL
SSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPC
PAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFN
WYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYK
CKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTC
LVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVD
KSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSQV
QLVESGGGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGLE
WVAVIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDT
AVYYCARDLWGWYFDYWGQGTLVTVSSGGGGSGGGGSGGGGSG
GGGSEIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQA
PRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQR
RNWPLTFGGGTKVEIK
Panitumumab HC DEC-205 509 QVQLVESGGGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGL mAb with EWVAVIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAED tumor mAb TAVYYCARDLWGWYFDYWGQGTLVTVSSASTKGPSVFPLAPSSKS ScFv TSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLY
SLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCP Antibody Region SEQ Sequence
ID
NO:
PCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKF
NWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEY
KCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLT
CLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTV
DKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSQ
VQLQESGPGLVKPSETLSLTCTVSGGSVSSGDYYWTWIRQSPGKGL
EWIGHIYYSGNTNYNPSLKSRLTISIDTSKTQFSLKLSSVTAADTAIY
YCVRDRVTGAFDIWGQGTMVTVSSGGGGSGGGGSGGGGSGGGGS
DIQMTQSPSSLSASVGDRVTITCQASQDISNYLNWYQQKPGKAPKL
LIYDASNLETGVPSRFSGSGSGTDFTFTISSLQPEDIATYFCQHFDHLP
LAFGGGTKVEIK
HC tumor 510 QVQLQESGPGLVKPSETLSLTCTVSGGSVSSGDYYWTWIRQSPGKG mAb with LEWIGHIYYSGNTNYNPSLKSRLTISIDTSKTQFSLKLSSVTAADTAI DEC-205 mAb YYCVRDRVTGAFDIWGQGTMVTVSSASTKGPSVFPLAPSSKSTSGG ScFv TAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSS
VVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCP
APELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNW
YVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKC
KVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCL
VKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDK
SRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSQVQ
LVESGGGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGLEW
VAVIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTA
VYYCARDLWGWYFDYWGQGTLVTVSSGGGGSGGGGSGGGGSGG
GGSEIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAP
RLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRR
NWPLTFGGGTKVEIK
Nimotuzumab HC DEC-205 511 QVQLVESGGGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGL mAb with EWVAVIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAED tumor mAb TAVYYCARDLWGWYFDYWGQGTLVTVSSASTKGPSVFPLAPSSKS ScFv TSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLY
SLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCP
PCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKF
NWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEY
KCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLT
CLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTV
DKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSQ
VQLQQSGAEVKKPGSSVKVSCKASGYTFTNYYIYWVRQAPGQGLE
WIGGINPTSGGSNFNEKFKTRVTITVDESTNTAYMELSSLRSEDTAF
YFCARQGLWFDSDGRGFDFWGQGSTVTVSSGGGGSGGGGSGGGG
SGGGGSDIQMTQSPSSLSASVGDRVTITCRSSQNIVHSNGNTYLDWY
QQTPGKAPKLLIYKVSNRFSGVPSRFSGSGSGTDFTFTISSLQPEDIAT
YYCFQYSHVPWTFGQGTKLEIK
HC tumor 512 QVQLQQSGAEVKKPGSSVKVSCKASGYTFTNYYIYWVRQAPGQGL mAb with EWIGGINPTSGGSNFNEKFKTRVTITVDESTNTAYMELSSLRSEDTAF DEC-205 mAb YFCARQGLWFDSDGRGFDFWGQGSTVTVSSASTKGPSVFPLAPSSK ScFv STSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGL
YSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTC
PPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVK
FNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKE
YKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSL
TCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTV
DKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSQ
VQLVESGGGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGLE
WVAVIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDT
AVYYCARDLWGWYFDYWGQGTLVTVSSGGGGSGGGGSGGGGSG
GGGSEIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQA
PRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQR
RNWPLTFGGGTKVEIK Antibody Region SEQ Sequence
ID
NO:
Zalutumumab HC DEC-205 513 QVQLVESGGGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGL mAb with EWVAVIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAED tumor mAb TAVYYCARDLWGWYFDYWGQGTLVTVSSASTKGPSVFPLAPSSKS ScFv TSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLY
SLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCP
PCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKF
NWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEY
KCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLT
CLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTV
DKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSQ
VQLVESGGGVVQPGRSLRLSCAASGFTFSTYGMHWVRQAPGKGLE
WVAVIWDDGSYKYYGDSVKGRFTISRDNSKNTLYLQMNSLRAEDT
AVYYCARDGITMVRGVMKDYFDYWGQGTLVTVSSGGGGSGGGGS
GGGGSGGGGSAIQLTQSPSSLSASVGDRVTITCRASQDISSALVWYQ
QKPGKAPKLLIYDASSLESGVPSRFSGSESGTDFTLTISSLQPEDFATY
YCQQFNSYPLTFGGGTKVEIK
HC tumor 514 QVQLVESGGGVVQPGRSLRLSCAASGFTFSTYGMHWVRQAPGKGL mAb with EWVAVIWDDGSYKYYGDSVKGRFTISRDNSKNTLYLQMNSLRAED DEC-205 mAb TAVYYCARDGITMVRGVMKDYFDYWGQGTLVTVSSASTKGPSVFP ScFv LAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAV
LQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSC
DKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVS
HEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQ
DWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREE
MTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGS
FFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGG
GGSGGGGSQVQLVESGGGVVQPGRSLRLSCAASGFTFSNYGMYWV
RQAPGKGLEWVAVIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQ
MNSLRAEDTAVYYCARDLWGWYFDYWGQGTLVTVSSGGGGSGG
GGSGGGGSGGGGSEIVLTQSPATLSLSPGERATLSCRASQSVSSYLA
WYQQKPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPED
FAVYYCQQRRNWPLTFGGGTKVEIK
Onartuzumab HC DEC-205 515 QVQLVESGGGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGL mAb with EWVAVIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAED tumor mAb TAVYYCARDLWGWYFDYWGQGTLVTVSSASTKGPSVFPLAPSSKS ScFv TSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLY
SLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCP
PCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKF
NWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEY
KCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLT
CLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTV
DKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSE
VQLVESGGGLVQPGGSLRLSCAASGYTFTSYWLHWVRQAPGKGLE
WVGMIDPSNSDTRFNPNFKDRFTISADTSKNTAYLQMNSLRAEDTA
VYYCATYRSYVTPLDYWGQGTLVTVSSGGGGSGGGGSGGGGSGG
GGSDIQMTQSPSSLSASVGDRVTITCKSSQSLLYTSSQKNYLAWYQQ
KPGKAPKLLIYWASTRESGVPSRFSGSGSGTDFTLTISSLQPEDFATY
YCQQYYAYPWTFGQGTKVEIK
HC tumor 516 EVQLVESGGGLVQPGGSLRLSCAASGYTFTSYWLHWVRQAPGKGL mAb with EWVGMIDPSNSDTRFNPNFKDRFTISADTSKNTAYLQMNSLRAEDT DEC-205 mAb AVYYCATYRSYVTPLDYWGQGTLVTVSSASTKGPSVFPLAPSSKST ScFv SGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS
LSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPP
CPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKF
NWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEY
KCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLT
CLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTV
DKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSQ
VQLVESGGGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGLE Antibody Region SEQ Sequence
ID
NO:
WVAVIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDT
AVYYCARDLWGWYFDYWGQGTLVTVSSGGGGSGGGGSGGGGSG
GGGSEIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQA
PRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQR
RNWPLTFGGGTKVEIK
Patritumab HC DEC-205 517 QVQLVESGGGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGL mAb with EWVAVIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAED tumor mAb TAVYYCARDLWGWYFDYWGQGTLVTVSSASTKGPSVFPLAPSSKS ScFv TSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLY
SLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCP
PCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKF
NWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEY
KCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLT
CLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTV
DKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSQ
VQLQQWGAGLLKPSETLSLTCAVYGGSFSGYYWSWIRQPPGKGLE
WIGEINHSGSTNYNPSLKSRVTISVETSKNQFSLKLSSVTAADTAVY
YCARDKWTWYFDLWGRGTLVTVSSGGGGSGGGGSGGGGSGGGGS
DIEMTQSPDSLAVSLGERATINCRSSQSVLYSSSNRNYLAWYQQNP
GQPPKLLIYWASTRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYY
CQQYYSTPRTFGQGTKVEIK
HC tumor 518 QVQLQQWGAGLLKPSETLSLTCAVYGGSFSGYYWSWIRQPPGKGL mAb with EWIGEINHSGSTNYNPSLKSRVTISVETSKNQFSLKLSSVTAADTAV DEC-205 mAb YYCARDKWTWYFDLWGRGTLVTVSSASTKGPSVFPLAPSSKSTSG ScFv GTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLS
SVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCP
APELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNW
YVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKC
KVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCL
VKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDK
SRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSQVQ
LVESGGGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGLEW
VAVIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTA
VYYCARDLWGWYFDYWGQGTLVTVSSGGGGSGGGGSGGGGSGG
GGSEIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAP
RLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRR
NWPLTFGGGTKVEIK
Clivatuzumab HC DEC-205 519 QVQLVESGGGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGL mAb with EWVAVIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAED tumor mAb TAVYYCARDLWGWYFDYWGQGTLVTVSSASTKGPSVFPLAPSSKS ScFv TSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLY
SLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCP
PCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKF
NWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEY
KCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLT
CLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTV
DKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSQ
VQLQQSGAEVKKPGASVKVSCEASGYTFPSYVLHWVKQAPGQGLE
WIGYINPYNDGTQYNEKFKGKATLTRDTSINTAYMELSRLRSDDTA
VYYCARGFGGSYGFAYWGQGTLVTVSSGGGGSGGGGSGGGGSGG
GGSDIQLTQSPSSLSASVGDRVTMTCSASSSVSSSYLYWYQQKPGK
APKLWIYSTSNLASGVPARFSGSGSGTDFTLTISSLQPEDSASYFCHQ
WNRYPYTFGGGTRLEIK
HC tumor 520 QVQLQQSGAEVKKPGASVKVSCEASGYTFPSYVLHWVKQAPGQGL mAb with EWIGYINPYNDGTQYNEKFKGKATLTRDTSINTAYMELSRLRSDDT DEC-205 mAb AVYYCARGFGGSYGFAYWGQGTLVTVSSASTKGPSVFPLAPSSKST ScFv SGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS
LSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPP
CPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKF Antibody Region SEQ Sequence
ID
NO:
NWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEY
KCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLT
CLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTV
DKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSQ
VQLVESGGGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGLE
WVAVIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDT
AVYYCARDLWGWYFDYWGQGTLVTVSSGGGGSGGGGSGGGGSG
GGGSEIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQA
PRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQR
RNWPLTFGGGTKVEIK
Sofituzumab HC DEC-205 521 QVQLVESGGGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGL mAb with EWVAVIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAED tumor mAb TAVYYCARDLWGWYFDYWGQGTLVTVSSASTKGPSVFPLAPSSKS ScFv TSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLY
SLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCP
PCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKF
NWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEY
KCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLT
CLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTV
DKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSE
VQLVESGGGLVQPGGSLRLSCAASGYSITNDYAWNWVRQAPGKGL
EWVGYISYSGYTTYNPSLKSRFTISRDTSKNTLYLQMNSLRAEDTAV
YYCARWTSGLDYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSD
IQMTQSPSSLSASVGDRVTITCKASDLIHNWLAWYQQKPGKAPKLLI
YGATSLETGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYWTTP
FTFGQGTKVEIK
HC tumor 522 EVQLVESGGGLVQPGGSLRLSCAASGYSITNDYAWNWVRQAPGKG mAb with LEWVGYISYSGYTTYNPSLKSRFTISRDTSKNTLYLQMNSLRAEDTA DEC-205 mAb VYYCARWTSGLDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGT ScFv AALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSV
VTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAP
ELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWY
VDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCK
VSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLV
KGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSR
WQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSQVQL
VESGGGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGLEWV
AVIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAV
YYCARDLWGWYFDYWGQGTLVTVSSGGGGSGGGGSGGGGSGGG
GSEIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPR
LLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRRN
WPLTFGGGTKVEIK
Edrecolomab HC DEC-205 523 QVQLVESGGGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGL mAb with EWVAVIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAED tumor mAb TAVYYCARDLWGWYFDYWGQGTLVTVSSASTKGPSVFPLAPSSKS ScFv TSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLY
SLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCP
PCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKF
NWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEY
KCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLT
CLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTV
DKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSQ
VQLQQSGAELVRPGTSVKVSCKASGYAFTNYLIEWVKQRPGQGLE
WIGVINPGSGGTNYNEKFKGKATLTADKSSSTAYMQLSSLTSDDSA
VYFCARDGPWFAYWGQGTLVTVSAGGGGSGGGGSGGGGSGGGGS
NIVMTQSPKSMSMSVGERVTLTCKASENVVTYVSWYQQKPEQSPK
LLIYGASNRYTGVPDRFTGSGSATDFTLTISSVQAEDLADYHCGQGY
SYPYTFGGGTKLEIK
HC tumor 524 QVQLQQSGAELVRPGTSVKVSCKASGYAFTNYLIEWVKQRPGQGL Antibody Region SEQ Sequence
ID
NO:
mAb with EWIGVINPGSGGTNYNEKFKGKATLTADKSSSTAYMQLSSLTSDDS DEC-205 mAb AVYFCARDGPWFAYWGQGTLVTVSAASTKGPSVFPLAPSSKSTSGG ScFv TAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSS
VVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCP
APELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNW
YVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKC
KVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCL
VKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDK
SRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSQVQ
LVESGGGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGLEW
VAVIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTA
VYYCARDLWGWYFDYWGQGTLVTVSSGGGGSGGGGSGGGGSGG
GGSEIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAP
RLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRR
NWPLTFGGGTKVEIK
Adecatumum HC DEC-205 525 QVQLVESGGGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGL ab mAb with EWVAVIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAED tumor mAb TAVYYCARDLWGWYFDYWGQGTLVTVSSASTKGPSVFPLAPSSKS ScFv TSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLY
SLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCP
PCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKF
NWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEY
KCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLT
CLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTV
DKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSE
VQLLESGGGVVQPGRSLRLSCAASGFTFSSYGMHWVRQAPGKGLE
WVAVISYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDT
AVYYCAKDMGWGSGWRPYYYYGMDVWGQGTTVTVSSGGGGSG
GGGSGGGGSGGGGSELQMTQSPSSLSASVGDRVTITCRTSQSISSYL
NWYQQKPGQPPKLLIYWASTRESGVPDRFSGSGSGTDFTLTISSLQP
EDSATYYCQQSYDIPYTFGQGTKLEIK
HC tumor 526 EVQLLESGGGVVQPGRSLRLSCAASGFTFSSYGMHWVRQAPGKGL mAb with EWVAVISYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAED DEC-205 mAb TAVYYCAKDMGWGSGWRPYYYYGMDVWGQGTTVTVSSASTKGP ScFv SVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTF
PAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEP
KSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVV
DVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVL
HQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSR
EEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSD
GSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG
GGGGSGGGGSQVQLVESGGGVVQPGRSLRLSCAASGFTFSNYGMY
WVRQAPGKGLEWVAVIWYDGSNKYYADSVKGRFTISRDNSKNTL
YLQMNSLRAEDTAVYYCARDLWGWYFDYWGQGTLVTVSSGGGG
SGGGGSGGGGSGGGGSEIVLTQSPATLSLSPGERATLSCRASQSVSS
YLAWYQQKPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSL
EPEDFAVYYCQQRRNWPLTFGGGTKVEIK
Anetumab HC DEC-205 527 QVQLVESGGGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGL mAb with EWVAVIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAED tumor mAb TAVYYCARDLWGWYFDYWGQGTLVTVSSASTKGPSVFPLAPSSKS ScFv TSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLY
SLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCP
PCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKF
NWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEY
KCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLT
CLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTV
DKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSQ
VELVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWVRQAPGKGLE
WMGIIDPGDSRTRYSPSFQGQVTISADKSISTAYLQWSSLKASDTAM Antibody Region SEQ Sequence
ID
NO:
YYCARGQLYGGTYMDGWGQGTLVTVSSGGGGSGGGGSGGGGSG GGGSDIALTQPASVSGSPGQSITISCTGTSSDIGGYNSVSWYQQHPGK APKLMIYGVNNRPSGVSNRFSGSKSGNTASLTISGLQAEDEADYYCS SYDIESATPVFGGGTKLTVL
HC tumor 528 QVELVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWVRQAPGKGLE mAb with WMGIIDPGDSRTRYSPSFQGQVTISADKSISTAYLQWSSLKASDTAM DEC-205 mAb YYCARGQLYGGTYMDGWGQGTLVTVSSASTKGPSVFPLAPSSKST ScFv SGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS
LSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPP
CPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKF
NWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEY
KCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLT
CLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTV
DKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSQ
VQLVESGGGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGLE
WVAVIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDT
AVYYCARDLWGWYFDYWGQGTLVTVSSGGGGSGGGGSGGGGSG
GGGSEIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQA
PRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQR
RNWPLTFGGGTKVEIK
huDS6 HC DEC-205 529 QVQLVESGGGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGL mAb with EWVAVIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAED tumor mAb TAVYYCARDLWGWYFDYWGQGTLVTVSSASTKGPSVFPLAPSSKS ScFv TSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLY
SLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCP
PCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKF
NWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEY
KCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLT
CLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTV
DKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSQ
AQLVQSGAEVVKPGASVKMSCKASGYTFTSYNMHWVKQTPGQGL
EWIGYIYPGNGATNYNQKFQGKATLTADPSSSTAYMQISSLTSEDSA
VYFCARGDSVPFAYWGQGTLVTVSAGGGGSGGGGSGGGGSGGGG
SEIVLTQSPATMSASPGERVTITCSAHSSVSFMHWFQQKPGTSPKLW
IYSTSSLASGVPARFGGSGSGTSYSLTISSMEAEDAATYYCQQRSSFP
LTFGAGTKLELK
HC tumor 530 QAQLVQSGAEVVKPGASVKMSCKASGYTFTSYNMHWVKQTPGQG mAb with LEWIGYIYPGNGATNYNQKFQGKATLTADPSSSTAYMQISSLTSEDS DEC-205 mAb AVYFCARGDSVPFAYWGQGTLVTVSAASTKGPSVFPLAPSSKSTSG ScFv GTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLS
SVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCP
APELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNW
YVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKC
KVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCL
VKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDK
SRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSQVQ
LVESGGGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGLEW
VAVIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTA
VYYCARDLWGWYFDYWGQGTLVTVSSGGGGSGGGGSGGGGSGG
GGSEIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAP
RLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRR
NWPLTFGGGTKVEIK
Lifastuzumab HC DEC-205 531 QVQLVESGGGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGL mAb with EWVAVIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAED tumor mAb TAVYYCARDLWGWYFDYWGQGTLVTVSSASTKGPSVFPLAPSSKS ScFv TSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLY
SLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCP
PCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKF
NWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEY Antibody Region SEQ Sequence
ID
NO:
KCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLT
CLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTV
DKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSE
VQLVESGGGLVQPGGSLRLSCAASGFSFSDFAMSWVRQAPGKGLE
WVATIGRVAFHTYYPDSMKGRFTISRDNSKNTLYLQMNSLRAEDT
AVYYCARHRGFDVGHFDFWGQGTLVTVSSGGGGSGGGGSGGGGS
GGGGSDIQMTQSPSSLSASVGDRVTITCRSSETLVHSSGNTYLEWYQ
QKPGKAPKLLIYRVSNRFSGVPSRFSGSGSGTDFTLTISSLQPEDFAT
YYCFQGSFNPLTFGQGTKVEIK
HC tumor 532 EVQLVESGGGLVQPGGSLRLSCAASGFSFSDFAMSWVRQAPGKGLE mAb with WVATIGRVAFHTYYPDSMKGRFTISRDNSKNTLYLQMNSLRAEDT DEC-205 mAb AVYYCARHRGFDVGHFDFWGQGTLVTVSSASTKGPSVFPLAPSSKS ScFv TSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLY
SLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCP
PCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKF
NWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEY
KCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLT
CLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTV
DKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSQ
VQLVESGGGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGLE
WVAVIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDT
AVYYCARDLWGWYFDYWGQGTLVTVSSGGGGSGGGGSGGGGSG
GGGSEIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQA
PRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQR
RNWPLTFGGGTKVEIK
Sacituzumab HC DEC-205 533 QVQLVESGGGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGL mAb with EWVAVIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAED tumor mAb TAVYYCARDLWGWYFDYWGQGTLVTVSSASTKGPSVFPLAPSSKS ScFv TSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLY
SLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCP
PCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKF
NWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEY
KCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLT
CLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTV
DKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSQ
VQLQQSGSELKKPGASVKVSCKASGYTFTNYGMNWVKQAPGQGL
KWMGWINTYTGEPTYTDDFKGRFAFSLDTSVSTAYLQISSLKADDT
AVYFCARGGFGSSYWYFDVWGQGSLVTVSSGGGGSGGGGSGGGG
SGGGGSDIQLTQSPSSLSASVGDRVSITCKASQDVSIAVAWYQQKPG
KAPKLLIYSASYRYTGVPDRFSGSGSGTDFTLTISSLQPEDFAVYYCQ
QHYITPLTFGAGTKVEIK
HC tumor 534 QVQLQQSGSELKKPGASVKVSCKASGYTFTNYGMNWVKQAPGQG mAb with LKWMGWINTYTGEPTYTDDFKGRFAFSLDTSVSTAYLQISSLKADD DEC-205 mAb TAVYFCARGGFGSSYWYFDVWGQGSLVTVSSASTKGPSVFPLAPSS ScFv KSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSG
LYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHT
CPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEV
KFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGK
EYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVS
LTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLT
VDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGS
QVQLVESGGGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGL
EWVAVIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAED
TAVYYCARDLWGWYFDYWGQGTLVTVSSGGGGSGGGGSGGGGS
GGGGSEIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQ
APRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQ
RRNWPLTFGGGTKVEIK
PR1A3 HC DEC-205 535 QVQLVESGGGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGL mAb with EWVAVIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAED Antibody Region SEQ Sequence
ID
NO:
tumor mAb TAVYYCARDLWGWYFDYWGQGTLVTVSSASTKGPSVFPLAPSSKS ScFv TSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLY
SLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCP
PCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKF
NWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEY
KCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLT
CLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTV
DKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSQ
VKLQQSGPELKKPGETVKISCKASGYTFTEFGMNWVKQAPGKGLK
WMGWINTKTGEATYVEEFKGRFAFSLETSATTAYLQINNLKNEDTA
KYFCARWDFYDYVEAMDYWGQGTTVTVSSGGGGSGGGGSGGGG
SGGGGSDIVMTQSQRFMSTSVGDRVSVTCKASQNVGTNVAWYQQ
KPGQSPKALIYSASYRYSGVPDRFTGSGSGTDFTLTISNVQSEDLAE
YFCHQYYTYPLFTFGSGTKLEMK
HC tumor 536 QVKLQQSGPELKKPGETVKISCKASGYTFTEFGMNWVKQAPGKGL mAb with KWMGWINTKTGEATYVEEFKGRFAFSLETSATTAYLQINNLKNEDT DEC-205 mAb AKYFCARWDFYDYVEAMDYWGQGTTVTVSSASTKGPSVFPLAPSS ScFv KSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSG
LYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHT
CPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEV
KFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGK
EYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVS
LTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLT
VDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGS
QVQLVESGGGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGL
EWVAVIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAED
TAVYYCARDLWGWYFDYWGQGTLVTVSSGGGGSGGGGSGGGGS
GGGGSEIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQ
APRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQ
RRNWPLTFGGGTKVEIK
Humanized HC DEC-205 818 QVQLVESGGGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGL PR1A3 mAb with EWVAVIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAED tumor mAb TAVYYCARDLWGWYFDYWGQGTLVTVSSASTKGPSVFPLAPSSKS ScFv TSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLY
SLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCP
PCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVK
FNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKE
YKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSL
TCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTV
DKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGS
QVQLVQSGAEVKKPGASVKVSCKASGYTFTEFGMNWVRQAPGQG
LEWMGWINTKTGEATYVEEFKGRVTFTTDTSTSTAYMELRSLRSD
DTAVYYCARWDFAYYVEAMDYWGQGTTVTVSSGGGGSGGGGSG
GGGSGGGGSDIQMTQSPSSLSASVGDRVTITCKASAAVGTYVAWY
QQKPGKAPKLLIYSASYRKRGVPSRFSGSGSGTDFTLTISSLQPEDFA
TYYCHQYYTYPLFTFGQGTKLEIK
HC tumor 819 QVQLVQSGAEVKKPGASVKVSCKASGYTFTEFGMNWVRQAPGQG mAb with LEWMGWINTKTGEATYVEEFKGRVTFTTDTSTSTAYMELRSLRSD DEC-205 mAb DTAVYYCARWDFAYYVEAMDYWGQGTTVTVSSASTKGPSVFPLA ScFv PSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQ
SSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDK
THTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHE
DPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDW
LNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTK
NQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFL
YSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGS
GGGGSQVQLVESGGGVVQPGRSLRLSCAASGFTFSNYGMYWVRQ
APGKGLEWVAVIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQM
NSLRAEDTAVYYCARDLWGWYFDYWGQGTLVTVSSGGGGSGGG Antibody Region SEQ Sequence
ID
NO:
GSGGGGSGGGGSEIVLTQSPATLSLSPGERATLSCRASQSVSSYLAW YQQKPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDF AVYYCQQRRNWPLTFGGGTKVEIK
Humanized HC DEC-205 836 QVQLVESGGGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGL Ab2-3 mAb with EWVAVIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAED tumor mAb TAVYYCARDLWGWYFDYWGQGTLVTVSSASTKGPSVFPLAPSSKS ScFv TSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLY
SLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCP
PCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVK
FNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKE
YKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSL
TCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTV
DKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSE
VQLQESGPGLVKPGGSLSLSCAASGFVFSSYDMSWVRQTPERGLEW
VAYISSGGGITYAPSTVKGRFTVSRDNAKNTLYLQMNSLTSEDTAV
YYCAAHYFGSSGPFAYWGQGTLVTVSSGGGGSGGGGSGGGGSGG
GGSDIQMTQSPASLSASVGDRVTITCRASENIFSYLAWYQQKPGKSP
KLLVYNTRTLAEGVPSRFSGSGSGTDFSLTISSLQPEDFATYYCQHH
YGTPFTFGSGTKLEIK
HC tumor 837 EVQLQESGPGLVKPGGSLSLSCAASGFVFSSYDMSWVRQTPERGLE mAb with WVAYISSGGGITYAPSTVKGRFTVSRDNAKNTLYLQMNSLTSEDTA DEC-205 mAb VYYCAAHYFGSSGPFAYWGQGTLVTVSSASTKGPSVFPLAPSSKST ScFv SGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS
LSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPP
CPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKF
NWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEY
KCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLT
CLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTV
DKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGS
QVQLVESGGGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGL
EWVAVIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAED
TAVYYCARDLWGWYFDYWGQGTLVTVSSGGGGSGGGGSGGGGS
GGGGSEIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQ
APRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQ
RRNWPLTFGGGTKVEIK
IMAB362, HC DEC-205 537 QVQLVESGGGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGL
CLAUDIXIM mAb with EWVAVIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAED
AB tumor mAb TAVYYCARDLWGWYFDYWGQGTLVTVSSASTKGPSVFPLAPSSKS
ScFv TSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLY
SLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCP
PCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKF
NWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEY
KCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLT
CLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTV
DKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSQ
VQLQQPGAELVRPGASVKLSCKASGYTFTSYWINWVKQRPGQGLE
WIGNIYPSDSYTNYNQKFKDKATLTVDKSSSTAYMQLSSPTSEDSA
VYYCTRSWRGNSFDYWGQGTTLTVSSGGGGSGGGGSGGGGSGGG
GSDIVMTQSPSSLTVTAGEKVTMSCKSSQSLLNSGNQKNYLTWYQQ
KPGQPPKLLIYWASTRESGVPDRFTGSGSGTDFTLTISSVQAEDLAV
YYCQNDYSYPFTFGSGTKLEIK
HC tumor 538 QVQLQQPGAELVRPGASVKLSCKASGYTFTSYWINWVKQRPGQGL mAb with EWIGNIYPSDSYTNYNQKFKDKATLTVDKSSSTAYMQLSSPTSEDSA DEC-205 mAb VYYCTRSWRGNSFDYWGQGTTLTVSSASTKGPSVFPLAPSSKSTSG ScFv GTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLS
SVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCP
APELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNW
YVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKC
KVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCL Antibody Region SEQ Sequence
ID
NO:
VKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDK
SRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSQVQ
LVESGGGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGLEW
VAVIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTA
VYYCARDLWGWYFDYWGQGTLVTVSSGGGGSGGGGSGGGGSGG
GGSEIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAP
RLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRR
NWPLTFGGGTKVEIK
AMG595 HC DEC-205 539 QVQLVESGGGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGL mAb with EWVAVIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAED tumor mAb TAVYYCARDLWGWYFDYWGQGTLVTVSSASTKGPSVFPLAPSSKS ScFv TSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLY
SLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCP
PCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKF
NWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEY
KCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLT
CLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTV
DKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSQ
VQLVESGGGVVQSGRSLRLSCAASGFTFRNYGMHWVRQAPGKGLE
WVAVIWYDGSDKYYADSVRGRFTISRDNSKNTLYLQMNSLRAEDT
AVYYCARDGYDILTGNPRDFDYWGQGTLVTVSSGGGGSGGGGSG
GGGSGGGGSDTVMTQTPLSSHVTLGQPASISCRSSQSLVHSDGNTY
LSWLQQRPGQPPRLLIYRISRRFSGVPDRFSGSGAGTDFTLEISRVEA
EDVGVYYCMQSTHVPRTFGQGTKVEIK
HC tumor 540 QVQLVESGGGVVQSGRSLRLSCAASGFTFRNYGMHWVRQAPGKG mAb with LEWVAVIWYDGSDKYYADSVRGRFTISRDNSKNTLYLQMNSLRAE DEC-205 mAb DTAVYYCARDGYDILTGNPRDFDYWGQGTLVTVSSASTKGPSVFPL ScFv APSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVL
QSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCD
KTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHE
DPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDW
LNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTK
NQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFL
YSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGS
GGGGSQVQLVESGGGVVQPGRSLRLSCAASGFTFSNYGMYWVRQ
APGKGLEWVAVIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQMN
SLRAEDTAVYYCARDLWGWYFDYWGQGTLVTVSSGGGGSGGGGS
GGGGSGGGGSEIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQ
QKPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAV
YYCQQRRNWPLTFGGGTKVEIK
ABT806 HC DEC-205 541 QVQLVESGGGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGL mAb with EWVAVIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAED tumor mAb TAVYYCARDLWGWYFDYWGQGTLVTVSSASTKGPSVFPLAPSSKS ScFv TSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLY
SLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCP
PCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKF
NWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEY
KCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLT
CLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTV
DKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSE
VQLQESGPGLVKPSQTLSLTCTVSGYSISRDFAWNWIRQPPGKGLE
WMGYISYNGNTRYQPSLKSRITISRDTSKNQFFLKLNSVTAADTATY
YCVTASRGFPYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSDIQ
MTQSPSSMSVSVGDRVTITCHSSQDINSNIGWLQQKPGKSFKGLIYH
GTNLDDGVPSRFSGSGSGTDYTLTISSLQPEDFATYYCVQYAQFPWT
FGGGTKLEIK
HC tumor 542 EVQLQESGPGLVKPSQTLSLTCTVSGYSISRDFAWNWIRQPPGKGLE mAb with WMGYISYNGNTRYQPSLKSRITISRDTSKNQFFLKLNSVTAADTATY DEC-205 mAb YCVTASRGFPYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAAL Antibody Region SEQ Sequence
ID
NO:
ScFv GCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTV
PSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELL
GGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDG
VEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSN
KALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGF
YPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQ
QGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSQVQLVES
GGGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGLEWVAVI
WYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYC
ARDLWGWYFDYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSEI
VLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLIY
DASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRRNWPL
TFGGGTKVEIK
Sibrotuzumab HC DEC-205 543 QVQLVESGGGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGL mAb with EWVAVIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAED tumor mAb TAVYYCARDLWGWYFDYWGQGTLVTVSSASTKGPSVFPLAPSSKS ScFv TSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLY
SLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCP
PCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKF
NWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEY
KCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLT
CLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTV
DKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSQ
VQLVQSGAEVKKPGASVKVSCKTSRYTFTEYTIHWVRQAPGQRLE
WIGGINPNNGIPNYNQKFKGRVTITVDTSASTAYMELSSLRSEDTAV
YYCARRRIAYGYDEGHAMDYWGQGTLVTVSSGGGGSGGGGSGGG
GSGGGGSDIVMTQSPDSLAVSLGERATINCKSSQSLLYSRNQKNYL
AWYQQKPGQPPKLLIFWASTRESGVPDRFSGSGFGTDFTLTISSLQA
EDVAVYYCQQYFSYPLTFGQGTKVEIK
HC tumor 544 QVQLVQSGAEVKKPGASVKVSCKTSRYTFTEYTIHWVRQAPGQRL mAb with EWIGGINPNNGIPNYNQKFKGRVTITVDTSASTAYMELSSLRSEDTA DEC-205 mAb VYYCARRRIAYGYDEGHAMDYWGQGTLVTVSSASTKGPSVFPLAP ScFv SSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQS
SGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKT
HTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDP
EVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLN
GKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQ
VSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSK
LTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGG
GSQVQLVESGGGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGK
GLEWVAVIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRA
EDTAVYYCARDLWGWYFDYWGQGTLVTVSSGGGGSGGGGSGGG
GSGGGGSEIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKP
GQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYC
QQRRNWPLTFGGGTKVEIK
DS-8895a HC DEC-205 545 QVQLVESGGGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGL variant 1 mAb with EWVAVIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAED tumor mAb TAVYYCARDLWGWYFDYWGQGTLVTVSSASTKGPSVFPLAPSSKS ScFv TSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLY
SLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCP
PCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKF
NWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEY
KCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLT
CLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTV
DKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSQ
VQLVQSGAEVKKPGASVKVSCKASGYTFIDYSMHWVRQAPGQGLE
WMGWINTYTGEPTYSDDFKGRVTITADTSTSTAYLELSSLRSEDTA
VYYCATYYRYERDFDYWGQGTLVTVSSGGGGSGGGGSGGGGSGG
GGSDIVMTQSPLSLPVTPGEPASISCRSSQSIVHSSGITYLEWYLQKP Antibody Region SEQ Sequence
ID
NO:
GQSPQLLIYKVSNRFSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYY CFQGSHVPYTFGQGTKVEIK
HC tumor 546 QVQLVQSGAEVKKPGASVKVSCKASGYTFIDYSMHWVRQAPGQG mAb with LEWMGWINTYTGEPTYSDDFKGRVTITADTSTSTAYLELSSLRSEDT DEC-205 mAb AVYYCATYYRYERDFDYWGQGTLVTVSSASTKGPSVFPLAPSSKST ScFv SGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS
LSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPP
CPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKF
NWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEY
KCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLT
CLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTV
DKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSQ
VQLVESGGGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGLE
WVAVIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDT
AVYYCARDLWGWYFDYWGQGTLVTVSSGGGGSGGGGSGGGGSG
GGGSEIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQA
PRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQR
RNWPLTFGGGTKVEIK
DS-8895a HC DEC-205 547 QVQLVESGGGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGL variant 2 mAb with EWVAVIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAED tumor mAb TAVYYCARDLWGWYFDYWGQGTLVTVSSASTKGPSVFPLAPSSKS ScFv TSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLY
SLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCP
PCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKF
NWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEY
KCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLT
CLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTV
DKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSQ
IQLVQSGAEVKKPGASVKVSCKASGYTFIDYSMHWVRQAPGQGLK
WMGWINTYTGEPTYSDDFKGRFAFSLDTSTSTAYLELSSLRSEDTA
VYYCATYYRYERDFDYWGQGTLVTVSSGGGGSGGGGSGGGSGGG
GSDVLMTQSPLSLPVTPGEPASISCRSSQSIVHSSGITYLEWYLQKPG
QSPQLLIYKVSNRFSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYC
FQGSHVPYTFGQGTKVEIK
HC tumor 548 QIQLVQSGAEVKKPGASVKVSCKASGYTFIDYSMHWVRQAPGQGL mAb with KWMGWINTYTGEPTYSDDFKGRFAFSLDTSTSTAYLELSSLRSEDT DEC-205 mAb AVYYCATYYRYERDFDYWGQGTLVTVSSASTKGPSVFPLAPSSKST ScFv SGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS
LSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPP
CPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKF
NWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEY
KCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLT
CLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTV
DKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSQ
VQLVESGGGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGLE
WVAVIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDT
AVYYCARDLWGWYFDYWGQGTLVTVSSGGGGGSGGGGSGGGSG
GGGSEIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQA
PRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQR
RNWPLTFGGGTKVEIK
MEDI-547 HC DEC-205 549 QVQLVESGGGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGL mAb with EWVAVIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAED tumor mAb TAVYYCARDLWGWYFDYWGQGTLVTVSSASTKGPSVFPLAPSSKS ScFv TSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLY
SLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCP
PCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKF
NWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEY
KCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLT
CLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTV Antibody Region SEQ Sequence
ID
NO:
DKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSE
VQLLESGGGLVQPGGSLRLSCAASGFTFSHYMMAWVRQAPGKGLE
WVSRIGPSGGPTHYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTA
VYYCAGYDSGYDYVAVAGPAEYFQHWGQGTLVTVSSGGGGSGGG
GSGGGGSGGGGSDIQMTQSPSSLSASVGDRVTITCRASQSISTWLAW
YQQKPGKAPKLLIYKASNLHTGVPSRFSGSGSGTEFSLTISGLQPDDF
ATYYCQQYNSYSRTFGQGTKVEIK
HC tumor 550 EVQLLESGGGLVQPGGSLRLSCAASGFTFSHYMMAWVRQAPGKGL mAb with EWVSRIGPSGGPTHYADSVKGRFTISRDNSKNTLYLQMNSLRAEDT DEC-205 mAb AVYYCAGYDSGYDYVAVAGPAEYFQHWGQGTLVTVSSASTKGPS ScFv VFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFP
AVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPK
SCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVD
VSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLH
QDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRE
EMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDG
SFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGG
GGGSGGGGSQVQLVESGGGVVQPGRSLRLSCAASGFTFSNYGMYW
VRQAPGKGLEWVAVIWYDGSNKYYADSVKGRFTISRDNSKNTLYL
QMNSLRAEDTAVYYCARDLWGWYFDYWGQGTLVTVSSGGGGSG
GGGSGGGGSGGGGSEIVLTQSPATLSLSPGERATLSCRASQSVSSYL
AWYQQKPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPE
DFAVYYCQQRRNWPLTFGGGTKVEIK
Narnatumab HC DEC-205 551 QVQLVESGGGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGL mAb with EWVAVIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAED tumor mAb TAVYYCARDLWGWYFDYWGQGTLVTVSSASTKGPSVFPLAPSSKS ScFv TSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLY
SLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCP
PCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKF
NWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEY
KCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLT
CLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTV
DKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSE
VQLVESGGGLVQPGGSLRLSCAASGFTFSSYLMTWVRQAPGKGLE
WVANIKQDGSEKYYVDSVKGRFTISRDNAKNSLNLQMNSLRAEDT
AVYYCTRDGYSSGRHYGMDVWGQGTTVIVSSGGGGSGGGGSGGG
GSGGGGSEIVLTQSPATLSLSPGERATLSCRASQSVSRYLAWYQQKP
GQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYC
QQRSNWPRTFGQGTKVEIK
HC tumor 552 EVQLVESGGGLVQPGGSLRLSCAASGFTFSSYLMTWVRQAPGKGLE mAb with WVANIKQDGSEKYYVDSVKGRFTISRDNAKNSLNLQMNSLRAEDT DEC-205 mAb AVYYCTRDGYSSGRHYGMDVWGQGTTVIVSSASTKGPSVFPLAPSS ScFv KSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSG
LYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHT
CPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEV
KFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGK
EYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVS
LTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLT
VDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGS
QVQLVESGGGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGL
EWVAVIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAED
TAVYYCARDLWGWYFDYWGQGTLVTVSSGGGGSGGGGSGGGGS
GGGGSEIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQ
APRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQ
RRNWPLTFGGGTKVEIK
RG7841 HC DEC-205 553 QVQLVESGGGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGL mAb with EWVAVIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAED tumor mAb TAVYYCARDLWGWYFDYWGQGTLVTVSSASTKGPSVFPLAPSSKS ScFv TSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLY Antibody Region SEQ Sequence
ID
NO:
SLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCP
PCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKF
NWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEY
KCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLT
CLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTV
DKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSE
VQLVESGPALVKPTQTLTLTCTVSGFSLTGYSVNWIRQPPGKALEW
LGMIWGDGSTDYNSALKSRLTISKDTSKNQVVLTMTNMDPVDTAT
YYCARDYYFNYASWFAYWGQGTLVTVSSGGGGSGGGGSGGGGSG
GGGSDIQMTQSPSSLSASVGDRVTITCSASQGISNYLNWYQQKPGKT
VKLLIYYTSNLHSGVPSRFSGSGSGTDYTLTISSLQPEDFATYYCQQY
SELPWTFGQGTKVEIK
HC tumor 554 EVQLVESGPALVKPTQTLTLTCTVSGFSLTGYSVNWIRQPPGKALE mAb with WLGMIWGDGSTDYNSALKSRLTISKDTSKNQVVLTMTNMDPVDTA DEC-205 mAb TYYCARDYYFNYASWFAYWGQGTLVTVSSASTKGPSVFPLAPSSKS ScFv TSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLY
SLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCP
PCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKF
NWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEY
KCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLT
CLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTV
DKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSQ
VQLVESGGGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGLE
WVAVIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDT
AVYYCARDLWGWYFDYWGQGTLVTVSSGGGGSGGGGSGGGGSG
GGGSEIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQA
PRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQR
RNWPLTFGGGTKVEIK
Farletuzumab HC DEC-205 555 QVQLVESGGGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGL mAb with EWVAVIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAED tumor mAb TAVYYCARDLWGWYFDYWGQGTLVTVSSASTKGPSVFPLAPSSKS ScFv TSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLY
SLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCP
PCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKF
NWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEY
KCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLT
CLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTV
DKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSE
VQLVESGGGVVQPGRSLRLSCSASGFTFSGYGLSWVRQAPGKGLE
WVAMISSGGSYTYYADSVKGRFAISRDNAKNTLFLQMDSLRPEDTG
VYFCARHGDDPAWFAYWGQGTPVTVSSGGGGSGGGGSGGGGSGG
GGSDIQLTQSPSSLSASVGDRVTITCSVSSSISSNNLHWYQQKPGKAP
KPWIYGTSNLASGVPSRFSGSGSGTDYTFTISSLQPEDIATYYCQQW
SSYPYMYTFGQGTKVEIK
HC tumor 556 EVQLVESGGGVVQPGRSLRLSCSASGFTFSGYGLSWVRQAPGKGLE mAb with WVAMISSGGSYTYYADSVKGRFAISRDNAKNTLFLQMDSLRPEDTG DEC-205 mAb VYFCARHGDDPAWFAYWGQGTPVTVSSASTKGPSVFPLAPSSKSTS ScFv GGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSL
SSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPC
PAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFN
WYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYK
CKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTC
LVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVD
KSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSQV
QLVESGGGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGLE
WVAVIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDT
AVYYCARDLWGWYFDYWGQGTLVTVSSGGGGSGGGGSGGGGSG
GGGSEIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQA
PRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQR Antibody Region SEQ Sequence
ID
NO:
RNWPLTFGGGTKVEIK
Mirvetuximab HC DEC-205 557 QVQLVESGGGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGL mAb with EWVAVIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAED tumor mAb TAVYYCARDLWGWYFDYWGQGTLVTVSSASTKGPSVFPLAPSSKS ScFv TSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLY
SLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCP
PCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKF
NWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEY
KCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLT
CLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTV
DKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSQ
VQLVQSGAEVVKPGASVKISCKASGYTFTGYFMNWVKQSPGQSLE
WIGRIHPYDGDTFYNQKFQGKATLTVDKSSNTAHMELLSLTSEDFA
VYYCTRYDGSRAMDYWGQGTTVTVSSGGGGSGGGGSGGGGSGGG
GSDIVLTQSPLSLAVSLGQPAIISCKASQSVSFAGTSLMHWYHQKPG
QQPRLLIYRASNLEAGVPDRFSGSGSKTDFTLTISPVEAEDAATYYC
QQSREYPYTFGGGTKLEIK
HC tumor 558 QVQLVQSGAEVVKPGASVKISCKASGYTFTGYFMNWVKQSPGQSL mAb with EWIGRIHPYDGDTFYNQKFQGKATLTVDKSSNTAHMELLSLTSEDF DEC-205 mAb AVYYCTRYDGSRAMDYWGQGTTVTVSSASTKGPSVFPLAPSSKSTS ScFv GGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSL
SSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPC
PAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFN
WYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYK
CKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTC
LVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVD
KSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSQV
QLVESGGGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGLE
WVAVIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDT
AVYYCARDLWGWYFDYWGQGTLVTVSSGGGGSGGGGSGGGGSG
GGGSEIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQA
PRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQR
RNWPLTFGGGTKVEIK
J591 variantl HC DEC-205 559 QVQLVESGGGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGL mAb with EWVAVIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAED tumor mAb TAVYYCARDLWGWYFDYWGQGTLVTVSSASTKGPSVFPLAPSSKS ScFv TSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLY
SLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCP
PCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKF
NWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEY
KCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLT
CLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTV
DKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSE
VQLQQSGPELKKPGTSVRISCKTSGYTFTEYTIHWVKQSHGKSLEWI
GNINPNNGGTTYNQKFEDKATLTVDKSSSTAYMELRSLTSEDSAVY
YCAAGWNFDYWGQGTTLTVSSGGGGSGGGGSGGGGSGGGGSDIV
MTQSHKFMSTSVGDRVSIICKASQDVGTAVDWYQQKPGQSPKLLIY
WASTRHTGVPDRFTGSGSGTDFTLTITNVQSEDLADYFCQQYNSYP
LTFGAGTMLDLK
HC tumor 560 EVQLQQSGPELKKPGTSVRISCKTSGYTFTEYTIHWVKQSHGKSLE mAb with WIGNINPNNGGTTYNQKFEDKATLTVDKSSSTAYMELRSLTSEDSA DEC-205 mAb VYYCAAGWNFDYWGQGTTLTVSSASTKGPSVFPLAPSSKSTSGGTA ScFv ALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVV
TVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPE
LLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYV
DGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKV
SNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVK
GFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSR
WQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSQVQL Antibody Region SEQ Sequence
ID
NO:
VESGGGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGLEWV
AVIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAV
YYCARDLWGWYFDYWGQGTLVTVSSGGGGSGGGGSGGGGSGGG
GSEIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPR
LLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRRN
WPLTFGGGTKVEIK
J591 variant 2 HC DEC-205 561 QVQLVESGGGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGL mAb with EWVAVIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAED tumor mAb TAVYYCARDLWGWYFDYWGQGTLVTVSSASTKGPSVFPLAPSSKS ScFv TSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLY
SLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCP
PCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKF
NWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEY
KCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLT
CLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTV
DKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSE
VQLQQSGPELVKPGTSVRISCKTSGYTFTEYTIHWVKQSHGKSLEWI
GNINPNNGGTTYNQKFEDKATLTVDKSSSTAYMELRSLTSEDSAVY
YCAAGWNFDYWGQGTTLTVSSGGGGSGGGGSGGGGSGGGGSNIV
MTQSPKSMSMSVGERVTLTCKASENVVTYVSWYQQKPEQSPKLLI
YGASNRYTGVPDRFTGSGSATDFTLTISSVQAEDLADYHCGQGYSY
PYTFGGGTKLEIK
HC tumor 562 EVQLQQSGPELVKPGTSVRISCKTSGYTFTEYTIHWVKQSHGKSLE mAb with WIGNINPNNGGTTYNQKFEDKATLTVDKSSSTAYMELRSLTSEDSA DEC-205 mAb VYYCAAGWNFDYWGQGTTLTVSSASTKGPSVFPLAPSSKSTSGGTA ScFv ALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVV
TVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPE
LLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYV
DGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKV
SNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVK
GFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSR
WQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSQVQL
VESGGGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGLEWV
AVIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAV
YYCARDLWGWYFDYWGQGTLVTVSSGGGGSGGGGSGGGGSGGG
GSEIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPR
LLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRRN
WPLTFGGGTKVEIK
Rovalpituzum HC DEC-205 563 QVQLVESGGGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGL ab mAb with EWVAVIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAED tumor mAb TAVYYCARDLWGWYFDYWGQGTLVTVSSASTKGPSVFPLAPSSKS ScFv TSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLY
SLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCP
PCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKF
NWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEY
KCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLT
CLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTV
DKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSQ
VQLVQSGAEVKKPGASVKVSCKASGYTFTNYGMNWVRQAPGQGL
EWMGWINTYTGEPTYADDFKGRVTMTTDTSTSTAYMELRSLRSDD
TAVYYCARIGDSSPSDYWGQGTLVTVSSGGGGSGGGGSGGGGSGG
GGSEIVMTQSPATLSVSPGERATLSCKASQSVSNDVVWYQQKPGQA
PRLLIYYASNRYTGIPARFSGSGSGTEFTLTISSLQSEDFAVYYCQQD
YTSPWTFGQGTKLEIK
HC tumor 564 QVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGMNWVRQAPGQG mAb with LEWMGWINTYTGEPTYADDFKGRVTMTTDTSTSTAYMELRSLRSD DEC-205 mAb DTAVYYCARIGDSSPSDYWGQGTLVTVSSASTKGPSVFPLAPSSKST ScFv SGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS
LSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPP Antibody Region SEQ Sequence
ID
NO:
CPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKF
NWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEY
KCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLT
CLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTV
DKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSQ
VQLVESGGGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGLE
WVAVIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDT
AVYYCARDLWGWYFDYWGQGTLVTVSSGGGGSGGGGSGGGGSG
GGGSEIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQA
PRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQR
RNWPLTFGGGTKVEIK
PF-06647020 HC DEC-205 565 QVQLVESGGGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGL mAb with EWVAVIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAED tumor mAb TAVYYCARDLWGWYFDYWGQGTLVTVSSASTKGPSVFPLAPSSKS ScFv TSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLY
SLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCP
PCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKF
NWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEY
KCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLT
CLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTV
DKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSQ
VQLVQSGPEVKKPGASVKVSCKASGYTFTDYAVHWVRQAPGKRLE
WIGVISTYNDYTYNNQDFKGRVTMTRDTSASTAYMELSRLRSEDTA
VYYCARGNSYFYALDYWGQGTSVTVSSGGGGSGGGGSGGGGSGG
GGSEIVLTQSPATLSLSPGERATLSCRASESVDSYGKSFMHWYQQKP
GQAPRLLIYRASNLESGIPARFSGSGSGTDFTLTISSLEPEDFAVYYC
QQSNEDPWTFGGGTKLEIK
HC tumor 566 QVQLVQSGPEVKKPGASVKVSCKASGYTFTDYAVHWVRQAPGKR mAb with LEWIGVISTYNDYTYNNQDFKGRVTMTRDTSASTAYMELSRLRSED DEC-205 mAb TAVYYCARGNSYFYALDYWGQGTSVTVSSASTKGPSVFPLAPSSKS ScFv TSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLY
SLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCP
PCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKF
NWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEY
KCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLT
CLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTV
DKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSQ
VQLVESGGGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGLE
WVAVIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDT
AVYYCARDLWGWYFDYWGQGTLVTVSSGGGGSGGGGSGGGGSG
GGGSEIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQA
PRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQR
RNWPLTFGGGTKVEIK
Antibody to HC DEC-205 567 QVQLVESGGGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGL PTK7 mAb with EWVAVIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAED tumor mAb TAVYYCARDLWGWYFDYWGQGTLVTVSSASTKGPSVFPLAPSSKS ScFv TSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLY
SLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCP
PCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKF
NWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEY
KCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLT
CLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTV
DKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSQ
VQLVESGGGVVQPGRSLRLSCAASGFTFSSYAFHWVRQAPGKGLE
WVAVISYDGSIKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTA
VYYCARTYYFDYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSEI
VLTQSPDFQSVTPKEKVTITCRASQSIGSSLHWYQQKPDQSPKLLIK
YASQSFSGVPSRFSGSGSGTDFTLTINSLEAEDAAAYYCHQSSSLPIT
FGQGTRLEIK Antibody Region SEQ Sequence
ID
NO:
HC tumor 568 QVQLVESGGGVVQPGRSLRLSCAASGFTFSSYAFHWVRQAPGKGL mAb with EWVAVISYDGSIKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDT DEC-205 mAb AVYYCARTYYFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGT ScFv AALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSV
VTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAP
ELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWY
VDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCK
VSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLV
KGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSR
WQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSQVQL
VESGGGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGLEWV
AVIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAV
YYCARDLWGWYFDYWGQGTLVTVSSGGGGSGGGGSGGGGSGGG
GSEIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPR
LLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRRN
WPLTFGGGTKVEIK
Ladiratuzuma HC DEC-205 569 QVQLVESGGGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGL b mAb with EWVAVIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAED tumor mAb TAVYYCARDLWGWYFDYWGQGTLVTVSSASTKGPSVFPLAPSSKS ScFv TSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLY
SLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCP
PCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKF
NWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEY
KCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLT
CLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTV
DKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSQ
VQLVQSGAEVKKPGASVKVSCKASGLTIEDYYMHWVRQAPGQGL
EWMGWIDPENGDTEYGPKFQGRVTMTRDTSINTAYMELSRLRSDD
TAVYYCAVHNAHYGTWFAYWGQGTLVTVSSGGGGSGGGGSGGG
GSGGGGSDVVMTQSPLSLPVTLGQPASISCRSSQSLLHSSGNTYLEY
FQQRPGQSPRPLIYKISTRFSGVPDRFSGSGSGTDFTLKISRVEAEDV
GVYYCFQGSHVPYTFGGGTKVEIK
HC tumor 570 QVQLVQSGAEVKKPGASVKVSCKASGLTIEDYYMHWVRQAPGQG mAb with LEWMGWIDPENGDTEYGPKFQGRVTMTRDTSINTAYMELSRLRSD DEC-205 mAb DTAVYYCAVHNAHYGTWFAYWGQGTLVTVSSASTKGPSVFPLAPS ScFv SKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSS
GLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKT
HTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDP
EVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLN
GKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQ
VSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSK
LTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGG
GSQVQLVESGGGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGK
GLEWVAVIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRA
EDTAVYYCARDLWGWYFDYWGQGTLVTVSSGGGGSGGGGSGGG
GSGGGGSEIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKP
GQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYC
QQRRNWPLTFGGGTKVEIK
Cirmtuzumab HC DEC-205 571 QVQLVESGGGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGL mAb with EWVAVIWYDGSNKYYADSVKGRFTISRDNSKNTLY tumor mAb LQMNSLRAEDTAVYYCARDLWGWYFDYWGQGTLVTVSSASTKGP ScFv SVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTF
PAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEP
KSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVV
DVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVL
HQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSR
EEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSD
GSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG
GGGGSGGGGSQVQLQESGPGLVKPSQTLSLTCTVSGYAFTAYNIHW Antibody Region SEQ Sequence
ID
NO:
VRQAPGQGLEWMGSFDPYDGGSSYNQKFKDRLTISKDTSKNQVVL TMTNMDPVDTATYYCARGWYYFDYWGHGTLVTVSSGGGGSGGG GSGGGGSGGGGSDIVMTQTPLSLPVTPGEPASISCRASKSISKYLAW YQQKPGQAPRLLIYSGSTLQSGIPPRFSGSGYGTDFTLTINNIESEDA AYYFCQQHDESPY TFGEGTKVEIK
HC tumor 572 QVQLQESGPGLVKPSQTLSLTCTVSGYAFTAYNIHWVRQAPGQGLE mAb with WMGSFDPYDGGSSYNQKFKDRLTISKDTSKNQVVLTMTNMDPVDT DEC-205 mAb ATYYCARGWYYFDYWGHGTLVTVSSASTKGPSVFPLAPSSKSTSG ScFv GTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLS
SVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCP
APELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNW
YVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKC
KVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCL
VKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDK
SRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSQVQ
LVESGGGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGLEW
VAVIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTA
VYYCARDLWGWYFDYWGQGTLVTVSSGGGGSGGGGSGGGGSGG
GGSEIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAP
RLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRR
NWPLTFGGGTKVEIK
Antibody to HC DEC-205 573 QVQLVESGGGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGL MAGE-A3 mAb with EWVAVIWYDGSNKYYADSVKGRFTISRDNSKNTLY
tumor mAb LQMNSLRAEDTAVYYCARDLWGWYFDYWGQGTLVTVSSASTKGP ScFv SVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTF
PAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEP
KSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVV
DVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVL
HQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSR
EEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSD
GSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG
GGGGSGGGGSQVQLVESGGGVVQPGRSLRLSCTASGFRFRSHGMH
WVRQAPGKGLEWVAVISYDGNNKLYADSVKGRITISRDNSKNTLFL
QMNNVRAEDTAVYYCASPYTSDWQYFQYWGQGTLVIVSSGGGGS
GGGGSGGGGSGGGGSEIVMTQSPATLSVSPGERATFSCRASQNISTT
LAWYQQKPGQAPRLLIYDTSTRATGIPARFSGSGSGTEFTLTISSLQS
EDLAVYYCQQSNSWPLTFGGGTKVEIK
HC tumor 574 QVQLVESGGGVVQPGRSLRLSCTASGFRFRSHGMHWVRQAPGKGL mAb with EWVAVISYDGNNKLYADSVKGRITISRDNSKNTLFLQMNNVRAEDT DEC-205 mAb AVYYCASPYTSDWQYFQYWGQGTLVIVSSASTKGPSVFPLAPSSKS ScFv TSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLY
SLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCP
PCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKF
NWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEY
KCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLT
CLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTV
DKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSQ
VQLVESGGGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGLE
WVAVIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDT
AVYYCARDLWGWYFDYWGQGTLVTVSSGGGGSGGGGSGGGGSG
GGGSEIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQA
PRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQR
RNWPLTFGGGTKVEIK
Antibody to HC DEC-205 575 QVQLVESGGGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGL NY-ESO-1 mAb with EWVAVIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAED tumor mAb TAVYYCARDLWGWYFDYWGQGTLVTVSSASTKGPSVFPLAPSSKS ScFv TSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLY
SLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCP PCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKF Antibody Region SEQ Sequence
ID
NO:
NWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEY
KCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLT
CLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTV
DKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSQ
VQLVQSGGGVVRPGGSLRLSCAASGFSFIDYGMSWVRQVPGKGLE
WVAGMNWSGDKKGHAESVKGRFIISRDNAKNTLYLEMSSLRVEDT
ALYFCARGEYSNRFDPRGRGTLVTVSSGGGGSGGGGSGGGGSGGG
GSDIVMTQTPLSLPVTLGQPASLSCRSSQSLVFTDGNTYLNWFQQRP
GQSPRRLIYKVSSRDPGVPDRFSGTGSGTDFTLEISRVEAEDIGVYYC
MQGTHWPPIFGQGTKVEIK
HC tumor 576 QVQLVQSGGGVVRPGGSLRLSCAASGFSFIDYGMSWVRQVPGKGL mAb with EWVAGMNWSGDKKGHAESVKGRFIISRDNAKNTLYLEMSSLRVED DEC-205 mAb TALYFCARGEYSNRFDPRGRGTLVTVSSASTKGPSVFPLAPSSKSTS ScFv GGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSL
SSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPC
PAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFN
WYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYK
CKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTC
LVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVD
KSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSQV
QLVESGGGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGLE
WVAVIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDT
AVYYCARDLWGWYFDYWGQGTLVTVSSGGGGSGGGGSGGGGSG
GGGSEIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQA
PRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQR
RNWPLTFGGGTKVEIK
Trastuzumab HC DEC-205 686 QVQLVESGGGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGL mAb with EWVAVIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAED tumor mAb TAVYYCARDLWGWYFDYWGQGTLVTVSSASTKGPSVFPLAPSSKS ScFv TSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLY
SLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCP
PCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKF
NWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEY
KCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLT
CLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTV
DKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSE
VQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLE
WVARIYPTNGYTRYADSVKGRFTISADTSKNTAYLQMNSLRAEDT
AVYYCSRWGGDGFYAMDYWGQGTLVTVSSGGGGSGGGGSGGGG
SGGGGSDIQMTQSPSSLSASVGDRVTITCRASQDVNTAVAWYQQKP
GKAPKLLIYSASFLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYC
QQHYTTPPTFGQGTKVEIK
HC tumor 687 EVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLE mAb with WVARIYPTNGYTRYADSVKGRFTISADTSKNTAYLQMNSLRAEDT DEC-205 mAb AVYYCSRWGGDGFYAMDYWGQGTLVTVSSASTKGPSVFPLAPSSK ScFv STSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGL
YSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTC
PPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVK
FNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKE
YKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSL
TCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTV
DKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSQ
VQLVESGGGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGLE
WVAVIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDT
AVYYCARDLWGWYFDYWGQGTLVTVSSGGGGSGGGGSGGGGSG
GGGSEIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQA
PRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQR
RNWPLTFGGGTKVEIK
HC DEC205 797 QVQLVESGGGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGL Antibody Region SEQ Sequence
ID
NO:
mAb with EWVAVIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAED tumor mAb TAVYYCARDLWGWYFDYWGQGTLVTVSSASTKGPSVFPLAPSSKS
ScFv TSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLY (LH,25mer) SLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCP
PCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKF
NWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEY
KCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLT
CLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTV
DKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSD
IQMTQSPSSLSASVGDRVTITCRASQDVNTAVAWYQQKPGKAPKLL
IYSASFLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQHYTTPP
TFGQGTKVEIKGGGGSGGGGSGGGGSGGGGSGGGGSEVQLVESGG
GLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTN
GYTRYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRWG
GDGFYAMDYWGQGTLVTVSS
TABLE 9. Fusion Sequences - CD40 fusions via the light chain
Antibody Region SEQ Sequence
ID
NO:
Pertuzumab LC CD40 766 DIQMTQSPSSVSASVGDRVTITCRASQGIYSWLAWYQQKPGKAPNL mAb LIYTASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQANIFP containing LTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRE tumor mAb AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK
ScFv HKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSEVQLVESGG
GLVQPGGSLRLSCAASGFTFTDYTMDWVRQAPGKGLEWVADVNP
NSGGSIYNQRFKGRFTLSVDRSKNTLYLQMNSLRAEDTAVYYCAR
NLGPSFYFDYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSDIQM
TQSPSSLSASVGDRVTITCKASQDVSIGVAWYQQKPGKAPKLLIYSA
SYRYTGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYYIYPYTFG
QGTKVEIK
LC tumor 23 DIQMTQSPSSLSASVGDRVTITCKASQDVSIGVAWYQQKPGKAPKL mAb LIYSASYRYTGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYYIY containing PYTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYP CD40 mAb REAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADY
ScFv EKHKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLVQS
GAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLEWMGW
INPDSGGTNYAQKFQGRVTMTRDTSISTAYMELNRLRSDDTAVYYC
ARDQPLGYCTNGVCSYFDYWGQGTLVTVSSGGGGSGGGGSGGGG
SGGGGSDIQMTQSPSSVSASVGDRVTITCRASQGIYSWLAWYQQKP
GKAPNLLIYTASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYC
QQANIFPLTFGGGTKVEIK
Cetuximab LC CD40 703 DIQMTQSPSSVSASVGDRVTITCRASQGIYSWLAWYQQKPGKAPNL mAb LIYTASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQANIFP containing LTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRE tumor mAb AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK
ScFv HKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLKQSGP
GLVQPSQSLSITCTVSGFSLTNYGVHWVRQSPGKGLEWLGVIWSGG
NTDYNTPFTSRLSINKDNSKSQVFFKMNSLQSNDTAIYYCARALTY
YDYEFAYWGQGTLVTVSAGGGGSGGGGSGGGGSGGGGSDILLTQS
PVILSVSPGERVSFSCRASQSIGTNIHWYQQRTNGSPRLLIKYASESIS
GIPSRFSGSGSGTDFTLSINSVESEDIADYYCQQNNNWPTTFGAGTK
LELK
LC tumor 36 DILLTQSPVILSVSPGERVSFSCRASQSIGTNIHWYQQRTNGSPRLLIK mAb YASESISGIPSRFSGSGSGTDFTLSINSVESEDIADYYCQQNNNWPTT containing FGAGTKLELKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREA CD40 mAb KVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKH
ScFv KVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLVQSGAE Antibody Region SEQ Sequence
ID
NO:
VKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLEWMGWINP
DSGGTNYAQKFQGRVTMTRDTSISTAYMELNRLRSDDTAVYYCAR
DQPLGYCTNGVCSYFDYWGQGTLVTVSSGGGGSGGGGSGGGGSG
GGGSDIQMTQSPSSVSASVGDRVTITCRASQGIYSWLAWYQQKPGK
APNLLIYTASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQ
ANIFPLTFGGGTKVEIK
Panitumumab LC CD40 760 DIQMTQSPSSVSASVGDRVTITCRASQGIYSWLAWYQQKPGKAPNL mAb LIYTASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQANIFP containing LTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRE tumor mAb AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK
ScFv HKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLQESGP
GLVKPSETLSLTCTVSGGSVSSGDYYWTWIRQSPGKGLEWIGfflYY
SGNTNYNPSLKSRLTISIDTSKTQFSLKLSSVTAADTAIYYCVRDRVT
GAFDIWGQGTMVTVSSGGGGSGGGGSGGGGSGGGGSDIQMTQSPS
SLSASVGDRVTITCQASQDISNYLNWYQQKPGKAPKLLIYDASNLE
TGVPSRFSGSGSGTDFTFTISSLQPEDIATYFCQHFDHLPLAFGGGTK
VEIK
LC tumor 49 DIQMTQSPSSLSASVGDRVTITCQASQDISNYLNWYQQKPGKAPKL mAb LIYDASNLETGVPSRFSGSGSGTDFTFTISSLQPEDIATYFCQHFDHLP containing LAFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPR CD40 mAb EAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYE
ScFv KHKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLVQSG
AEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLEWMGWI
NPDSGGTNYAQKFQGRVTMTRDTSISTAYMELNRLRSDDTAVYYC
ARDQPLGYCTNGVCSYFDYWGQGTLVTVSSGGGGSGGGGSGGGG
SGGGGSDIQMTQSPSSVSASVGDRVTITCRASQGIYSWLAWYQQKP
GKAPNLLIYTASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYC
QQ ANIFPLTFGGGTKVEIK
Nimotuzumab LC CD40 751 DIQMTQSPSSVSASVGDRVTITCRASQGIYSWLAWYQQKPGKAPNL mAb LIYTASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQANIFP containing LTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRE tumor mAb AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK
ScFv HKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLQQSGA
EVKKPGSSVKVSCKASGYTFTNYYIYWVRQAPGQGLEWIGGINPTS
GGSNFNEKFKTRVTITVDESTNTAYMELSSLRSEDTAFYFCARQGL
WFDSDGRGFDFWGQGSTVTVSSGGGGSGGGGSGGGGSGGGGSDIQ
MTQSPSSLSASVGDRVTITCRSSQNIVHSNGNTYLDWYQQTPGKAP
KLLIYKVSNRFSGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCFQYSH
VPWTFGQGTKLEIK
LC tumor 62 DIQMTQSPSSLSASVGDRVTITCRSSQNIVHSNGNTYLDWYQQTPG mAb KAPKLLIYKVSNRFSGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCFQ containing YSHVPWTFGQGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLN CD40 mAb NFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSK
ScFv ADYEKHKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQL
VQSGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLEW
MGWINPDSGGTNYAQKFQGRVTMTRDTSISTAYMELNRLRSDDTA
VYYCARDQPLGYCTNGVCSYFDYWGQGTLVTVSSGGGGSGGGGS
GGGGSGGGGSDIQMTQSPSSVSASVGDRVTITCRASQGIYSWLAWY
QQKPGKAPNLLIYTASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFA
TYYCQQ ANIFPLTFGGGTKVEIK
Zalutumumab LC CD40 802 DIQMTQSPSSVSASVGDRVTITCRASQGIYSWLAWYQQKPGKAPNL mAb LIYTASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQANIFP containing LTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRE tumor mAb AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK
ScFv HKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLVESGG
GVVQPGRSLRLSCAASGFTFSTYGMHWVRQAPGKGLEWVAVIWD
DGSYKYYGDSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAR
DGITMVRGVMKDYFDYWGQGTLVTVSSGGGGSGGGGSGGGGSGG
GGSAIQLTQSPSSLSASVGDRVTITCRASQDISSALVWYQQKPGKAP Antibody Region SEQ Sequence
ID
NO:
KLLIYDASSLESGVPSRFSGSESGTDFTLTISSLQPEDFATYYCQQFNS YPLTFGGGTKVEIK
LC tumor 75 AIQLTQSPSSLSASVGDRVTITCRASQDISSALVWYQQKPGKAPKLLI mAb YDASSLESGVPSRFSGSESGTDFTLTISSLQPEDFATYYCQQFNSYPL containing TFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRE CD40 mAb AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK
ScFv HKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLVQSGA
EVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLEWMGWIN
PDSGGTNYAQKFQGRVTMTRDTSISTAYMELNRLRSDDTAVYYCA
RDQPLGYCTNGVCSYFDYWGQGTLVTVSSGGGGSGGGGSGGGGS
GGGGSDIQMTQSPSSVSASVGDRVTITCRASQGIYSWLAWYQQKPG
KAPNLLIYTASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQ
QANIFPLTFGGGTKVEIK
Onartuzumab LC CD40 757 DIQMTQSPSSVSASVGDRVTITCRASQGIYSWLAWYQQKPGKAPNL mAb LIYTASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQANIFP containing LTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRE tumor mAb AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK
ScFv HKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSEVQLVESGG
GLVQPGGSLRLSCAASGYTFTSYWLHWVRQAPGKGLEWVGMIDPS
NSDTRFNPNFKDRFTISADTSKNTAYLQMNSLRAEDTAVYYCATYR
SYVTPLDYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSDIQMTQ
SPSSLSASVGDRVTITCKSSQSLLYTSSQKNYLAWYQQKPGKAPKLL
IYWASTRESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYYAY
PWTFGQGTKVEIK
LC tumor 88 DIQMTQSPSSLSASVGDRVTITCKSSQSLLYTSSQKNYLAWYQQKP mAb GKAPKLLIYWASTRESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYC containing QQYYAYPWTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVC CD40 mAb LLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLT
ScFv LSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQ
VQLVQSGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGL
EWMGWINPDSGGTNYAQKFQGRVTMTRDTSISTAYMELNRLRSDD
TAVYYCARDQPLGYCTNGVCSYFDYWGQGTLVTVSSGGGGSGGG
GSGGGGSGGGGSDIQMTQSPSSVSASVGDRVTITCRASQGIYSWLA
WYQQKPGKAPNLLIYTASTLQSGVPSRFSGSGSGTDFTLTISSLQPED
FATYYCQQANIFPLTFGGGTKVEIK
Patritumab LC CD40 763 DIQMTQSPSSVSASVGDRVTITCRASQGIYSWLAWYQQKPGKAPNL mAb LIYTASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQANIFP containing LTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRE tumor mAb AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK
ScFv HKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLQQWG
AGLLKPSETLSLTCAVYGGSFSGYYWSWIRQPPGKGLEWIGEINHS
GSTNYNPSLKSRVTISVETSKNQFSLKLSSVTAADTAVYYCARDKW
TWYFDLWGRGTLVTVSSGGGGSGGGGSGGGGSGGGGSDIEMTQSP
DSLAVSLGERATINCRSSQSVLYSSSNRNYLAWYQQNPGQPPKLLIY
WASTRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQYYSTP
RTFGQGTKVEIK
LC tumor 101 DIEMTQSPDSLAVSLGERATINCRSSQSVLYSSSNRNYLAWYQQNP mAb GQPPKLLIYWASTRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYY containing CQQYYSTPRTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVC CD40 mAb LLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLT
ScFv LSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQ
VQLVQSGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGL
EWMGWINPDSGGTNYAQKFQGRVTMTRDTSISTAYMELNRLRSDD
TAVYYCARDQPLGYCTNGVCSYFDYWGQGTLVTVSSGGGGSGGG
GSGGGGSGGGGSDIQMTQSPSSVSASVGDRVTITCRASQGIYSWLA
WYQQKPGKAPNLLIYTASTLQSGVPSRFSGSGSGTDFTLTISSLQPED
FATYYCQQANIFPLTFGGGTKVEIK
Clivatuzumab LC CD40 709 DIQMTQSPSSVSASVGDRVTITCRASQGIYSWLAWYQQKPGKAPNL mAb LIYTASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQANIFP Antibody Region SEQ Sequence
ID
NO:
containing LTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRE tumor mAb AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK ScFv HKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLQQSGA
EVKKPGASVKVSCEASGYTFPSYVLHWVKQAPGQGLEWIGYINPY
NDGTQYNEKFKGKATLTRDTSINTAYMELSRLRSDDTAVYYCARG
FGGSYGFAYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSDIQLT
QSPSSLSASVGDRVTMTCSASSSVSSSYLYWYQQKPGKAPKLWIYS
TSNLASGVPARFSGSGSGTDFTLTISSLQPEDSASYFCHQWNRYPYT
FGGGTRLEIK
LC tumor 114 DIQLTQSPSSLSASVGDRVTMTCSASSSVSSSYLYWYQQKPGKAPK mAb LWIYSTSNLASGVPARFSGSGSGTDFTLTISSLQPEDSASYFCHQWN containing RYPYTFGGGTRLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNF CD40 mAb YPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKAD
ScFv YEKHKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLVQ
SGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLEWMG
WINPDSGGTNYAQKFQGRVTMTRDTSISTAYMELNRLRSDDTAVY
YCARDQPLGYCTNGVCSYFDYWGQGTLVTVSSGGGGSGGGGSGG
GGSGGGGSDIQMTQSPSSVSASVGDRVTITCRASQGIYSWLAWYQQ
KPGKAPNLLIYTASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATY
YCQQANIFPLTFGGGTKVEIK
Sofituzumab LC CD40 790 DIQMTQSPSSVSASVGDRVTITCRASQGIYSWLAWYQQKPGKAPNL mAb LIYTASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQANIFP containing LTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRE tumor mAb AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK
ScFv HKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSEVQLVESGG
GLVQPGGSLRLSCAASGYSITNDYAWNWVRQAPGKGLEWVGYISY
SGYTTYNPSLKSRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARWT
SGLDYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSDIQMTQSPS
SLSASVGDRVTITCKASDLIHNWLAWYQQKPGKAPKLLIYGATSLE
TGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYWTTPFTFGQGT
KVEIK
LC tumor 127 DIQMTQSPSSLSASVGDRVTITCKASDLIHNWLAWYQQKPGKAPKL mAb LIYGATSLETGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYWTT containing PFTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPR CD40 mAb EAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYE
ScFv KHKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLVQSG
AEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLEWMGWI
NPDSGGTNYAQKFQGRVTMTRDTSISTAYMELNRLRSDDTAVYYC
ARDQPLGYCTNGVCSYFDYWGQGTLVTVSSGGGGSGGGGSGGGG
SGGGGSDIQMTQSPSSVSASVGDRVTITCRASQGIYSWLAWYQQKP
GKAPNLLIYTASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYC
QQANIFPLTFGGGTKVEIK
Edrecolomab LC CD40 718 DIQMTQSPSSVSASVGDRVTITCRASQGIYSWLAWYQQKPGKAPNL mAb LIYTASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQANIFP containing LTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRE tumor mAb AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK
ScFv HKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLQQSGA
ELVRPGTSVKVSCKASGYAFTNYLIEWVKQRPGQGLEWIGVINPGS
GGTNYNEKFKGKATLTADKSSSTAYMQLSSLTSDDSAVYFCARDG
PWFAYWGQGTLVTVSAGGGGSGGGGSGGGGSGGGGSNIVMTQSP
KSMSMSVGERVTLTCKASENVVTYVSWYQQKPEQSPKLLIYGASN
RYTGVPDRFTGSGSATDFTLTISSVQAEDLADYHCGQGYSYPYTFG
GGTKLEIK
LC tumor 140 NIVMTQSPKSMSMSVGERVTLTCKASENVVTYVSWYQQKPEQSPK mAb LLIYGASNRYTGVPDRFTGSGSATDFTLTISSVQAEDLADYHCGQG containing YSYPYTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNF CD40 mAb YPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKAD
ScFv YEKHKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLVQ
SGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLEWMG Antibody Region SEQ Sequence
ID
NO:
WINPDSGGTNYAQKFQGRVTMTRDTSISTAYMELNRLRSDDTAVY
YCARDQPLGYCTNGVCSYFDYWGQGTLVTVSSGGGGSGGGGSGG
GGSGGGGSDIQMTQSPSSVSASVGDRVTITCRASQGIYSWLAWYQQ
KPGKAPNLLIYTASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATY
YCQQANIFPLTFGGGTKVEIK
Adecatumum LC CD40 694 DIQMTQSPSSVSASVGDRVTITCRASQGIYSWLAWYQQKPGKAPNL ab mAb LIYTASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQANIFP containing LTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRE tumor mAb AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK
ScFv HKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSEVQLLESGG
GVVQPGRSLRLSCAASGFTFSSYGMHWVRQAPGKGLEWVAVISYD
GSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKD
MGWGSGWRPYYYYGMDVWGQGTTVTVSSGGGGSGGGGSGGGGS
GGGGSELQMTQSPSSLSASVGDRVTITCRTSQSISSYLNWYQQKPGQ
PPKLLIYWASTRESGVPDRFSGSGSGTDFTLTISSLQPEDSATYYCQQ
SYDIPYTFGQGTKLEIK
LC tumor 153 ELQMTQSPSSLSASVGDRVTITCRTSQSISSYLNWYQQKPGQPPKLLI mAb YWASTRESGVPDRFSGSGSGTDFTLTISSLQPEDSATYYCQQSYDIP containing YTFGQGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRE CD40 mAb AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK
ScFv HKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLVQSGA
EVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLEWMGWIN
PDSGGTNYAQKFQGRVTMTRDTSISTAYMELNRLRSDDTAVYYCA
RDQPLGYCTNGVCSYFDYWGQGTLVTVSSGGGGSGGGGSGGGGS
GGGGSDIQMTQSPSSVSASVGDRVTITCRASQGIYSWLAWYQQKPG
KAPNLLIYTASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQ
QANIFPLTFGGGTKVEIK
Anetumab LC CD40 700 DIQMTQSPSSVSASVGDRVTITCRASQGIYSWLAWYQQKPGKAPNL mAb LIYTASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQANIFP containing LTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRE tumor mAb AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK
ScFv HKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVELVQSGA
EVKKPGESLKISCKGSGYSFTSYWIGWVRQAPGKGLEWMGIIDPGD
SRTRYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCARGQL
YGGTYMDGWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSDIALT
QPASVSGSPGQSITISCTGTSSDIGGYNSVSWYQQHPGKAPKLMIYG
VNNRPSGVSNRFSGSKSGNTASLTISGLQAEDEADYYCSSYDIESAT
PVFGGGTKLTVL
LC tumor 166 DIALTQPASVSGSPGQSITISCTGTSSDIGGYNSVSWYQQHPGKAPKL mAb MIYGVNNRPSGVSNRFSGSKSGNTASLTISGLQAEDEADYYCSSYDI containing ESATPVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLIS CD40 mAb DFYPGAVTVAWKGDSSPVKAGVETTTPSKQSNNKYAASSYLSLTPE
ScFv QWKSHRSYSCQVTHEGSTVEKTVAPTECSGGGGSGGGGSQVQLVQ
SGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLEWMG
WINPDSGGTNYAQKFQGRVTMTRDTSISTAYMELNRLRSDDTAVY
YCARDQPLGYCTNGVCSYFDYWGQGTLVTVSSGGGGSGGGGSGG
GGSGGGGSDIQMTQSPSSVSASVGDRVTITCRASQGIYSWLAWYQQ
KPGKAPNLLIYTASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATY
YCQQANIFPLTFGGGTKVEIK
huDS6 LC CD40 724 DIQMTQSPSSVSASVGDRVTITCRASQGIYSWLAWYQQKPGKAPNL mAb LIYTASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQANIFP containing LTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRE tumor mAb AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK
ScFv HKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQAQLVQSGA
EVVKPGASVKMSCKASGYTFTSYNMHWVKQTPGQGLEWIGYIYPG
NGATNYNQKFQGKATLTADPSSSTAYMQISSLTSEDSAVYFCARGD
SVPFAYWGQGTLVTVSAGGGGSGGGGSGGGGSGGGGSEIVLTQSP
ATMSASPGERVTITCSAHSSVSFMHWFQQKPGTSPKLWIYSTSSLAS
GVPARFGGSGSGTSYSLTISSMEAEDAATYYCQQRSSFPLTFGAGTK Antibody Region SEQ Sequence
ID
NO:
LELK
LC tumor 179 EIVLTQSPATMSASPGERVTITCSAHSSVSFMHWFQQKPGTSPKLWI mAb YSTSSLASGVPARFGGSGSGTSYSLTISSMEAEDAATYYCQQRSSFP containing LTFGAGTKLELKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPR CD40 mAb EAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYE
ScFv KHKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLVQSG
AEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLEWMGWI
NPDSGGTNYAQKFQGRVTMTRDTSISTAYMELNRLRSDDTAVYYC
ARDQPLGYCTNGVCSYFDYWGQGTLVTVSSGGGGSGGGGSGGGG
SGGGGSDIQMTQSPSSVSASVGDRVTITCRASQGIYSWLAWYQQKP
GKAPNLLIYTASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYC
QQANIFPLTFGGGTKVEIK
Lifastuzumab LC CD40 736 DIQMTQSPSSVSASVGDRVTITCRASQGIYSWLAWYQQKPGKAPNL mAb LIYTASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQANIFP containing LTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRE tumor mAb AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK
ScFv HKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSEVQLVESGG
GLVQPGGSLRLSCAASGFSFSDFAMSWVRQAPGKGLEWVATIGRV
AFHTYYPDSMKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARH
RGFDVGHFDFWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSDIQ
MTQSPSSLSASVGDRVTITCRSSETLVHSSGNTYLEWYQQKPGKAP
KLLIYRVSNRFSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCFQGS
FNPLTFGQGTKVEIK
LC tumor 192 DIQMTQSPSSLSASVGDRVTITCRSSETLVHSSGNTYLEWYQQKPGK mAb APKLLIYRVSNRFSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCFQ containing GSFNPLTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNN CD40 mAb FYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKA
ScFv DYEKHKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLV
QSGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLEWM
GWINPDSGGTNYAQKFQGRVTMTRDTSISTAYMELNRLRSDDTAV
YYCARDQPLGYCTNGVCSYFDYWGQGTLVTVSSGGGGSGGGGSG
GGGSGGGGSDIQMTQSPSSVSASVGDRVTITCRASQGIYSWLAWYQ
QKPGKAPNLLIYTASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFAT
YYCQQANIFPLTFGGGTKVEIK
Sacituzumab LC CD40 781 DIQMTQSPSSVSASVGDRVTITCRASQGIYSWLAWYQQKPGKAPNL mAb LIYTASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQANIFP containing LTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRE tumor mAb AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK
ScFv HKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLQQSGS
ELKKPGASVKVSCKASGYTFTNYGMNWVKQAPGQGLKWMGWIN
TYTGEPTYTDDFKGRFAFSLDTSVSTAYLQISSLKADDTAVYFCARG
GFGSSYWYFDVWGQGSLVTVSSGGGGSGGGGSGGGGSGGGGSDIQ
LTQSPSSLSASVGDRVSITCKASQDVSIAVAWYQQKPGKAPKLLIYS
ASYRYTGVPDRFSGSGSGTDFTLTISSLQPEDFAVYYCQQHYITPLTF
GAGTKVEIK
LC tumor 205 DIQLTQSPSSLSASVGDRVSITCKASQDVSIAVAWYQQKPGKAPKLL mAb IYSASYRYTGVPDRFSGSGSGTDFTLTISSLQPEDFAVYYCQQHYITP containing LTFGAGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRE CD40 mAb AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK
ScFv HKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLVQSGA
EVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLEWMGWIN
PDSGGTNYAQKFQGRVTMTRDTSISTAYMELNRLRSDDTAVYYCA
RDQPLGYCTNGVCSYFDYWGQGTLVTVSSGGGGSGGGGSGGGGS
GGGGSDIQMTQSPSSVSASVGDRVTITCRASQGIYSWLAWYQQKPG
KAPNLLIYTASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQ
QANIFPLTFGGGTKVEIK
PR1A3 LC CD40 772 DIQMTQSPSSVSASVGDRVTITCRASQGIYSWLAWYQQKPGKAPNL mAb LIYTASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQANIFP containing LTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRE Antibody Region SEQ Sequence
ID
NO:
tumor mAb AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK ScFv HKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVKLQQSGP
ELKKPGETVKISCKASGYTFTEFGMNWVKQAPGKGLKWMGWINT
KTGEATYVEEFKGRFAFSLETSATTAYLQINNLKNEDTAKYFCARW
DFYDYVEAMDYWGQGTTVTVSSGGGGSGGGGSGGGGSGGGGSDI
VMTQSQRFMSTSVGDRVSVTCKASQNVGTNVAWYQQKPGQSPKA
LIYSASYRYSGVPDRFTGSGSGTDFTLTISNVQSEDLAEYFCHQYYT
YPLFTFGSGTKLEMK
LC tumor 218 DIVMTQSQRFMSTSVGDRVSVTCKASQNVGTNVAWYQQKPGQSP mAb KALIYSASYRYSGVPDRFTGSGSGTDFTLTISNVQSEDLAEYFCHQY containing YTYPLFTFGSGTKLEMKRTVAAPSVFIFPPSDEQLKSGTASVVCLLN CD40 mAb NFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSK
ScFv ADYEKHKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQL
VQSGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLEW
MGWINPDSGGTNYAQKFQGRVTMTRDTSISTAYMELNRLRSDDTA
VYYCARDQPLGYCTNGVCSYFDYWGQGTLVTVSSGGGGSGGGGS
GGGGSGGGGSDIQMTQSPSSVSASVGDRVTITCRASQGIYSWLAWY
QQKPGKAPNLLIYTASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFA
TYYCQQANIFPLTFGGGTKVEIK
Humanized LC CD40 820 DIQMTQSPSSVSASVGDRVTITCRASQGIYSWLAWYQQKPGKAPNL PR1A3 mAb LIYTASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQANIFP containing LTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRE tumor mAb AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK
ScFv HKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLVQSGA
EVKKPGASVKVSCKASGYTFTEFGMNWVRQAPGQGLEWMGWINT
KTGEATYVEEFKGRVTFTTDTSTSTAYMELRSLRSDDTAVYYCAR
WDFAYYVEAMDYWGQGTTVTVSSGGGGSGGGGSGGGGSGGGGS
DIQMTQSPSSLSASVGDRVTITCKASAAVGTYVAWYQQKPGKAPK
LLIYSASYRKRGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCHQYYT
YPLFTFGQGTKLEIK
LC tumor 817 DIQMTQSPSSLSASVGDRVTITCKASAAVGTYVAWYQQKPGKAPK mAb LLIYSASYRKRGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCHQYYT containing YPLFTFGQGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFY CD40 mAb PREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADY
ScFv EKHKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLVQS
GAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLEWMGW
INPDSGGTNYAQKFQGRVTMTRDTSISTAYMELNRLRSDDTAVYYC
ARDQPLGYCTNGVCSYFDYWGQGTLVTVSSGGGGSGGGGSGGGG
SGGGGSDIQMTQSPSSVSASVGDRVTITCRASQGIYSWLAWYQQKP
GKAPNLLIYTASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYC
QQANIFPLTFGGGTKVEIK
LC tumor 844 DIQMTQSPSSLSASVGDRVTITCKASAAVGTYVAWYQQKPGKAPK mAb LLIYSASYRKRGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCHQYYT containing YPLFTFGQGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFY CD40 mab PREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADY scFv (LH) EKHKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSDIQMTQS
PSSVSASVGDRVTITCRASQGIYSWLAWYQQKPGKAPNLLIYTASTL
QSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQANIFPLTFGGGT
KVEIKGGGGSGGGGSGGGGSGGGGSQVQLVQSGAEVKKPGASVK
VSCKASGYTFTGYYMHWVRQAPGQGLEWMGWINPDSGGTNYAQ
KFQGRVTMTRDTSISTAYMELNRLRSDDTAVYYCARDQPLGYCTN
GVCSYFDYWGQGTLVTVSS
Humanized LC CD40 838 DIQMTQSPSSVSASVGDRVTITCRASQGIYSWLAWYQQKPGKAPNL Ab2-3 mAb LIYTASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQANIFP containing LTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRE tumor mAb AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK
ScFv HKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSEVQLQESGP
GLVKPGGSLSLSCAASGFVFSSYDMSWVRQTPERGLEWVAYISSGG
GITYAPSTVKGRFTVSRDNAKNTLYLQMNSLTSEDTAVYYCAAHY Antibody Region SEQ Sequence
ID
NO:
FGSSGPFAYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSDIQMT QSPASLSASVGDRVTITCRASENIFSYLAWYQQKPGKSPKLLVYNTR TLAEGVPSRFSGSGSGTDFSLTISSLQPEDFATYYCQHHYGTPFTFGS GTKLEIK
LC tumor 835 DIQMTQSPASLSASVGDRVTITCRASENIFSYLAWYQQKPGKSPKLL mAb VYNTRTLAEGVPSRFSGSGSGTDFSLTISSLQPEDFATYYCQHHYGT containing PFTFGSGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPR CD40 mAb EAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYE
ScFv KHKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLVQSG
AEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLEWMGWI
NPDSGGTNYAQKFQGRVTMTRDTSISTAYMELNRLRSDDTAVYYC
ARDQPLGYCTNGVCSYFDYWGQGTLVTVSSGGGGSGGGGSGGGG
SGGGGSDIQMTQSPSSVSASVGDRVTITCRASQGIYSWLAWYQQKP
GKAPNLLIYTASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYC
QQANIFPLTFGGGTKVEIK
LC tumor 842 DIQMTQSPASLSASVGDRVTITCRASENIFSYLAWYQQKPGKSPKLL mAb VYNTRTLAEGVPSRFSGSGSGTDFSLTISSLQPEDFATYYCQHHYGT containing PFTFGSGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPR CD40 mab EAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYE scFv (LH) KHKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSDIQMTQSPS
SVSASVGDRVTITCRASQGIYSWLAWYQQKPGKAPNLLIYTASTLQ
SGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQANIFPLTFGGGTK
VEIKGGGGSGGGGSGGGGSGGGGSQVQLVQSGAEVKKPGASVKVS
CKASGYTFTGYYMHWVRQAPGQGLEWMGWINPDSGGTNYAQKF
QGRVTMTRDTSISTAYMELNRLRSDDTAVYYCARDQPLGYCTNGV
CSYFDYWGQGTLVTVSS
IMAB362, LC CD40 727 DIQMTQSPSSVSASVGDRVTITCRASQGIYSWLAWYQQKPGKAPNL
CLAUDIXIM mAb LIYTASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQANIFP
AB containing LTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRE tumor mAb AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK
ScFv HKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLQQPGA
ELVRPGASVKLSCKASGYTFTSYWINWVKQRPGQGLEWIGNIYPSD
SYTNYNQKFKDKATLTVDKSSSTAYMQLSSPTSEDSAVYYCTRSW
RGNSFDYWGQGTTLTVSSGGGGSGGGGSGGGGSGGGGSDIVMTQS
PSSLTVTAGEKVTMSCKSSQSLLNSGNQKNYLTWYQQKPGQPPKLL
IYWASTRESGVPDRFTGSGSGTDFTLTISSVQAEDLAVYYCQNDYS
YPFTFGSGTKLEIK
LC tumor 231 DIVMTQSPSSLTVTAGEKVTMSCKSSQSLLNSGNQKNYLTWYQQK mAb PGQPPKLLIYWASTRESGVPDRFTGSGSGTDFTLTISSVQAEDLAVY containing YCQNDYSYPFTFGSGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVC CD40 mAb LLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLT
ScFv LSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQ
VQLVQSGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGL
EWMGWINPDSGGTNYAQKFQGRVTMTRDTSISTAYMELNRLRSDD
TAVYYCARDQPLGYCTNGVCSYFDYWGQGTLVTVSSGGGGSGGG
GSGGGGSGGGGSDIQMTQSPSSVSASVGDRVTITCRASQGIYSWLA
WYQQKPGKAPNLLIYTASTLQSGVPSRFSGSGSGTDFTLTISSLQPED
FATYYCQQANIFPLTFGGGTKVEIK
AMG595 LC CD40 697 DIQMTQSPSSVSASVGDRVTITCRASQGIYSWLAWYQQKPGKAPNL mAb LIYTASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQANIFP containing LTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRE tumor mAb AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK
ScFv HKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLVESGG
GVVQSGRSLRLSCAASGFTFRNYGMHWVRQAPGKGLEWVAVIWY
DGSDKYYADSVRGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAR
DGYDILTGNPRDFDYWGQGTLVTVSSGGGGSGGGGSGGGGSGGG
GSDTVMTQTPLSSHVTLGQPASISCRSSQSLVHSDGNTYLSWLQQRP
GQPPRLLIYRISRRFSGVPDRFSGSGAGTDFTLEISRVEAEDVGVYYC
MQSTHVPRTFGQGTKVEIK Antibody Region SEQ Sequence
ID
NO:
LC tumor 270 DTVMTQTPLSSHVTLGQPASISCRSSQSLVHSDGNTYLSWLQQRPG mAb QPPRLLIYRISRRFSGVPDRFSGSGAGTDFTLEISRVEAEDVGVYYC containing MQSTHVPRTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCL CD40 mAb LNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTL
ScFv SKADYEKHKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQV
QLVQSGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLE
WMGWINPDSGGTNYAQKFQGRVTMTRDTSISTAYMELNRLRSDDT
AVYYCARDQPLGYCTNGVCSYFDYWGQGTLVTVSSGGGGSGGGG
SGGGGSGGGGSDIQMTQSPSSVSASVGDRVTITCRASQGIYSWLAW
YQQKPGKAPNLLIYTASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDF
ATYYCQQANIFPLTFGGGTKVEIK
ABT806 LC CD40 691 DIQMTQSPSSVSASVGDRVTITCRASQGIYSWLAWYQQKPGKAPNL mAb LIYTASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQANIFP containing LTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRE tumor mAb AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK
ScFv HKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSEVQLQESGP
GLVKPSQTLSLTCTVSGYSISRDFAWNWIRQPPGKGLEWMGYISYN
GNTRYQPSLKSRITISRDTSKNQFFLKLNSVTAADTATYYCVTASRG
FPYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSDIQMTQSPSSM
SVSVGDRVTITCHSSQDINSNIGWLQQKPGKSFKGLIYHGTNLDDGV
PSRFSGSGSGTDYTLTISSLQPEDFATYYCVQYAQFPWTFGGGTKLE
IK
LC tumor 283 DIQMTQSPSSMSVSVGDRVTITCHSSQDINSNIGWLQQKPGKSFKGL mAb IYHGTNLDDGVPSRFSGSGSGTDYTLTISSLQPEDFATYYCVQYAQF containing PWTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYP CD40 mAb REAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADY
ScFv EKHKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLVQS
GAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLEWMGW
INPDSGGTNYAQKFQGRVTMTRDTSISTAYMELNRLRSDDTAVYYC
ARDQPLGYCTNGVCSYFDYWGQGTLVTVSSGGGGSGGGGSGGGG
SGGGGSDIQMTQSPSSVSASVGDRVTITCRASQGIYSWLAWYQQKP
GKAPNLLIYTASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYC
QQANIFPLTFGGGTKVEIK
Sibrotuzumab LC CD40 787 DIQMTQSPSSVSASVGDRVTITCRASQGIYSWLAWYQQKPGKAPNL mAb LIYTASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQANIFP containing LTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRE tumor mAb AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK
ScFv HKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLVQSGA
EVKKPGASVKVSCKTSRYTFTEYTIHWVRQAPGQRLEWIGGINPNN
GIPNYNQKFKGRVTITVDTSASTAYMELSSLRSEDTAVYYCARRRIA
YGYDEGHAMDYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSDI
VMTQSPDSLAVSLGERATINCKSSQSLLYSRNQKNYLAWYQQKPG
QPPKLLIFWASTRESGVPDRFSGSGFGTDFTLTISSLQAEDVAVYYC
QQYFSYPLTFGQGTKVEIK
LC tumor 296 DIVMTQSPDSLAVSLGERATINCKSSQSLLYSRNQKNYLAWYQQKP mAb GQPPKLLIFWASTRESGVPDRFSGSGFGTDFTLTISSLQAEDVAVYY containing CQQYFSYPLTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVC CD40 mAb LLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLT
ScFv LSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQ
VQLVQSGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGL
EWMGWINPDSGGTNYAQKFQGRVTMTRDTSISTAYMELNRLRSDD
TAVYYCARDQPLGYCTNGVCSYFDYWGQGTLVTVSSGGGGSGGG
GSGGGGSGGGGSDIQMTQSPSSVSASVGDRVTITCRASQGIYSWLA
WYQQKPGKAPNLLIYTASTLQSGVPSRFSGSGSGTDFTLTISSLQPED
FATYYCQQANIFPLTFGGGTKVEIK
DS-8895a LC CD40 712 DIQMTQSPSSVSASVGDRVTITCRASQGIYSWLAWYQQKPGKAPNL variant 1 mAb LIYTASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQANIFP containing LTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRE tumor mAb AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK Antibody Region SEQ Sequence
ID
NO:
ScFv HKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLVQSGA
EVKKPGASVKVSCKASGYTFIDYSMHWVRQAPGQGLEWMGWINT
YTGEPTYSDDFKGRVTITADTSTSTAYLELSSLRSEDTAVYYCATYY
RYERDFDYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSDIVMT
QSPLSLPVTPGEPASISCRSSQSIVHSSGITYLEWYLQKPGQSPQLLIY
KVSNRFSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCFQGSHVP
YTFGQGTKVEIK
LC tumor 309 DIVMTQSPLSLPVTPGEPASISCRSSQSIVHSSGITYLEWYLQKPGQSP mAb QLLIYKVSNRFSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCFQG containing SHVPYTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNN CD40 mAb FYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKA
ScFv DYEKHKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLV
QSGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLEWM
GWINPDSGGTNYAQKFQGRVTMTRDTSISTAYMELNRLRSDDTAV
YYCARDQPLGYCTNGVCSYFDYWGQGTLVTVSSGGGGSGGGGSG
GGGSGGGGSDIQMTQSPSSVSASVGDRVTITCRASQGIYSWLAWYQ
QKPGKAPNLLIYTASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFAT
YYCQQANIFPLTFGGGTKVEIK
DS-8895a LC CD40 715 DIQMTQSPSSVSASVGDRVTITCRASQGIYSWLAWYQQKPGKAPNL variant 2 mAb LIYTASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQANIFP containing LTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRE tumor mAb AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK
ScFv HKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQIQLVQSGA
EVKKPGASVKVSCKASGYTFIDYSMHWVRQAPGQGLKWMGWINT
YTGEPTYSDDFKGRFAFSLDTSTSTAYLELSSLRSEDTAVYYCATYY
RYERDFDYWGQGTLVTVSSGGGGSGGGGSGGGSGGGGSDVLMTQ
SPLSLPVTPGEPASISCRSSQSIVHSSGITYLEWYLQKPGQSPQLLIYK
VSNRFSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCFQGSHVPYT
FGQGTKVEIK
LC tumor 322 DVLMTQSPLSLPVTPGEPASISCRSSQSIVHSSGITYLEWYLQKPGQS mAb PQLLIYKVSNRFSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCFQ containing GSHVPYTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLN CD40 mAb NFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSK
ScFv ADYEKHKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQL
VQSGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLEW
MGWINPDSGGTNYAQKFQGRVTMTRDTSISTAYMELNRLRSDDTA
VYYCARDQPLGYCTNGVCSYFDYWGQGTLVTVSSGGGGSGGGGS
GGGGSGGGGSDIQMTQSPSSVSASVGDRVTITCRASQGIYSWLAWY
QQKPGKAPNLLIYTASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFA
TYYCQQANIFPLTFGGGTKVEIK
MEDI-547 LC CD40 742 DIQMTQSPSSVSASVGDRVTITCRASQGIYSWLAWYQQKPGKAPNL mAb LIYTASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQANIFP containing LTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRE tumor mAb AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK
ScFv HKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSEVQLLESGG
GLVQPGGSLRLSCAASGFTFSHYMMAWVRQAPGKGLEWVSRIGPS
GGPTHYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAGY
DSGYDYVAVAGPAEYFQHWGQGTLVTVSSGGGGSGGGGSGGGGS
GGGGSDIQMTQSPSSLSASVGDRVTITCRASQSISTWLAWYQQKPG
KAPKLLIYKASNLHTGVPSRFSGSGSGTEFSLTISGLQPDDFATYYCQ
QYNSYSRTFGQGTKVEIK
LC tumor 335 DIQMTQSPSSLSASVGDRVTITCRASQSISTWLAWYQQKPGKAPKLL mAb IYKASNLHTGVPSRFSGSGSGTEFSLTISGLQPDDFATYYCQQYNSY containing SRTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPR CD40 mAb EAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYE
ScFv KHKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLVQSG
AEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLEWMGWI
NPDSGGTNYAQKFQGRVTMTRDTSISTAYMELNRLRSDDTAVYYC
ARDQPLGYCTNGVCSYFDYWGQGTLVTVSSGGGGSGGGGSGGGG Antibody Region SEQ Sequence
ID
NO:
SGGGGSDIQMTQSPSSVSASVGDRVTITCRASQGIYSWLAWYQQKP GKAPNLLIYTASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYC QQANIFPLTFGGGTKVEIK
Narnatumab LC CD40 748 DIQMTQSPSSVSASVGDRVTITCRASQGIYSWLAWYQQKPGKAPNL mAb LIYTASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQANIFP containing LTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRE tumor mAb AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK
ScFv HKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSEVQLVESGG
GLVQPGGSLRLSCAASGFTFSSYLMTWVRQAPGKGLEWVANIKQD
GSEKYYVDSVKGRFTISRDNAKNSLNLQMNSLRAEDTAVYYCTRD
GYSSGRHYGMDVWGQGTTVIVSSGGGGSGGGGSGGGGSGGGGSEI
VLTQSPATLSLSPGERATLSCRASQSVSRYLAWYQQKPGQAPRLLIY
DASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRSNWPR
TFGQGTKVEIK
LC tumor 348 EIVLTQSPATLSLSPGERATLSCRASQSVSRYLAWYQQKPGQAPRLL mAb IYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRSNWP containing RTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPR CD40 mAb EAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYE
ScFv KHKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLVQSG
AEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLEWMGWI
NPDSGGTNYAQKFQGRVTMTRDTSISTAYMELNRLRSDDTAVYYC
ARDQPLGYCTNGVCSYFDYWGQGTLVTVSSGGGGSGGGGSGGGG
SGGGGSDIQMTQSPSSVSASVGDRVTITCRASQGIYSWLAWYQQKP
GKAPNLLIYTASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYC
QQANIFPLTFGGGTKVEIK
RG7841 LC CD40 775 DIQMTQSPSSVSASVGDRVTITCRASQGIYSWLAWYQQKPGKAPNL mAb LIYTASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQANIFP containing LTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRE tumor mAb AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK
ScFv HKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSEVQLVESGP
ALVKPTQTLTLTCTVSGFSLTGYSVNWIRQPPGKALEWLGMIWGD
GSTDYNSALKSRLTISKDTSKNQVVLTMTNMDPVDTATYYCARDY
YFNYASWFAYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSDIQ
MTQSPSSLSASVGDRVTITCSASQGISNYLNWYQQKPGKTVKLLIYY
TSNLHSGVPSRFSGSGSGTDYTLTISSLQPEDFATYYCQQYSELPWT
FGQGTKVEIK
LC tumor 361 DIQMTQSPSSLSASVGDRVTITCSASQGISNYLNWYQQKPGKTVKLL mAb IYYTSNLHSGVPSRFSGSGSGTDYTLTISSLQPEDFATYYCQQYSELP containing WTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPR CD40 mAb EAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYE
ScFv KHKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLVQSG
AEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLEWMGWI
NPDSGGTNYAQKFQGRVTMTRDTSISTAYMELNRLRSDDTAVYYC
ARDQPLGYCTNGVCSYFDYWGQGTLVTVSSGGGGSGGGGSGGGG
SGGGGSDIQMTQSPSSVSASVGDRVTITCRASQGIYSWLAWYQQKP
GKAPNLLIYTASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYC
QQANIFPLTFGGGTKVEIK
Farletuzumab LC CD40 721 DIQMTQSPSSVSASVGDRVTITCRASQGIYSWLAWYQQKPGKAPNL mAb LIYTASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQANIFP containing LTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRE tumor mAb AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK
ScFv HKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSEVQLVESGG
GVVQPGRSLRLSCSASGFTFSGYGLSWVRQAPGKGLEWVAMISSG
GSYTYYADSVKGRFAISRDNAKNTLFLQMDSLRPEDTGVYFCARH
GDDPAWFAYWGQGTPVTVSSGGGGSGGGGSGGGGSGGGGSDIQL
TQSPSSLSASVGDRVTITCSVSSSISSNNLHWYQQKPGKAPKPWIYG
TSNLASGVPSRFSGSGSGTDYTFTISSLQPEDIATYYCQQWSSYPYM
YTFGQGTKVEIK
LC tumor 374 DIQLTQSPSSLSASVGDRVTITCSVSSSISSNNLHWYQQKPGKAPKP Antibody Region SEQ Sequence
ID
NO:
mAb WIYGTSNLASGVPSRFSGSGSGTDYTFTISSLQPEDIATYYCQQWSS containing YPYMYTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNN CD40 mAb FYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKA
ScFv DYEKHKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLV
QSGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLEWM
GWINPDSGGTNYAQKFQGRVTMTRDTSISTAYMELNRLRSDDTAV
YYCARDQPLGYCTNGVCSYFDYWGQGTLVTVSSGGGGSGGGGSG
GGGSGGGGSDIQMTQSPSSVSASVGDRVTITCRASQGIYSWLAWYQ
QKPGKAPNLLIYTASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFAT
YYCQQANIFPLTFGGGTKVEIK
Mirvetuximab LC CD40 745 DIQMTQSPSSVSASVGDRVTITCRASQGIYSWLAWYQQKPGKAPNL mAb LIYTASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQANIFP containing LTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRE tumor mAb AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK
ScFv HKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLVQSGA
EVVKPGASVKISCKASGYTFTGYFMNWVKQSPGQSLEWIGRIHPYD
GDTFYNQKFQGKATLTVDKSSNTAHMELLSLTSEDFAVYYCTRYD
GSRAMDYWGQGTTVTVSSGGGGSGGGGSGGGGSGGGGSDIVLTQ
SPLSLAVSLGQPAIISCKASQSVSFAGTSLMHWYHQKPGQQPRLLIY
RASNLEAGVPDRFSGSGSKTDFTLTISPVEAEDAATYYCQQSREYPY
TFGGGTKLEIK
LC tumor 387 DIVLTQSPLSLAVSLGQPAIISCKASQSVSFAGTSLMHWYHQKPGQQ mAb PRLLIYRASNLEAGVPDRFSGSGSKTDFTLTISPVEAEDAATYYCQQ containing SREYPYTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNN CD40 mAb FYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKA
ScFv DYEKHKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLV
QSGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLEWM
GWINPDSGGTNYAQKFQGRVTMTRDTSISTAYMELNRLRSDDTAV
YYCARDQPLGYCTNGVCSYFDYWGQGTLVTVSSGGGGSGGGGSG
GGGSGGGGSDIQMTQSPSSVSASVGDRVTITCRASQGIYSWLAWYQ
QKPGKAPNLLIYTASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFAT
YYCQQANIFPLTFGGGTKVEIK
J591 variantl LC CD40 730 DIQMTQSPSSVSASVGDRVTITCRASQGIYSWLAWYQQKPGKAPNL mAb LIYTASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQANIFP containing LTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRE tumor mAb AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK
ScFv HKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSEVQLQQSGP
ELKKPGTSVRISCKTSGYTFTEYTIHWVKQSHGKSLEWIGNINPNNG
GTTYNQKFEDKATLTVDKSSSTAYMELRSLTSEDSAVYYCAAGWN
FDYWGQGTTLTVSSGGGGSGGGGSGGGGSGGGGSDIVMTQSHKF
MSTSVGDRVSIICKASQDVGTAVDWYQQKPGQSPKLLIYWASTRHT
GVPDRFTGSGSGTDFTLTITNVQSEDLADYFCQQYNSYPLTFGAGT
MLDLK
LC tumor 400 DIVMTQSHKFMSTSVGDRVSIICKASQDVGTAVDWYQQKPGQSPK mAb LLIYWASTRHTGVPDRFTGSGSGTDFTLTITNVQSEDLADYFCQQY containing NSYPLTFGAGTMLDLKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNN CD40 mAb FYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKA
ScFv DYEKHKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLV
QSGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLEWM
GWINPDSGGTNYAQKFQGRVTMTRDTSISTAYMELNRLRSDDTAV
YYCARDQPLGYCTNGVCSYFDYWGQGTLVTVSSGGGGSGGGGSG
GGGSGGGGSDIQMTQSPSSVSASVGDRVTITCRASQGIYSWLAWYQ
QKPGKAPNLLIYTASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFAT
YYCQQANIFPLTFGGGTKVEIK
J591 variant 2 LC CD40 733 DIQMTQSPSSVSASVGDRVTITCRASQGIYSWLAWYQQKPGKAPNL mAb LIYTASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQANIFP containing LTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRE tumor mAb AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK
ScFv HKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSEVQLQQSGP Antibody Region SEQ Sequence
ID
NO:
ELVKPGTSVRISCKTSGYTFTEYTIHWVKQSHGKSLEWIGNINPNNG
GTTYNQKFEDKATLTVDKSSSTAYMELRSLTSEDSAVYYCAAGWN
FDYWGQGTTLTVSSGGGGSGGGGSGGGGSGGGGSNIVMTQSPKSM
SMSVGERVTLTCKASENVVTYVSWYQQKPEQSPKLLIYGASNRYT
GVPDRFTGSGSATDFTLTISSVQAEDLADYHCGQGYSYPYTFGGGT
KLEIK
LC tumor 413 NIVMTQSPKSMSMSVGERVTLTCKASENVVTYVSWYQQKPEQSPK mAb LLIYGASNRYTGVPDRFTGSGSATDFTLTISSVQAEDLADYHCGQG containing YSYPYTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNF CD40 mAb YPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKAD
ScFv YEKHKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLVQ
SGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLEWMG
WINPDSGGTNYAQKFQGRVTMTRDTSISTAYMELNRLRSDDTAVY
YCARDQPLGYCTNGVCSYFDYWGQGTLVTVSSGGGGSGGGGSGG
GGSGGGGSDIQMTQSPSSVSASVGDRVTITCRASQGIYSWLAWYQQ
KPGKAPNLLIYTASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATY
YCQQANIFPLTFGGGTKVEIK
Rovalpituzum LC CD40 778 DIQMTQSPSSVSASVGDRVTITCRASQGIYSWLAWYQQKPGKAPNL ab mAb LIYTASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQANIFP containing LTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRE tumor mAb AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK
ScFv HKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLVQSGA
EVKKPGASVKVSCKASGYTFTNYGMNWVRQAPGQGLEWMGWIN
TYTGEPTYADDFKGRVTMTTDTSTSTAYMELRSLRSDDTAVYYCA
RIGDSSPSDYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSEIVM
TQSPATLSVSPGERATLSCKASQSVSNDVVWYQQKPGQAPRLLIYY
ASNRYTGIPARFSGSGSGTEFTLTISSLQSEDFAVYYCQQDYTSPWTF
GQGTKLEIK
LC tumor 426 EIVMTQSPATLSVSPGERATLSCKASQSVSNDVVWYQQKPGQAPRL mAb LIYYASNRYTGIPARFSGSGSGTEFTLTISSLQSEDFAVYYCQQDYTS containing PWTFGQGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYP CD40 mAb REAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADY
ScFv EKHKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLVQS
GAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLEWMGW
INPDSGGTNYAQKFQGRVTMTRDTSISTAYMELNRLRSDDTAVYYC
ARDQPLGYCTNGVCSYFDYWGQGTLVTVSSGGGGSGGGGSGGGG
SGGGGSDIQMTQSPSSVSASVGDRVTITCRASQGIYSWLAWYQQKP
GKAPNLLIYTASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYC
QQANIFPLTFGGGTKVEIK
PF-06647020 LC CD40 769 DIQMTQSPSSVSASVGDRVTITCRASQGIYSWLAWYQQKPGKAPNL mAb LIYTASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQANIFP containing LTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRE tumor mAb AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK
ScFv HKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLVQSGP
EVKKPGASVKVSCKASGYTFTDYAVHWVRQAPGKRLEWIGVISTY
NDYTYNNQDFKGRVTMTRDTSASTAYMELSRLRSEDTAVYYCAR
GNSYFYALDYWGQGTSVTVSSGGGGSGGGGSGGGGSGGGGSEIVL
TQSPATLSLSPGERATLSCRASESVDSYGKSFMHWYQQKPGQAPRL
LIYRASNLESGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQSNEDP
WTFGGGTKLEIK
LC tumor 439 EIVLTQSPATLSLSPGERATLSCRASESVDSYGKSFMHWYQQKPGQ mAb APRLLIYRASNLESGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQS containing NEDPWTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNN CD40 mAb FYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKA
ScFv DYEKHKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLV
QSGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLEWM
GWINPDSGGTNYAQKFQGRVTMTRDTSISTAYMELNRLRSDDTAV
YYCARDQPLGYCTNGVCSYFDYWGQGTLVTVSSGGGGSGGGGSG
GGGSGGGGSDIQMTQSPSSVSASVGDRVTITCRASQGIYSWLAWYQ Antibody Region SEQ Sequence
ID
NO:
QKPGKAPNLLIYTASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFAT YYCQQANIFPLTFGGGTKVEIK
Antibody to LC CD40 688 DIQMTQSPSSVSASVGDRVTITCRASQGIYSWLAWYQQKPGKAPNL PTK7 mAb LIYTASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQANIFP containing LTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRE tumor mAb AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK
ScFv HKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLVESGG
GVVQPGRSLRLSCAASGFTFSSYAFHWVRQAPGKGLEWVAVISYD
GSIKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARTY
YFDYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSEIVLTQSPDF
QSVTPKEKVTITCRASQSIGSSLHWYQQKPDQSPKLLIKYASQSFSG
VPSRFSGSGSGTDFTLTINSLEAEDAAAYYCHQSSSLPITFGQGTRLE
IK
LC tumor 452 EIVLTQSPDFQSVTPKEKVTITCRASQSIGSSLHWYQQKPDQSPKLLI mAb KYASQSFSGVPSRFSGSGSGTDFTLTINSLEAEDAAAYYCHQSSSLPI containing TFGQGTRLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREA CD40 mAb KVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKH
ScFv KVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLVQSGAE
VKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLEWMGWINP
DSGGTNYAQKFQGRVTMTRDTSISTAYMELNRLRSDDTAVYYCAR
DQPLGYCTNGVCSYFDYWGQGTLVTVSSGGGGSGGGGSGGGGSG
GGGSDIQMTQSPSSVSASVGDRVTITCRASQGIYSWLAWYQQKPGK
APNLLIYTASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQ
ANIFPLTFGGGTKVEIK
Ladiratuzuma LC CD40 784 DIQMTQSPSSVSASVGDRVTITCRASQGIYSWLAWYQQKPGKAPNL b mAb LIYTASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQANIFP containing LTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRE tumor mAb AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK
ScFv HKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLVQSGA
EVKKPGASVKVSCKASGLTIEDYYMHWVRQAPGQGLEWMGWIDP
ENGDTEYGPKFQGRVTMTRDTSINTAYMELSRLRSDDTAVYYCAV
HNAHYGTWFAYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSD
VVMTQSPLSLPVTLGQPASISCRSSQSLLHSSGNTYLEYFQQRPGQSP
RPLIYKISTRFSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCFQGS
HVPYTFGGGTKVEIK
LC tumor 465 DVVMTQSPLSLPVTLGQPASISCRSSQSLLHSSGNTYLEYFQQRPGQ mAb SPRPLIYKISTRFSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCFQ containing GSHVPYTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLN CD40 mAb NFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSK
ScFv ADYEKHKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQL
VQSGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLEW
MGWINPDSGGTNYAQKFQGRVTMTRDTSISTAYMELNRLRSDDTA
VYYCARDQPLGYCTNGVCSYFDYWGQGTLVTVSSGGGGSGGGGS
GGGGSGGGGSDIQMTQSPSSVSASVGDRVTITCRASQGIYSWLAWY
QQKPGKAPNLLIYTASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFA
TYYCQQ ANIFPLTFGGGTKVEIK
Cirmtuzumab LC CD40 706 DIQMTQSPSSVSASVGDRVTITCRASQGIYSWLAWYQQKPGKAPNL mAb LIYTASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQANIFP containing LTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRE tumor mAb AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK
ScFv HKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLQESGP
GLVKPSQTLSLTCTVSGYAFTAYNIHWVRQAPGQGLEWMGSFDPY
DGGSSYNQKFKDRLTISKDTSKNQVVLTMTNMDPVDTATYYCARG
WYYFDYWGHGTLVTVSSGGGGSGGGGSGGGGSGGGGSDIVMTQT
PLSLPVTPGEPASISCRASKSISKYLAWYQQKPGQAPRLLIYSGSTLQ
SGIPPRFSGSGYGTDFTLTINNIESEDAAYYFCQQHDESPY
LC tumor 478 DIVMTQTPLSLPVTPGEPASISCRASKSISKYLAWYQQKPGQAPRLLI mAb YSGSTLQSGIPPRFSGSGYGTDFTLTINNIESEDAAYYFCQQHDESPY containing TFGEGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREA Antibody Region SEQ Sequence
ID
NO:
CD40 mAb KVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKH ScFv KVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLVQSGAE
VKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLEWMGWINP
DSGGTNYAQKFQGRVTMTRDTSISTAYMELNRLRSDDTAVYYCAR
DQPLGYCTNGVCSYFDYWGQGTLVTVSSGGGGSGGGGSGGGGSG
GGGSDIQMTQSPSSVSASVGDRVTITCRASQGIYSWLAWYQQKPGK
APNLLIYTASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQ
ANIFPLTFGGGTKVEIK
Antibody to LC CD40 739 DIQMTQSPSSVSASVGDRVTITCRASQGIYSWLAWYQQKPGKAPNL MAGE-A3 mAb LIYTASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQANIFP containing LTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRE tumor mAb AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK
ScFv HKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLVESGG
GVVQPGRSLRLSCTASGFRFRSHGMHWVRQAPGKGLEWVAVISYD
GNNKLYADSVKGRITISRDNSKNTLFLQMNNVRAEDTAVYYCASP
YTSDWQYFQYWGQGTLVIVSSGGGGSGGGGSGGGGSGGGGSEIV
MTQSPATLSVSPGERATFSCRASQNISTTLAWYQQKPGQAPRLLIYD
TSTRATGIPARFSGSGSGTEFTLTISSLQSEDLAVYYCQQSNSWPLTF
GGGTKVEIK
LC tumor 491 EIVMTQSPATLSVSPGERATFSCRASQNISTTLAWYQQKPGQAPRLL mAb IYDTSTRATGIPARFSGSGSGTEFTLTISSLQSEDLAVYYCQQSNSWP containing LTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRE CD40 mAb AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK
ScFv HKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLVQSGA
EVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLEWMGWIN
PDSGGTNYAQKFQGRVTMTRDTSISTAYMELNRLRSDDTAVYYCA
RDQPLGYCTNGVCSYFDYWGQGTLVTVSSGGGGSGGGGSGGGGS
GGGGSDIQMTQSPSSVSASVGDRVTITCRASQGIYSWLAWYQQKPG
KAPNLLIYTASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQ
QANIFPLTFGGGTKVEIK
Antibody to LC CD40 754 DIQMTQSPSSVSASVGDRVTITCRASQGIYSWLAWYQQKPGKAPNL NY-ESO-1 mAb LIYTASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQANIFP containing LTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRE tumor mAb AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK
ScFv HKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLVQSGG
GVVRPGGSLRLSCAASGFSFIDYGMSWVRQVPGKGLEWVAGMNW
SGDKKGHAESVKGRFIISRDNAKNTLYLEMSSLRVEDTALYFCARG
EYSNRFDPRGRGTLVTVSSGGGGSGGGGSGGGGSGGGGSDIVMTQ
TPLSLPVTLGQPASLSCRSSQSLVFTDGNTYLNWFQQRPGQSPRRLI
YKVSSRDPGVPDRFSGTGSGTDFTLEISRVEAEDIGVYYCMQGTHW
PPIFGQGTKVEIK
LC tumor 504 DIVMTQTPLSLPVTLGQPASLSCRSSQSLVFTDGNTYLNWFQQRPG mAb QSPRRLIYKVSSRDPGVPDRFSGTGSGTDFTLEISRVEAEDIGVYYC containing MQGTHWPPIFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCL CD40 mAb LNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTL
ScFv SKADYEKHKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQV
QLVQSGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLE
WMGWINPDSGGTNYAQKFQGRVTMTRDTSISTAYMELNRLRSDDT
AVYYCARDQPLGYCTNGVCSYFDYWGQGTLVTVSSGGGGSGGGG
SGGGGSGGGGSDIQMTQSPSSVSASVGDRVTITCRASQGIYSWLAW
YQQKPGKAPNLLIYTASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDF
ATYYCQQ ANIFPLTFGGGTKVEIK
Trastuzumab LC CD40 793 DIQMTQSPSSVSASVGDRVTITCRASQGIYSWLAWYQQKPGKAPNL mAb LIYTASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDF ATYYCQQ ANIFP containing LTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRE tumor mAb AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK
ScFv HKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSEVQLVESGG
GLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTN
GYTRYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRWG Antibody Region SEQ Sequence
ID
NO:
GDGFYAMDYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSDIQM TQSPSSLSASVGDRVTITCRASQDVNTAVAWYQQKPGKAPKLLIYS ASFLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQHYTTPPTF GQGTKVEIK
LC tumor 685 DIQMTQSPSSLSASVGDRVTITCRASQDVNTAVAWYQQKPGKAPKL mAb LIYSASFLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQHYTT containing PPTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPR CD40 mAb EAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYE
ScFv KHKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLVQSG
AEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLEWMGWI
NPDSGGTNYAQKFQGRVTMTRDTSISTAYMELNRLRSDDTAVYYC
ARDQPLGYCTNGVCSYFDYWGQGTLVTVSSGGGGSGGGGSGGGG
SGGGGSDIQMTQSPSSVSASVGDRVTITCRASQGIYSWLAWYQQKP
GKAPNLLIYTASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYC
QQANIFPLTFGGGTKVEIK
LC CD40 798 DIQMTQSPSSVSASVGDRVTITCRASQGIYSWLAWYQQKPGKAPNL mAb LIYTASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQANIFP containing LTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRE tumor mAb AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK scFv HKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSDIQMTQSPSS (LH,25mer) LSASVGDRVTITCRASQDVNTAVAWYQQKPGKAPKLLIYSASFLYS
GVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQHYTTPPTFGQGTK
VEIKGGGGSGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVQPGG
SLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYAD
SVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAM
DYWGQGTLVTVSS
TABLE 10. Fusion Sequences - DEC-205 fusions via the light chain
Antibody Region SEQ Sequence
ID
NO:
Pertuzumab LC DEC-205 767 EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLI mAb YDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRRNWP containing LTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRE tumor mAb AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK ScFv HKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSEVQLVESGG
GLVQPGGSLRLSCAASGFTFTDYTMDWVRQAPGKGLEWVADVNP
NSGGSIYNQRFKGRFTLSVDRSKNTLYLQMNSLRAEDTAVYYCAR
NLGPSFYFDYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSDIQM
TQSPSSLSASVGDRVTITCKASQDVSIGVAWYQQKPGKAPKLLIYSA
SYRYTGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYYIYPYTFG
QGTKVEIK
LC tumor 768 DIQMTQSPSSLSASVGDRVTITCKASQDVSIGVAWYQQKPGKAPKL mAb LIYSASYRYTGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYYIY containing PYTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPR DEC-205 mAb EAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYE ScFv KHKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLVESG
GGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGLEWVAVIW
YDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCA
RDLWGWYFDYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSEIV
LTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLIYD
ASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRRNWPLTF
GGGTKVEIK
Cetuximab LC DEC-205 704 EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLI mAb YDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRRNWP containing LTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRE tumor mAb AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK ScFv HKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLKQSGP Antibody Region SEQ Sequence
ID
NO:
GLVQPSQSLSITCTVSGFSLTNYGVHWVRQSPGKGLEWLGVIWSGG
NTDYNTPFTSRLSINKDNSKSQVFFKMNSLQSNDTAIYYCARALTY
YDYEFAYWGQGTLVTVSAGGGGSGGGGSGGGGSGGGGSDILLTQS
PVILSVSPGERVSFSCRASQSIGTNIHWYQQRTNGSPRLLIKYASESIS
GIPSRFSGSGSGTDFTLSINSVESEDIADYYCQQNNNWPTTFGAGTK
LELK
LC tumor 705 DILLTQSPVILSVSPGERVSFSCRASQSIGTNIHWYQQRTNGSPRLLIK mAb YASESISGIPSRFSGSGSGTDFTLSINSVESEDIADYYCQQNNNWPTTF containing GAGTKLELKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAK DEC-205 mAb VQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHK ScFv VYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLVESGGGV
VQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGLEWVAVIWYDG
SNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDL
WGWYFDYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSEIVLTQ
SPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLIYDASN
RATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRRNWPLTFGG
GTKVEIK
Panitumumab LC DEC-205 761 EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLI mAb YDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRRNWP containing LTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRE tumor mAb AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK ScFv HKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLQESGP
GLVKPSETLSLTCTVSGGSVSSGDYYWTWIRQSPGKGLEWIGfflYYS
GNTNYNPSLKSRLTISIDTSKTQFSLKLSSVTAADTAIYYCVRDRVTG
AFDIWGQGTMVTVSSGGGGSGGGGSGGGGSGGGGSDIQMTQSPSS
LSASVGDRVTITCQASQDISNYLNWYQQKPGKAPKLLIYDASNLET
GVPSRFSGSGSGTDFTFTISSLQPEDIATYFCQHFDHLPLAFGGGTKV
EIK
LC tumor 762 DIQMTQSPSSLSASVGDRVTITCQASQDISNYLNWYQQKPGKAPKL mAb LIYDASNLETGVPSRFSGSGSGTDFTFTISSLQPEDIATYFCQHFDHLP containing LAFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPR DEC-205 mAb EAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYE ScFv KHKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLVESG
GGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGLEWVAVIW
YDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCA
RDLWGWYFDYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSEIV
LTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLIYD
ASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRRNWPLTF
GGGTKVEIK
Nimotuzumab LC DEC-205 752 EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLI mAb YDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRRNWP containing LTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRE tumor mAb AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK ScFv HKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLQQSGA
EVKKPGSSVKVSCKASGYTFTNYYIYWVRQAPGQGLEWIGGINPTS
GGSNFNEKFKTRVTITVDESTNTAYMELSSLRSEDTAFYFCARQGL
WFDSDGRGFDFWGQGSTVTVSSGGGGSGGGGSGGGGSGGGGSDIQ
MTQSPSSLSASVGDRVTITCRSSQNIVHSNGNTYLDWYQQTPGKAP
KLLIYKVSNRFSGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCFQYSH
VPWTFGQGTKLEIK
LC tumor 753 DIQMTQSPSSLSASVGDRVTITCRSSQNIVHSNGNTYLDWYQQTPGK mAb APKLLIYKVSNRFSGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCFQY containing SHVPWTFGQGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNN DEC-205 mAb FYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKA ScFv DYEKHKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLV
ESGGGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGLEWVA
VIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVY
YCARDLWGWYFDYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGG
SEIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRL Antibody Region SEQ Sequence
ID
NO:
LIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRRN WPLTFGGGTKVEIK
Zalutumumab LC DEC-205 803 EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLI mAb YDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRRNWP containing LTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRE tumor mAb AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK ScFv HKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLVESGG
GVVQPGRSLRLSCAASGFTFSTYGMHWVRQAPGKGLEWVAVIWD
DGSYKYYGDSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAR
DGITMVRGVMKDYFDYWGQGTLVTVSSGGGGSGGGGSGGGGSGG
GGSAIQLTQSPSSLSASVGDRVTITCRASQDISSALVWYQQKPGKAP
KLLIYDASSLESGVPSRFSGSESGTDFTLTISSLQPEDFATYYCQQFNS
YPLTFGGGTKVEIK
LC tumor 804 AIQLTQSPSSLSASVGDRVTITCRASQDISSALVWYQQKPGKAPKLLI mAb YDASSLESGVPSRFSGSESGTDFTLTISSLQPEDFATYYCQQFNSYPL containing TFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRE DEC-205 mAb AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK ScFv HKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLVESGG
GVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGLEWVAVIWY
DGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAR
DLWGWYFDYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSEIVL
TQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLIYDA
SNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRRNWPLTFG
GGTKVEIK
Onartuzumab LC DEC-205 758 EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLI mAb YDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRRNWP containing LTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRE tumor mAb AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK ScFv HKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSEVQLVESGG
GLVQPGGSLRLSCAASGYTFTSYWLHWVRQAPGKGLEWVGMIDPS
NSDTRFNPNFKDRFTISADTSKNTAYLQMNSLRAEDTAVYYCATYR
SYVTPLDYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSDIQMTQ
SPSSLSASVGDRVTITCKSSQSLLYTSSQKNYLAWYQQKPGKAPKLL
IYWASTRESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYYAY
PWTFGQGTKVEIK
LC tumor 759 DIQMTQSPSSLSASVGDRVTITCKSSQSLLYTSSQKNYLAWYQQKPG mAb KAPKLLIYWASTRESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQ containing QYYAYPWTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLL DEC-205 mAb NNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLS ScFv KADYEKHKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQ
LVESGGGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGLEW
VAVIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTA
VYYCARDLWGWYFDYWGQGTLVTVSSGGGGSGGGGSGGGGSGG
GGSEIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAP
RLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRR
NWPLTFGGGTKVEIK
Patritumab LC DEC-205 764 EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLI mAb YDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRRNWP containing LTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRE tumor mAb AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK ScFv HKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLQQWG
AGLLKPSETLSLTCAVYGGSFSGYYWSWIRQPPGKGLEWIGEINHSG
STNYNPSLKSRVTISVETSKNQFSLKLSSVTAADTAVYYCARDKWT
WYFDLWGRGTLVTVSSGGGGSGGGGSGGGGSGGGGSDIEMTQSPD
SLAVSLGERATINCRSSQSVLYSSSNRNYLAWYQQNPGQPPKLLIY
WASTRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQYYSTPR
TFGQGTKVEIK
LC tumor 765 DIEMTQSPDSLAVSLGERATINCRSSQSVLYSSSNRNYLAWYQQNP mAb GQPPKLLIYWASTRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYY Antibody Region SEQ Sequence
ID
NO:
containing CQQYYSTPRTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVC DEC-205 mAb LLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLT ScFv LSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQ
VQLVESGGGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGLE
WVAVIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDT
AVYYCARDLWGWYFDYWGQGTLVTVSSGGGGSGGGGSGGGGSG
GGGSEIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQA
PRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQR
RNWPLTFGGGTKVEIK
Clivatuzumab LC DEC-205 710 EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLI mAb YDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRRNWP containing LTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRE tumor mAb AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK ScFv HKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLQQSGA
EVKKPGASVKVSCEASGYTFPSYVLHWVKQAPGQGLEWIGYINPY
NDGTQYNEKFKGKATLTRDTSINTAYMELSRLRSDDTAVYYCARG
FGGSYGFAYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSDIQLT
QSPSSLSASVGDRVTMTCSASSSVSSSYLYWYQQKPGKAPKLWIYS
TSNLASGVPARFSGSGSGTDFTLTISSLQPEDSASYFCHQWNRYPYT
FGGGTRLEIK
LC tumor 711 DIQLTQSPSSLSASVGDRVTMTCSASSSVSSSYLYWYQQKPGKAPKL mAb WIYSTSNLASGVPARFSGSGSGTDFTLTISSLQPEDSASYFCHQWNR containing YPYTFGGGTRLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYP DEC-205 mAb REAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYE ScFv KHKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLVESG
GGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGLEWVAVIW
YDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCA
RDLWGWYFDYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSEIV
LTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLIYD
ASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRRNWPLTF
GGGTKVEIK
Sofituzumab LC DEC-205 791 EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLI mAb YDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRRNWP containing LTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRE tumor mAb AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK ScFv HKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSEVQLVESGG
GLVQPGGSLRLSCAASGYSITNDYAWNWVRQAPGKGLEWVGYISY
SGYTTYNPSLKSRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARWT
SGLDYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSDIQMTQSPS
SLSASVGDRVTITCKASDLIHNWLAWYQQKPGKAPKLLIYGATSLE
TGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYWTTPFTFGQGT
KVEIK
LC tumor 792 DIQMTQSPSSLSASVGDRVTITCKASDLIHNWLAWYQQKPGKAPKL mAb LIYGATSLETGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYWTT containing PFTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPR DEC-205 mAb EAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYE ScFv KHKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLVESG
GGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGLEWVAVIW
YDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCA
RDLWGWYFDYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSEIV
LTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLIYD
ASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRRNWPLTF
GGGTKVEIK
Edrecolomab LC DEC-205 719 EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLI mAb YDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRRNWP containing LTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRE tumor mAb AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK ScFv HKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLQQSGA
ELVRPGTSVKVSCKASGYAFTNYLIEWVKQRPGQGLEWIGVINPGS Antibody Region SEQ Sequence
ID
NO:
GGTNYNEKFKGKATLTADKSSSTAYMQLSSLTSDDSAVYFCARDG PWFAYWGQGTLVTVSAGGGGSGGGGSGGGGSGGGGSNIVMTQSP KSMSMSVGERVTLTCKASENVVTYVSWYQQKPEQSPKLLIYGASN RYTGVPDRFTGSGSATDFTLTISSVQAEDLADYHCGQGYSYPYTFG GGTKLEIK
LC tumor 720 NIVMTQSPKSMSMSVGERVTLTCKASENVVTYVSWYQQKPEQSPK mAb LLIYGASNRYTGVPDRFTGSGSATDFTLTISSVQAEDLADYHCGQGY containing SYPYTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFY DEC-205 mAb PREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADY ScFv EKHKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLVES
GGGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGLEWVAVI
WYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYC
ARDLWGWYFDYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSEI
VLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLIY
DASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRRNWPL
TFGGGTKVEIK
Adecatumum LC DEC-205 695 EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLI ab mAb YDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRRNWP containing LTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRE tumor mAb AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK ScFv HKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSEVQLLESGG
GVVQPGRSLRLSCAASGFTFSSYGMHWVRQAPGKGLEWVAVISYD
GSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKD
MGWGSGWRPYYYYGMDVWGQGTTVTVSSGGGGSGGGGSGGGGS
GGGGSELQMTQSPSSLSASVGDRVTITCRTSQSISSYLNWYQQKPGQ
PPKLLIYWASTRESGVPDRFSGSGSGTDFTLTISSLQPEDSATYYCQQ
SYDIPYTFGQGTKLEIK
LC tumor 696 ELQMTQSPSSLSASVGDRVTITCRTSQSISSYLNWYQQKPGQPPKLLI mAb YWASTRESGVPDRFSGSGSGTDFTLTISSLQPEDSATYYCQQSYDIP containing YTFGQGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRE DEC-205 mAb AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK ScFv HKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLVESGG
GVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGLEWVAVIWY
DGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAR
DLWGWYFDYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSEIVL
TQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLIYDA
SNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRRNWPLTFG
GGTKVEIK
Anetumab LC DEC-205 701 EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLI mAb YDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRRNWP containing LTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRE tumor mAb AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK ScFv HKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVELVQSGA
EVKKPGESLKISCKGSGYSFTSYWIGWVRQAPGKGLEWMGIIDPGD
SRTRYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCARGQL
YGGTYMDGWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSDIALT
QPASVSGSPGQSITISCTGTSSDIGGYNSVSWYQQHPGKAPKLMIYG
VNNRPSGVSNRFSGSKSGNTASLTISGLQAEDEADYYCSSYDIESAT
PVFGGGTKLTVL
LC tumor 702 DIALTQPASVSGSPGQSITISCTGTSSDIGGYNSVSWYQQHPGKAPKL mAb MIYGVNNRPSGVSNRFSGSKSGNTASLTISGLQAEDEADYYCSSYDI containing ESATPVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLIS DEC-205 mAb DFYPGAVTVAWKGDSSPVKAGVETTTPSKQSNNKYAASSYLSLTPE ScFv QWKSHRSYSCQVTHEGSTVEKTVAPTECSGGGGSGGGGSQVQLVE
SGGGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGLEWVAVI
WYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYC
ARDLWGWYFDYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSEI
VLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLIY
DASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRRNWPL Antibody Region SEQ Sequence
ID
NO:
TFGGGTKVEIK
huDS6 LC DEC-205 725 EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLI mAb YDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRRNWP containing LTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRE tumor mAb AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK ScFv HKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQAQLVQSGA
EVVKPGASVKMSCKASGYTFTSYNMHWVKQTPGQGLEWIGYIYPG
NGATNYNQKFQGKATLTADPSSSTAYMQISSLTSEDSAVYFCARGD
SVPFAYWGQGTLVTVSAGGGGSGGGGSGGGGSGGGGSEIVLTQSP
ATMSASPGERVTITCSAHSSVSFMHWFQQKPGTSPKLWIYSTSSLAS
GVPARFGGSGSGTSYSLTISSMEAEDAATYYCQQRSSFPLTFGAGTK
LELK
LC tumor 726 EIVLTQSPATMSASPGERVTITCSAHSSVSFMHWFQQKPGTSPKLWI mAb YSTSSLASGVPARFGGSGSGTSYSLTISSMEAEDAATYYCQQRSSFP containing LTFGAGTKLELKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPR DEC-205 mAb EAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYE ScFv KHKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLVESG
GGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGLEWVAVIW
YDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCA
RDLWGWYFDYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSEIV
LTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLIYD
ASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRRNWPLTF
GGGTKVEIK
Lifastuzumab LC DEC-205 737 EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLI mAb YDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRRNWP containing LTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRE tumor mAb AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK ScFv HKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSEVQLVESGG
GLVQPGGSLRLSCAASGFSFSDFAMSWVRQAPGKGLEWVATIGRV
AFHTYYPDSMKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARH
RGFDVGHFDFWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSDIQ
MTQSPSSLSASVGDRVTITCRSSETLVHSSGNTYLEWYQQKPGKAP
KLLIYRVSNRFSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCFQGSF
NPLTFGQGTKVEIK
LC tumor 738 DIQMTQSPSSLSASVGDRVTITCRSSETLVHSSGNTYLEWYQQKPGK mAb APKLLIYRVSNRFSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCFQ containing GSFNPLTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNN DEC-205 mAb FYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKA ScFv DYEKHKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLV
ESGGGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGLEWVA
VIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVY
YCARDLWGWYFDYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGG
SEIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRL
LIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRRN
WPLTFGGGTKVEIK
Sacituzumab LC DEC-205 782 EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLI mAb YDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRRNWP containing LTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRE tumor mAb AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK ScFv HKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLQQSGS
ELKKPGASVKVSCKASGYTFTNYGMNWVKQAPGQGLKWMGWIN
TYTGEPTYTDDFKGRFAFSLDTSVSTAYLQISSLKADDTAVYFCARG
GFGSSYWYFDVWGQGSLVTVSSGGGGSGGGGSGGGGSGGGGSDIQ
LTQSPSSLSASVGDRVSITCKASQDVSIAVAWYQQKPGKAPKLLIYS
ASYRYTGVPDRFSGSGSGTDFTLTISSLQPEDFAVYYCQQHYITPLTF
GAGTKVEIK
LC tumor 783 DIQLTQSPSSLSASVGDRVSITCKASQDVSIAVAWYQQKPGKAPKLL mAb IYSASYRYTGVPDRFSGSGSGTDFTLTISSLQPEDFAVYYCQQHYITP containing LTFGAGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRE Antibody Region SEQ Sequence
ID
NO:
DEC-205 mAb AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK ScFv HKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLVESGG
GVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGLEWVAVIWY
DGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAR
DLWGWYFDYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSEIVL
TQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLIYDA
SNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRRNWPLTFG
GGTKVEIK
PR1A3 LC DEC-205 773 EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLI mAb YDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRRNWP containing LTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRE tumor mAb AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK ScFv HKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVKLQQSGP
ELKKPGETVKISCKASGYTFTEFGMNWVKQAPGKGLKWMGWINT
KTGEATYVEEFKGRFAFSLETSATTAYLQINNLKNEDTAKYFCARW
DFYDYVEAMDYWGQGTTVTVSSGGGGSGGGGSGGGGSGGGGSDI
VMTQSQRFMSTSVGDRVSVTCKASQNVGTNVAWYQQKPGQSPKA
LIYSASYRYSGVPDRFTGSGSGTDFTLTISNVQSEDLAEYFCHQYYT
YPLFTFGSGTKLEMK
LC tumor 774 DIVMTQSQRFMSTSVGDRVSVTCKASQNVGTNVAWYQQKPGQSPK mAb ALIYSASYRYSGVPDRFTGSGSGTDFTLTISNVQSEDLAEYFCHQYY containing TYPLFTFGSGTKLEMKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNN DEC-205 mAb FYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKA ScFv DYEKHKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLV
ESGGGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGLEWVA
VIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVY
YCARDLWGWYFDYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGG
SEIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRL
LIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRRN
WPLTFGGGTKVEIK
Humanized LC DEC-205 821 EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLL PR1A3 mAb IYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRRNW containing PLTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPR tumor mAb EAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYE ScFv KHKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLVQSG
AEVKKPGASVKVSCKASGYTFTEFGMNWVRQAPGQGLEWMGWIN
TKTGEATYVEEFKGRVTFTTDTSTSTAYMELRSLRSDDTAVYYCAR
WDFAYYVEAMDYWGQGTTVTVSSGGGGSGGGGSGGGGSGGGGS
DIQMTQSPSSLSASVGDRVTITCKASAAVGTYVAWYQQKPGKAPK
LLIYSASYRKRGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCHQYYT
YPLFTFGQGTKLEIK
LC tumor 822 DIQMTQSPSSLSASVGDRVTITCKASAAVGTYVAWYQQKPGKAPK mAb LLIYSASYRKRGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCHQYYT containing YPLFTFGQGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFY DEC-205 mAb PREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADY ScFv EKHKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLVES
GGGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGLEWVAVI
WYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYC
ARDLWGWYFDYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSEI
VLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLIY
DASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRRNWPL
TFGGGTKVEIK
Humanized LC DEC-205 839 EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLL Ab2-3 mAb IYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRRNW containing PLTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPR tumor mAb EAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYE ScFv KHKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSEVQLQESG
PGLVKPGGSLSLSCAASGFVFSSYDMSWVRQTPERGLEWVAYISSG
GGITYAPSTVKGRFTVSRDNAKNTLYLQMNSLTSEDTAVYYCAAH Antibody Region SEQ Sequence
ID
NO:
YFGSSGPFAYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSDIQM TQSPASLSASVGDRVTITCRASENIFSYLAWYQQKPGKSPKLLVYNT RTLAEGVPSRFSGSGSGTDFSLTISSLQPEDFATYYCQHHYGTPFTFG SGTKLEIK
LC tumor 840 DIQMTQSPASLSASVGDRVTITCRASENIFSYLAWYQQKPGKSPKLL mAb VYNTRTLAEGVPSRFSGSGSGTDFSLTISSLQPEDFATYYCQHHYGT containing PFTFGSGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPR DEC-205 mAb EAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYE ScFv KHKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLVESG
GGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGLEWVAVIW
YDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCA
RDLWGWYFDYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSEIV
LTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLIYD
ASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRRNWPLT
FGGGTKVEIK
IMAB362, LC DEC-205 728 EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLI
CLAUDIXIM mAb YDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRRNWP
AB containing LTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRE tumor mAb AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK ScFv HKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLQQPGA
ELVRPGASVKLSCKASGYTFTSYWINWVKQRPGQGLEWIGNIYPSD
SYTNYNQKFKDKATLTVDKSSSTAYMQLSSPTSEDSAVYYCTRSW
RGNSFDYWGQGTTLTVSSGGGGSGGGGSGGGGSGGGGSDIVMTQS
PSSLTVTAGEKVTMSCKSSQSLLNSGNQKNYLTWYQQKPGQPPKLL
IYWASTRESGVPDRFTGSGSGTDFTLTISSVQAEDLAVYYCQNDYSY
PFTFGSGTKLEIK
LC tumor 729 DIVMTQSPSSLTVTAGEKVTMSCKSSQSLLNSGNQKNYLTWYQQKP mAb GQPPKLLIYWASTRESGVPDRFTGSGSGTDFTLTISSVQAEDLAVYY containing CQNDYSYPFTFGSGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCL DEC-205 mAb LNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTL ScFv SKADYEKHKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQV
QLVESGGGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGLE
WVAVIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDT
AVYYCARDLWGWYFDYWGQGTLVTVSSGGGGSGGGGSGGGGSG
GGGSEIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQA
PRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQR
RNWPLTFGGGTKVEIK
AMG595 LC DEC-205 698 EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLI mAb YDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRRNWP containing LTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRE tumor mAb AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK ScFv HKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLVESGG
GVVQSGRSLRLSCAASGFTFRNYGMHWVRQAPGKGLEWVAVIWY
DGSDKYYADSVRGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAR
DGYDILTGNPRDFDYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGG
SDTVMTQTPLSSHVTLGQPASISCRSSQSLVHSDGNTYLSWLQQRPG
QPPRLLIYRISRRFSGVPDRFSGSGAGTDFTLEISRVEAEDVGVYYCM
QSTHVPRTFGQGTKVEIK
LC tumor 699 DTVMTQTPLSSHVTLGQPASISCRSSQSLVHSDGNTYLSWLQQRPG mAb QPPRLLIYRISRRFSGVPDRFSGSGAGTDFTLEISRVEAEDVGVYYCM containing QSTHVPRTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLL DEC-205 mAb NNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLS ScFv KADYEKHKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQ
LVESGGGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGLEW
VAVIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTA
VYYCARDLWGWYFDYWGQGTLVTVSSGGGGSGGGGSGGGGSGG
GGSEIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAP
RLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRR
NWPLTFGGGTKVEIK Antibody Region SEQ Sequence
ID
NO:
ABT806 LC DEC-205 692 EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLI mAb YDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRRNWP containing LTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRE tumor mAb AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK ScFv HKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSEVQLQESGP
GLVKPSQTLSLTCTVSGYSISRDFAWNWIRQPPGKGLEWMGYISYN
GNTRYQPSLKSRITISRDTSKNQFFLKLNSVTAADTATYYCVTASRG
FPYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSDIQMTQSPSSM
SVSVGDRVTITCHSSQDINSNIGWLQQKPGKSFKGLIYHGTNLDDGV
PSRFSGSGSGTDYTLTISSLQPEDFATYYCVQYAQFPWTFGGGTKLE
IK
LC tumor 693 DIQMTQSPSSMSVSVGDRVTITCHSSQDINSNIGWLQQKPGKSFKGL mAb IYHGTNLDDGVPSRFSGSGSGTDYTLTISSLQPEDFATYYCVQYAQF containing PWTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYP DEC-205 mAb REAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYE ScFv KHKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLVESG
GGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGLEWVAVIW
YDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCA
RDLWGWYFDYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSEIV
LTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLIYD
ASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRRNWPLTF
GGGTKVEIK
Sibrotuzumab LC DEC-205 788 EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLI mAb YDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRRNWP containing LTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRE tumor mAb AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK ScFv HKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLVQSGA
EVKKPGASVKVSCKTSRYTFTEYTIHWVRQAPGQRLEWIGGINPNN
GIPNYNQKFKGRVTITVDTSASTAYMELSSLRSEDTAVYYCARRRIA
YGYDEGHAMDYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSDI
VMTQSPDSLAVSLGERATINCKSSQSLLYSRNQKNYLAWYQQKPG
QPPKLLIFWASTRESGVPDRFSGSGFGTDFTLTISSLQAEDVAVYYC
QQYFSYPLTFGQGTKVEIK
LC tumor 789 DIVMTQSPDSLAVSLGERATINCKSSQSLLYSRNQKNYLAWYQQKP mAb GQPPKLLIFWASTRESGVPDRFSGSGFGTDFTLTISSLQAEDVAVYY containing CQQYFSYPLTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCL DEC-205 mAb LNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTL ScFv SKADYEKHKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQV
QLVESGGGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGLE
WVAVIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDT
AVYYCARDLWGWYFDYWGQGTLVTVSSGGGGSGGGGSGGGGSG
GGGSEIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQA
PRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQR
RNWPLTFGGGTKVEIK
DS-8895a LC DEC-205 713 EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLI variant 1 mAb YDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRRNWP containing LTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRE tumor mAb AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK ScFv HKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLVQSGA
EVKKPGASVKVSCKASGYTFIDYSMHWVRQAPGQGLEWMGWINT
YTGEPTYSDDFKGRVTITADTSTSTAYLELSSLRSEDTAVYYCATYY
RYERDFDYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSDIVMT
QSPLSLPVTPGEPASISCRSSQSIVHSSGITYLEWYLQKPGQSPQLLIY
KVSNRFSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCFQGSHVP
YTFGQGTKVEIK
LC tumor 714 DIVMTQSPLSLPVTPGEPASISCRSSQSIVHSSGITYLEWYLQKPGQSP mAb QLLIYKVSNRFSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCFQG containing SHVPYTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNN DEC-205 mAb FYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKA Antibody Region SEQ Sequence
ID
NO:
ScFv DYEKHKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLV
ESGGGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGLEWVA
VIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVY
YCARDLWGWYFDYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGG
SEIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRL
LIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRRN
WPLTFGGGTKVEIK
DS-8895a LC DEC-205 716 EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLI variant 2 mAb YDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRRNWP containing LTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRE tumor mAb AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK ScFv HKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQIQLVQSGAE
VKKPGASVKVSCKASGYTFIDYSMHWVRQAPGQGLKWMGWINTY
TGEPTYSDDFKGRFAFSLDTSTSTAYLELSSLRSEDTAVYYCATYYR
YERDFDYWGQGTLVTVSSGGGGSGGGGSGGGSGGGGSDVLMTQS
PLSLPVTPGEPASISCRSSQSIVHSSGITYLEWYLQKPGQSPQLLIYKV
SNRFSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCFQGSHVPYTF
GQGTKVEIK
LC tumor 717 DVLMTQSPLSLPVTPGEPASISCRSSQSIVHSSGITYLEWYLQKPGQS mAb PQLLIYKVSNRFSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCFQ containing GSHVPYTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLN DEC-205 mAb NFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSK ScFv ADYEKHKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQL
VESGGGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGLEWV
AVIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAV
YYCARDLWGWYFDYWGQGTLVTVSSGGGGGSGGGGSGGGSGGG
GSEIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPR
LLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRRN
WPLTFGGGTKVEIK
MEDI-547 LC DEC-205 743 EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLI mAb YDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRRNWP containing LTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRE tumor mAb AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK ScFv HKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSEVQLLESGG
GLVQPGGSLRLSCAASGFTFSHYMMAWVRQAPGKGLEWVSRIGPS
GGPTHYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAGY
DSGYDYVAVAGPAEYFQHWGQGTLVTVSSGGGGSGGGGSGGGGS
GGGGSDIQMTQSPSSLSASVGDRVTITCRASQSISTWLAWYQQKPG
KAPKLLIYKASNLHTGVPSRFSGSGSGTEFSLTISGLQPDDFATYYCQ
QYNSYSRTFGQGTKVEIK
LC tumor 744 DIQMTQSPSSLSASVGDRVTITCRASQSISTWLAWYQQKPGKAPKLL mAb IYKASNLHTGVPSRFSGSGSGTEFSLTISGLQPDDFATYYCQQYNSYS containing RTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRE DEC-205 mAb AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK ScFv HKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLVESGG
GVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGLEWVAVIWY
DGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAR
DLWGWYFDYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSEIVL
TQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLIYDA
SNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRRNWPLTFG
GGTKVEIK
Narnatumab LC DEC-205 749 EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLI mAb YDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRRNWP containing LTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRE tumor mAb AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK ScFv HKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSEVQLVESGG
GLVQPGGSLRLSCAASGFTFSSYLMTWVRQAPGKGLEWVANIKQD
GSEKYYVDSVKGRFTISRDNAKNSLNLQMNSLRAEDTAVYYCTRD
GYSSGRHYGMDVWGQGTTVIVSSGGGGSGGGGSGGGGSGGGGSEI Antibody Region SEQ Sequence
ID
NO:
VLTQSPATLSLSPGERATLSCRASQSVSRYLAWYQQKPGQAPRLLIY
DASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRSNWPR
TFGQGTKVEIK
LC tumor 750 EIVLTQSPATLSLSPGERATLSCRASQSVSRYLAWYQQKPGQAPRLL mAb IYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRSNWP containing RTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRE DEC-205 mAb AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK ScFv HKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLVESGG
GVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGLEWVAVIWY
DGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAR
DLWGWYFDYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSEIVL
TQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLIYDA
SNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRRNWPLTFG
GGTKVEIK
RG7841 LC DEC-205 776 EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLI mAb YDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRRNWP containing LTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRE tumor mAb AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK ScFv HKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSEVQLVESGP
ALVKPTQTLTLTCTVSGFSLTGYSVNWIRQPPGKALEWLGMIWGDG
STDYNSALKSRLTISKDTSKNQVVLTMTNMDPVDTATYYCARDYY
FNYASWFAYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSDIQM
TQSPSSLSASVGDRVTITCSASQGISNYLNWYQQKPGKTVKLLIYYT
SNLHSGVPSRFSGSGSGTDYTLTISSLQPEDFATYYCQQYSELPWTF
GQGTKVEIK
LC tumor 777 DIQMTQSPSSLSASVGDRVTITCSASQGISNYLNWYQQKPGKTVKLL mAb IYYTSNLHSGVPSRFSGSGSGTDYTLTISSLQPEDFATYYCQQYSELP containing WTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPR DEC-205 mAb EAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYE ScFv KHKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLVESG
GGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGLEWVAVIW
YDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCA
RDLWGWYFDYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSEIV
LTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLIYD
ASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRRNWPLTF
GGGTKVEIK
Farletuzumab LC DEC-205 722 EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLI mAb YDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRRNWP containing LTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRE tumor mAb AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK ScFv HKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSEVQLVESGG
GVVQPGRSLRLSCSASGFTFSGYGLSWVRQAPGKGLEWVAMISSGG
SYTYYADSVKGRFAISRDNAKNTLFLQMDSLRPEDTGVYFCARHG
DDPAWFAYWGQGTPVTVSSGGGGSGGGGSGGGGSGGGGSDIQLT
QSPSSLSASVGDRVTITCSVSSSISSNNLHWYQQKPGKAPKPWIYGT
SNLASGVPSRFSGSGSGTDYTFTISSLQPEDIATYYCQQWSSYPYMY
TFGQGTKVEIK
LC tumor 723 DIQLTQSPSSLSASVGDRVTITCSVSSSISSNNLHWYQQKPGKAPKP mAb WIYGTSNLASGVPSRFSGSGSGTDYTFTISSLQPEDIATYYCQQWSS containing YPYMYTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNN DEC-205 mAb FYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKA ScFv DYEKHKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLV
ESGGGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGLEWVA
VIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVY
YCARDLWGWYFDYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGG
SEIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRL
LIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRRN
WPLTFGGGTKVEIK
Mirvetuximab LC DEC-205 746 EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLI Antibody Region SEQ Sequence
ID
NO:
mAb YDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRRNWP containing LTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRE tumor mAb AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK
ScFv HKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLVQSGA
EVVKPGASVKISCKASGYTFTGYFMNWVKQSPGQSLEWIGRIHPYD
GDTFYNQKFQGKATLTVDKSSNTAHMELLSLTSEDFAVYYCTRYD
GSRAMDYWGQGTTVTVSSGGGGSGGGGSGGGGSGGGGSDIVLTQS
PLSLAVSLGQPAIISCKASQSVSFAGTSLMHWYHQKPGQQPRLLIYR
ASNLEAGVPDRFSGSGSKTDFTLTISPVEAEDAATYYCQQSREYPYT
FGGGTKLEIK
LC tumor 747 DIVLTQSPLSLAVSLGQPAIISCKASQSVSFAGTSLMHWYHQKPGQQ mAb PRLLIYRASNLEAGVPDRFSGSGSKTDFTLTISPVEAEDAATYYCQQ containing SREYPYTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNN DEC-205 mAb FYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKA ScFv DYEKHKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLV
ESGGGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGLEWVA
VIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVY
YCARDLWGWYFDYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGG
SEIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRL
LIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRRN
WPLTFGGGTKVEIK
J591 variantl LC DEC-205 731 EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLI mAb YDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRRNWP containing LTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRE tumor mAb AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK ScFv HKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSEVQLQQSGP
ELKKPGTSVRISCKTSGYTFTEYTIHWVKQSHGKSLEWIGNINPNNG
GTTYNQKFEDKATLTVDKSSSTAYMELRSLTSEDSAVYYCAAGWN
FDYWGQGTTLTVSSGGGGSGGGGSGGGGSGGGGSDIVMTQSHKF
MSTSVGDRVSIICKASQDVGTAVDWYQQKPGQSPKLLIYWASTRHT
GVPDRFTGSGSGTDFTLTITNVQSEDLADYFCQQYNSYPLTFGAGT
MLDLK
LC tumor 732 DIVMTQSHKFMSTSVGDRVSIICKASQDVGTAVDWYQQKPGQSPKL mAb LIYWASTRHTGVPDRFTGSGSGTDFTLTITNVQSEDLADYFCQQYNS containing YPLTFGAGTMLDLKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFY DEC-205 mAb PREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADY ScFv EKHKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLVES
GGGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGLEWVAVI
WYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYC
ARDLWGWYFDYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSEI
VLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLIY
DASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRRNWPL
TFGGGTKVEIK
J591 variant 2 LC DEC-205 734 EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLI mAb YDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRRNWP containing LTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRE tumor mAb AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK ScFv HKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSEVQLQQSGP
ELVKPGTSVRISCKTSGYTFTEYTIHWVKQSHGKSLEWIGNINPNNG
GTTYNQKFEDKATLTVDKSSSTAYMELRSLTSEDSAVYYCAAGWN
FDYWGQGTTLTVSSGGGGSGGGGSGGGGSGGGGSNIVMTQSPKSM
SMSVGERVTLTCKASENVVTYVSWYQQKPEQSPKLLIYGASNRYT
GVPDRFTGSGSATDFTLTISSVQAEDLADYHCGQGYSYPYTFGGGT
KLEIK
LC tumor 735 NIVMTQSPKSMSMSVGERVTLTCKASENVVTYVSWYQQKPEQSPK mAb LLIYGASNRYTGVPDRFTGSGSATDFTLTISSVQAEDLADYHCGQGY containing SYPYTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFY DEC-205 mAb PREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADY ScFv EKHKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLVES Antibody Region SEQ Sequence
ID
NO:
GGGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGLEWVAVI
WYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYC
ARDLWGWYFDYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSEI
VLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLIY
DASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRRNWPL
TFGGGTKVEIK
Rovalpituzum LC DEC-205 779 EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLI ab mAb YDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRRNWP containing LTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRE tumor mAb AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK ScFv HKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLVQSGA
EVKKPGASVKVSCKASGYTFTNYGMNWVRQAPGQGLEWMGWIN
TYTGEPTYADDFKGRVTMTTDTSTSTAYMELRSLRSDDTAVYYCA
RIGDSSPSDYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSEIVMT
QSPATLSVSPGERATLSCKASQSVSNDVVWYQQKPGQAPRLLIYYA
SNRYTGIPARFSGSGSGTEFTLTISSLQSEDFAVYYCQQDYTSPWTFG
QGTKLEIK
LC tumor 780 EIVMTQSPATLSVSPGERATLSCKASQSVSNDVVWYQQKPGQAPRL mAb LIYYASNRYTGIPARFSGSGSGTEFTLTISSLQSEDFAVYYCQQDYTS containing PWTFGQGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYP DEC-205 mAb REAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYE ScFv KHKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLVESG
GGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGLEWVAVIW
YDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCA
RDLWGWYFDYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSEIV
LTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLIYD
ASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRRNWPLTF
GGGTKVEIK
PF-06647020 LC DEC-205 770 EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLI mAb YDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRRNWP containing LTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRE tumor mAb AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK ScFv HKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLVQSGP
EVKKPGASVKVSCKASGYTFTDYAVHWVRQAPGKRLEWIGVISTY
NDYTYNNQDFKGRVTMTRDTSASTAYMELSRLRSEDTAVYYCARG
NSYFYALDYWGQGTSVTVSSGGGGSGGGGSGGGGSGGGGSEIVLT
QSPATLSLSPGERATLSCRASESVDSYGKSFMHWYQQKPGQAPRLLI
YRASNLESGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQSNEDPW
TFGGGTKLEIK
LC tumor 771 EIVLTQSPATLSLSPGERATLSCRASESVDSYGKSFMHWYQQKPGQ mAb APRLLIYRASNLESGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQS containing NEDPWTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNN DEC-205 mAb FYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKA ScFv DYEKHKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLV
ESGGGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGLEWVA
VIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVY
YCARDLWGWYFDYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGG
SEIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRL
LIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRRN
WPLTFGGGTKVEIK
Antibody to LC DEC-205 689 EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLI PTK7 mAb YDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRRNWP containing LTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRE tumor mAb AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK ScFv HKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLVESGG
GVVQPGRSLRLSCAASGFTFSSYAFHWVRQAPGKGLEWVAVISYD
GSIKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARTY
YFDYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSEIVLTQSPDF
QSVTPKEKVTITCRASQSIGSSLHWYQQKPDQSPKLLIKYASQSFSG Antibody Region SEQ Sequence
ID
NO:
VPSRFSGSGSGTDFTLTINSLEAEDAAAYYCHQSSSLPITFGQGTRLEI K
LC tumor 690 EIVLTQSPDFQSVTPKEKVTITCRASQSIGSSLHWYQQKPDQSPKLLI mAb KYASQSFSGVPSRFSGSGSGTDFTLTINSLEAEDAAAYYCHQSSSLPI containing TFGQGTRLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREA DEC-205 mAb KVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKH ScFv KVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLVESGGG
VVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGLEWVAVIWYD
GSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARD
LWGWYFDYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSEIVLT
QSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLIYDAS
NRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRRNWPLTFG
GGTKVEIK
Ladiratuzuma LC DEC-205 785 EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLI b mAb YDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRRNWP containing LTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRE tumor mAb AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK ScFv HKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLVQSGA
EVKKPGASVKVSCKASGLTIEDYYMHWVRQAPGQGLEWMGWIDP
ENGDTEYGPKFQGRVTMTRDTSINTAYMELSRLRSDDTAVYYCAV
HNAHYGTWFAYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSD
VVMTQSPLSLPVTLGQPASISCRSSQSLLHSSGNTYLEYFQQRPGQSP
RPLIYKISTRFSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCFQGS
HVPYTFGGGTKVEIK
LC tumor 786 DVVMTQSPLSLPVTLGQPASISCRSSQSLLHSSGNTYLEYFQQRPGQ mAb SPRPLIYKISTRFSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCFQ containing GSHVPYTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLN DEC-205 mAb NFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSK ScFv ADYEKHKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQL
VESGGGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGLEWV
AVIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAV
YYCARDLWGWYFDYWGQGTLVTVSSGGGGSGGGGSGGGGSGGG
GSEIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPR
LLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRRN
WPLTFGGGTKVEIK
Cirmtuzumab LC DEC-205 707 EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLI mAb YDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRRNWP containing LTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRE tumor mAb AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK ScFv HKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLQESGP
GLVKPSQTLSLTCTVSGYAFTAYNIHWVRQAPGQGLEWMGSFDPY
DGGSSYNQKFKDRLTISKDTSKNQVVLTMTNMDPVDTATYYCARG
WYYFDYWGHGTLVTVSSGGGGSGGGGSGGGGSGGGGSDIVMTQT
PLSLPVTPGEPASISCRASKSISKYLAWYQQKPGQAPRLLIYSGSTLQ
SGIPPRFSGSGYGTDFTLTINNIESEDAAYYFCQQHDESPY
LC tumor 708 DIVMTQTPLSLPVTPGEPASISCRASKSISKYLAWYQQKPGQAPRLLI mAb YSGSTLQSGIPPRFSGSGYGTDFTLTINNIESEDAAYYFCQQHDESPY containing TFGEGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREA DEC-205 mAb KVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKH ScFv KVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLVESGGG
VVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGLEWVAVIWYD
GSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARD
LWGWYFDYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSEIVLT
QSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLIYDAS
NRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRRNWPLTFG
GGTKVEIK
Antibody to LC DEC-205 740 EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLI MAGE-A3 mAb YDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRRNWP containing LTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRE Antibody Region SEQ Sequence
ID
NO:
tumor mAb AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK ScFv HKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLVESGG
GVVQPGRSLRLSCTASGFRFRSHGMHWVRQAPGKGLEWVAVISYD
GNNKLYADSVKGRITISRDNSKNTLFLQMNNVRAEDTAVYYCASP
YTSDWQYFQYWGQGTLVIVSSGGGGSGGGGSGGGGSGGGGSEIVM
TQSPATLSVSPGERATFSCRASQNISTTLAWYQQKPGQAPRLLIYDT
STRATGIPARFSGSGSGTEFTLTISSLQSEDLAVYYCQQSNSWPLTFG
GGTKVEIK
LC tumor 741 EIVMTQSPATLSVSPGERATFSCRASQNISTTLAWYQQKPGQAPRLL mAb IYDTSTRATGIPARFSGSGSGTEFTLTISSLQSEDLAVYYCQQSNSWP containing LTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRE DEC-205 mAb AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK ScFv HKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLVESGG
GVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGLEWVAVIWY
DGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAR
DLWGWYFDYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSEIVL
TQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLIYDA
SNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRRNWPLTFG
GGTKVEIK
Antibody to LC DEC-205 755 EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLI NY-ESO-1 mAb YDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRRNWP containing LTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRE tumor mAb AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK ScFv HKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLVQSGG
GVVRPGGSLRLSCAASGFSFIDYGMSWVRQVPGKGLEWVAGMNW
SGDKKGHAESVKGRFIISRDNAKNTLYLEMSSLRVEDTALYFCARG
EYSNRFDPRGRGTLVTVSSGGGGSGGGGSGGGGSGGGGSDIVMTQ
TPLSLPVTLGQPASLSCRSSQSLVFTDGNTYLNWFQQRPGQSPRRLI
YKVSSRDPGVPDRFSGTGSGTDFTLEISRVEAEDIGVYYCMQGTHW
PPIFGQGTKVEIK
LC tumor 756 DIVMTQTPLSLPVTLGQPASLSCRSSQSLVFTDGNTYLNWFQQRPGQ mAb SPRRLIYKVSSRDPGVPDRFSGTGSGTDFTLEISRVEAEDIGVYYCM containing QGTHWPPIFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLL DEC-205 mAb NNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLS ScFv KADYEKHKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQ
LVESGGGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGLEW
VAVIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTA
VYYCARDLWGWYFDYWGQGTLVTVSSGGGGSGGGGSGGGGSGG
GGSEIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAP
RLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRR
NWPLTFGGGTKVEIK
Trastuzumab LC DEC-205 794 EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLI mAb YDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRRNWP containing LTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRE tumor mAb AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK ScFv HKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSEVQLVESGG
GLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTN
GYTRYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRWG
GDGFYAMDYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSDIQM
TQSPSSLSASVGDRVTITCRASQDVNTAVAWYQQKPGKAPKLLIYS
ASFLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQHYTTPPTF
GQGTKVEIK
LC tumor 795 DIQMTQSPSSLSASVGDRVTITCRASQDVNTAVAWYQQKPGKAPKL mAb LIYSASFLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQHYTTP containing PTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRE DEC-205 mAb AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK ScFv HKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLVESGG
GVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGLEWVAVIWY
DGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAR Antibody Region SEQ Sequence
ID
NO:
DLWGWYFDYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSEIVL TQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLIYDA SNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRRNWPLTFG GGTKVEIK
LC DEC205 799 EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLI mAb YDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRRNWP containing LTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRE tumor mAb AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK scFv HKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSDIQMTQSPSS (LH,25mer) LSASVGDRVTITCRASQDVNTAVAWYQQKPGKAPKLLIYSASFLYS
GVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQHYTTPPTFGQGTKV
EIKGGGGSGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVQPGGS
LRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYADS
VKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAM
DYWGQGTLVTVSS
TABLE 11. Fusion Sequences - CD40 fusion with DEC205
Antibody Region SEQ Sequence
ID
NO:
DEC205 HC DEC205 243 QVQLVESGGGVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGL variant 1 mAb with EWVAVIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAED
CD40 mAb TAVYYCARDLWGWYFDYWGQGTLVTVSSASTKGPSVFPLAPSSKS ScFv TSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLY
SLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCP
PCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVK
FNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKE
YKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSL
TCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTV
DKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGS
QVQLVQSGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQG
LEWMGWINPDSGGTNYAQKFQGRVTMTRDTSISTAYMELNRLRSD
DTAVYYCARDQPLGYCTNGVCSYFDYWGQGTLVTVSSGGGGSGG
GGSGGGGSGGGGSDIQMTQSPSSVSASVGDRVTITCRASQGIYSWL
AWYQQKPGKAPNLLIYTASTLQSGVPSRFSGSGSGTDFTLTISSLQPE
DFATYYCQQANIFPLTFGGGTKVEIK
HC CD40 242 QVQLVQSGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQG mAb with LEWMGWINPDSGGTNYAQKFQGRVTMTRDTSISTAYMELNRLRSD DEC205 mAb DTAVYYCARDQPLGYCTNGVCSYFDYWGQGTLVTVSSASTKGPSV ScFv FPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPA
VLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKS
CDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDV
SHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQ
DWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREE
MTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGS
FFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGG
GGSGGGGSQVQLVESGGGVVQPGRSLRLSCAASGFTFSNYGMYWV
RQAPGKGLEWVAVIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQ
MNSLRAEDTAVYYCARDLWGWYFDYWGQGTLVTVSSGGGGSGG
GGSGGGGSGGGGSEIVLTQSPATLSLSPGERATLSCRASQSVSSYLA
WYQQKPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPED
FAVYYCQQRRNWPLTFGGGTKVEIK
LC DEC205 244 EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLL mAb IYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRRNW containing PLTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPR CD40 mAb EAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYE scFv KHKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLVQSG
AEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLEWMGWI Antibody Region SEQ Sequence
ID
NO:
NPDSGGTNYAQKFQGRVTMTRDTSISTAYMELNRLRSDDTAVYYC ARDQPLGYCTNGVCSYFDYWGQGTLVTVSSGGGGSGGGGSGGGG SGGGGSDIQMTQSPSSVSASVGDRVTITCRASQGIYSWLAWYQQKP GKAPNLLIYTASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYC QQANIFPLTFGGGTKVEIK
LC CD40 800 DIQMTQSPSSVSASVGDRVTITCRASQGIYSWLAWYQQKPGKAPNL mAb LIYTASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQANIFP containing LTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRE DEC205 mAb AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK scFv HKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLVESGG
GVVQPGRSLRLSCAASGFTFSNYGMYWVRQAPGKGLEWVAVIWY
DGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAR
DLWGWYFDYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSEIVL
TQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLIYDA
SNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRRNWPLTF
GGGTKVEIK
DEC205 HC DEC205 256 EVQLVQSGAEVKKPGESLRISCKGSGDSFTTYWIGWVRQMPGKGL variant 2 mAb with EWMGIIYPGDSDTIYSPSFQGQVTISADKSISTAYLQWSSLKASDTA
CD40 mAb MYYCTRGDRGVDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGG ScFv TAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSS
VVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCP
APELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNW
YVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKC
KVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCL
VKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDK
SRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSQV
QLVQSGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLE
WMGWINPDSGGTNYAQKFQGRVTMTRDTSISTAYMELNRLRSDDT
AVYYCARDQPLGYCTNGVCSYFDYWGQGTLVTVSSGGGGSGGGG
SGGGGSGGGGSDIQMTQSPSSVSASVGDRVTITCRASQGIYSWLAW
YQQKPGKAPNLLIYTASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDF
ATYYCQQANIFPLTFGGGTKVEIK
HC CD40 255 QVQLVQSGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQG mAb with LEWMGWINPDSGGTNYAQKFQGRVTMTRDTSISTAYMELNRLRSD DEC205 mAb DTAVYYCARDQPLGYCTNGVCSYFDYWGQGTLVTVSSASTKGPSV ScFv FPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPA
VLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKS
CDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDV
SHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQ
DWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREE
MTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGS
FFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGG
GGSGGGGSEVQLVQSGAEVKKPGESLRISCKGSGDSFTTYWIGWVR
QMPGKGLEWMGIIYPGDSDTIYSPSFQGQVTISADKSISTAYLQWSS
LKASDTAMYYCTRGDRGVDYWGQGTLVTVSSGGGGSGGGGSGGG
GSGGGGSDIQMTQSPSSLSASVGDRVTITCRASQGISRWLAWYQQK
PEKAPKSLIYAASSLQSGVPSRFSGSGSGTDFTLTISGLQPEDFATYY
CQQYNSYPRTFGQGTKVEIK
LC DEC205 257 DIQMTQSPSSLSASVGDRVTITCRASQGISRWLAWYQQKPEKAPKS mAb LIYAASSLQSGVPSRFSGSGSGTDFTLTISGLQPEDFATYYCQQYNSY containing PRTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPR CD40 mAb EAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYE scFv KHKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSQVQLVQSG
AEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLEWMGWI
NPDSGGTNYAQKFQGRVTMTRDTSISTAYMELNRLRSDDTAVYYC
ARDQPLGYCTNGVCSYFDYWGQGTLVTVSSGGGGSGGGGSGGGG
SGGGGSDIQMTQSPSSVSASVGDRVTITCRASQGIYSWLAWYQQKP
GKAPNLLIYTASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYC
QQANIFPLTFGGGTKVEIK Antibody Region SEQ Sequence
ID
NO:
LC CD40 801 DIQMTQSPSSVSASVGDRVTITCRASQGIYSWLAWYQQKPGKAPNL mAb LIYTASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQANIFP containing LTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRE DEC205 mAb AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK scFv HKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSEVQLVQSGA
EVKKPGESLRISCKGSGDSFTTYWIGWVRQMPGKGLEWMGIIYPGD
SDTIYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCTRGDRG
VDYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSDIQMTQSPSSL
SASVGDRVTITCRASQGISRWLAWYQQKPEKAPKSLIYAASSLQSG
VPSRFSGSGSGTDFTLTISGLQPEDFATYYCQQYNSYPRTFGQGTKV
EIK
Immune-Stimulatory Compounds
[0304] In some embodiments, the immune- stimulatory conjugates described herein further comprise an immune- stimulatory compound. An immune- stimulatory compound can be a small molecule, a compound or molecule that binds to a protein target and can activate the target protein' s function, or a compound that binds to a protein target and can inhibit the protein target's function, resulting in immune stimulation or modulation. In certain embodiments, an immune- stimulatory compound of a conjugate (i.e., attached to an antibody construct either directly or via a linker) can stimulate or activate its protein target with no or minimal processing of the conjugate. In certain embodiments, an immune- stimulatory compound of a conjugate (i.e., attached to an antibody construct either directly or via a linker) can inhibit its protein target with no or minimal processing of the conjugate. In this context, processing refers to degradation of the antibody construct or cleavage of the linker to liberate the immune- stimulatory compound or degredation product of the conjugate containing the immune- stimulatory compound. In certain embodiments, the protein target of the immune- stimulatory compound is an extracellular protein target and is located on the cell surface membrane or cellular compartments in communication with the cell surface, such as in the endoplasmic reticulum (ER). In certain embodiments, the protein target is intracellular, such as in the cytoplasm.
[0305] In some embodiments, an immune- stimulatory compound can activate immune cells. In some embodiments, an immune- stimulatory compound can reduce inhibition of immune cells. In some embodiments, an immune- stimulatory compound can stimulate immune activation by triggering degradation of a protein target.
[0306] In some embodiments, an immune- stimulatory compound can be a molecule or
compound whose action on its target can lead to immune stimulation by direct immune cell activation. In some embodiments, the immune activation can be indirect by alteration of the immune suppressive microenvironment of a tumor (e.g., removing an immunosuppressive signal or altering an immunosuppressive state). In some embodiments, the immune- stimulatory compound's activity can be both direct and indirect. In certain embodiments, an immune- stimulatory conjugate can alter the activity of its protein target in cells having an antigen bound by the conjugate as compared to activity of the protein target in non- antigen bearing cells (i.e., the immune- stimulatory activity is antigen-specific).
[0307] In certain embodiments, the immune- stimulatory compound can be coupled to an Fc domain or other portion of an antibody construct via a linker. In each of the embodiments described herein, the immune- stimulatory compound can be coupled to the antibody construct via a linker.
[0308] An immune- stimulatory compound can be a Pattern recognition receptor (PRR) agonist. Pattern recognition receptors (PRRs) can recognize pathogen-associated molecular patterns (PAMPs) and damage-associated molecular patterns (DAMPs). A PRR can be membrane bound. A PRR can be cytosolic. A PAMP molecule can be a toll-like receptor agonist. A PRR can be a to 11- like receptor (TLR). A PRR can be RIG-I-like receptor. A PRR can be a receptor kinase. A PRR can be a C-type lectin receptor. A PRR can be a NOD-like receptor. A PRR can be TLR1, TLR2, TLR3, TLR4, TLR5, TLR6, TLR7, TLR8, TLR9, TLR 10, TLR11, TLR 12, or TLR13.
[0309] A PRR agonist can be a damage-associated molecular pattern (DAMP) molecule. A DAMP molecule can be an intracellular protein. A DAMP molecule can be a heat-shock protein. A DAMP molecule can be an HMGB 1 protein. A DAMP molecule can be a protein derived from the extracellular matrix that is generated after tissue injury. A DAMP molecule can be a hyaluronan fragment. A DAMP molecule can be DNA. A DAMP molecule can be RNA. A DAMP molecule can be an S 100 molecule. A DAMP molecule can be a nucleotide(s). A DAMP molecule can be ATP. A DAMP molecule can be a nucleoside(s). A DAMP molecule can be an adenosine. A DAMP molecule can be uric acid.
[0310] In some embodiments, the immune- stimulatory compound can be a Toll-like receptor agonist, a RIG-I agonist, a STING agonist, a GPCR agonist, an ion channel agonist, a membrane transporter agonist, an ER protein agonist, a beta-catenin pathway inhibitor, a kinase inhibitor, a TNIK inhibitor, a Tankyrase inhibitor, a GPCR antagonist, an HSP90 inhibitor, or an AAA- ATPase p97 inhibitor. In some embodiments, the immune- stimulatory compound can be a Tolllike receptor agonist, a RIG-I agonist or a STING agonist. In some embodiments, the immune- stimulatory compound can be a beta-catenin pathway inhibitor, such as a TNIK inhibitor or a Tankyrase inhibitor.
[0311] In some embodiments, the immune- stimulatory compound is a Toll-like receptor agonist.
A toll- like receptor agonist can be any molecule that acts as an agonist to at least one toll- like receptor. In some embodiments, the Toll- like receptor agonist can be a molecule selected from a
CpG oligonucleotide, Poly G10, Poly G3, Poly I:C, a lipopolysaccharide, a zymosan, a bacterial flagella protein (e.g., flagellin), PamjCSIQ, PamCysPamSK4, dsRNA, ssRNA, a diacylated lipopeptide, a bacterial lipoprotein, a triacylated lipoprotein, lipoteichoic acid, or a peptidoglycan (such as a bacterial peptidoglycan).
[0312] In some embodiments, an immune- stimulatory compound is not a toll-like receptor agonist. In some embodiments, a toll- like receptor agonist is not a naturally occurring molecule, such as CpG oligonucleotide, a lipopolysaccharide, a zymosan, a bacterial flagella protein (e.g., flagellin), PamjCSIQ, PamCysPamSIQ, dsRNA, ssRNA, a diacylated lipopeptide, a bacterial lipoprotein, a triacylated lipoprotein, lipoteichoic acid, or a peptidoglycan (such as a bacterial peptidoglycan). In some further embodiments, a toll- like receptor agonist is not a synthetic nucleic acid, such as Poly G10, Poly G3 or Poly I:C.
[0313] In some embodiments, a toll- like receptor agonist can be a synthetic small molecule. A to 11- like receptor agonist can be imiquimod. A to 11- like receptor agonist can be CL307. A to 11- like receptor agonist can be S-27609. A toll-like receptor agonist can be resiquimod. A toll-like receptor agonist can be UC-IV150. A to 11- like receptor agonist can be gardiquimod. A to 11- like receptor agonist can be motolimod. A toll-like receptor agonist can be a motolimod analog. A to 11- like receptor agonist can be VTX-1463. A to 11- like receptor agonist can be GS-9620. A tolllike receptor agonist can be GSK2245035. A toll- like receptor agonist can be TMX-101. A tolllike receptor agonist can be TMX-201. A toll- like receptor agonist can be TMX-202. A toll- like receptor agonist can be isatoribine. A toll-like receptor agonist can be AZD8848. A toll-like receptor agonist can be MEDI9197. A to 11- like receptor agonist can be 3M-051. A to 11- like receptor agonist can be 3M-852. A toll-like receptor agonist can be 3M-052. A toll-like receptor agonist can be 3M-854A. A toll-like receptor agonist can be S-34240. A toll-like receptor agonist can be CL663. A to 11- like receptor agonist can be KU34B.
[0314] A RIG-I agonist can be KIN1148. A RIG-I agonist can be SB-9200. A RIG-I agonist comprises a 5'ppp-dsRNA.
[0315] In some embodiments, the immune- stimulatory compound can comprise a non-naturally occurring chemotype, such as a substituted pyrimidine, a substituted purine, a substituted guanine nucleoside, a substituted 8-oxoadenine, a substituted imidazoquinoline, a substituted
thiazoquinoline, a substituted 2- amino imidazole, a substituted furo[2,3-c]pyridine, a substituted pyrazine, a substituted furo[2,3-c]quinoline, a substituted 2-aminobenzimidazole, a substituted 2- aminoquinoline, or a substituted 2-aminobenzazepine.
[0316] In certain embodiments, the immune- stimulatory compound can comprise S-27609, CL307, UC-IV150, imiquimod, gardiquimod, resiquimod, motolimod, VTS-1463, GS-9620, GSK2245035, TMX-101, TMX-201, TMX-202, isatoribine, AZD8848, MEDI9197, 3M-051, 3M-852, 3M-052, 3M-854A, S-34240, KU34B, SB9200, SB 11285, 8-substituted imidazo[l,5- a]pyridine, or CL663.
[0317] An immune- stimulatory compound can comprise an inhibitor of TGFbeta, Beta-Catenin, PI3K-beta, STAT3, IL-10, IDO or TDO. The immune- stimulatory compound can comprise LY2109761, GSK263771, iCRT3, iCRT5, iCRT14, LY2090314, CGX-1321, PRI-724, BC21, ZINCO2092166, LGK974, IWP2, LY3022859, LY364947, SB431542, AZD8186, SD-208, indoximod (NLG8189), F001287, GDC-0919, epacadostat (INCB024360), RG70099, 1-methyl- L-tryptophan, methylthiohydantoin tryptophan, brassinin, annulin B, exiguamine A, PIM, LM10, 8-substituted 2-amino-3H-benzo[b]azepine-4-carboxamide, or INCB023843.
[0318] Additionally, stimulator of interferon genes (STING) can act as a cytosolic DNA sensor wherein cytosolic DNA and unique bacterial nucleic acids called cyclic dinucleo tides are recognized by STING, and therefore STING agonists. In certain embodiments, the STING agonist can comprise a cyclic dinucleotide. Other non- limiting examples of STING agonists include:
[0319] , wherein in some embodiments, Xi=X2=0; X3=G; X4=G; X6=2 TEAH; in some embodiments, X1=X2=S [Rp,Rp]; X3=G; X4=A; X5=H; X6=2 TEAH; in some embodiments, Xi=X2=S [RP,RP] ; X3=A; X4=A; X5=H; X6=2 Na; in some embodiments, X1=X2=S [RP,RP] ; X3=A; X4=A; X5=H; X6=2 NH4; and in some embodiments, X1=X2=0 ; X3=G; X4=A; X5=H; X6=2 TEAH;
, wherein Ri=R2=H; R2=H; Ri=H, R2=propargyl; Ri=allyl, R2=H; Ri=H, R2=allyl; Ri=methyl, R2=H; Ri=H, R2=methyl; Ri=ethyl, R2=H; Ri=H, R2=ethyl; Ri=propyl, R2=H; Ri=H, R2=propyl; Ri=benzyl, R2=H; Ri=H, R2=benzyl; Ri=myristoyl, R2=H; Ri=H, R2=myristoyl; Ri=R2=heptanoyl; Ri=R2=hexanoyl; or Ri=R2=pentanoyl;
[0320] wherein Ri=R2=H; Ri=propargyl, R2=H; Ri=H, R2=propargyl; Ri=allyl, R2=H; Ri=H, R2=allyl; Ri=methyl, R2=H; Ri=H, R2=methyl; Ri=ethyl, R2=H; Ri=H, R2=ethyl; Ri=propyl, R2=H; Ri=H, R2=propyl; Ri=benzyl, R2=H; Ri=H, R2=benzyl; Ri=myristoyl, R2=H; Ri=H, R2=myristoyl; Ri=R2=heptanoyl; Ri=R2=hexanoyl; or Ri=R2=pentanoyl;
[0321] wherein Ri=R2=H; Ri=propargyl, R2=H; Ri=H, R2=propargyl; Ri=allyl, R2=H; Ri=H, R2=allyl; Ri=methyl, R2=H; Ri=H, R2=methyl; Ri=ethyl, R2=H; Ri=H, R2=ethyl; Ri=propyl, R2=H; Ri=H, R2=propyl; Ri=benzyl, R2=H; Ri=H, R2=benzyl; Ri=myristoyl, R2=H; Ri=H, R2=myristoyl; Ri=R2=heptanoyl; Ri=R2=hexanoyl; or Ri=R2=pentanoyl;
, wherein each X is independently O or S, and R3 and R4 are each independently H or an optionally substituted straight chain alkyl of from 1 to 18 carbons and from 0 to 3 heteroatoms, an optionally substituted alkenyl of from 1-9 carbons, an optionally substituted alkynyl of from 1-9 carbons, or an optionally substituted aryl, wherein substitution(s), when present, may be independently selected from the group consisting of Ci-6 alkyl straight or branched chain, benzyl, halogen, trihalo methyl, Ci-6 alkoxy,— N02,— NH2,— OH, =0,— COOR ' where R ' is H or lower alkyl,— CH2OH, and— CONH2, wherein R3 and R4 are not both H;
0; X1=X2=S; or Xi=0 and X2=S,
-198- [0322] In some embodiments, an immune- stimulatory compound can be a kinase inhibitor. An immune- stimulatory compound can inhibit one or more kinases.
[0323] An immune- stimulatory compound can be an inhibitor of ALK, Bcr-Abl, BRAF, BTK, c- KIT, EGFR, ErbB2, JAK, MEK, MET, PDGFRB, RET, ROS 1, Syk, SRC, TGFpR2, TNIK, TNKS, TNKS2, TrkA, TrkB, TrkC, VEGFR1, VEGFR2, VEGFR3, FGFR1, FGFR2, FGFR3, FGFR4, CSFIR, RON/MSTIR, TYR03, MERTK, AXL, ΡΙ3Κδ, ΡΙ3Κγ, MAP4K1, PERK, KIT, or any combination thereof. An immune- stimulatory compound can be an inhibitor of TGFpRl, TGFpR2, TNIK, TNKS, ΡΙ3Κ-β, STAT3, IL-10, IDO, or TDO.
[0324] In various embodiments, the immune- stimulatory compound comprises LY2109761, GSK263771, iCRT3, iCRT5, iCRT14, LY2090314, CGX-1321, PRI-724, BC21,
ZINCO2092166, LGK974, IWP2, LY3022859, LY364947, SB431542, AZD8186, SD-208, indoximod (NLG8189), F001287, GDC-0919, epacadostat (INCB024360), RG70099, 1-methyl- L-tryptophan, methylthiohydantoin tryptophan, brassinin, annulin B, exiguamine A, PIM, LM10, INCB023843, or 8-substituted imidazo[ 1,5 -a] pyridine.
[0325] An immune- stimulatory compound can be an agonist of a GPCR, an ion channel, a membrane transporter, or an ER protein.
[0326] An immune- stimulatory compound can be an antagonist of the GPCR A2aR, the sphingosine 1 -phosphate receptor 1, prostaglandin receptor EP3, prostanglandin receptor E2, Frizzled, CXCR4, or an LPA receptor.
[0327] An immune- stimulatory compound can be an ion channel agonist for CRAC, Kvl.3, or KCa3.1.
[0328] An immune- stimulatory compound can be an inhibitor of HSP90 or AAA-ATPase p97. Immune-Stimulatory Conjugate Properties
[0329] In certain embodiments, an immune- stimulatory compound of a conjugate (i.e, attached to an antibody construct either directly or via a linker) has a biological potency no less than at least about 0.33%, about 1%, about 5%, about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 85%, about 90%, about 95%, or about 100% of the potency of the control (free) immune- stimulatory compound (i.e., not attached to an antibody construct).
[0330] The specificity of the antigen-binding domain (of a conjugate) for an antigen can be influenced by the attachment of an immune- stimulatory compound to an antibody construct. In various embodiments, an antigen-binding domain of the conjugate can bind to its antigen with at least about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 85%, about 90%, about 95%, or about 100% of the specificity of the antigen-binding domain for the antigen in the absence of attachment of the immune- stimulatory compound.
[0331] The specificity of the Fc domain (of a conjugate) for an Fc receptor can be influenced by the attachment site of an immune- stimulatory compound (directly or via a linker). In some embodiments, the Fc domain of a conjugate retains the specificity of the unconjugated Fc domain to bind to an Fc receptor. In specific embodiments, the Fc domain of the conjugate can bind to an Fc receptor with at least about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 85%, about 90%, about 95%, or about 100% of the specificity of the Fc domain to the Fc receptor in the absence of attachment of the immune- stimulatory compound.
[0332] In some embodiments, the Fc domain of a conjugate has an altered specificity for an Fc receptor relative to the corresponding unconjugated Fc domain. In specific embodiments, the Fc domain of the conjugate can bind to an Fc receptor with at least about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 85%, about 90%, about 95%, or about 100% loss of specificity of the Fc domain to the Fc receptor as compared to an Fc domain of a conjugate not attached to an immune- stimulatory compound (in the absence of the immune- stimulatory compound). In some embodiments, the Fc domain is an Fc null.
[0333] The affinity of the antigen-binding domain of a conjugate to an antigen can be influenced by the attachment of an immune- stimulatory compound attached to the antibody construct. In some embodiments, the affinity of the antigen-binding domain of the conjugate for binding to an antigen is at least about 1%, about 5%, about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 85%, about 90%, about 95%, or about 100% of the affinity of the antigen-binding domain to the antigen in the absence of the immune- stimulatory compound (not attached to the antibody construct).
[0334] The affinity of the Fc domain to an Fc receptor of a conjugate can be influenced by attachment of an immune- stimulatory compound to the antibody construct (either directly or indirectly). In some embodiments, the Fc domain of the conjugate can bind to an Fc receptor with at least about 1%, about 5%, about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 85%, about 90%, about 95%, or about 100% of the affinity of the Fc domain to the Fc receptor in the absence of the immune- stimulatory compound attached to the antibody construct. In some embodiments, the Fc domain of the conjugate can bind to an Fc receptor with a reduced affinity of at least about 1%, about 5%, about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 85%, about 90%, about 95%, or about 100% of the affinity of the Fc domain to the Fc receptor in the absence of attachment the immune- stimulatory compound (directly or via a linker). In some embodiments, the Fc domain is an Fc null.
[0335] The Kd for binding of an antigen-binding domain to an antigen when an immune- stimulatory compound is attached to the antibody construct can be about 2 times, about 3 times, about 4 times, about 5 times, about 6 times, about 7 times, about 8 times, about 9 times, about 10 times, about 15 times, about 20 times, about 25 times, about 30 times, about 35 times, about 40 times, about 45 times, about 50 times, about 60 times, about 70 times, about 80 times, about 90 times, about 100 times, about 110 times, or about 120 times greater than the Kd for binding of the antigen binding domain to the antigen in the absence of the immune- stimulatory compound attached to the antibody construct.
[0336] In some embodiments, the Kd for binding of an Fc domain to an Fc receptor when an immune- stimulatory compound is attached to an antibody construct can be about 2 times, about 3 times, about 4 times, about 5 times, about 6 times, about 7 times, about 8 times, about 9 times, about 10 times, about 15 times, about 20 times, about 25 times, about 30 times, about 35 times, about 40 times, about 45 times, about 50 times, about 60 times, about 70 times, about 80 times, about 90 times, about 100 times, about 110 times, or about 120 times greater than the Kd for binding of the Fc domain to the Fc receptor without the immune- stimulatory compound attached to the antibody construct.
[0337] In some embodiments, the Kd for binding of an Fc domain to a Fc receptor when the immune- stimulatory compound is attached to the antibody construct can be about 2 times, about 3 times, about 4 times, about 5 times, about 6 times, about 7 times, about 8 times, about 9 times, about 10 times, about 15 times, about 20 times, about 25 times, about 30 times, about 35 times, about 40 times, about 45 times, about 50 times, about 60 times, about 70 times, about 80 times, about 90 times, about 100 times, about 110 times, or about 120 times less than the Kd for binding of the Fc domain to the Fc receptor when the immune- stimulatory compound is not attached to the antibody construct.
[0338] Affinity is the strength of the sum total of noncovalent interactions between a single binding site of a molecule, for example, an antibody, and the binding partner of the molecule, for example, an antigen. The affinity can also measure the strength of an interaction between an Fc domain of an antibody and an Fc receptor. Unless indicated otherwise, as used herein, "binding affinity" refers to intrinsic binding affinity which reflects a 1: 1 interaction between members of a binding pair (e.g., antibody and antigen or Fc domain and Fc receptor). The affinity of a molecule
X for its partner Y can generally be represented by the dissociation constant (Kd). Affinity can be measured by common methods known in the art. Specific illustrative and exemplary
embodiments for measuring binding affinity are described in the following. [0339] In some embodiments, an antibody construct (e.g., an antibody or antigen-binding fragment thereof) can have a dissociation constant (Kd) for an antigen or Fc receptor of about 1 μΜ, about 100 nM, about 10 nM, about 5 nM, about 2 nM, about 1 nM, about 0.5 nM, about 0.1 nM, about 0.05 nM, about 0.01 nM, or about 0.001 nM or less (e.g., 10"6 or less, 10"8 M or less, from 10"8 M to 10"13 M, or from 10"9 M to 10"13 M). An affinity matured antibody can be an antibody with one or more alterations in one or more complementarity determining regions (CDRs), compared to a parent antibody, which may not possess such alterations, such alterations resulting in an improvement in the affinity of the antibody for antigen. These antibodies can bind to their antigen with a Kd of about 5xl0"9 M, about 2xl0"9 M, about lxlO"9 M, about 5xl0"10 M, about lxlO"10 M, about 5xl0"u M, about lxlO"11 M, about 5xl0"12 M, about lxlO"12 M, or less. In some embodiments, the antibody construct (e.g., affinity matured antibody) can have an increased affinity of at least 1.5-fold, 2-fold, 2.5-fold, 3-fold, 4-fold, 5-fold, 10-fold, 20-fold, or greater as compared to an antibody construct without alterations in one or more CDRs.
[0340] Kd can be measured by any suitable assay. For example, Kd can be measured by a radiolabeled antigen binding assay (RIA). For example, Kd can be measured using surface plasmon resonance assays (e.g., using a BIACORE®-2000 or a BIACORE®-3000).
[0341] The molar ratio or drug-antibody ratio of a conjugate refers to the number of immune- stimulatory compounds attached to an antibody construct in a conjugate or preparation of immune- stimulatory conjugates. The molar ratio can refer to the number of immune- stimulatory compounds attached (e.g., conjugated) to an antibody construct of a particular conjugate and is an integer, such as from 0-8 or 0 to 20. The molar ratio can also refer to the average number of immune- stimulatory compounds attached to antibody constructs in a mixture of conjugates, such as in a pharmaceutical composition.
[0342] The molar ratio can be determined, for example, by Liquid Chromatography/Mas s Spectrometry (LC/MS), in which the number of immune- stimulatory compounds attached to the antibody construct can be directly determined. Additionally, as non- limiting examples, the molar ratio can be determined based on hydrophobic interaction chromatography (HIC) peak area, by liquid chromatography coupled to electrospray ionization mass spectrometry (LC-ESI-MS), by UV/Vis spectroscopy, by reversed-phase-HPLC (RP-HPLC), or by matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS).
[0343] In some embodiments, the molar ratio of immune- stimulatory compound to antibody construct can be less than 8 or less than 20. In other embodiments, the molar ratio of immune- stimulatory compound to an antibody construct can be 8, 7, 6, 5, 4, 3, 2, or 1. In some embodiments, the average molar ratio of immune- stimulatory compounds to antibody constructs in a composition can be less than 8, such as about 3 to 5 or about 2. In other embodiments, the average molar ratio of immune- stimulatory compounds to antibody constructs in a composition can be 8, 7, 6, 5, 4, 3, 2, or 1 or fractions hereof, or such as about 3.5 or about 1.8.
[0344] In a conjugate, an antibody construct (e.g., an antibody) can be linked to an immune- stimulatory compound in such a way that the antibody construct can still bind to an antigen and the Fc domain of the antibody construct can still bind to an FcR. In a conjugate, an antibody construct can be linked to an immune- stimulatory compound in such a way that the linking does not interfere with the ability of the antigen binding domain of the antibody construct to bind to its antigen, the ability of the Fc domain of the antibody construct to bind to an FcR, or FcR- mediated signaling resulting from the Fc domain of the antibody construct binding to an FcR. In a conjugate, an immune- stimulatory compound can be linked to an antibody construct in such a way that the linking does not interfere with the ability of the immune- stimulatory compound to bind to its receptor or otherwise can induce a biological effect. In some embodiments, a conjugate can produce stronger immune stimulation and a greater therapeutic window than components of the conjugate alone. For example, in an anti-tumor or anti-CD40 antibody linked to a TLR agonist, the combination of CD40 agonism, TLR agonism and an accessible Fc domain of the anti-CD40 antibody resulting in FcR-mediated signaling can produce stronger immune stimulation and a greater therapeutic window than the CD40 agonism, TLR agonism, or the FcR- mediated signaling alone.
[0345] In some embodiments, a conjugate can comprise a first binding domain, wherein the first binding domain contributes to immune- stimulatory activity; a first and second binding domain, wherein the second binding domain contributes to immune stimulatory activity; or a first binding domain and second binding domain, wherein the first binding domain and the second binding domain contribute to immune- stimulatory activity. The first binding domain and the second binding domain can contribute to the same immune- stimulatory activity. The first binding domain and the second binding domain can contribute to different immune- stimulatory activities.
[0346] In some embodiments, a conjugate in which the first binding domain contributes to immune- stimulatory activity can increase the immune- stimulation of the conjugate.
Immune-Stimulatory Compound Potency and Binding Activity
[0347] In certain embodiments, an immune- stimulatory compound has similar activity when bound to the antibody construct as when not bound to the antibody construct. In certain embodiments, the immune- stimulatory compound maintains the same level of potency and/or binding affinity when bound to an antibody construct as compared to the unbound immune- stimulatory compound. [0348] For many known antibody-drug conjugates, the payload/drug of the conjugate is a cytotoxic agent that acts on an intracellular target. The antibody generally targets a certain tumor marker on the surface of a cancer cell and upon binding of the antibody to the tumor cell, the cancer cell then internalizes the antibody-cytotoxic agent conjugate. The cytotoxic agent is released within the cell through enzymatic cleavage or other cleavage of the agent's linkage to the antibody or through enzymatic degradation of the antibody. The released cytotoxic agent or degredation product acts on the intracellular target of the cytotoxic agent to kill the cancer cell. Important to the mechanism of action of many antibody-cytotoxic agent conjugates is: (1) that the cytotoxic agent is bound to the antibody with a linker, wherein the linker is not cleaved until exposed to enzymes or conditions inside a target cell, e.g., a cancer cell; (2) that the cytotoxic agent is released from the antibody inside of the cell to perform its cytotoxic function; and (3) that the cytotoxic agent is not active or minimally active (i.e., in a prodrug form) when bound to the antibody such that the cytotoxic agent does not indiscriminately kill cells and harm organ systems distributed thorught the body on the path to the cancer cell.
[0349] In certain aspects, the immune- stimulatory conjugates of the disclosure operate under a different paradigm from such antibody-cytotoxic agent conjugates. The immune- stimulatory conjugates of the disclosure can be designed in a way that the payload immune- stimulatory compound has the same potency, similar potency, or increased potency when bound to the antibody construct as compared to the unbound immune- stimulatory compound and in contrast to antibody-cytotoxic agent conjugates. The immune- stimulatory conjugates of the disclosure may perform, and in some cases preferably perform, the intended function of the compound, i.e., stimulate or modulate the function of immune cells or other target cells, without prior release of the compound from the conjugate (i.e., while attached to the antibody construct). The features of the immune- stimulatory compound-conjugates and assays that enable these functions and others are described further herein.
[0350] The potency of the immune- stimulatory compound of the conjugate may not be significantly reduced relative to the potency of the un-attached (free) immune- stimulatory compound. In particular, the potency of the immune- stimulatory compound of the conjugate (i.e., as part of the conjugate or covalently bound to the antibody construct) is preferably no greater than 500-fold less, no greater than 400-fold less, no greater than 300-fold less, no greater than 200-fold less, no greater than 100-fold less, no greater than 50-fold less, or no greater than 10- fold less than the potency of a control compound, wherein the control compound is the unbound immune- stimulatory compound. For example, for an immune- stimulatory conjugate represented by the structure: Ab - L - C, wherein A is an antibody construct, L is a linker, and C is an immune- stimulatory compound, the immune- stimulatory activity of C of the conjugate A - L - C is preferably no greater than 500-fold less, no greater than 400-fold less, no greater than 300-fold less, no greater than 200-fold less, no greater than 100-fold less, no greater than 50-fold less, or no greater than 10-fold less than the potency of the immune- stimulatory compound, C, unbound from the immune- stimulatory conjugate, A - L - C, in the absence of processing of the immune- stimulatory conjugate in a cell.
[0351] In particular embodiments, the potency of an immune- stimulatory compound of the conjugate (i.e., covalently bound to the antibody construct) is near or equivalent to the potency of the unbound immune- stimulatory compound, such as within about 10-fold, within about 8-fold, within about 5-fold, or within about 2-fold of the potency of the unbound immune- stimulatory compound. The potency of the immune- stimulatory compound of the conjugate may be greater than the potency of the unbound immune- stimulatory compound, such as about 2-fold or greater, 5-fold or greater, 10-fold or greater, 100-fold or greater, 200-fold or greater, 300-fold or greater, 400-fold or greater, or even 500-fold or greater than the potency of the unbound immune- stimulatory compound. In certain embodiments, the tolerability of an immune- stimulatory compound when part of a construct is greater than the tolerability of the unbound immune- stimulatory compound in a subject.
[0352] In certain embodiments, the potency of the immune- stimulatory compound when bound to a 5-500 atom linker is the same, similar, or increased as compared to the potency of the immune- stimulatory compound not bound to the 5-500 atom linker. In certain embodiments, the tolerability of the immune- stimulatory compound in a subject when bound to a 5-500 atom linker is the same, similar, or increased as compared to the tolerability of the immune- stimulatory compound not bound to the 5-500 atom linker.
[0353] The "5-500 atom linker" referred to herein has 5 to 500 consecutive atoms from end to end. When attached to an immune- stimulatory compound and to an antibody construct, a 5-500 atom linker has 5-500 consecutive atoms between the point of attachment to the immune- stimulatory compound and the point of attachment to the antibody construct. A 5-500 atom linker can have, for example, from about 50 to about 500 atoms, such as about 50 to about 300 atoms or such as about 50 to about 200 atoms. A 5-500 atom linker can have, for example, from about 25 to about 500 atoms, such as about 25 to about 300 atoms or such as about 25 to about 200 atoms. In certain embodiments, the linker includes one or more peptide bonds. In certain embodiments, the linker includes one or more ethylene glycol groups. In certain embodiments, the linker includes a peptide backbone and one or more side chains. In certain embodiments, the linker is not cleaved from the immune- stimulatory compound in the assay evaluating potency.
[0354] Exemplary 5-500 atom linkers include Fleximer linkers, linkers with one or more carbamate or amide linkages and linkers represented by the formula:
antibody construct, R is hydrogen, Ci-ioalkyl, sulfonate and methyl sulfonate and the wavy line indicates an attachment to the rest of the linker or to the immune- stimulatory compound.
[0355] In some embodiments, the linker is a non-cleavable linker. Examples of non-cleavable linkers include the following:
wherein Rx is a reactive moiety for attachment to the antibody construct, R is hydrogen, Q. loalkyl, sulfonate and methyl sulfonate and the wavy line indicates an attachment to the rest of the linker or to the immune- stimulatory compound.
[0356] Further examples of non-cleavable linkers include:
wherein the wavy line indicates an attachment to the rest of the linker or to the immune- stimulatory compound.
[0357] In some embodiments, the potency of an immune- stimulatory compound when bound to a 5-500 atom linker can be no greater than 500-fold less, no greater than 400-fold less, no greater than 300-fold less, no greater than 200-fold less, no greater than 100-fold less, no greater than 50-fold less, or no greater than 10-fold less than the potency of a control compound, wherein the control compound is the unbound immune- stimulatory compound. For example, for compounds suitable for use in the conjugates of the disclosure, the immune- stimulatory activity of the immune- stimulatory compound bound to a 5-500 atom linker, represented by the formula: C - L* , wherein L* is a 5-500 atom linker, is preferably no greater than 500-fold less, no greater than 400-fold less, no greater than 300-fold less, no greater than 200-fold less, no greater than 100- fold less, no greater than 50-fold less, or no greater than 10-fold less than the potency of the immune- stimulatory compound, C, unbound from linker L*.
[0358] In particular embodiments, the potency of the immune- stimulatory compound bound to a 5-500 atom linker is equivalent to the potency of the unbound immune- stimulatory compound, such as within about 10-fold, within about 8-fold, within about 5-fold, or within about 2-fold of the potency of the unbound immune- stimulatory compound. One exemplary embodiment is depicted in FIGURE 16, wherein the designed inhibitor bound to a linker surrogate, L* as discussed herein, displays a similar EC50 value relative to the compound unbound from the linker. In certain embodiments, the potency of the immune- stimulatory compound bound to a 5- 500 atom linker may be greater than the potency of the unbound immune- stimulatory compound, such as about 2-fold or greater, 5-fold or greater, 10-fold or greater, 100-fold or greater, 200-fold or greater, 300-fold or greater, 400-fold or greater than 500-fold than the potency of the unbound immune- stimulatory compound. In certain embodiments, the tolerability of the immune- stimulatory compound bound to an antibody construct by a 5-500 atom linker may be greater than the potency of the unbound immune- stimulatory compound, such as about 2-fold or greater, 5-fold or greater, 10-fold or greater, 100-fold or greater, 200-fold or greater, 300-fold or greater, 400-fold or greater than 500-fold than the potency of the unbound immune- stimulatory compound.
[0359] "Unbound immune- stimulatory compound" or reference to an immune- stimulatory compound without specifying its connection to a linker or antibody, as used herein, refers to a compound with immune- stimulatory activity that is not bound to a linker (such as the linkers described herein) and not part of an immune- stimulatory conjugate, as described herein. In certain embodiments, the "control compound" as compared to a conjugate or linker-bound immune- stimulatory compound is the unbound immune- stimulatory compound. Unbound immune- stimulatory compounds are generally not suitable for direct attachment to an antibody construct (such as an antibody) without a linker, such as those described herein. An unbound immune- stimulatory compound may be a synthetic precursor to an immune- stimulatory conjugate, wherein the immune- stimulatory compound may be modified by attaching a linker such as the linkers described herein and optionally further bound to an antibody to form an immune- stimulatory conjugate.
[0360] In certain aspects, the potency of the immune- stimulatory compound of the conjugate, the potency of the immune- stimulatory compound bound to a 5-500 atom linker, and the potency of the unbound immune- stimulatory compound may be determined using a PBMC assay as described in Example 7.
[0361] The immune- stimulatory conjugates of the disclosure can be engineered to maintain, not significantly reduce, or increase the potency of the immune- stimulatory compound in the conjugate relative to the unbound immune- stimulatory compound. In certain embodiments, a highly potent immune- stimulatory compound, e.g., the unbound immune- stimulatory agent has an EC50 of less than 500 nM, such as less than 400 nM, such as less than 300 nM, such as less than 200 nM, such as less than 100 nM, such as less than 50 nM, or such as less than 10 nM, is used as an immune- stimulatory compound of the conjugates herein.
[0362] In designing an immune- stimulatory conjugate of the disclosure, an unbound immune- stimulatory compound can be bound to a linker at a position on the immune- stimulatory compound that does not interfere with the ability of the immune- stimulatory compound to interact with its protein binding site. For example, a linker can be attached to an immune- stimulatory compound at a solvent accessible site on the compound when the compound would be bound to its protein target. For example, as depicted in FIGURE 20A, an antibody 3405 bound to a linker 3410 is bound to an immune- stimulatory compound 3415. The linker 3410 is bound to a position of the immune- stimulatory compound 3415 such that the linker 3410 and/or antibody 3405 does not interfere with the interaction between the immune- stimulatory compound 3415 and the active site 3425 of a target protein 3420 (FIGURE 20B). The linker 3410 may extend out from the active site 3425 of the target protein 3420 at the solvent/active site interface 3430.
[0363] The potency of the unbound immune- stimulatory compound may also be evaluated relative to the potency of the immune- stimulatory compound bound to a 5-500 atom linker as a surrogate for the immune- stimulatory conjugate. Evaluating the potency of the immune- stimulatory compound bound to a 5-500 atom linker, as described herein, can be predictive of the activity of the immune- stimulatory compound in the conjugate. For example, as depicted in
FIGURE 20C, a linker 3410 is bound to an immune- stimulatory compound 3415. The linker 3410 is bound to a portion of the immune- stimulatory compound 3415 such that the linker 3410 does not interfere with the interaction between the immune- stimulatory compound 3415 and the active site 3425 of a target protein 3420. FIGURE 20D depicts the unbound immune- stimulatory compound 3415 in the active site 3425 of a target protein 3420.
[0364] In certain embodiments, the binding position of the linker 3410 on the immune- stimulatory compound 3415 may be determined using target protein crystal structures and active site protein modeling. As depicted in FIGURE 15B and FIGURE 15C an immune- stimulatory compound, such as the left-hand compound of FIGURE 15A, may be modeled in an active site of a target protein and the positioning of the linker attachment to the immune- stimulatory compound may be selected based on this modeling. For example, the modeling may indicate that positioning of a linker on the 4-position of the benzimidazole of the left-hand compound of
FIGURE 15A would extend from the active site and not interfere with the active site interactions of the compound as depicted in FIGURE 15B and FIGURE 15C.
[0365] The linker can be covalently bound to the immune- stimulatory compound at a position on the immune- stimulatory compound that is at or near the solvent interface of the protein active site, as determined by modeling of the immune- stimulatory compound in the protein active site. In certain embodiments, the linker is covalently bound to the immune- stimulatory compound at a position on the immune- stimulatory compound such that when the immune- stimulatory compound is positioned in the protein active site, the linker extends out from the protein active site into the solvent, as determined by modeling of the immune- stimulatory compound in the protein active site. FIGURE 17B and FIGURE 17C depict binding of the left-hand compound of FIGURE 17A. A linker may be positioned on the phenyl ring to extend out of the protein active site, e.g., see left portion of right-hand compound of FIGURE 17A. FIGURE 18B and FIGURE 18C depict binding of the left-hand compound of FIGURE 18A. The trifluoromethyl substituent on the phenyl group may be replaced with a linker at this position to extend out of the protein active site, e.g., see left portion of right-hand compound of FIGURE 18A. FIGURE 19B and FIGURE 19C depict binding of the left-hand compound of FIGURE 19A with the linker (left side of right hand molecule) positioned to extend out of the protein active site.
[0366] In certain embodiments, the immune- stimulatory compounds have similar binding affinity to a protein active site when bound to the antibody construct as when not bound to the antibody construct. In certain embodiments, the immune- stimulatory compounds maintain the binding affinity to a protein active site when bound to an antibody construct as compared to the binding affinity of the unbound immune- stimulatory compound.
[0367] In certain embodiments, the Kd for binding of the immune- stimulatory compound of the conjugate to a protein active site is no greater than 50 times the Kd for binding of a control compound to the protein active site, wherein the control compound is the unbound immune- stimulatory compound. (As used herein, the phrase "of the conjugate" refers to the immune- stimulatory compounds while attached to the antibody construct, usually via a linker(s).) For example, the Kd for binding of the immune- stimulatory compound of the conjugate to the protein active site may be no greater than 40 times, no greater than 30 times, no greater than 20 times, or no greater than 10 times the Kd for binding of the unbound immune- stimulatory compound to the protein active site. In certain embodiments, the Kd for binding of the immune- stimulatory compound of the conjugate to the protein active site is no greater than 10 times the Kd for binding of a control compound to the protein active site, wherein the control compound is the unbound immune- stimulatory compound. The Kd for binding of the immune- stimulatory compound of the conjugate to the protein active site may be no greater than 9 times, no greater than 8 times, no greater than 7 times, no greater than 6 times, no greater than 5 times, no greater than 4 times, no greater than 3 times, or no greater than 2 times the Kd for binding of the unbound immune- stimulatory compound to the protein active site.
[0368] In certain embodiments, the Kd for binding of the immune- stimulatory compound of the conjugate to the protein active site is close to or equivalent to the Kd for binding of a control compound, such as within about 10 times, within about 8 times, within about 6 times, within about 5 times, within about 4 times, within about 3 times or within about 2 times the binding affinity of the unbound immune- stimulatory compound. In certain embodiments, the Kd for binding of the immune- stimulatory compound of the conjugate to the protein active site is less than the Kd for binding of the unbound immune- stimulatory compound.
[0369] In certain embodiments, the binding affinity to a protein active site of an immune- stimulatory compound bound to a 5-500 atom linker, as described herein, is the same, similar, or increased as compared to the binding affinity of the immune- stimulatory compound not bound to the 5-500 atom linker. In particular embodiments, the binding affinity of the immune- stimulatory compound when bound to a 5-500 atom linker is no greater than 40 times, no greater than 30 times, no greater than 20 times, or no greater than 10 times the Kd for binding of the unbound immune- stimulatory compound to the protein active site.
[0370] In certain embodiments, the binding affinity to a protein active site of the immune- stimulatory compound bound to a 5-500 atom linker is no greater than 10 times the Kd for binding of a control compound to the protein active site, wherein the control compound is the unbound immune- stimulatory compound. The binding affinity to a protein active site of the immune- stimulatory compound bound to a 5-500 atom linker may be no greater than 9 times, no greater than 8 times, no greater than 7 times, no greater than 6 times, no greater than 5 times, no greater than 4 times, no greater than 3 times, or no greater than 2 times the Kd for binding of the unbound immune- stimulatory compound to the protein active site.
[0371] In certain embodiments, the binding affinity to a protein active site of the immune- stimulatory compound bound to a 5-500 atom linker is close to or equivalent to the Kd for binding of a control compound, such as within about 10 times, within about 8 times, within about 6 times, within about 5 times, within about 4 times, within about 3 times or within about 2 times the binding affinity of the unbound immune- stimulatory compound. In certain embodiments, the binding affinity to a protein active site of the immune- stimulatory compound bound to a 5-500 atom linker is less than the Kd for binding of the unbound immune- stimulatory compound.
[0372] In certain aspects, the binding affinity of the immune- stimulatory compound of the conjugate, the binding affinity of the immune- stimulatory compound bound to a 5-500 atom linker, and the binding affinity of the unbound immune- stimulatory compound may be determined using an assay such as a Bio-layer Interferometry (BLI) as described in Example 8.
[0373] In certain embodiments, the immune- stimulatory compound binds to a cell membrane or ER protein target. In certain embodiments, the immune- stimulatory compound binds to a cell membrane or ER protein target and is other than a TLR agonist. In some embodiments, the immune- stimulatory compound is a kinase inhibitor and the protein active site is a kinase active site. Kinase inhibitors are a type of enzyme inhibitor that block the action of at least one protein kinase, which are enzymes that can add at least one phosphate group to a protein and thereby modulate its function or activity. In certain embodiments, the immune- stimulatory compound is a kinase inhibitor. In some embodiments, the kinase inhibitor targets a protein kinase that acts on both serine and threonine residues. In certain embodiments, the kinase inhibitor targets protein kinases that act on tyrosine residues. In certain embodiments, the kinase inhibitor targets protein kinases that act on serine, threonine and tyrosine residues. In other embodiments, the kinase inhibitor targets protein kinases that act on amino acids other than serine, threonine and tyrosine, such as histine kinases. The kinase inhibitors may target ALK, Bcr-Abl, BRAF, BTK, c-KIT, EGFR, ErbB2, JAK, MEK, MET, PDGFRB, RET, ROS 1, Syk, SRC, TGFpR2, TNIK, TNKS, TNKS2, TrkA, TrkB, TrkC, VEGFR1, VEGFR2, VEGFR3, PI3K, AKT, mTOR, and
combinations thereof.
[0374] In various embodiments, when the immune- stimulatory compound is a kinase inhibitor, the linker is covalently bound to the kinase inhibitor at a position on the kinase inhibitor that is at or near the solvent interface of the kinase active site when the inhibitor is bound to the protein target, as determined by modeling of the kinase inhibitor in the kinase active site. In certain embodiments, the linker is covalently bound to the kinase inhibitor at a position on the kinase inhibitor such that when the kinase inhibitor is bound to the active site, the linker extends out from the kinase active site into the solvent, as determined by modeling of the kinase inhibitor in the kinase active site.
[0375] In some aspects, the present disclosure provides an immune- stimulatory conjugate comprising: (a) an immune- stimulatory compound that binds to a protein active site and stimulates an immune response; (b) an antibody construct comprising an antigen binding domain and an Fc domain, wherein the antigen binding domain binds to a first antigen and wherein the Fc domain binds to an Fc receptor; and (c) a linker, wherein the linker is covalently bound to the antibody construct and the linker is covalently bound to the immune- stimulatory compound; wherein the Kd for binding of the immune- stimulatory compound of the conjugate (attached to the antibody construct via the linker) to the protein active site is no greater than 100 times the Kd for binding of a control compound to the protein active site or wherein the EC50 or IC50 of the immune- stimulatory compound of the conjugate is no greater than 300-fold higher the EC50 or IC50 of a control compound, wherein the control compound is the unbound immune- stimulatory compound and optionally wherein the Kd for binding of the Fc domain of the conjugate to an Fc receptor is no greater than about 100 times the Kd for binding of a control antibody to the Fc receptor, wherein the control antibody is the antibody construct itself.
[0376] In various embodiments, the Kd for binding of the immune- stimulatory compound of the conjugate (attached to the antibody construct via the linker) to the protein active site is no greater than 50 times the Kd for binding of a control compound to the protein active site, wherein the control compound is the unbound immune- stimulatory compound. In certain embodiments, the Kd for binding of the immune- stimulatory compound of the conjugate to the protein active site is no greater than 10 times the Kd for binding of a control compound to the protein active site, wherein the control compound is the unbound immune- stimulatory compound. In an exemplary embodiment, the Kd for binding of the immune- stimulatory compound of the conjugate to the protein active site is equivalent to or less than the Kd for binding of a control compound to the protein active site, wherein the control compound is the unbound immune- stimulatory compound.
[0377] In some embodiments, the EC50 or IC50 of the immune- stimulatory compound of the conjugate is no greater than 100-fold higher than the EC50 or IC50 of a control compound, wherein the control compound is the unbound immune- stimulatory compound. In certain embodiments, the EC50 or IC50 of the immune- stimulatory compound of the conjugate is no greater than 10-fold higher than the EC50 or IC50 of a control compound, wherein the control compound is the unbound immune- stimulatory compound. In an exemplary embodiment, the potency of the immune- stimulatory compound of the conjugate is equivalent to or greater than the potency of a control compound, wherein the control compound is the unbound immune- stimulatory compound.
[0378] In some embodiments, the EC50 or IC50 of the immune- stimulatory compound-linker construct is no greater than 100-fold higher than the EC50 or IC50 of a control compound, wherein the control compound is the unbound immune- stimulatory compound. In certain embodiments, the EC50 or IC50 of the immune- stimulatory compound-linker construct is no greater than 10-fold higher than the EC50 or IC50 of a control compound, wherein the control compound is the unbound immune- stimulatory compound. In an exemplary embodiment, the potency of the immune- stimulatory compound-linker construct is equivalent to or greater than the potency of a control compound, wherein the control compound is the unbound immune- stimulatory compound.
[0379] In some aspects, the present disclosure provides an immune- stimulatory conjugate comprising: (a) an immune- stimulatory compound that binds to a protein active site and stimulates an immune response through target inhibition (e.g., inhibition of a protein); (b) an antibody construct comprising an antigen binding domain and an Fc domain, wherein the antigen binding domain binds to a first antigen and wherein the Fc domain binds to an Fc receptor; and (c) a linker, wherein the linker is covalently bound to the antibody construct and the linker is covalently bound to the immune- stimulatory compound; wherein the Kd for binding of the immune- stimulatory compound of the conjugate to the protein active site is no greater than 100 times the Kd for binding of a control compound to the protein active site or wherein the IC50 of the immune- stimulatory compound of the conjugate is no greater than 300-fold the IC50 of a control compound, wherein the control compound is the unbound immune- stimulatory compound, and optionally wherein the Kd for binding of the Fc domain of the conjugate to an Fc receptor is no greater than about 100 times the Kd for binding of a control antibody to the Fc receptor, wherein the control antibody is the antibody construct itself.
[0380] A first antigen, second antigen, or first and second antigens of the antibody construct binding domains are chosen to deliver the immune- stimulatory compound into tumor cells, immune cells, or both, but not into non-antigen bearing cell types, thereby selectively increasing immune activation and lowering systemic toxicity.
[0381] In some embodiments, the immune- stimulatory conjugate targets the activity of the immune- stimulatory compound to within an immune cell, such as an immune cell like an APC, to within tumor cells, or within both.
[0382] In some embodiments, the immune stimulatory conjugate targets the activity of the immune- stimulatory compound to cells within a tissue or within the tumor microenvironment.
[0383] In some embodiments, the immune- stimulatory conjugate systemically increases antitumor immunity with lower toxicity than an effective systemic dose of the immune- stimulatory compound itself.
[0384] In some aspects, the present disclosure provides an immune- stimulatory conjugate comprising: (a) an immune- stimulatory compound that binds to a protein active site and stimulates or otherwise modulates an immune response; (b) an antibody construct comprising an antigen binding domain and an Fc domain, wherein the antigen binding domain binds to a first antigen and wherein the Fc domain binds to an Fc receptor; and (c) a linker, wherein the linker is covalently bound to the antibody construct and the linker is covalently bound to the immune- stimulatory compound; wherein the Kd for binding of the immune- stimulatory compound, when bound to a 5-100 atom linker, to the protein active site is no greater than 100 times the Kd for binding of a control compound to the protein active site or wherein the EC50 or IC50 of the immune- stimulatory compound when bound to a 5-100 atom linker is no greater than 300-fold EC50 or IC50 of a control compound, wherein the control compound is the immune- stimulatory compound, and optionally wherein the Kd for binding of the Fc domain of the conjugate to an Fc receptor is no greater than about 100 times the Kd for binding of a control antibody to the Fc receptor, wherein the control antibody is the antibody construct.
[0385] In various embodiments, the potency of the immune- stimulatory compound when bound to a 5-100 atom linker is no greater than 100-fold less than the potency of a control compound, wherein the control compound is the unbound immune- stimulatory compound. In certain embodiments, the potency of the immune- stimulatory compound when bound to a 5-100 atom linker is no greater than 10-fold less than the potency of a control compound, wherein the control compound is the immune- stimulatory compound. In an exemplary embodiment, the potency of the immune- stimulatory compound when bound to a 5-100 atom linker is equivalent to or greater than the potency of a control compound, wherein the control compound is the unbound immune- stimulatory compound.
[0386] In some embodiments, the Kd for binding of the immune- stimulatory compound when bound to a 5-100 atom linker to the protein active site is no greater than 50 times the Kd for binding of a control compound to the protein active site, wherein the control compound is the unbound immune- stimulatory compound. In certain embodiments, the Kd for binding of the of the immune- stimulatory compound when bound to a 5-100 atom linker to the protein active site is no greater than 10 times the Kd for binding of a control compound to the protein active site. In an exemplary embodiment, the Kd for binding of the immune- stimulatory compound when bound to a 5-100 atom linker to the protein active site is equivalent to or less than the Kd for binding of a control compound to the protein active site.
Linkers
[0387] The immune- stimulatory compounds and salts thereof described herein may be attached, i.e., covalently attached, to a linker, e.g., a cleavable linker or a non-cleavable linker, and to an antibody construct and referred to as an immune- stimulatory compound conjugate, an immune- stimulatory conjugate or a conjugate. Linkers of the conjugates described herein may not affect the binding of active portions of a conjugate, e.g., the first binding domains, Fc domains, second antigen binding domains, antibodies, immune- stimulatory compounds or the like, to a target molecule, which can be a cognate binding partner such as an antigen. A conjugate can comprise multiple linkers. These linkers can be the same linkers or different linkers. A linker described herein can be a multi-functional linker linking two small molecule binding moieties and linking the linked small molecules to an antibody. A linker also may be referred to as a linker of a linker moiety.
[0388] A linker can be short, flexible, rigid, cleavable, non-cleavable, hydrophilic, or hydrophobic. A linker can contain segments that have different characteristics, such as segments of flexibility or segments of rigidity. The linker can be chemically stable to extracellular environments, for example, chemically stable in the blood stream, or may include linkages that are not stable. The linker can include linkages that are designed to cleave and/or immolate or otherwise breakdown specifically or non- specifically inside cells. A cleavable linker can be sensitive to enzymes. A cleavable linker can be cleaved by enzymes such as proteases. A cleavable linker can have a valine-citrulline or a valine- alanine dipeptide. A valine-citrulline- or valine-alanine-containing linker also can contain a pentafluorophenyl group. A valine-citrulline- or valine-alanine-containing linker also can contain a succinimide group or a maleimide group. A valine-citrulline- or valine-alanine-containing linker can also contain a para aminobenzoic acid (PABA) group. A valine-citrulline or valine- alanine (VA)-containing linker or a glycine-glycine- phenylalanine-glycine (GGFG) tetrapeptide-containing linker also can contain a PABA group and a pentafluorophenyl group. A peptide-based valine-citrulline or valine- alanine linker can contain a PABA group and a succinimide group. [0389] A non-cleavable linker is protease insensitive and insensitive to cleavage by other intracellular processes. A non-cleavable linker can be maleimidocaproyl linker. A
maleimidocaproyl linker can comprise N-maleimidomethylcyclohexane-l-carboxylate. A maleimidocaproyl linker can contain a succinimide group. A linker can be a combination of a maleimidocaproyl group and one or more polyethylene glycol molecules. A linker can be a maleimide-PEGx linker, where x = 2-12. A linker can be a combination of a maleimidocaproyl linker containing a succinimide group and one or more polyethylene glycol molecules. A linker can contain a maleimide(s) linked to polyethylene glycol molecules in which the polyethylene glycol can allow for more linker flexibility or can be used.
[0390] A linker can also include an alkylene, alkenylene, alkynylene, polyether, polyester, polyamide, polyamino acids, polypeptides, cleavable peptides, or amino benzylcarbamates. A linker can contain a maleimide at one end and an N-hydroxysuccinimidyl ester at the other end. A linker can contain a lysine with an N-terminal amine acetylated, and a valine-citrulline peptide cleavage site. A linker can be a link created by a microbial transglutaminase, wherein the link can be created between an amine-containing moiety and a moiety engineered to contain glutamine as a result of the enzyme catalyzing a bond formation between the acyl group of a glutamine side chain and the primary amine of a lysine chain. A linker can contain a reactive primary amine. A linker can be a Sortase A linker. A Sortase A linker can be created by a Sortase A enzyme fusing an LPXTG recognition motif (SEQ ID NO: 672) to an N-terminal GGG motif to regenerate a native amide bond. A linker created can therefore link a moiety attached to the LPXTG recognition motif (SEQ ID NO: 672) with a moiety attached to the N-terminal GGG motif.
[0391] In the conjugates described herein, an immune- stimulatory compound is linked to the antibody construct by way of a linker. The linker attaching the compound or a salt thereof to the antibody construct may be short, long, hydrophobic, hydrophilic, flexible or rigid, or may be composed of segments that each independently have one or more of the above-mentioned properties such that the linker may include segments having different properties. A linker may be polyvalent such that it covalently links more than one compound or salt to a single site on the antibody construct, or monovalent such that covalently it links a single compound or salt to a single site on the antibody construct.
[0392] As will be appreciated by skilled artisans, the linkers attach the immune- stimulatory compounds to the antibody construct by covalent linkages between the linker(s) and the antibody construct and compound. As used herein, the expression "linker" is intended to include (i) unconjugated forms of the linker that include a functional group capable of covalently linking the linker to an immune- stimulatory compound and a functional group capable of covalently linking the linker to an antibody; (ii) partially conjugated forms of the linker that include a functional group capable of covalently linking the linker to an antibody and that is covalently linked to an immune- stimulatory compound, or vice versa; and (iii) fully conjugated forms of the linker that is covalently linked to both an immune- stimulatory compound and an antibody.
[0393] Exemplary polyvalent linkers that may be used to link many immune- stimulatory compounds to an antibody construct are described. In some embodiments, the conjugate uses any linker as disclosed in U.S. Patent No. 9,254,339, U.S. Patent No. 9,144,615, U.S. Patent No. 8,821,850, U.S. Patent No. 8,808,679, U.S. Patent No. 8,685,383, U.S. Patent No. 8,524,214, or Published U.S. Publication No. 2011/0243892, in which each of these references are herein incorporated by reference in their entirety. For example, Fleximer® linker technology has the potential to enable high-DAR conjugates with good physicochemical properties. As shown below, the Fleximer® linker technology is based on incorporating drug molecules into a solubilizing poly-acetal backbone via a sequence of ester bonds. The methodology renders highly-loaded conjugates (DAR up to 20) whilst maintaining good physicochemical properties.
add Fleximer linker
[0394] To utilize the Fleximer® linker technology depicted in the scheme above, an aliphatic alcohol can be present or introduced into the immune- stimulatory compound. The alcohol moiety is then attached to an alanine moiety, which is then synthetically incorporated into the Fleximer® linker. Liposomal processing of the conjugate in vitro releases the parent alcohol-containing drug.
[0395] Cleavable linkers can be cleavable in vitro and in vivo. Cleavable linkers can include chemically or enzymatically unstable or degradable linkages. Cleavable linkers can rely on processes inside the cell to liberate an immune- stimulatory compound, such as reduction in the cytoplasm, exposure to acidic conditions in the lysosome, or cleavage by specific proteases or other enzymes within the cell. Cleavable linkers can incorporate one or more chemical bonds that are either chemically or enzymatically cleavable while the remainder of the linker can be non- cleavable. [0396] A linker can contain a chemically labile group such as a hydrazone and/or disulfide group. Linkers comprising chemically labile groups can exploit differential properties between the plasma and some cytoplasmic compartments. The intracellular conditions that can facilitate immune- stimulatory compound release for hydrazone-containing linkers can be the acidic environment of endosomes and lysosomes, while the disulfide containing linkers can be reduced in the cytosol, which can contain high thiol concentrations, e.g., glutathione. The plasma stability of a linker containing a chemically labile group can be increased by introducing steric hindrance using substituents near the chemically labile group.
[0397] Acid-labile groups, such as hydrazone, can remain intact during systemic circulation in the blood's neutral pH environment (pH 7.3-7.5) and can undergo hydrolysis and can release the immune- stimulatory compound once the immune- stimulatory conjugate is internalized into mildly acidic endosomal (pH 5.0-6.5) and lysosomal (pH 4.5-5.0) compartments of the cell. This pH dependent release mechanism can be associated with nonspecific release of the compound. To increase the stability of the hydrazone group of the linker, the linker can be varied by chemical modification, e.g., substitution, allowing tuning to achieve more efficient release in the lysosome with a minimized loss in circulation.
[0398] Hydrazone-containing linkers can contain additional cleavage sites, such as additional acid-labile cleavage sites and/or enzymatically labile cleavage sites. Immune- stimulatory conjugates can include exemplary hydrazone-containing linkers can include, for example, the following structures:
wherein D is an immune- stimulatory compound, Ab is an antibody construct, -NH- or -S-
(connected to Ab) are part of the antibody construct, and n represents the number of compounds bound to linkers bound to the antibody construct. In certain linkers, such as linker (la), the linker can comprise two cleavable groups- a disulfide and a hydrazone moiety. For such linkers, effective release of the unmodified free immune- stimulatory compound can require acidic pH or disulfide reduction and acidic pH. Linkers such as (lb) and (Ic) can be effective with a single hydrazone cleavage site.
[0399] Other acid-labile groups that can be included in linkers include czs-aconityl-containing linkers, cis- Aconityl chemistry can use a carboxylic acid juxtaposed to an amide bond to accelerate amide hydrolysis under acidic conditions.
[0400] Cleavable linkers can also include a disulfide group. Disulfides can be
thermodynamically stable at physiological pH and can be designed to release the immune- stimulatory compound upon internalization inside cells, wherein the cytosol can provide a significantly more reducing environment compared to the extracellular environment. Scission of disulfide bonds can require the presence of a cytoplasmic thiol cofactor, such as (reduced) glutathione (GSH), such that disulfide-containing linkers can be reasonably stable in circulation, selectively releasing the immune- stimulatory compound in the cytosol. The intracellular enzyme protein disulfide isomerase, or similar enzymes capable of cleaving disulfide bonds, can also contribute to the preferential cleavage of disulfide bonds inside cells. GSH can be present in cells in the concentration range of 0.5-10 mM compared with a significantly lower concentration of GSH or cysteine, the most abundant low-molecular weight thiol, in circulation at approximately 5 μΜ. Tumor cells, where irregular blood flow can lead to a hypoxic state, can result in enhanced activity of reductive enzymes and therefore even higher glutathione concentrations. The in vivo stability of a disulfide-containing linker can be enhanced by chemical modification of the linker, e.g., use of steric hindrance adjacent to the disulfide bond.
[0401] Immune- stimulatory conjugates can include exemplary disulfide-containing linkers having the following structures:
wherein D is an immune- stimulatory compound, Ab is an antibody construct, -NH- or -S- (connected to Ab) are part of the antibody construct, n represents the number of compounds bound to linkers bound to the antibody construct and R is independently selected at each occurrence from hydrogen or Ci-ioalkyl, for example. Increasing steric hindrance adjacent to the disulfide bond can increase the stability of the linker. Structures such as (Ila) and (lie) can show increased in vivo stability when one or more R groups is selected from a lower alkyl such as methyl.
[0402] Another type of linker that can be used is a linker that is specifically cleaved by an enzyme. For example, the linker can be cleaved by a lysosomal enzyme. Such linkers can be peptide-based or can include peptidic regions that can act as substrates for enzymes. Peptide based linkers can be more stable in plasma and extracellular milieu than chemically labile linkers.
[0403] Peptide bonds can have good serum stability, as lysosomal proteolytic enzymes can have very low activity in blood due to endogenous inhibitors and the unfavorably high pH value of blood compared to lysosomes. Release of an immune- stimulatory compound from an antibody construct can occur due to the action of lysosomal proteases, e.g., cathepsin and plasmin. These proteases can be present at elevated levels in certain tumor tissues. The linker can be cleavable by a lysosomal enzyme. The lysosomal enzyme can be, for example, cathepsin B, β- glucuronidase, or β-galactosidase.
[0404] The peptide can be selected from tetrapeptides such as Gly-Phe-Leu-Gly, Ala- Leu- Ala- Leu, Gly-Gly-Phe-Gly or dipeptides such as Val-Cit, Val-Ala, and Phe-Lys. Dipeptides can have lower hydrophobicity compared to longer peptides, depending on amino acid composition. A peptide can also include non-natural amino acids or a mix of natural and non-natural amino acids.
[0405] A variety of dipeptide-based cleavable linkers can be used in the immune- stimulatory conjugates described herein.
[0406] Enzymatically cleavable linkers can include a self-immolative spacer to spatially separate the immune- stimulatory compound from the site of enzymatic cleavage. The direct attachment of an immune- stimulatory compound to a peptide linker can result in proteolytic release of an amino acid adduct of the immune- stimulatory compound, thereby impairing its activity. The use of a self-immolative spacer can allow for the elimination of the fully active, chemically unmodified immune- stimulatory compound upon amide bond hydrolysis.
[0407] One self-immolative spacer can be a bifunctional /?ara-aminobenzyl alcohol group, which can link to the peptide through an amino group of the immune- stimulatory compound, forming an amide bond, while amine containing immune- stimulatory compounds can be attached through carbamate functionalities to the benzylic hydroxyl group of the linker (to give a p- amidobenzylcarbamate, PABC). The resulting pro-immune-stimulatory compound can be activated upon protease-mediated cleavage, leading to a 1,6-elimination reaction releasing the unmodified immune- stimulatory compound, carbon dioxide, and remnants of the linker. The following scheme depicts the fragmentation of p- amidobenzyl carbamate and release of the immune- stimulator compound:
X-D
wherein X-D represents the unmodified immune- stimulatory compound. Heterocyclic variants of this self-immolative group have also been described.
[0408] The enzymatically cleavable linker can be a B-glucuronic acid-based linker. Facile release of the immune- stimulatory compound can be realized through cleavage of the B-glucuronide glycosidic bond by the lysosomal enzyme B-glucuronidase. This enzyme can be abundantly present within lysosomes and can be overexpressed in some tumor types, while the enzyme activity outside cells can be low. B-Glucuronic acid-based linkers can be used to circumvent the tendency of an immune- stimulatory conjugate to undergo aggregation due to the hydrophilic nature of B-glucuronides. In certain embodiments, B-glucuronic acid-based linkers can link an antibody construct to a hydrophobic immune- stimulatory compound. The following scheme depicts the release of an immune- stimulatory compound (D) from an immune- stimulatory con ugate containing a B-glucuronic acid-based linker and an antibody construct (Ab):
[0409] A variety of cleavable β-glucuronic acid-based linkers useful for linking drugs such as auristatins, camptothecin and doxorubicin analogues, CBI minor-groove binders, and psymberin to antibodies have been described. These β-glucuronic acid-based linkers may be used in the conjugates described herein. In certain embodiments, the enzymatically cleavable linker is a β- galacto side-based linker. β-Galactoside is present abundantly within lysosomes, while the enzyme activity outside cells is low.
[0410] Additionally, immune- stimulatory compounds containing a phenol group can be covalently bonded to a linker through the phenolic oxygen. One such linker relies on a methodology in which a diamino-ethane "Space Link" is used in conjunction with traditional "PABO" -based self-immolative groups to deliver phenols. Methylene carbamate linkers have also been described that allow linkages to hydroxyl groups on compounds.
[0411] Immune- stimulatory compounds containing a tertiary amine can be covalently bond to a linker to the tertiary amine by creating a quaternary amine linkage.
[0412] Cleavable linkers can include non-cleavable portions or segments, and/or cleavable segments or portions can be included in an otherwise non-cleavable linker to render it cleavable. By way of example only, polyethylene glycol (PEG) and related polymers can include cleavable groups in the polymer backbone. For example, a polyethylene glycol or polymer linker can include one or more cleavable groups such as a disulfide, a hydrazone or a dipeptide.
[0413] Other degradable linkages that can be included in linkers can include ester linkages formed by the reaction of PEG carboxylic acids or activated PEG carboxylic acids with alcohol groups on an immune- stimulatory compound, wherein such ester groups can hydrolyze under physiological conditions to release the immune- stimulatory compound. Hydrolytically degradable linkages can include, but are not limited to, carbonate linkages; imine linkages resulting from reaction of an amine and an aldehyde; phosphate ester linkages formed by reacting an alcohol with a phosphate group; acetal linkages that are the reaction product of an aldehyde and an alcohol; orthoester linkages that are the reaction product of a formate and an alcohol; and oligonucleotide linkages formed by a phosphoramidite group, including but not limited to, at the end of a polymer, and a 5' hydroxyl group of an oligonucleotide.
[0414] A linker can contain an enzymatically cleavable peptide moiety, for example, a linker comprising structural formula (Ilia), (Mb), (IIIc), or (Hid):
or a salt thereof, wherein: peptide represents a peptide (illustrated N→C, wherein peptide includes the amino and carboxy "termini") cleavable by a lysosomal enzyme; T represents a polymer comprising one or more ethylene glycol units or an alkylene chain, or combinations thereof; R is selected from hydrogen, alkyl, sulfonate and methyl sulfonate; Ry is hydrogen or Ci.4 alkyl-(0)r-(Ci-4 alkylene)s-G1 or Ci-4 alkyl-(N)-[(Ci-4 alkylene)^1 ]2; Rz is C14 alkyl-(0)r- (Ci-4 alkylene)s-G2; G1 is S03H, C02H, PEG 4-32, or sugar moiety; G2 is S03H, C02H, or PEG 4-32 moiety; r is 0 or 1; s is 0 or 1; p is an integer ranging from 0 to 5; q is 0 or 1; x is 0 or 1; y is
i
0 or 1; represents the point of attachment of the linker to an immune- stimulatory compound; and * represents the point of attachment to the remainder of the linker, such as to a reactive group (Rx).
[0415] In certain embodiments, the peptide can be selected from a tripeptide or a dipeptide. In particular embodiments, the dipeptide can be selected from: Val-Cit; Cit-Val; Ala- Ala; Ala-Cit; Cit-Ala; Asn-Cit; Cit-Asn; Cit-Cit; Val-Glu; Glu-Val; Ser-Cit; Cit-Ser; Lys-Cit; Cit-Lys; Asp-Cit; Cit-Asp; Ala-Val; Val-Ala; Phe-Lys; Lys-Phe; Val-Lys; Lys-Val; Ala-Lys; Lys-Ala; Phe-Cit; Cit-Phe; Leu- Cit; Cit-Leu; Ile-Cit; Cit-Ile; Phe-Arg; Arg-Phe; Cit-Trp; and Trp-Cit, or salts thereof.
[0416] In certain embodiments, the peptide can be selected from a di-peptide or tri-peptide of non-natural amino acids, a mixture of natural and non-natural amino acids, amino acid analogs or a mixture of amino acids and amino acid analogs.
[0417] Exemplary embodiments of linkers according to structural formula (Ilia) that can be included in the conjugates described herein can include the linkers illustrated below (as illustrated, the linkers include a group suitable for covalently linking the linker to an antibody construct): wherein the wavy line indicates point to the immune- stimulatory compound.
[0418] Exemplary embodiments of linkers according to structural formula (Illb), (IIIc), or (Hid) that can be included in the conjugates described herein can include the linkers illustrated below (as illustrated, the linkers can include a group suitable for covalently linking the linker to an antibody construct):
-227-
-229-
wherein the wavy line indicates point to the immune- stimulatory compound.
[0419] The linker can contain an enzymatically cleavable sugar moiety, for example, a linker comprising structural formula (IVa), (IVb), (IVc), (IVd), or (IVe):
or a salt thereof, wherein: q is 0 or 1; r is 0 or 1; X l is C¾, O, or NH; * * represents the point of attachment of the linker to an immune- stimulatory compound; and * represents the point of attachment to the remainder of the linker.
[0420] Exemplary embodiments of linkers according to structural formula (IVa) that may be included in the immune- stimulatory conjugates described herein can include the linkers illustrated below (as illustrated, the linkers include a group suitable for covalently linking the linker to an antibody construct):
-234-
wherein the wavy line indicates a point of attachment to the immune- stimulatory compound.
[0421] Exemplary embodiments of linkers according to structural formula (IVb) that may be included in the conjugates described herein include the linkers illustrated below (as illustrated, the linkers include a group suitable for covalently linking the linker to an antibody construct):
-236-
-237-
wherein the wavy line indicates a point of attachment to the immune- stimulatory compound.
[0422] Exemplary embodiments of linkers according to structural formula (IVc) that may be included in the conjugates described herein include the linkers illustrated below (as illustrated, the linkers include a group suitable for covalently linking the linker to an antibody construct):
-239-
wherein the wavy line indicates a point of attachment to the immune- stimulatory compound.
[0423] Exemplary embodiments of linkers according to structural formula (IVd) that may be included in the conjugates described herein include the linkers illustrated below (as illustrated, the linkers include a group suitable for covalently linking the linker to an antibody construct):
wherein the wavy line indicates a point of attachment to the immune- stimulatory compound. [0424] Exemplary embodiments of linkers according to structural formula (IVe) that may be included in the conjugates described herein include the linkers illustrated below (as illustrated, the linkers include a group suitable for covalently linking the linker to an antibody construct):
wherein the wavy line indicates a point of attachment to the immune- stimulatory compound.
[0425] Although cleavable linkers can provide certain advantages, the linkers comprising the conjugate described herein need not be cleavable. For non-cleavable linkers, the immune- stimulatory compound release may not depend on the differential properties between the plasma and some cytoplasmic compartments. The release of an active form of the immune- stimulatory compound can occur after internalization of the immune- stimulatory conjugate via antigen- mediated endocytosis and delivery to lysosomal compartment, where the antibody construct can be degraded to the level of amino acids through intracellular proteolytic degradation. This process can release an immune- stimulatory compound derivative, which is formed by the immune- stimulatory compound, the linker, and the amino acid residue to which the linker was covalently attached (e.g., a cysteine residue). The immune- stimulatory compound derivative from immune- stimulatory conjugates with non-cleavable linkers can be more hydrophilic and less membrane permeable, which can lead to less bystander effects and less nonspecific toxicities compared to immune- stimulatory conjugates with a cleavable linker. Immune- stimulatory conjugates with non-cleavable linkers can have greater stability in circulation than immune- stimulatory conjugates with cleavable linkers. Non-cleavable linkers can be alkylene chains, or can be polymeric, such as, for example, based upon polyalkylene glycol polymers, amide polymers, or can include segments of alkylene chains, polyalkylene glycols and/or amide polymers. The linker can contain a polyethylene glycol segment having from 1 to 6 ethylene glycol units.
[0426] The linker can be non-cleavable in vivo, for example, a linker according to the
formulations below:
thereof, wherein: R is selected from hydrogen, Ci-ioalkyl, sulfonate and methyl sulfonate; Rx is a moiety including a functional group capable of covalently linking the linker to an antibody construct; and ^ represents the point of attachment of the linker to an immune- stimulatory compound.
[0427] Exemplary embodiments of linkers according to structural formula (Va)-(Ve) that may be included in the conjugates described herein include the linkers illustrated below (as illustrated, the linkers include a group suitable for covalently linking the linker to an antibody construct, and represents the point of attachment to an immune- stimulatory compound):
0428]
[0429] Attachment groups (also referred to as reactive groups herein) that are used to attach the linkers to an antibody construct can be electrophilic in nature and include, for example, maleimide groups, activated disulfides, active esters such as NHS esters and HOBt esters, haloformates, acid halides, alkyl, and benzyl halides such as haloacetamides. There are also emerging technologies related to "self- stabilizing" maleimides and "bridging disulfides" that can be used in accordance with the disclosure.
[0430] One example of a "self- stabilizing" maleimide group that hydrolyzes spontaneously under conjugation conditions to give a conjugate species with improved stability is depicted in the schematic below. Thus, the maleimide attachment group is reacted with a sulfhydryl of an antibody to give an intermediate succinimide ring, after which the succinmide ring hydrolizes opens to form a ring opened form. The hydro lyzed form of the attachment group is resistant to deconjugation in the presence of plasma proteins.
Leads to "DAR loss" over time
Self-stabilizing attachment:
contains maleimide contains succinumide hydrolized forms of succinumide ring rin9 r'n9 hydrolized forms are stable in plasma
[0431] A method for bridging a pair of sulfhydryl groups derived from reduction of a native hinge disulfide bond has been disclosed and is depicted in the schematic below. An advantage of this methodology is the ability to synthesize homogenous DAR4 conjugates by full reduction of
IgGls (to give 4 pairs of reactive sulfhydryls) followed by reaction with 4 equivalents of the alkylating agent. Conjugates containing "bridged disulfides" are also claimed to have increased stability.
"bridged disulfide"
[0432] Similarly, as depicted below, a maleimide derivative that is capable of bridging a pair of sulfhydryl groups has been developed.
[0433] The attachment moiety can contain the following structural formulas (Via), (VIb), or (Vic):
or salts thereof, wherein: Rq is H or-0-(CH2CH20)n-CH3; x is 0 or 1; y is 0 or 1; G2 is- CH2CH2CH2S03H or-CH2CH20-(CH2CH20)n-CH3; Rw is-0-CH2CH2S03H or-NH(CO)- CH2CH20-(CH2CH20)i2-CH3; and * represents the point of attachment to the remainder of the linker.
[0434] Exemplary embodiments of linkers according to structural formulae (Via) and (VIb) that can be included in the conjugates described herein can include the linkers illustrated below (as illustrated, the linkers can include a group suitable for covalently linking the linker to an antibody construct):
-247-
-248-
-249- wherein the wavy line indicates a point of attachment to the immune- stimulatory compound.
[0435] Exemplary embodiments of linkers according to structural formula (Vic) that can be included in the immune- stimulatory conjugates described herein can include the linkers illustrated below (as illustrated, the linkers can include a group suitable for covalently linking the linker to an antibod construct):
wherein the wavy line indicates a point of attachment to the immune- stimulatory compound.
[0436] As is known by skilled artisans, the linker selected for a particular immune- stimulatory conjugate may be influenced by a variety of factors, including but not limited to, the site of attachment to the antibody construct (e.g., lysine, cysteine, glutamine, glutamate or other amino acid residue(s)), structural constraints of the drug pharmacophore and the lipophilicity of the drug. The specific linker selected for a conjugate should seek to balance these different factors for the specific antibody construct/immune stimulatory compound combination.
[0437] For example, ADCs with cytotoxic agents have been observed to effect killing of bystander antigen-negative cells present in the vicinity of the antigen-positive tumor cells. The mechanism of bystander cell killing by ADCs has indicated that metabolic products formed during intracellular processing of the ADCs may play a role. Neutral and/or hydrophobic cytotoxic metabolites generated by metabolism of the ADCs in antigen-positive cells appear to play a role in bystander cell killing while charged or hydrophilic metabolites may be prevented from diffusing across the membrane into the medium and therefore cannot affect bystander killing. In certain embodiments as described herein, the linker for an immune- stimulatory conjugate is selected to attenuate the bystander effect caused by cellular metabolites of the conjugate. In certain embodiments, the linker is selected to increase the bystander effect.
[0438] The properties of the linker may also affect aggregation of the conjugate under conditions of use and/or storage. Typically, ADCs reported in the literature contain no more than 3-4 drug molecules per antibody molecule. Attempts to obtain higher drug-to-antibody ratios ("DAR") often failed, particularly if both the drug and the linker were hydrophobic, due to aggregation of the ADC. In many instances, DARs higher than 3-4 could be beneficial as a means of increasing potency. In instances where the drug is hydrophobic in nature, it may be desirable to select linkers that are relatively hydrophilic as a means of reducing aggregation, especially in instances where DARs greater than 3-4 are desired. Thus, in certain embodiments, the linker of an immune- stimulatory conjugate incorporates chemical moieties that reduce aggregation of the conjugate during storage and/or use. A linker may incorporate polar or hydrophilic groups such as charged groups or groups that become charged under physiological pH to reduce the aggregation of the conjugate. For example, a linker may incorporate charged groups such as salts or groups that deprotonate, e.g., carboxylates, or protonate, e.g., amines, at physiological pH.
[0439] In particular embodiments, the aggregation of the conjugates during storage or use is less than about 40% as determined by size-exclusion chromatography (SEC). In particular
embodiments, the aggregation of the conjugates during storage or use is less than 35%, such as less than about 30%, such as less than about 25%, such as less than about 20%, such as less than about 15%, such as less than about 10%, such as less than about 5%, such as less than about 4%, or even less, as determined by size-exclusion chromatography (SEC).
General method for interchain cysteine-based bioconjugations
[0440] An antibody construct can be conjugated to a linker via cysteine-based bioconjugation.
An antibody construct can be exchanged into an appropriate buffer, for example, phosphate, borate, PBS, histidine, Tris-Acetate at a concentration of about 2 mg/mL to about 10 mg/mL with an appropriate number of equivalents of a reducing agent, for example, dithiothreitol or tris(2- carboxyethyl)phosphine. The resultant solution can be stirred for an appropriate amount of time and temperature to effect the desired reduction. A construct of an amino-pyrazinecarboxamide compound and a linker can be added as a solution with stirring. Dependent on the physical properties of the linker-pay load, a co-solvent can be introduced prior to the addition of the linker- payload to facilitate solubility. The reaction can be stirred at room temperature for about 1 hour to about 12 hours depending on the observed reactivity. The progression of the reaction can be monitored by liquid chromatography-mass spectrometry (LC-MS). Once the reaction is deemed complete, the remaining free linker-pay load can be removed by applicable methods and the antibody conjugate can be exchanged into the desired formulation buffer. Such cysteine-based conjugates can be synthesized starting with an antibody (rnAb) and linker-payload, e.g., 7 equivalents, using the conditions described in Scheme A below (Conjugate = antibody conjugate). Monomer content and drug-antibody ratios can be determined by methods described herein.
Scheme A.
1 . reducing agent
rnAb Conjugate
7 eq of compound-linker construct
sodium phosphate
pH = 8
20% v/v DMSO
General method for lysine-based bioconjugations
[0441] An antibody construct can be conjugated to a linker via lysine-based bioconjugation. An antibody construct can be exchanged into an appropriate buffer, for example, phosphate, borate, PBS, histidine, Tris-Acetate at a concentration of about 2 mg/mL to about 10 mg/mL. An appropriate number of equivalents of a construct of an amino-pyrazinecarboxamide compound, and a linker, linker-payload, as described herein, can be added as a solution with stirring.
Dependent on the physical properties of the linker-payload, a co-solvent can be introduced prior to the addition of the linker-payload to facilitate solubility. The reaction can be stirred at room temperature for 2 hours to about 12 hours depending on the observed reactivity. The progression of the reaction can be monitored by LC-MS. Once the reaction is deemed complete, the remaining linker-payloads can be removed by applicable methods and the antibody conjugate can be exchanged into the desired formulation buffer. Lysine-linked conjugates can be synthesized starting with ab antibody (rnAb) and linker-payload, e.g., 10 equivalents, following Scheme B below (Conjugate = antibody conjugate). Monomer content and drug-antibody construct ratios (molar ratios) can be determined by methods described herein.
Scheme B.
10 eq of compound-linker construct
sodium phosphate
rnAb Conjugate
pH = 8
20% v/v DMSO Conjugates of PROTACS
[0442] The conjugates described herein may also comprise antibody constructs and constructs having immune-modulatory activity through targeted degradation of proteins. As used herein, such conjugates are termed immune-modulatory conjugates and have a direct or indirect effect on the immune system. In some embodiments, the conjugates can target proteins involved in immune activation, inhibition or regulation. In some embodiments, the conjugates can target proteins in tumor cells and cause cell death or apoptois and indirectly stimulate the immune system. In various embodiments, proteolysis targeting modules (PTM) are attached (e.g., conjugated) through a linker to an antibody construct to form immune-modulatory conjugates.
[0443] In certain embodiments, an immune-modulatory conjugate comprises an antibody construct, a linker, and two binding moieties. The linker can be a multifunctional linker (W) that can covalently attach two binding moieties (X and Y) to form a proteolysis targeting module (X- W-Y) designed to induce degradation of a protein target. A PTM can comprise a first binding moiety (X) that can bind to a protein target (the moiety also referred to as a target protein binding moiety or protein targeting moiety) and a second binding moiety (Y) that can bind to an E3 ubiquitin ligase. The multi-functional linker also can covalently attach the PTM to a residue(s) (e.g., lysine, cysteine or engineered residue) on an antibody construct (Z) (e.g., an antibody) to form an immune-modulatory conjugate, as shown below:
W = multifunctional linker; X = Binding moeity 1 ; Y = Binding moiety 2; Z = antibody
[0444] In some aspects, an immune-modulatory conjugate comprises a proteolysis targeting module that can bind to a protein active site and can stimulate an immune response through protein target degradation; an antibody construct comprising an antigen binding domain and an Fc domain, wherein the antigen binding domain can bind to a first antigen; and a multifunctional linker, wherein the linker is covalently bound to the antibody construct and to protein targeting moiety (a first binding moiety) and an E3 ubiquitin ligase binding moiety (second binding moiety). In some embodiments, the Fc domain is an Fc null. In some embodiments, the Fc domain is a wild-type IgG that can bind to Fey receptors. In some embodiments, the Fc domain can bind to an Fc receptor, wherein the IQ for binding of the Fc domain of the conjugate to an Fc receptor is no greater than about 100 times the Kd for binding of a control antibody construct to the Fc receptor, wherein the control antibody construct is the unconjugated antibody construct. In some embodiments, the Kd for binding of the first binding moiety of the conjugate to the protein active site is no greater than 100 times the Kd for binding of a control compound to the protein active site or wherein the IC50 of the first binding moiety of the conjugate is no greater than 300-fold the IC50 of a control compound, wherein the control compound is the free first binding moiety. In some embodiments, the proteolysis targeting module can be comprised of: a) a small molecule that can bind a protein target, degredation or inhibition of which is immune stimulatory, b) a multi-functional linker that can be covalently bonded to elements a) and c); and c) a small molecule that can bind to an E3 ubiquitin ligase.
[0445] In some embodiments, the proteolysis targeting module can be comprised of a) a small molecule that can bind to a protein target, inhibition or degredation of which is immune stimulatory, b) a multi-functional linker that can be covalently bonded to elements a) and c); and c) a small molecule that can bind an E3 ubiquitin ligase, wherein the proteolysis targeting module can increase degradation of the protein target within a cell.
[0446] In some embodiments, the proteolysis targeting module can be comprised of a) a first binding moiety that can bind an immune inhibitory target, b) a linker that can attach the first binding moiety to element c), and c) a compound that can bind an E3 ubiquitin ligase, wherein the PTM can increase degradation of the immune-inhibitory target in cells expressing antigen of the first binding domain or second binding domain of the conjugate (containing the PTM) compared to degradation of the immune-inhibitory target in cells not expressing, or expressing lower levels of, antigen of the first binding domain or second binding domain of the conjugate. The PTM is attached to the antibody construct via another linker, as further described below.
[0447] In other embodiments, the proteolysis targeting module can be comprised of a) a first binding moiety that can bind to an immune stimulatory target, b) a second linker that can attach the first binding moiety to element c), and c) a compound that can bind to an E3 ubiquitin ligase, wherein the PTM can increase degradation of the immune- stimulatory target in cells expressing antigen of the first binding domain or second binding domain of the conjugate (containing the PTM) compared to degradation of the immune- stimulatory target in cells not expressing, or expressing lower levels of, antigen of the first binding domain or second binding domain of the conjugate.
[0448] These conjugates can be made by various methods. It is understood that one skilled in the art may be able to make these compounds by similar methods or by combining other methods known to one skilled in the art. It is also understood that one skilled in the art would be able to make, in a similar manner as described herein by using the appropriate starting materials and modifying the synthetic route as needed. Starting materials and reagents can be obtained from commercial vendors or synthesized according to sources known to those skilled in the art or prepared as described herein.
[0449] In other embodiments, immune-modulatory conjugates have the following general formula:
wherein Ab is an antibody construct, L is a linker, D is an immune-modulatory compound, x may be from 1 to 20 (wherein each x denotes a distinct compound), n may be from 1 to 20, and z may be from 1 to 20.
[0450] In some embodiments, x is 1, n is 1 and z may be from 1 to 10, such as from 1 to 9, such as from 1 to 8, such as from 2 to 8, such as from 1 to 6, such as from 3 to 5 or such as about 2.
[0451] In some embodiments x is 1, n is 2, and z may be from 1 to 10, such as from 1 to 9, such as from 1 to 8, such as from 2 to 8, such as from 1 to 6 or such as from 3 to 5. In certain embodiments, z is 4.
[0452] In certain embodiments, con ugates are represented by the following formula: wherein Ab is an antibody construct, L is a linker having the structure -Aa-Ww-Yy-, where A is a spacer, a is 0 or 1, W is a cleavable unit, w may be from 0 to 10, Y is a stretcher, y may be from
0 to 3, D is an immune-modulatory compound, x may be from 1 to 20 (wherein each x denotes a distinct compound), n may be from 1 to 20, and z may be from 1 to 20.
[0453] In some embodiments, a is 1, w is 0, y is 0, x is 1, n is 1, and z may be from 1 to 20, 1 to 10, 1 to 9, 1 to 8, such as from 2 to 8, 1 to 6 or 3 to 5. In certain embodiments, z is 4 or 2.
[0454] In some embodiments, a isl, w is 1, y is 1, x is 1, n is 1 and z may be from 1 to 10, 1 to 9,
1 to 8, 2 to 8, 1 to 6, 3 to 5, or 4.
[0455] In some embodiments, a isl, w is 1, y is from 0, x is 1, n is 1 and z may be from 1 to 10, 1 to 9, 1 to 8, 2 to 8, 1 to 6, 3 to 5, or 4.
[0456] In some embodiments, a is 0 or 1, w is from 0 to 10, y is from 0-3, where at least one of A, W or Y is present, x may be from 1 to 20, n may be from 1-20, and z may be from 1 to 20.
[0457] In certain embodiments, an immune-modulatory conjugate can be designed to increase ubiquitin-mediated target protein destruction via the ubiquitin pathway. The process of attaching ubiquitin molecules to a protein target typically involves 3 enzymates and steps: 1) an El enzyme that can activate ubiquitin, 2) an E2 enzyme that can transfer activated ubiquitin, and 3) a multi- subunit E3 enzyme ligase that can receive the activated ubiquitin and catalyze a ubiquitin attachment to the target protein.
[0458] In some embodiments, an immune-modulatory conjugate is provided that includes a proteolysis targeting module (PTM; also referred to as a proteolysis-targetting chimera or
PROTAC). PTMs can comprise a small molecule, a target binding moiety that binds a protein target and can be covalently attached, directly or by a spacer, to the small molecule that can bind an E3 ubiquitin ligase subunit. In some embodiments, a PTM includes a protein targeting moiety, such as an immune-modulatory compound (IMC), that is covalently attached to an E3 ubiquitin ligase binding moiety (ULM) through a spacer (S) and a linker (L) that is covalently attached to the PTM and to the antibody construct (Ab), as represented by the formula Ab-(L-PTMn)z, where n is from 1-20 and z is from 1 to 20. In some embodiments, L is a cleavable linker. The cleavable linker can be a peptide linker or other cleavable linker described above in the Section on Linkers. In some embodiments, L is a non-cleavable linker. In some embodiments, the Fc domain of the conjugate is an Fc null. In some embodiments, the Fc domain is a wild-type IgG that can bind to Fey receptors. In some embodiments, the Fc domain can bind to an Fc receptor, wherein the Kd for binding of the Fc domain of the conjugate to an Fc receptor is no greater than about 100 times the Kd for binding of a control antibody construct to the Fc receptor, wherein the control antibody construct is the unconjugated antibody construct. In some embodiments, the Kd for binding of the IMC of the conjugate to the protein active site is no greater than 100 times the Kd for binding of a control compound to the protein active site or wherein the IC50 of the IMC of the conjugate is no greater than 300-fold the IC50 of a control compound, wherein the control compound is the free IMC.
[0459] In some embodiments, a protein targeting moiety, such as an immune-modulatory compound (IMC), is covalently attached to an E3 ubiquitin ligase binding moiety (ULM) through a spacer (S) and a linker (L) is covalently attached to the spacer (s), n is from 1-20 and z is from 1 to 20 as represented by the formula:
VII [0460] In some embodiments, L is a cleavable linker. The cleavable linker can be a peptide linker or other cleavable linker described above in the Section on Linkers. In some embodiments, L is a non-cleavable linker. In some embodiments, the Fc domain of the conjugate is an Fc null. In some embodiments, the Fc domain is a wild-type IgG that can bind to Fey receptors. In some embodiments, the Fc domain can bind to an Fc receptor, wherein the Kd for binding of the Fc domain of the conjugate to an Fc receptor is no greater than about 100 times the Kd for binding of a control antibody construct to the Fc receptor, wherein the control antibody construct is the unconjugated antibody construct. In some embodiments, the Kd for binding of the IMC of the conjugate to the protein active site is no greater than 100 times the Kd for binding of a control compound to the protein active site or wherein the IC50 of the IMC of the conjugate is no greater than 300-fold the IC50 of a control compound, wherein the control compound is the free IMC.
[0461] In some embodiments, a protein targeting moiety, such as an immune-modulatory compound (IMC), is covalently attached to an E3 ubiquitin ligase binding moiety (ULM) through a spacer (S) and a linker (L) is covalently attached to the protein targeting moiety, n is from 1-20 and z is from 1 to 20 as represented by the formula:
[0462] In some embodiments, L is a cleavable linker. The cleavable linker can be a peptide linker or other cleavable linker described above in the Section on Linkers. In some embodiments, L is a non-cleavable linker. In some embodiments, the Fc domain of the conjugate is an Fc null. In some embodiments, the Fc domain is a wild-type IgG that can bind to Fey receptors. In some embodiments, the Fc domain can bind to an Fc receptor, wherein the Kd for binding of the Fc domain of the conjugate to an Fc receptor is no greater than about 100 times the Kd for binding of a control antibody construct to the Fc receptor, wherein the control antibody construct is the unconjugated antibody construct. In some embodiments, the Kd for binding of the IMC of the conjugate to the protein active site is no greater than 100 times the Kd for binding of a control compound to the protein active site or wherein the IC50 of the IMC of the conjugate is no greater than 300-fold the IC50 of a control compound, wherein the control compound is the free IMC.
[0463] In some embodiments, a protein targeting moiety, such as an immune-modulatory compound (IMC), is covalently attached to an E3 ubiquitin ligase binding moiety (ULM) through a spacer (S) and linker L is covalently attached to the ubiquitin E3 ligase moiety (ULM), n is from 1-20 and z is from 1 to 20 as represented by the formula:
(IX)
[0464] In some embodiments, L is a cleavable linker. The cleavable linker can be a peptide linker or other cleavable linker described above in the Section on Linkers. In some embodiments, L is a non-cleavable linker. In some embodiments, the Fc domain of the conjugate is an Fc null. In some embodiments, the Fc domain is a wild-type IgG that can bind to Fey receptors. In some embodiments, the Fc domain can bind to an Fc receptor, wherein the Kd for binding of the Fc domain of the conjugate to an Fc receptor is no greater than about 100 times the Kd for binding of a control antibody construct to the Fc receptor, wherein the control antibody construct is the unconjugated antibody construct. In some embodiments, the Kd for binding of the IMC of the conjugate to the protein active site is no greater than 100 times the Kd for binding of a control compound to the protein active site or wherein the IC50 of the IMC of the conjugate is no greater than 300-fold the IC50 of a control compound, wherein the control compound is the free IMC.
[0465] In certain embodiments, the E3 ubiquitin ligase binding moiety is linked to a protein targeting moiety, such as an immune-modulatory compound, in the conjugate as described herein, via a spacer. In certain embodiments, the E3 ubiquitin ligase binding moiety can be linked to the protein targeting moiety via a spacer having a linear non-hydrogen atom number in the range of 1 to 25 or 1 to 20. In certain embodiments, the spacer has 5 to 20 or 5 to 15 linear non- hydrogen atoms. The spacer is typically non-cleavable.
[0466] The E3 ubiquitin ligase binding moiety can be linked to the spacer of the protein targeting moiety with a functional group such as an ether, amide, alkane, alkene, alkyne, ketone, hydroxyl, carboxylic acid, thioether, sulfoxide, and sulfone. The E3 ubiquitin ligase binding moiety can be linked to the spacer of the protein targeting moiety via a spacer comprising an aromatic, heteroaromatic, cyclic, bicyclic, and/or tricyclic moiety.
[0467] Spacer length can be varied to optimize the activity of the protein targeting moiety for its target protein. In some embodiments, the spacer is non-cleavable and comprises segments of alkylene, alkenylene, alkynylene, -(CH20)-, -CH2CH20)-, -(CH2OCH2)-, -C(O)-, -NH-, and -0-, having a length of from 1-25, 1-20, 1-15, 5-25, 5-20, or 5-15 linear non-hydrogen atoms. A spacer may be optionally substituted with Ci-C8alkyl, C2-C8alkenyl, C2-C8alkynyl, -(CH20)niH, - (CH2CH20)niH, -(CH20)niCH3, -C(0)OH or -NH2, wherein nl is from 1 to 8, and may further optionally comprise a reactive group, Rx, to form a functional group, such as an ether, amide, alkane, alkene, alkyne, ketone, hydroxyl, carboxylic acid, thioether, sulfoxide, and sulfone, forming an attachment to a linker (L). In some embodiments, the spacer is not unsubstituted. In some embodiments, the spacer is substituted with Rx.
[0468] A spacer may be a Ci-25alkylene or optionally substituted Q-25 heteroalkylene, wherein the heteroalkylene is a Q-24 alkylene chain interspersed with one or more groups independently selected from: -0-, -S-, -NH2-, and -C(0)NH-. The spacer may also be optionally substituted with a reactive group, R , that can form a functional group, such as an amide bond, an ester bond, an ether bond, a carbonate bond, a carbamate bond, or a thioether bond; such reactive groups can be, for example, amino groups; carboxyl groups; aldehyde groups; azide groups; alkyne and alkene groups; ketones; carbonates; carbonyl functionalities bonded to leaving groups such as cyano and succinimidyl and hydroxyl groups. In some embodiments, Rx can be -NH2, -S or a maleimide. In some embodiments, Rx is -NH2. The spacer may also be optionally substituted with Ci-C8alkyl, C2-C8alkenyl, C2-C8alkynyl, -(CH20)niH, -(CH2CH20)niH, - (CH20)niCH3, -C(0)OH or -NH2, wherein nl is from 1 to 8. In some embodiments, the spacer is not unsubstituted. In some embodiments, the spacer is substituted with Rx.
[0469] In certain embodiments, the spacer (S) has the formula -C(O)N(R100)R101C(O)N(R100)-, - C(O)R101C(O)-, -C(0)R101N(RiooK -N(Ri00)R101C(O)-, -N(R100)C(O)R101C(O)-, - N(R100)C(0)R101N(RiooK -N(Ri00)R101C(O)N(R100)-, -N(R100)C(O)R101C(O)N(R100)-, - N(R100)C(0)R101N(Rioo)C(0)-, and -C(O)N(Ri00)R101C(O)N(R100)-; wherein each R100 is independently selected from H or C1-C3 alkyl and R101 is -Ci-C25alkylene-, -Ci-C25alkenylene-, - Ci-C25alkynlene-, -Ci-Ci2alkylene(CH20)nCi-Ci5alkylene-, -Ci-Ci2alkylene((CH2OCH2)nCi- Ci2alkylene-, -Ci-Ci2alkylene(CH2CH20)nCi-Ci2alkylene-, -Ci-Ci2alkenylene-((CH20)nCi- C i2alkylene- , -C 1 -C i2alkenylene-(CH2CH20)nC 1 -C i2alkylene- , -C 1 -C i2alkenylene- ((CH2OCH2)nCi-Ci2alkylene-, -Ci-Ci2alkylene-(CH20)nCi-Ci2alkenylene-, -Ci-Ci2alkylene- (CH2CH20)nCi-Ci2alkenylene-, -Ci-Ci2alkylene-(CH2OCH2)nCi-Ci2alkenylene-, -Q- Ci2alkynylene-(CH20)nCi-Ci2alkylene-, -Ci-Ci2alkynylene-(CH2CH20)nCi-Ci2alkylene-, -Q- Ci2alkynylene-(CH2OCH2)nCi-Ci2alkylene-, -Ci-Ci2alkynylene-(CH20)nCi-Ci2alkenylene-, -Q- Ci2alkynylene-(CH2CH20)nCi-Ci2alkenylene-, -Ci-Ci2alkynylene-(CH2OCH2)nCi- Ci2alkenylene-,-Ci-Ci2alkynylene-(CH20)nCi-Ci2alkynylene-, -Ci-Ci2alkynylene- (CH2CH20)nCi-Ci2alkynylene-, -Ci-Ci2alkynylene-(CH2OCH2)nCi-Ci2alkynylene-, in each case optionally substituted with a reactive moiety Rx for attachment to the linker (L), and n is 0 to 8. R can be a reactive group that can form an amide bond, an ester bond, an ether bond, a carbonate bond, a carbamate bond, or a thioether bond; such reactive groups can be, for example, amino groups; carboxyl groups; aldehyde groups; azide groups; alkyne and alkene groups;
ketones; carbonates; carbonyl functionalities bonded to leaving groups such as cyano and succinimidyl and hydroxyl groups. In some embodiments, Rx can be -N¾, -S or a maleimide. In some embodiments, Rx is -N¾.
[0470] In certain embodiments, the spacer (S) comprises glutamate, a glycine-glutamate dipeptide, glycine-PEGl -glutamate, glycine-PEG2-glutamate, glycine-PEG3 -glutamate, glycine-PEG4-glutamate or glycine-PEG5-glutamate, wherein the E3 ubquitin ligase binding moiety and the protein targeting moiety are attached to the spacer via amide bonds.
[0471] An E3 ubiquitin ligase binding moiety can bind to an E3 ubiquitin ligase, such as Von Hippel-Lindaue E3 ubiquitin ligase (VHL), cereblon, mouse double minute 2 homolog (MDM2), AMFR, APC/Cdc20, APC/Cdhl, C6orfl57, Cbl, CBLL1, CHFR, CHIP, DTL (Cdt2), E6-AP, HACEl, HECTDl, HECTD2, HECTD3, HECWl, HECW2, HERC2, HERC3, HERC4, HERC5, HUWE1, HYD, ITCH, LNX1, mahogunin, MARCH-I, MARCH-II, MARCH-III, MARCH-IV, MARCH- VI, MARCH- VII, MARCH- VIII, MARCH-X, MEKK1, MIB1, MIB2, MycBP2, NEDD4, NEDD4L, Parkin, PELI1, Pirh2, PJA1, PJA2, RFFL, RFWD2, Rictor, RNF5, RNF8, RNF19, RNF190, RNF20, RNF34, RNF40, RNF125, RNF128, RNF138, RNF168, SCF/p-TrCP, SCF/FBW7, SCF/Skp2, SHPRH, SIAH1, SIAH2, SMURF1, SMURF2, TOPORS, TRAF6, TRAF7, TRIM63, UBE3B, UBE3C, UBR1, UBR2, UHRF2, WWP1, WWP2, or ZNRF1.
[0472] In other embodiments, an E3 ubiquitin ligase binding moiety can be selected from an E3 ubiquitin ligase selected from von Rippel-Lindau (VHL), cereblon, XIAP, E3A, MDM2, Anaphase-promoting complex (APC), UBR5 (EDDI), SOCS/ BC-box/ eloBC/ CUL5/ RING, LNXp80, CBX4, CBLLI, HACEl, HECTDl, HECTD2, HECTD3, HECWl, HECW2, HERCI, HERC2, HERC3, HERC4, HUWEI, ITCH, NEDD4, NEDD4L, PPIL2, PRPFI9, PIASI, PIAS2, PI AS 3, PI AS 4, RANBP2, RNF4, RBXI, SMURFI, SMURF2, STUB I, TOPORS, TRIPI2, UBE3A, UBE3B, UBE3C, UBE4A, UBE4B, UBOX5, UBR5, WWPI, WWP2, Parkin,
A20/TNFAIP3, AMFR/gp78, ARA54, beta-TrCPI/BTRC, BRCAI, CBL, CHIP/STUB I, E6, E6AP/UBE3A, F-box protein I5/FBXOI5, FBXW7/Cdc4, GRAIL/RNFI28, HOIP/RNF3 I, cIAP-I/HIAP-2, cIAP-2/HIAP-I, cIAP (pan), ITCH/AIP4, KAPI, MARCH8, Mind Bomb I/MIBI, Mind Bomb 2/MIB2, MuRFI/TRIM63, NDFIPI, NEDD4, NleL, Parkin, RNF2, RNF4, RNF8, RNFI68, RNF43, SARTI, Skp2, SMURF2, TRAF-I, TRAF-2, TRAF-3, TRAF-4, TRAF- 5, TRAF-6, TRIMS, TRIM2I, TRIM32, UBR5, and ZNRF3.
[0473] In further embodiments, an E3 ubiquitin ligase can be selected from the following types: HECT type, RING-type, PARKIN-finger type, RING-variant type, U-box type, A20-finger type, PIAS-finger type, PHD-finger type, Skpl-like type, Cullin-type, F-box type, SOCS-box type, BTB-type, DDBl-like type and APC/cyclosome type.
[0474] An E3 ubiquitin ligase binding moiety can be a VHL binding moiety such as (S)-2- amino-Nl-(4-(5-amino-6-((4-morpholinopyridin-3-yl)carbamoyl)pyrazin-2-yl)benzyl)-N5-(2-(3- (((S)-l-((2S,4R)-4-hydroxy-2-((4-(4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidin-l-yl)-^ dimethyl- l-oxobutan-2-yl)amino)-3-oxopropoxy)ethyl)pentanediamide (Example 1) or a cereblon binding moiety such as 3-amino-6-(4-(2-((2S)-2-amino-6-(2-((2-(2,6-dioxopiperidin-3- yl)-l,3-dioxoisoindolin-4-yl)oxy)acetamido)hexanamido)ethyl)phenyl)-N-(4-morpholinopyridin- 3-yl)pyrazine-2-carboxamide (Example 2). Other compounds that bind VHL may be
hydroxyproline compounds such as those disclosed in WO 2013/106643, and other compounds described in US 2016/0045607, WO 2014187777, US 2014/0356322, and U.S. 9,249,153. Other compounds that bind to cereblon include thalidomide, lenalidomide, pomalidomide and analogs thereof. Other small molecule compounds that bind to cereblon are also known, e.g., the compounds disclosed as an in US 2016/0058872 and US 2015/0291562.
[0475] In certain embodiments, the linker (L) is attached to the PTM at a reactive site Rx in the spacer. In certain embodiments, the linker (L) is attached to the PTM via an attachment site in the E3 ubiquitin ligase binding moiety. In certain embodiments, the linker (L) is attached to the PTM via an attachment site in the protein targeting moiety.
[0476] The linker (L) and/or covalent attachment site(s) of the linker (L) to the proteolysis targeting module can be cleavable or non-cleavable. In certain embodiments, the linker is cleavable. In certain embodiments, the linker is non-cleavable linker. In some embodiments, the linker is non-cleavable and is attached to the proteolysis targeting module at site wherein the protein targeting moiety can bind to its protein target, and, if active, does not lose immune- modulatory activity, as determined by Kd measurement, by altered target protein function in a cell-based assay, or both. Linker length can be varied to optimize the activity of the protein targeting moiety for its target protein. Such linkers can be short, flexible, rigid, hydrophilic, or hydrophobic. The linker can contain segments that have different characteristics, such as segments of flexibility or segments of rigidity. The linker can be chemically stable to
extracellular environments. Non-limiting examples can be maleimidocaproyl linkers. A maleimidocaproyl linker can comprise N-maleimidomethylcyclohexane-l-carboxylate.
[0477] In some embodiments, the linker (L) is a cleavable linker and can be selected from the linkers of formulae Ila, lib, lie, Ilia, Mb, IIIc, Hid, IVa, IVb, IVc, IVd, and IVe and specific structures therein, as shown above.
[0478] A linker (L) can be a combination of a maleimidocaproyl group and one or more polyethylene glycol molecules.
[0479] A linker (L) may comprise from 5 to 100 linear non-hydrogen atoms that may be covalently attached to an antibody construct.
[0480] In some embodiments, the protein targeting moiety of the proteolysis targeting module is an immune-modulatory compound, such as those disclosed herein. In some embodiments, For example, a protein targeting moiety can be an antagonist of an immune-modulatory or immune- inhibitory protein, such as a beta-catenin pathway inhibitor, a kinase inhibitor, a TNIK inhibitor, a Tankyrase (TNKS) inhibitor, TGFpRl, TGFpR2, ΡΙ3Κ-β, STAT3, IL-10, IDO, or TDO. In some embodiments, the protein targeting moiety is a kinase inhibitor that binds to ALK, Bcr-Abl, BRAF, BTK, c-KIT, EGFR, ErbB2, JAK, MEK, MET, PDGFRB, RET, ROS 1, Syk, SRC, TGFpR2, TNIK, TNKS, TNKS 2, TrkA, TrkB, TrkC, VEGFR1, VEGFR2, VEGFR3, FGFR1, FGFR2, FGFR3, FGFR4, CSF1R, RON/MS T1R, TYR03, MERTK, AXL, ΡΙ3Κδ, ΡΙ3Κγ, MAP4K1, PERK, KIT, or any combination thereof.
[0481] In some embodiments, the protein targeting moiety binds to aryl hydrocarbon receptor, androgen receptor, estrogen receptor, FK506-binding protein 12, fibroblast growth factor receptor substrate 2, phosphatidylinositol -4,5-biphosphate 3-kinase, SMAD family member 3, bromodomain and extra-territorial family of proteins (BET), bromodomain-containing protein 4 member of the BET family, Abelson tyrosine kinase, receptor- interacting serine/threo nine- protein kinase 1, estrogen-related receptor, TRAF2 and NCK- interacting protein kinases (TNIK), and transforming growth factor beta. The protein targeting moiety can be an antagonist or inhibitor of any of these protein targeting moieties.
[0482] In some embodiment, the protein targeting moiety binds to TGFPR2, TGFpRl, SMAD2, SMAD3, SMAD4, beta-catenin, TNIK, CREBB2, Beta catenin/TCF4, beta catenin/LEF, beta catenin/CREBBP, YAP, TAZ, YAP/TAZ, TNKS 1 , TNKS 2, MST1, MST2, NRAS, HRAS, KRAS, RASmutl2, RASmutl3, PERK (EIF2AK3), RON/MS T1R, PARP1,2, MTOR, STAT3, MCTl, MCT2, or MCT4. The protein targeting moiety can be an antagonist or inhibitor of any of these protein targeting moieties.
[0483] In some embodiments, the protein targeting moiety is CSFR1, RON/MST1, PI3Kd, PI3Kg, PARP1, PD-L1, PP2A, A2ar, TYR03, AXL, or MER. The protein targeting moiety can be an antagonist or inhibitor of any of these protein targeting moieties.
[0484] In other embodiments, the protein targeting moiety can be a Pattern recognition receptor (PRR) agonist, such a PAMP molecule or a DAMP molecule. In some embodiments, the protein targeting moiety can be a Toll- like receptor agonist, a RIG-I agonist, a STING agonist, a GPCR agonist, an ion channel agonist, a membrane transporter agonist, or an ER protein agonist.
Conjugates
[0485] In some embodiments, the conjugates described herein comprise an antibody construct, at least one an immune- stimulatory compound, and at least one linker. A conjugate can comprise an antibody construct, at least one pattern recognition receptor (PRR) agonist, and at least one linker. A conjugate can comprise an antibody construct, at least one pattern-associated molecular pattern (PAMP) molecule, and at least one linker. A conjugate can comprise an antibody construct, at least one damage-associated molecular pattern (DAMP) molecule, and at least one linker. A conjugate can comprise an antibody construct, at least one STING agonist, and at least one linker. A conjugate can comprise an antibody construct, at least one toll-like receptor agonist, and at least one linker. A conjugate can comprise an antibody construct, at least one kinase inhibitor and at least one linker. An antibody construct of any of the conjugates described herein can have a modified Fc domain.
[0486] In some embodiments, an Fc domain or region can exhibit reduced binding affinity to one or more Fc receptors. In some embodiments, an Fc domain or region can exhibit reduced binding affinity to one or more Fcgamma receptors. In some embodiments, an Fc domain or region can exhibit reduced binding affinity to FcRn receptors. In some embodiments, an Fc domain or region can exhibit reduced binding affinity to Fcgamma and FcRn receptors. In some embodiments, an Fc domain is an Fc null domain or region. As used herein, an "Fc null" refers to a domain that exhibits weak to no binding to any of the Fcgamma receptors. In some embodiments, an Fc null domain or region exhibits a reduction in binding affinity (e.g., increase in Kd) to Fc gamma receptors of at least 1000-fold.
[0487] The Fc domain may have one or more, two or more, three or more, or four or more amino acid substitutions that decrease binding of the Fc domain to an Fc receptor. In certain
embodiments, an Fc domain exhibits decreased binding to FcyRI (CD64), FcyRIIA (CD32), FcyRIIIA (CD 16a), FcyRIIIB (CD 16b), or any combination thereof. In order to decrease binding affinity of an Fc domain or region to an Fc receptor, the Fc domain or region may comprise one or more substitutions that has the effect of reducing the affinity of the Fc domain or region to an Fc receptor. In certain embodiments, the one or more substitutions comprise any one or more of IgGl heavy chain mutations corresponding to E233P, L234V, L234A, L235A, L235E, AG236, G237A, E318A, K320A, K322A, A327G, A330S, or P331S according to the EU index of Kabat numbering.
[0488] In some embodiments, the Fc domain or region can comprise a sequence of the IgGl isoform that has been modified from the wild-type IgGl sequence. A modification can comprise a substitution at more than one amino acid residue, such as at 5 different amino acid residues including L235V/F243L/R292P/Y300L/P396L (IgGlVLPLL) according to the EU index of
Kabat numbering. A modification can comprise a substitution at more than one amino acid residue such as at 2 different amino acid residues including S239D/I332E (IgGlDE) according to the EU index of Kabat numbering. A modification can comprise a substitution at more than one amino acid residue such as at 3 different amino acid residues including S298A/E333A/K334A
(IgGl AAA) according to the EU index of Kabat numbering. [0489] In some embodiments, the Fc domain or region can comprise a sequence of an IgG isoform that has been modified from the wild-type IgG sequence. In some embodiments, the Fc domain or region can comprise a sequence of the IgGl isoform that has been modified from the wild-type IgGl sequence. In some embodiments, the modification comprises substitution of one or more amino acids that reduce binding affinity of an IgG Fc domain or region to all Fey receptors. A modification can be substitution of E233, L234, and L235, such as
E233P/L234V/L235A or E233P/L234V/L235A/AG236, according to the EU index of Kabat. A modification can be substitution of L235, F243, R292, Y300 and P396, such as
L235V/F243L/R292P/Y300L/P396L (IgGlVLPLL) according to the EU index of Kabat. A modification can be a substitution of P238, such as P238A, according to the EU index of Kabat. A modification can be a substitution of D265, such as D265A, according to the EU index of Kabat. A modification can be a substitution of N297, such as N297A, according to the EU index of Kabat. A modification can be a substitution of A327, such as A327Q, according to the EU index of Kabat. A modification can be a substitution of P329, such as P239A, according to the EU index of Kabat.
[0490] In some embodiments, an IgG Fc domain or region comprises at least one amino acid substitution that reduces its binding affinity to FcyRl, as compared to a wild-type or reference IgG Fc domain. A modification can comprise a substitution at F241, such as F241A, according to the EU index of Kabat. A modification can comprise a substitution at F243, such as F243A, according to the EU index of Kabat. A modification can comprise a substitution at V264, such as V264A, according to the EU index of Kabat. A modification can comprise a substitution at D265, such as D265A according to the EU index of Kabat.
[0491] In some embodiments, an IgG Fc domain or region comprises at least one amino acid substitution that increases its binding affinity to FcyRl, as compared to a wild-type or reference IgG Fc domain. A modification can comprise a substitution at A327 and P329, such as
A327Q/P329A, according to the EU index of Kabat.
[0492] In some embodiments, the modification comprises substitution of one or more amino acids that reduce binding affinity of an IgG Fc domain or region to FcyRII and FcyRIIIA receptors. A modification can be a substitution of D270, such as D270A, according to the EU index of Kabat. A modification can be a substitution of Q295, such as Q295A, according to the EU index of Kabat. A modification can be a substitution of A327, such as A237S, according to the EU index of Kabat.
[0493] In some embodiments, the modification comprises substitution of one or more amino acids that increases binding affinity of an IgG Fc domain or region to FcyRII and FcyRIIIA receptors. A modification can be a substitution of T256, such as T256A, according to the EU index of Kabat. A modification can be a substitution of K290, such as K290A, according to the EU index of Kabat.
[0494] In some embodiments, the modification comprises substitution of one or more amino acids that increases binding affinity of an IgG Fc domain or region to FcyRII receptor. A modification can be a substitution of R255, such as R255A, according to the EU index of Kabat. A modification can be a substitution of E258, such as E258A, according to the EU index of Kabat. A modification can be a substitution of S267, such as S267A, according to the EU index of Kabat. A modification can be a substitution of E272, such as E272A, according to the EU index of Kabat. A modification can be a substitution of N276, such as N276A, according to the EU index of Kabat. A modification can be a substitution of D280, such as D280A, according to the EU index of Kabat. A modification can be a substitution of H285, such as H285A, according to the EU index of Kabat. A modification can be a substitution of N286, such as N286A, according to the EU index of Kabat. A modification can be a substitution of T307, such as T307A, according to the EU index of Kabat. A modification can be a substitution of L309, such as L309A, according to the EU index of Kabat. A modification can be a substitution of N315, such as N315A, according to the EU index of Kabat. A modification can be a substitution of K326, such as K326A, according to the EU index of Kabat. A modification can be a substitution of P331, such as P331A, according to the EU index of Kabat. A modification can be a substitution of S337, such as S337A, according to the EU index of Kabat. A modification can be a substitution of A378, such as A378A, according to the EU index of Kabat. A modification can be a substitution of E430, such as E430, according to the EU index of Kabat.
[0495] In some embodiments, the modification comprises substitution of one or more amino acids that increases binding affinity of an IgG Fc domain or region to FcyRII receptor and reduces the binding affinity to FcyRIIIA receptor. A modification can be a substitution of H268, such as H268A, according to the EU index of Kabat. A modification can be a substitution of R301, such as R301A, according to the EU index of Kabat. A modification can be a substitution of K322, such as K322A, according to the EU index of Kabat.
[0496] In some embodiments, the modification comprises substitution of one or more amino acids that decreases binding affinity of an IgG Fc domain or region to FcyRII receptor but does not affect the binding affinity to FcyRIIIA receptor. A modification can be a substitution of R292, such as R292A, according to the EU index of Kabat. A modification can be a substitution of K414, such as K414A, according to the EU index of Kabat.
[0497] In some embodiments, the modification comprises substitution of one or more amino acids that decreases binding affinity of an IgG Fc domain or region to FcyRII receptor and increases the binding affinity to FcyRIIIA receptor. A modification can be a substitution of S298, such as S298A, according to the EU index of Kabat. A modification can be substitution of S239, 1332 and A330, such as S239D/I332E/A330L. A modification can be substitution of S239 and 1332, such as S239D/I332E.
[0498] In some embodiments, the modification comprises substitution of one or more amino acids that decreases binding affinity of an IgG Fc domain or region to FcyRIIIA receptor and does not affect the binding affinity to FcyRII receptor. A modification can be a substitution of S239, such as S239A, according to the EU index of Kabat. A modification can be a substitution of E269, such as E269A, according to the EU index of Kabat. A modification can be a substitution of E293, such as E293A, according to the EU index of Kabat. A modification can be a substitution of Y296, such as Y296F, according to the EU index of Kabat. A modification can be a substitution of V303, such as V303A, according to the EU index of Kabat. A modification can be a substitution of A327, such as A327G, according to the EU index of Kabat. A
modification can be a substitution of K338, such as K338A, according to the EU index of Kabat. A modification can be a substitution of D376, such as D376A, according to the EU index of Kabat.
[0499] In some embodiments, the modification comprises substitution of one or more amino acids that increases binding affinity of an IgG Fc domain or region to FcyRIIIA receptor and does not affect the binding affinity to FcyRII receptor. A modification can be a substitution of E333, such as E333A, according to the EU index of Kabat. A modification can be a substitution of K334, such as K334A, according to the EU index of Kabat. A modification can be a substitution of A339, such as A339T, according to the EU index of Kabat. A modification can be substitution of S239 and 1332, such as S239D/I332E.
[0500] In some embodiments, an IgG Fc domain or region comprises at least one amino acid substitution that reduces the binding affinity to FcRn, as compared to a wild-type or reference
IgG Fc domain. A modification can comprise a substitution at H435, such as H435A according to the EU index of Kabat. A modification can comprise a substitution at 1253, such as 1253 A according to the EU index of Kabat. A modification can comprise a substitution at H310, such as
H310A according to the EU index of Kabat. A modification can comprise substitutions at 1253,
H310 and H435, such as I253A/H310A/H435A according to the EU index of Kabat.
[0501] A modification can comprise a substitution of one amino acid residue that increases the binding affinity of an IgG Fc domain for FcRn, relative to a wildtype or reference IgG Fc domain. A modification can comprise a substitution at V308, such as V308P according to the
EU index of Kabat. A modification can comprise a substitution at M428, such as M428L according to the EU index of Kabat. A modification can comprise a substitution at N434, such as
N434A according to the EU index of Kabat or N434H according to the EU index of Kabat. A modification can comprise substitutions at T250 and M428, such as T250Q and M428L according to the EU index of Kabat. A modification can comprise substitutions at M428 and N434, such as M428L and N434S, N434A or N434H according to the EU index of Kabat. A modification can comprise substitutions at M252, S254 and T256, such as M252Y/S254T/T256E according to the EU index of Kabat. A modification can be a substitution of one or more amino acids selected from P257L, P257N, P257I, V279E, V279Q, V279Y, A281S, E283F, V284E, L306Y, T307V, V308F, Q311V, D376V, and N434H. Other substitutions in an IgG Fc domain that affect its interaction with FcRn are disclosed in U.S. Patent No. 9,803,023 (the disclosure of which is incorporated by reference herein).
[0502] The antibody construct of a conjugate can be an anti-tumor antigen construct. The antibody construct can be an anti-tumor antigen antibody. An antigen recognized by the antibody construct can be CD5, CD19, CD20, CD25, CD37, CD30, CD33, CD45, CAMPATH-1, BCMA, CS-1, PD-L1, B7-H3, B7-DC (PD-L2), HLD-DR, carcinoembryonic antigen (CEA), TAG-72, EpCAM, MUC1, MUC15, MUC16, fo late-binding protein, A33, G250, pro state- specific membrane antigen (PSMA), ferritin, GD2, GD3, GM2, Ley, CA-125, CA19-9, epidermal growth factor, pl85HER2, IL-2 receptor, EGFRvIII (de2-7 EGFR), fibroblast activation protein (FAP), tenascin, a metalloproteinase, endosialin, vascular endothelial growth factor, ανβ3, WT1, LMP2, HPV E6 E7, Her-2/neu, p53 nonmutant, NY-ESO-1, MelanA/M ART 1 , Ras mutant, gplOO, p53 mutant, PR1, bcr-abl, tyrosinase, survivin, PSA, hTERT, a Sarcoma translocation breakpoint protein, EphA2, PAP, ML-IAP, AFP, ERG, NA17, PAX3, ALK, androgen receptor, cyclin Bl, polysialic acid, MYCN, RhoC, TRP-2, fucosyl GM1, mesothelin (MSLN), PSCA, MAGE Al, MAGE A3, sLe(animal), CYPIBI, PLAVl, GM3, BORIS, Tn, GloboH, ETV6-AML, NY-BR-1, RGS5, SART3, STn, Carbonic anhydrase IX, PAX5, OY-TES 1, Sperm protein 17, LCK, HMWMAA, AKAP-4, SSX2, XAGE 1, Legumain, Tie 3, Page4, VEGFR2, MAD-CT-1, PDGFR-B, MAD-CT-2, ROR2, TRAIL 1, MAGE A4, MAGE C2, GAGE, EGFR, CMET, HER3, EPCAM, CA6, NAPI2B, TROP2, Claudin-6 (CLDN6), Claudin-16 (CLDN16),
CLDN18.2, RON, LY6E, FRA, DLL3, PTK7, Uroplakin-IB (UPK1B), LIV1, ROR1, STRA6, TMPRSS3, TMPRSS4, TMEM238, Clorfl86, Fos-related antigen 1, VEGFR1, endoglin, PD- Ll, VTCN1 (B7-H4), VISTA, or any fragment thereof.
[0503] The antibody construct can recognize an antigen that can be expressed on a cell. The antibody construct can recognize an antigen that can be expressed by a cell. The antibody construct can recognize an antigen that can be expressed in the context of a Major
Histocompatibility Complex. The antibody construct can recognize an antigen that can stimulate activity of a cell, such as an immune cell. The antibody construct can recognize an antigen that can stimulate an immune response. The antibody construct can recognize an antigen that can reduce an immune response. The antibody construct can recognize an antigen that can reduce activity of a cell. The antibody construct can recognize an antigen that can be expressed on an immune cell. The antibody construct can recognize an antigen that can be expressed by an immune cell. The antibody construct can recognize an antigen that can be in the context of a Major Histocompatibility Complex. The antibody construct can recognize an antigen on a cell wherein the antigen can be involved in stimulating activity of a cell. The antibody construct can recognize an antigen on an immune cell that can be involved in the costimulation of an immune cell. The antibody construct can recognize an antigen on an immune cell that can be involved in the costimulation of an immune cell during an immune response. The antibody construct can recognize a receptor. The antibody construct can recognize a receptor on a cell. The antibody construct can recognize a receptor ligand. The antibody construct can recognize a receptor on a cell wherein the receptor can be involved in stimulating activity of a cell. The antibody construct can recognize a receptor on an immune cell. The antibody construct can recognize a receptor on an immune cell that can be involved in stimulating activity of an immune cell. The antibody construct can recognize a receptor on an immune cell that can be involved in the costimulation of an immune cell. The antibody construct can recognize a receptor on an immune cell that can be involved in the costimulation of an immune cell during an immune response. The antibody construct can recognize an antigen that can be expressed on an immune cell and that can stimulate activity of an immune cell. The antibody construct can recognize an antigen that can be expressed on an immune that can reduce activity of an immune cell.
[0504] The antibody construct can be an antibody that specifically binds to CEA, HER2, TROP2, Claudin-6 (CLDN6), Claudin-16 (CLDN16), CLDN18.2, RON, LY6E, FRA, DLL3, PTK7, Uroplakin-IB (UPK1B), LIV1, ROR1, STRA6, TMPRSS3, TMPRSS4, TMEM238, Clorfl86, Fos-related antigen 1, VEGFR1, endoglin, VTCN1 (B7-H4), VISTA, or gpNMB.
[0505] The antibody construct can be capable of recognizing a single antigen. The antibody construct can be capable of recognizing two or more antigens. The antibody construct can be capable of recognizing three or more antigens. The IQ for binding of a second binding domain of an immune- stimulatory conjugate to an antigen in the presence of an immune- stimulatory compound can be about 2 times, about 3 times, about 4 times, about 5 times, about 6 times, about 7 times, about 8 times, about 9 times, about 10 times, about 15 times, about 20 times, about 25 times, about 30 times, about 35 times, about 40 times, about 45 times, about 50 times, about 60 times, about 70 times, about 80 times, about 90 times, about 100 times, about 110 times, or about 120 times greater than the IQ for binding of the second binding domain to the antigen of an antibody construct in the absence of the immune- stimulatory compound. The Kd for binding of a second binding domain of an immune- stimulatory conjugate to an antigen in the presence of the immune- stimulatory compound can be less than 10 nM. The Kd for binding of a second binding domain of an immune- stimulatory conjugate to an antigen in the presence of the immune- stimulatory compound can be less than 100 nM, less than 50 nM, less than 20 nM, less than 5 nM, less than 1 nM, or less than 0.1 nM. In contrast, the Kd for binding of a second binding domain of an immune- stimulatory conjugate to an antigen in the presence of the immune- stimulatory compound when the first binding domain is bound to the first binding domain's antigen can be greater than 100 nM. The Kd for binding of a second binding domain of an immune- stimulatory conjugate to an antigen in the presence of the immune- stimulatory compound when the first binding domain is bound to the first binding domain's antigen can be greater than 100 nM, greater than 200 nM, greater than 300 nM, greater than 400 nM, greater than 500 nM, or greater than 1000 nM. The Kd for binding of a first binding domain of an immune- stimulatory conjugate to an antigen in the presence of an immune- stimulatory compound can be about 2 times, about 3 times, about 4 times, about 5 times, about 6 times, about 7 times, about 8 times, about 9 times, about 10 times, about 15 times, about 20 times, about 25 times, about 30 times, about 35 times, about 40 times, about 45 times, about 50 times, about 60 times, about 70 times, about 80 times, about 90 times, about 100 times, about 110 times, or about 120 times greater than the Kd for binding of the first binding domain to the antigen of an antibody construct in the absence of the immune- stimulatory compound. The Kd for binding of a first binding domain of an immune- stimulatory conjugate to an antigen in the presence of the immune- stimulatory compound can be less than 10 nM. The Kd for binding of a first binding domain of an immune- stimulatory conjugate to an antigen in the presence of the immune- stimulatory compound can be less than 100 nM, less than 50 nM, less than 20 nM, less than 5 nM, less than 1 nM, or less than 0.1 nM.
[0506] In some embodiments, an engineered cysteine is introduced in an antibody construct so that a linker can be attached at such engineered cysteine. For example an engineered cysteine can be introduced into an IgG (typically an IgGl) at Tl 14 (heavy chain), A 140 (heavy chain), L174 (heavy chain), L179 (heavy chain), T187 (heavy chain), T209 (heavy chain), S239 (heavy chain), V262 (heavy chain), G371 (heavy chain), Y373 (heavy chain), E382 (heavy chain), S400 (heavy chain), S424 (heavy chain), N434 (heavy chain), Q438 (heavy chain), 1106 (light chain), R108 (light chain), A118 (heavy chain), R142 (light chain), K149 (light chain) and/or V205 (light chain), according to the EU numbering of Kabat.
[0507] In certain embodiments, a linker or linker bound to an immune-modulatory compound disclosed herein may not be attached to an amino acid residue of an IgG Fc domain disclosed herein selected from: 221, 224, 227, 228, 230, 231, 223, 233, 234, 235, 236, 237, 238, 239, 240,
241, 243, 244, 245, 247, 249, 250, 258, 262, 263, 264, 265, 266, 267, 268, 269, 270, 271, 272, 273, 275, 276, 278, 280, 281, 283, 285, 286, 291, 292, 293, 294, 295, 296, 297, 298, 299, 300, 302, 305, 313, 318, 323, 324, 325, 327, 328, 329, 330, 331, 332, 333, 335, 336, 396, or 428, wherein numbering of amino acid residues in the Fc domain is according to the EU index as in Kabat.
[0508] In certain embodiments, a linker or linker bound to an immune-modulatory compound disclosed herein may be attached to an amino acid residue of an IgG Fc domain disclosed herein selected from: 221, 224, 227, 228, 230, 231, 223, 233, 234, 235, 236, 237, 238, 239, 240, 241, 243, 244, 245, 247, 249, 250, 258, 262, 263, 264, 265, 266, 267, 268, 269, 270, 271, 272, 273, 275, 276, 278, 280, 281, 283, 285, 286, 291, 292, 293, 294, 295, 296, 297, 298, 299, 300, 302, 305, 313, 318, 323, 324, 325, 327, 328, 329, 330, 331, 332, 333, 335, 336, 396, or 428, wherein numbering of amino acid residues in the Fc domain is according to the EU index as in Kabat.
[0509] The conjugates can have an Fc domain that can bind to an FcR when linked to an immune- stimulatory compound. The conjugate can have an Fc domain that can bind to an FcR to initiate FcR-mediated signaling when linked to an immune stimulatory compound. The conjugate can bind to its antigen when linked to an immune- stimulatory compound. The conjugate can bind to its antigen when linked to an immune- stimulatory compound and the Fc domain of the conjugate can bind to an FcR when linked to an immune- stimulatory compound. The conjugate can bind to its antigen when linked to an immune- stimulatory compound and the Fc domain of the antibody construct can bind to an FcR to initiate FcR-mediated signaling when linked to an immune stimulatory compound. The Fc domain linked to an immune- stimulatory compound can be a modified Fc domain as described above.
[0510] The Kd for binding of an Fc domain to a Fc receptor when the Fc domain is linked to an immune- stimulatory compound can be about 2 times, about 3 times, about 4 times, about 5 times, about 6 times, about 7 times, about 8 times, about 9 times, about 10 times, about 15 times, about
20 times, about 25 times, about 30 times, about 35 times, about 40 times, about 45 times, about
50 times, about 60 times, about 70 times, about 80 times, about 90 times, about 100 times, about
110 times, or about 120 times greater than the Kd for binding of the Fc domain to the Fc receptor in the absence of linking to the immune- stimulatory compound. The Kd for binding of an Fc domain to an Fc receptor when linked to an immune- stimulatory compound can be less than 10 nM. The Kd for binding of an Fc domain to an Fc receptor when linked to an immune- stimulatory compound can be less than 100 nM, less than 50 nM, less than 20 nM, less than 5 nM, less than 1 nM, or less than 0.1 nM. In contrast, the Kd for binding of an Fc domain to an Fc receptor when linked to an immune- stimulatory compound and when the first binding domain is bound to its antigen can be greater than 100 nM. The Kd for binding of an Fc domain to an Fc receptor when linked to an immune- stimulatory compound and when the first binding domain is bound to its antigen can be greater than 100 nM, greater than 200 nM, greater than 300 nM, greater than 400 nM, greater than 500 nM, or greater than 1000 nM.
[0511] The binding domain can be selected in order to recognize an antigen. For example, an antigen can be expressed on an immune cell. An antigen can be a peptide or fragment thereof. An antigen can be expressed on an antigen-presenting cell. An antigen can be expressed on a dendritic cell, a macrophage, or a B cell. An antigen can be CD40 and a binding domain can recognize a CD40 antigen. A binding domain can be a CD40 agonist. A binding domain can be CD40.
[0512] A conjugate can be formed by a linker that can connect an antibody construct to a molecule that binds a PRR. A conjugate can be formed by a linker that can connect an antibody construct to a PAMP molecule. A conjugate can be formed by a linker that can connect an antibody construct and a DAMP molecule. A conjugate can be formed by a linker that can connect an antibody construct to a molecule that binds a PRR, and a linker that can connect an antibody construct and a binding domain. A conjugate can be formed by a linker that can connect an antibody construct to a PAMP molecule, and a linker that can connect an antibody construct and a binding domain. A conjugate can be formed by a linker that can connect an antibody construct and a DAMP molecule, and a linker that can connect an antibody construct and a binding domain.
[0513] A conjugate can be formed by a linker that can attach an antibody construct to a kinase inhibitor, an antibody construct attached to a GPCR antagonist, an antibody construct attached to an ion channel antagonist, an antibody construct attached to a membrane transport inhibitor, an antibody construct to a phosphatase inhibitor, or an antibody construct attached to a kinase inhibitor.
[0514] A linker can be connected to an antibody construct by a direct linkage between the antibody construct and the linker. A linker can be connected to an anti-tumor antigen antibody construct by a direct linkage between the anti-tumor antigen antibody construct and the linker. A linker can be connected to an anti-tumor antigen antibody by a direct linkage between the antitumor antigen antibody and the linker. A direct linkage is a covalent bond. For example, a linker can be attached to a terminus of an amino acid sequence of an antibody construct, or could be attached to a side chain modification of the antibody construct, such as the side chain of a lysine, serine, threonine, cysteine, tyrosine, aspartic acid, a non-natural amino acid residue, glutamine, or glutamic acid residue. An attachment can be via any of a number of bonds, for example but not limited to, an amide bond, an ester bond, an ether bond, a carbon-nitrogen bond, a carbon- carbon single double or triple bond, a disulfide bond, or a thioether bond. A linker can have at least one functional group, which can be linked to the antibody construct. Non- limiting examples of the functional groups can include those which form an amide bond, an ester bond, an ether bond, a carbonate bond, a carbamate bond, or a thioether bond; such functional groups can be, for example, amino groups; carboxyl groups; aldehyde groups; azide groups; alkyne and alkene groups; ketones; carbonates; carbonyl functionalities bonded to leaving groups such as cyano and succinimidyl and hydroxyl groups. A linker can be attached to an antibody construct at an interchain disulfide. A linker can be connected to an antibody construct at a hinge cysteine. A linker can be connected to an antibody construct at an engineered cysteine. A linker can be connected to an antibody construct at a light chain constant domain lysine. A linker can be connected to an antibody construct at an engineered cysteine in the light chain. A linker can be connected to an antibody construct at an engineered light chain glutamine. A linker can be connected to an antibody construct at an unnatural amino acid engineered into the light chain. A linker can be connected to an antibody construct at a heavy chain constant domain lysine. A linker can be connected to an antibody construct at an engineered cysteine in the heavy chain. A linker can be connected to an antibody construct at an engineered heavy chain glutamine. A linker can be connected to an antibody construct at an unnatural amino acid engineered into the heavy chain. Amino acids can be engineered into an amino acid sequence of an antibody construct as described herein. Engineered amino acids can be added to a sequence of existing amino acids. Engineered amino acids can be substituted for one or more existing amino acids of a sequence of amino acids. A linker can be conjugated to an antibody construt via a sulfhydryl group. A linker can be conjugated to an antibody construct via a primary amine. A linker can be a link created between an unnatural amino acid on an antibody construct by reacting with oxime bond that was formed by modifying a ketone group with an alkoxyamine on an immune- stimulatory compound. When a linker is connected to an antibody construct at the sites described herein, an Fc domain of the construct can bind to Fc receptors. When a linker is connected to an antibody construct at the sites described herein, the antigen binding domain of the construct can bind its antigen. When a linker is connected to an antibody construct at the sites described herein, a binding domain of the construct can bind its antigen.
[0515] An antibody with engineered reactive cysteine residues (THIOMAB) can be used to link a binding domain to an antibody or to an antibody construct. A linker can connect an antibody construct to a binding domain via Sortase A linker. A Sortase A linker can be created by a Sortase A enzyme fusing an LPXTG recognition motif (SEQ ID NO: 672) to an N-terminal GGG motif to regenerate a native amide bond. The linker created can therefore link an antibody construct attached to the LPXTG recognition motif (SEQ ID NO: 672) with a binding domain attached to the N-terminal GGG motif. A binding domain can be connected to a linker by a direct linkage. A direct linkage can be a covalent bond. For example, a linker can be attached to a terminus of an amino acid sequence of a binding domain, or could be attached to a side chain modification to the binding domain, such as the side chain of a lysine, serine, threonine, cysteine, tyrosine, aspartic acid, a non-natural amino acid residue, glutamine, or glutamic acid residue. An attachment can be via any of a number of bonds, for example but not limited to, an amide bond, an ester bond, an ether bond, a carbon-nitrogen bond, a carbon-carbon single double or triple bond, a disulfide bond, or a thioether bond. A linker can have at least one functional group, which can be linked to the binding domain. Non- limiting examples of the functional groups can include those which form an amide bond, an ester bond, an ether bond, a carbonate bond, a carbamate bond, or a thioether bond; such functional groups can be, for example, amino groups; carboxyl groups; aldehyde groups; azide groups; alkyne and alkene groups; ketones; carbonates; and carbonyl functionalities bonded to leaving groups such as cyano and succinimidyl and hydroxyl groups. Amino acids can be engineered into an amino acid sequence of the binding domain. Engineered amino acids can be added to a sequence of existing amino acids. Engineered amino acids can be substituted for one or more existing amino acids of a sequence of amino acids. A linker can be conjugated to a binding domain via a sulfhydryl group. A linker can be conjugated to a binding domain via a primary amine. A binding domain can be conjugated to the C-terminal of an Fc domain of an antibody construct.
[0516] An antibody with engineered reactive cysteine residues (THIOMAB) can be used to link an immune- stimulatory compound to an antibody or antibody construct. A linker can connect an antibody construct to an immune- stimulatory compound via an mc-vc-PABC linker. A linker can connect an antibody construct to an immune- stimulatory compound via Sortase A linker. A
Sortase A linker can be created by a Sortase A enzyme fusing an LPXTG recognition motif (SEQ
ID NO: 672) to an N-terminal GGG motif to regenerate a native amide bond. The linker created can therefore link an antibody construct attached the LPXTG recognition motif (SEQ ID NO:
672) with an immune- stimulatory compound attached to the N-terminal GGG motif. A linker can be a link created between an unnatural amino acid on a construct by reacting with oxime bond that was formed by modifying a ketone group with an alkoxyamine on an immune- stimulatory compound. The immune- stimulatory compound can comprise one or more rings selected from carbocyclic and heterocyclic rings. The immune- stimulatory compound can be covalently bound to a linker by a bond to an exocyclic carbon or nitrogen atom on the immune- stimulatory compound. A linker can be attached to an immune- stimulatory compound via an exocyclic nitrogen or carbon atom of an immune- stimulatory compound. The Kd for binding of a binding domain of any of these conjugates to its antigen can be about 2 times, about 3 times, about 4 times, about 5 times, about 6 times, about 7 times, about 8 times, about 9 times, about 10 times, about 15 times, about 20 times, about 25 times, about 30 times, about 35 times, about 40 times, about 45 times, about 50 times, about 60 times, about 70 times, about 80 times, about 90 times, about 100 times, about 110 times, or about 120 times greater than the Kd for binding of the binding domain to its antigen in the absence of the immune- stimulatory compound. The Kd for binding of the binding domain of any of these conjugates to its antigen can be less than 10 nM. The Kd for binding of the binding domain of any of the conjugates to its antigen can be less than 100 nM, less than 50 nM, less than 20 nM, less than 5 nM, less than 1 nM, or less than 0.1 nM. The Kd for binding of the Fc domain of any of the conjugates to an Fc receptor can be about 2 times, about 3 times, about 4 times, about 5 times, about 6 times, about 7 times, about 8 times, about 9 times, about 10 times, about 15 times, about 20 times, about 25 times, about 30 times, about 35 times, about 40 times, about 45 times, about 50 times, about 60 times, about 70 times, about 80 times, about 90 times, about 100 times, about 110 times, or about 120 times greater than the Kd for binding of the Fc domain to the Fc receptor in the absence of the immune- stimulatory compound. The Kd for binding of the Fc domain of any of the conjugates to an Fc receptor can be less than 10 nM. The Kd for binding of the Fc domain of any of the conjugates to an Fc receptor can be less than 100 nM, less than 50 nM, less than 20 nM, less than 5 nM, less than 1 nM, or less than 0.1 nM.
[0517] In some embodiments, in a conjugate, an antibody can be linked to an immune- stimulatory compound in such a way that the antibody construct, such as an antibody, can still bind to an antigen and the Fc domain of the antibody construct, can still bind to an FcR. In a conjugate, an antibody construct is linked to an immune- stimulatory compound in such a way that the linking does not interfere with ability of the antigen binding domain of the antibody construct to bind to antigen, the ability of the Fc domain of the antibody construct to bind to an FcR, or FcR-mediated signaling resulting from the Fc domain of the antibody construct from binding to an FcR. In a conjugate, an immune- stimulatory compound can be linked to an antibody construct in such a way the linking does not interfere with the ability of the immune- stimulatory compound to bind to its receptor. A conjugate can produce stronger immune stimulation and a greater therapeutic window than components of the conjugate alone.
Pharmaceutical Formulations
[0518] The conjugates and methods described herein can be useful as pharmaceutical compositions for administration to a subject in need thereof. Pharmaceutical compositions can comprise at least the conjugates described herein and one or more pharmaceutically acceptable carriers, diluents, excipients, stabilizers, dispersing agents, suspending agents, and/or thickening agents. The pharmaceutical composition can comprise a conjugate having an antibody construct and an immune stimulatory compound, scuh as an agonist. A pharmaceutical composition can comprise a conjugate having an antibody construct having a binding domain, and an immune- stimulatory compound. A pharmaceutical composition can comprise any conjugate described herein. In some embodiments, a pharmaceutical composition can comprise an antibody to a tumor antigen and an immune stimulatory compound selected from a toll- like receptor agonist, a STING agonist, RIG-I agonist, PAMP agonist, DAMP agonist and a kinase inhibitor. In some embodiments, a pharmaceutical composition can comprise an antibody to a tumor antigen and an immune stimulatory compound selected from a tankyrase inhibitor and a TNIK inhibitor.
[0519] In some embodiments, the antibody construct is an anti-HER2 antibody, an anti-TROP2 antibody, an anti-CEA antibody, an anti-claudin-6 (CLDN6) antibody, an anti-Claudin-16 (CLD 16) antibody, an anti-CLD 18.2 antibody, an anti-RON antibody, an anti-LY6E antibody, an anti-FRA antibody, an anti-DLL3 antibody, an anti-TK7 antibody, an anti- Uroplakin-IB (UPK1B) antibody, an anti-LIVl antibody, an anti-RORl antibody, an anti- STRA6 antibody, an anti-TMPRSS3 antibody, an anti-TMPRSS4 antibody, an anti-TMEM238 antibody, an anti-Cl or fl86 antibody, an anti-Fos-related antigen 1 antibody, an anti-VEGFRl antibody, an anti-endoglin antibody, an anti-VTCNl (B7-H4) antibody, an anti- VISTA antibody or an anti-gpNMB antibody. A conjugate can comprise an antibody and a PAMP molecule. A conjugate can comprise an antibody and a DAMP molecule. A pharmaceutical composition can further comprise buffers, antibiotics, steroids, carbohydrates, drugs (e.g., chemotherapy drugs), radiation, polypeptides, chelators, adjuvants, and/or preservatives.
[0520] In some embodiments, a pharmaceutical composition comprises an immune-modulatory conjugate comprising an antibody construct, a proteolysis targeting module and a linker covalently linking the antibody construct and the proteolysis targeting module. The antibody construct can comprise an antibody to a tumor antigen or an immune cell antigen.
[0521] Pharmaceutical compositions can be formulated using one or more physiologically- acceptable carriers comprising excipients and auxiliaries. Formulation can be modified depending upon the route of administration chosen. Pharmaceutical compositions comprising a conjugate as described herein can be manufactured, for example, by lyophilizing the conjugate, mixing, dissolving, emulsifying, encapsulating, or entrapping the conjugate. The pharmaceutical compositions can also include the conjugates described herein in a free-base form or
pharmaceutically-acceptable salt form.
[0522] Methods for formulation of the conjugates described herein can include formulating any of the conjugates described herein with one or more inert, pharmaceutically-acceptable excipients or carriers to form a solid, semi-solid, or liquid composition. Solid compositions can include, for example, powders, tablets, dispersible granules and capsules, and in some aspects, the solid compositions further contain nontoxic, auxiliary substances, for example wetting or emulsifying agents, H buffering agents, and other pharmaceutically-acceptable additives. Alternatively, the pharmaceutical compositions described herein can be lyophilized or in powder form for re- constitution with a suitable vehicle, e.g., sterile pyrogen-free water, before use.
[0523] Pharmaceutical compositions of the conjugates described herein can comprise at least an active ingredient (i.e., a conjugate). The active ingredients can be entrapped in microcapsules prepared, for example, by coacervation techniques or by interfacial polymerization (e.g., hydroxymethylcellulose or gelatin microcapsules and poly-(methylmethacylate) microcapsules, respectively), in colloidal drug-delivery systems (e.g., liposomes, albumin microspheres, microemulsions, nano-particles and nanocapsules) or in macroemulsions.
[0524] Pharmaceutical compositions as described herein often further can comprise more than one active compound as necessary for the particular indication being treated. The active compounds can have complementary activities that do not adversely affect each other. For example, the composition can comprise a conjugate and a checkpoint inhibitor, chemotherapeutic agent, cytotoxic agent, cytokine, growth-inhibitory agent, anti-hormonal agent, anti-angiogenic agent, and/or cardioprotectant. Such molecules can be present in combination in amounts that are effective for the purpose intended.
[0525] The compositions and formulations can be sterilized. Sterilization can be accomplished by filtration through sterile filtration.
[0526] The pharmaceutical compositions described herein can be formulated for administration as an injection. Non-limiting examples of formulations for injection can include a sterile suspension, solution or emulsion in oily or aqueous vehicles. Suitable oily vehicles can include, but are not limited to, lipophilic solvents or vehicles such as fatty oils or synthetic fatty acid esters, or liposomes. Aqueous injection suspensions can contain substances which increase the viscosity of the suspension. The suspension can also contain suitable stabilizers. Injections can be formulated for bolus injection or continuous infusion. Alternatively, the compositions described herein can be lyophilized or in powder form for reconstitution with a suitable vehicle, e.g., sterile pyrogen-free water, before use.
[0527] For parenteral administration the conjugates can be formulated in a unit dosage injectable form (e.g., a solution, suspension or emulsion) in association with a pharmaceutically acceptable parenteral vehicle. Such vehicles can be inherently nontoxic, and no n- therapeutic. A vehicle can be water, saline, Ringer's solution, dextrose solution, and 5% human serum albumin.
Nonaqueous vehicles such as fixed oils and ethyl oleate can also be used. Liposomes can be used as carriers. The vehicle can contain minor amounts of additives such as substances that enhance isotonicity and chemical stability (e.g., buffers and preservatives). [0528] Sustained-release preparations can also be prepared. Examples of sustained-release preparations can include semipermeable matrices of solid hydrophobic polymers that can contain the antibody, and these matrices can be in the form of shaped articles (e.g., films or
microcapsules). Examples of sustained-release matrices can include polyesters, hydrogels (e.g., poly(2-hydroxyethyl-methacrylate), or poly(vinylalcohol)), polylactides, copolymers of L- glutamic acid and γ ethyl-L-glutamate, non-degradable ethylene- vinyl acetate, degradable lactic acid-glycolic acid copolymers such as the LUPRON DEPO™ (i.e., injectable microspheres composed of lactic acid-glycolic acid copolymer and leuprolide acetate), and poly-D-( - )-3- hydroxybutyric acid.
[0529] Pharmaceutical formulations of the compositions described herein can be prepared for storage by mixing a conjugate with a pharmaceutically acceptable carrier, excipient, and/or a stabilizer. This formulation can be a lyophilized formulation or an aqueous solution. Acceptable carriers, excipients, and/or stabilizers can be nontoxic to recipients at the dosages and
concentrations used. Acceptable carriers, excipients, and/or stabilizers can include buffers such as phosphate, citrate, and other organic acids; antioxidants including ascorbic acid and methionine; preservatives, polypeptides; proteins, such as serum albumin or gelatin; hydrophilic polymers; amino acids; monosaccharides, disaccharides, and other carbohydrates including glucose, mannose, or dextrins; chelating agents such as EDTA; sugars such as sucrose, mannitol, trehalose or sorbitol; salt-forming counter-ions such as sodium; metal complexes; and/or non- ionic surfactants or polyethylene glycol.
Therapeutic Applications
[0530] The pharmaceutical compositions, conjugates and methods of the present disclosure can be useful for a plurality of different subjects including, but are not limited to, a mammal, human, non-human mammal, a domesticated animal (e.g., laboratory animals, household pets, or livestock), non-domesticated animal (e.g., wildlife), dog, cat, rodent, mouse, hamster, cow, bird, chicken, fish, pig, horse, goat, sheep, rabbit, and any combination thereof.
[0531] The compositions, conjugates and methods described herein can be useful as a
therapeutic, for example a treatment that can be administered to a subject in need thereof. A therapeutic effect of the present disclosure can be obtained in a subject by reduction, suppression, remission, or eradication of a disease state, including, but not limited to, a symptom thereof. A therapeutic effect in a subject having a disease or condition, or pre-disposed to have or is beginning to have the disease or condition, can be obtained by a reduction, a suppression, a prevention, a remission, or an eradication of the condition or disease, or pre-condition or pre- disease state. [0532] In practicing the methods described herein, therapeutically-effective amounts of the compositions or conjugates described herein can be administered to a subject in need thereof, often for treating and/or preventing a condition or progression thereof. A pharmaceutical composition can affect the physiology of the subject, such as the immune system, inflammatory response, or other physiologic affect. A therapeutically-effective amount can vary widely depending on the severity of the disease, the age and relative health of the subject, the potency of the compounds used, and other factors.
[0533] Treat and/or treating can refer to any indicia of success in the treatment or amelioration of the disease or condition. Treating can include, for example, reducing, delaying or alleviating the severity of one or more symptoms of the disease or condition, or it can include reducing the frequency with which symptoms of a disease, defect, disorder, or adverse condition, and the like, are experienced by a patient. Treat can be used herein to refer to a method that results in some level of treatment or amelioration of the disease or condition, and can contemplate a range of results directed to that end, including but not restricted to prevention of the condition entirely.
[0534] Prevent, preventing and the like can refer to the prevention of the disease or condition, e.g. , tumor formation, in the patient. For example, if an individual at risk of developing a tumor or other form of cancer is treated with the methods of the present disclosure and does not later develop the tumor or other form of cancer, then the disease has been prevented, at least over a period of time, in that individual. Prevent can also refer to preventing relapse in a patient.
[0535] A therapeutically effective amount can be the amount of a composition or an active component (i.e., a conjugate) thereof sufficient to provide a beneficial effect or to otherwise reduce a detrimental non-beneficial event to the individual to whom the composition is administered. A therapeutically effective dose can be a dose that produces one or more desired or desirable (e.g., beneficial) effects for which it is administered, such administration occurring one or more times over a given period of time. An exact dose can depend on the purpose of the treatment, and can be ascertainable by one skilled in the art using known techniques.
[0536] The conjugates described herein that can be used in therapy can be formulated and dosages established in a fashion consistent with good medical practice taking into account the disorder to be treated, the condition of the individual patient, the site of delivery of the composition, the method of administration and other factors known to practitioners. The conjugates described herein can be prepared according to the description of preparation described herein.
[0537] Pharmaceutical compositions of the conjugates can be considerd useful with the methods described herein and can be administered to a subject in need thereof using a technique known to one of ordinary skill in the art which can be suitable as a therapy for the disease or condition affecting the subject. One of ordinary skill in the art would understand that the amount, duration and frequency of administration of a pharmaceutical composition described herein to a subject in need thereof depends on several factors including, for example but not limited to, the health of the subject, the specific disease or condition of the patient, the grade or level of a specific disease or condition of the patient, the additional therapeutics the subject is being or has been
administered, and the like.
[0538] The methods and compositions described herein can be for administration to a subject in need thereof. Often, administration of the compositions described herein can include routes of administration, non- limiting examples of administration routes include intravenous, intraarterial, subcutaneous, subdural, intramuscular, intracranial, intrasternal, intratumoral, or
intraperitoneally. Additionally, a pharmaceutical composition can be administered to a subject by additional routes of administration, for example, by inhalation, oral, dermal, intranasal, or intrathecal administration.
[0539] Compositions of the present disclosure can be administered to a subject in need thereof in a first administration, and in one or more additional administrations. The one or more additional administrations can be administered to the subject in need thereof minutes, hours, days, weeks or months following the first administration. Any one of the additional administrations can be administered to the subject in need thereof every 21 days, every 14 days, every 10 days, every 7 days, every 4 days or every day after the first administration over the course of a cycle. The additional administrations can also be administered to the subject in need thereof less than 21 days, or less than 14 days, less than 10 days, less than 7 days, less than 4 days, or less than 1 day after the first administration. The one or more administrations can occur more than once per day, more than once per week or more than once per month. Cycles of administration of a
pharmaceutical composition can be weekly, biweekly, every three weeks, monthly, every six weeks, every two months, or the like.
[0540] Suitable doses of an immune- stimulatory conjugate with a DAR of 1-8 can be from about 0.05 mg/kg to about 20 mg/kg or about 0.5 mg/kg to about 20 mg/kg, or about 1 mg/kg to about 10 mg/kg. Doses of conjugates with greater DARs can be scaled accordingly.
Increased dosages and reduced side-effects
[0541] In certain embodiments, the conjugates can comprise two binding domains, one of which binds to a tumor antigen and another of which binds to a target on an immune cell such as an antigen presenting cell and can be administered in a dosage that is about 10%, about 25%, about 50%, about 100% or greater than an antibody from which one of the tumor antigen binding domains or binding domains that bind to an antigen on an immune cell, such as an antigen presenting cell (APC), is derived. For example, a common regimen for administering pertuzumab comprises 840 mg intravenous (IV) administered as an initial dose over 60 minutes, followed every 3 weeks thereafter by 420 mg IV over 30 to 60 minutes. An initial dosage of a conjugate having a binding domain of pertuzumab and a second binding domain binding to an immune cell antigen can range from 900 mg to 1700 mg or more and a maintenance dose can range from 450 mg to 900 mg or more. An increased initial dose and/or maintenance dose can be used with an immune stimulatory conjugate or pharmaceutical composition of this disclosure, such as bispecific tumor targeting compositions comprising a tumor antigen binding domain that binds a tumor antigen with at least 80% or 100% sequence identity to CD5, CD19, CD20, CD25, CD37, CD30, CD33, CD45, CAMPATH-1, BCMA, CS-1, PD-L1, B7-H3, B7-DC (PD-L2), HLD-DR, carcinoembryonic antigen (CEA), TAG-72, EpCAM, MUC1, MUC15, MUC16, fo late-binding protein, A33, G250, pro state- specific membrane antigen (PSMA), ferritin, GD2, GD3, GM2, Ley, CA-125, CA19-9, epidermal growth factor, pl85HER2, IL-2 receptor, EGFRvIII (de2-7 EGFR), fibroblast activation protein (FAP), tenascin, a metalloproteinase, endosialin, vascular endothelial growth factor, ανβ3, WT1, LMP2, HPV E6 E7, Her-2/neu, p53 nonmutant, NY-ESO-1,
MelanA/MARTl, Ras mutant, gplOO, p53 mutant, PR1, bcr-abl, tyrosinase, survivin, PSA, hTERT, a Sarcoma translocation breakpoint protein, EphA2, PAP, ML-IAP, AFP, ERG, NA17, PAX3, ALK, androgen receptor, cyclin Bl, polysialic acid, MYCN, RhoC, TRP-2, fucosyl GMl, mesothelin (MSLN), PSCA, MAGE Al, MAGE A3, sLe(animal), CYP1B1, PLAV1, GM3, BORIS, Tn, GloboH, ETV6-AML, NY-BR-1, RGS5, SART3, STn, Carbonic anhydrase IX, PAX5, OY-TES 1, Sperm protein 17, LCK, HMWMAA, AKAP-4, SSX2, XAGE 1, Legumain, Tie 3, VEGFR2, MAD-CT-1, PDGFR-B, MAD-CT-2, ROR2, , TRAIL 1, MAGE A4, MAGE C2, GAGE, EGFR, CMET, HER3, EPCAM, CA6, NAPI2B, TROP2, Claudin-6 (CLDN6), Claudin-16 (CLDN16), CLDN18.2, RON, LY6E, FRA, DLL3, PTK7, Uroplakin-IB (UPK1B), LIV1, ROR1, STRA6, TMPRSS3, TMPRSS4, TMEM238, Clorfl86, Fos-related antigen 1, VEGFR1, endoglin, PD-L1, VTCN1 (B7-H4), VISTA, or any fragment thereof. Antibodies that bind costimulatory molecules or other cell surface molecules on APCs can have small therapeutic windows and high dose-limiting toxicity. For example, CP-870,893 can be shown to have a maximum tolerated dosage of 0.2 mg/kg to 0.3 mg/kg. Using a bispecific immune- stimulatory conjugate of this disclosure can allow administration of a conjugate at greater than 0.2 mg/kg to 0.3 mg/kg that comprises a binding domain derived from CP-870,893 or any binding domain that binds to an antigen on an APC with at least 80% or 100% sequence identity to CD40, CD47, TNFR2, DEC-205, PD-L1, CD36 mannose scavenger receptor 1, DC-SIGN, CLEC9A,
CLEC12A, BDCA-2, OX40L, 41BBL, CD204, MARCO, CLEC5A, Dectin 1, Dectin 2,
CLECIOA, CD206, CD64, CD32A, CD 16 A, HVEM, CD38, CD32B, or a fragment thereof. [0542] In certain embodiments, using an immune- stimulatory conjugate of this disclosure can allow administration of a conjugate that comprises a binding domain that binds to an antigen on an antigen presenting cell at greater levels than an antibody alone comprising a binding domain that binds to that molecule on an antigen presenting cell. In certain embodiments, a bispecific immune- stimulatory conjugate that comprises a binding domain that binds to an antigen on an antigen presenting cell can be administered at levels equivalent to that of an antibody from which the binding domain is derived.
[0543] In certain embodiments, the immune- stimulatory conjugate can be administered at a dose higher than the maximum tolerated dose for that immune- stimulatory compound administered in the absence of attachment to an antibody construct.
[0544] In certain embodiments, administration of an immune stimulatory conjugate can be associated with fewer side effects than an antibody from which one of binding domains is derived. In certain embodiments, administration of the immune stimulatory conjugate can be associated with fewer side effects than when the immune- stimulatory compound is administered alone. In certain embodiments, administration of immune- stimulatory conjugate with a chemotherapeutic agent can be associated with fewer side effects than when the
chemotherapeutic agent is administered with the immune- stimulatory compound. In certain embodiments, the chemotherapeutic agent can be an alkylating agent (e.g., cyclophosphamide, ifosfamide, chlorambucil, busulfan, melphalan, mechlorethamine, uramustine, thiotepa, nitrosoureas, or temozolomide), an anthracycline (e.g., doxorubicin, adriamycin, daunorubicin, epirubicin, or mitoxantrone), a cytoskeletal disruptor (e.g., paclitaxel or docetaxel), a histone deacetylase inhibitor (e.g., vorinostat or romidepsin), an inhibitor of topoisomerase (e.g., irinotecan, topotecan, amsacrine, etoposide, or teniposide), a kinase inhibitor (e.g., bortezomib, erlotinib, gefitinib, imatinib, vemurafenib, or vismodegib), a nucleoside analog or precursor analog (e.g., azacitidine, azathioprine, capecitabine, cytarabine, fluorouracil, gemcitabine, hydroxyurea, mercaptopurine, methotrexate, or thioguanine), a peptide antibiotic (e.g., actinomycin or bleomycin), a platinum-based agent (e.g., cisplatin, oxaloplatin, or carboplatin), or a plant alkaloid (e.g., vincristine, vinblastine, vinorelbine, vindesine, podophyllotoxin, paclitaxel, or docetaxel). In some embodiments, the chemotherapeutic agent can be a nucleoside analog. In some embodiments, the chemotherapeutic agent can be gemcitabine. In certain embodiments, the additional therapeutic agent can be radiation therapy.
Diseases, Conditions and the Like
[0545] The pharmaceutical compositions, conjugates and methods provided herein can be useful for the treatment of a plurality of diseases, conditions, preventing a disease or a condition in a subject, preventing relapse of a disease or condition or other therapeutic applications for subjects in need thereof. Often the compositions, conjugates and methods provided herein can be useful for treatment of hyperplastic conditions, including but not limited to, neoplasms, cancers, tumors and the like.
[0546] A condition, such as a cancer, can be associated with expression of an antigen on the cancer cells. Often, the molecule expressed by the cancer cells can comprise an extracellular portion capable of recognition by a binding domain (e.g., an antibody or antigen-binding portion thereof) of the conjugate. An antigen expressed by the cancer cells can be a tumor antigen. A binding domain of the conjugate can recognize a tumor antigen. A tumor antigen can include for example, CD5, CD19, CD20, CD25, CD37, CD30, CD33, CD45, CAMPATH-1, BCMA, CS-1, PD-Ll, B7-H3, B7-DC (PD-L2), HLD-DR, carcinoembryonic antigen (CEA), TAG-72, EpCAM, MUC1, MUC15, MUC16, fo late-binding protein, A33, G250, pro state- specific membrane antigen (PSMA), ferritin, GD2, GD3, GM2, Ley, CA-125, CA19-9, epidermal growth factor, pl85HER2, IL-2 receptor, EGFRvIII (de2-7 EGFR), fibroblast activation protein (FAP), tenascin, a metalloproteinase, endosialin, vascular endothelial growth factor, ανβ3, WT1, LMP2, HPV E6 E7, Her-2/neu, p53 nonmutant, NY-ESO-1, MelanA/M ART 1 , Ras mutant, gplOO, p53 mutant, PR1, bcr-abl, tyrosinase, survivin, PSA, hTERT, a Sarcoma translocation breakpoint protein, EphA2, PAP, ML-IAP, AFP, ERG, NA17, PAX3, ALK, androgen receptor, cyclin Bl, polysialic acid, MYCN, RhoC, TRP-2, fucosyl GM1, mesothelin (MSLN), PSCA, MAGE Al, MAGE A3, sLe(animal), CYPIBI, PLAVl, GM3, BORIS, Tn, GloboH, ETV6-AML, NY-BR-1, RGS5, SART3, STn, Carbonic anhydrase IX, PAX5, OY-TES 1, Sperm protein 17, LCK, HMWMAA, AKAP-4, SSX2, XAGE 1, Legumain, Tie 3, VEGFR2, MAD-CT-1, PDGFR-B, MAD-CT-2, ROR2, , TRAILl, MAGE A4, MAGE C2, GAGE, EGFR, CMET, HER3, EPCAM, CA6, NAPI2B, TROP2, Claudin-6 (CLDN6), Claudin-16 (CLDN16), CLDN18.2, RON, LY6E, FRA, DLL3, PTK7, Uroplakin-IB (UPK1B), LIV1, ROR1, STRA6, TMPRSS3, TMPRSS4, TMEM238, Clorfl86, Fos-related antigen 1, VEGFR1, endoglin, PD-Ll, VTCN1 (B7-H4), VISTA, gpNMB, or any fragment thereof.
[0547] As described herein, an antigen binding domain portion of the conjugate can be configured to recognize an antigen expressed by a cancer cell, such as for example, a disease antigen, tumor antigen or a cancer antigen. Such antigens are known to those of ordinary skill in the art, or can be newly found to be associated with such a disease or condition, to be commonly associated with, and/or, specific to, such disease or condition. For example, a disease antigen, tumor antigen or a cancer antigen is, but is not limited to, CD5, CD19, CD20, CD25, CD37, CD30, CD33, CD45, CAMPATH-1, BCMA, CS-1, PD-Ll, B7-H3, B7-DC (PD-L2), HLD-DR, carcinoembryonic antigen (CEA), TAG-72, EpCAM, MUC1, MUC15, MUC16, fo late-binding protein, A33, G250, pro state- specific membrane antigen (PSMA), ferritin, GD2, GD3, GM2, Ley, CA-125, CA19-9, epidermal growth factor, pl85HER2, IL-2 receptor, EGFRvIII (de2-7 EGFR), fibroblast activation protein (FAP), tenascin, a metalloproteinase, endosialin, vascular endothelial growth factor, ανβ3, WT1, LMP2, HPV E6 E7, Her-2/neu, p53 nonmutant, NY-ESO-1,
MelanA/MARTl, Ras mutant, gplOO, p53 mutant, PR1, bcr-abl, tyrosinase, survivin, PSA, hTERT, a Sarcoma translocation breakpoint protein, EphA2, PAP, ML-IAP, AFP, ERG, NA17, PAX3, ALK, androgen receptor, cyclin Bl, polysialic acid, MYCN, RhoC, TRP-2, fucosyl GMl, mesothelin (MSLN), PSCA, MAGE Al, MAGE A3, sLe(animal), CYP1B1, PLAV1, GM3, BORIS, Tn, GloboH, ETV6-AML, NY-BR-1, RGS5, SART3, STn, Carbonic anhydrase IX, PAX5, OY-TES 1, Sperm protein 17, LCK, HMWMAA, AKAP-4, SSX2, XAGE 1, Legumain, Tie 3, VEGFR2, MAD-CT-1, PDGFR-B, MAD-CT-2, ROR2, , TRAIL 1, MAGE A4, MAGE C2, GAGE, EGFR, CMET, HER3, EPCAM, CA6, NAPI2B, TROP2, Claudin-6 (CLDN6), Claudin-16 (CLDN16), CLDN18.2, RON, LY6E, FRA, DLL3, PTK7, Uroplakin-IB (UPK1B), LIV1, ROR1, STRA6, TMPRSS3, TMPRSS4, TMEM238, Clorfl86, Fos-related antigen 1, VEGFR1, endoglin, PD-L1, VTCN1 (B7-H4), VISTA, or any fragment thereof.
[0548] Additionally, such tumor antigens can be associated with the following specific conditions and/or families of diseases or conditions, including but not limited to, cancers such as brain cancers, skin cancers, lymphomas, sarcomas, lung cancer, liver cancer, leukemias, uterine cancer, breast cancer, ovarian cancer, cervical cancer, bladder cancer, kidney cancer,
hemangio sarcomas, bone cancers, blood cancers, testicular cancer, prostate cancer, stomach cancer, intestinal cancers, pancreatic cancer, and other types of cancers as well as pre-cancerous conditions such as hyperplasia or the like.
[0549] Non- limiting examples of cancers can include Acute lymphoblastic leukemia (ALL);
Acute myeloid leukemia; Adrenocortical carcinoma; Astrocytoma, childhood cerebellar or cerebral; Basal-cell carcinoma; Bladder cancer; Bone tumor, osteo sarcoma/malignant fibrous histiocytoma; Brain cancer; Brain tumors, such as, cerebellar astrocytoma, malignant glioma, ependymoma, medulloblastoma, visual pathway and hypothalamic glioma; Brainstem glioma;
Breast cancer; Bronchial adenomas/carcinoids; Burkitt's lymphoma; Cerebellar astrocytoma;
Cervical cancer; Cholangiocarcinoma; Chondrosarcoma; Chronic lymphocytic leukemia;
Chronic myelogenous leukemia; Chronic myeloproliferative disorders; Colon cancer; Cutaneous
T-cell lymphoma; Endometrial cancer; Ependymoma; Esophageal cancer; Eye cancers, such as, intraocular melanoma and retinoblastoma; Gallbladder cancer; Glioma; Hairy cell leukemia;
Head and neck cancer; Heart cancer; Hepatocellular (liver) cancer; Hodgkin lymphoma;
Hypopharyngeal cancer; Islet cell carcinoma (endocrine pancreas); Kaposi sarcoma; Kidney cancer (renal cell cancer); Laryngeal cancer; Leukaemia, such as, acute lymphoblastic, acute myeloid, chronic lymphocytic, chronic myelogenous and, hairy cell; Lip and oral cavity cancer; Liposarcoma; Lung cancer, such as, non-small cell and small cell; Lymphoma, such as, AIDS- related, Burkitt; Lymphoma, cutaneous T-Cell, Hodgkin and Non-Hodgkin, Macroglobulinemia, Malignant fibrous histiocytoma of bone/osteosarcoma; Melanoma; Merkel cell cancer;
Mesothelioma; Multiple myeloma/plasma cell neoplasm; Mycosis fungoides; Myelodysplastic syndromes; Myelodysplastic/myeloproliferative diseases; Myeloproliferative disorders, chronic; Nasal cavity and paranasal sinus cancer; Nasopharyngeal carcinoma; Neuroblastoma;
Oligodendroglioma; Oropharyngeal cancer; Osteosarcoma/malignant fibrous histiocytoma of bone; Ovarian cancer; Pancreatic cancer; Parathyroid cancer; Pharyngeal cancer;
Pheochromocytoma; Pituitary adenoma; Plasma cell neoplasia; Pleuropulmonary blastoma; Prostate cancer; Rectal cancer; Renal cell carcinoma (kidney cancer); Renal pelvis and ureter, transitional cell cancer; Rhabdomyosarcoma; Salivary gland cancer; Sarcoma, Ewing family of tumors; Sarcoma, Kaposi; Sarcoma, soft tissue; Sarcoma, uterine; Sezary syndrome; Skin cancer (non- melanoma); Skin carcinoma; Small intestine cancer; Soft tissue sarcoma; Squamous cell carcinoma; Squamous neck cancer with occult primary, metastatic; Stomach cancer; Testicular cancer; Throat cancer; Thymoma and thymic carcinoma; Thymoma,; Thyroid cancer; Thyroid cancer, childhood; Uterine cancer; Vaginal cancer; Waldenstrom macroglobulinemia; Wilms tumor; and any combination thereof.
[0550] The invention provides any therapeutic compound or conjugate disclosed herein for use in a method of treatment of the human or animal body by therapy. Therapy may be by any mechanism disclosed herein, such as by stimulation of the immune system. The invention provides any therapeutic compound or conjugate disclosed herein for use in stimulation of the immune system, vaccination or immunotherapy, including for example enhancing an immune response. The invention further provides any therapeutic compound or conjugate disclosed herein for prevention or treatment of any condition disclosed herein, for example cancer, autoimmune disease, inflammation, sepsis, allergy, asthma, graft rejection, graft-versus-host disease, immunodeficiency or infectious disease (typically caused by an infectious pathogen). The invention also provides any therapeutic compound or conjugate disclosed herein for obtaining any clinical outcome disclosed herein for any condition disclosed herein, such as reducing tumour cells in vivo. The invention also provides use of any therapeutic compound or conjugate disclosed herein in the manufacture of a medicament for preventing or treating any condition disclosed herein. Embodiments:
1. An immune- stimulatory conjugate comprising:
(a) an immune- stimulatory compound that optionally binds to a protein active site to stimulate an immune response;
(b) an antibody construct comprising an antigen binding domain and an Fc domain, wherein said antigen binding domain binds to at least a first antigen and wherein said Fc domain binds to an Fc receptor; and
(c) a linker, wherein said linker is covalently bound to said antibody construct and said linker is covalently bound to said immune- stimulatory compound; and
wherein the dissociation constant (Kd) for binding of said Fc domain of the conjugate to an Fc receptor is equal to, or up to no greater than about 100 times the Kd for binding of a control antibody to said Fc receptor, wherein the control antibody is the antibody construct; and wherein the Kd for binding of said immune- stimulatory compound of the conjugate to said protein active site is equal to, or up to no greater than 100 times the Kd for binding of a control compound to said protein active site or wherein the EC50 or IC50 of said immune- stimulatory compound of the conjugate is no greater than 300-fold the EC50 or IC50 of a control compound, wherein said control compound is the unbound immune- stimulatory compound.
2. An immune- stimulatory conjugate comprising:
(a) an immune- stimulatory compound that optionally binds to a protein active site of a binding protein to stimulate an immune response by inhibition of the activity of said binding protein;
(b) an antibody construct comprising an antigen binding domain and an Fc domain, wherein said antigen binding domain binds to at least a first antigen and wherein said Fc domain binds to an Fc receptor; and
(c) a linker, wherein said linker is covalently bound to said antibody construct and said linker is covalently bound to said immune- stimulatory compound; and wherein the dissociation constant (Kd) for binding of said Fc domain of the conjugate to an Fc receptor is equal to, or up to no greater than about 100 times the Kd for binding of a control antibody to said Fc receptor, wherein the control antibody is the antibody construct; and wherein the Kd for binding of said immune- stimulatory compound of the conjugate to said protein active site is equal to, or up to no greater than 100 times the Kd for binding of a control compound to said protein active site or wherein the IC50 of said immune- stimulatory compound of the conjugate is no greater than 300- fold the IC50 of a control compound, wherein said control compound is the unbound immune- stimulatory compound 3. An immune- stimulatory conjugate comprising:
(a) an immune- stimulatory compound that optionally binds to a protein active site of a binding protein to stimulate an immune response by degradation of said binding protein;
(b) an antibody construct comprising an antigen binding domain and an Fc domain, wherein said antigen binding domain binds to at least a first antigen and wherein said Fc domain binds to an Fc receptor; and
(c) a linker, wherein said linker is covalently bound to said antibody construct and said linker is covalently bound to said immune- stimulatory compound; wherein the dissociation constant (Kd) for binding of said Fc domain of the conjugate to an Fc receptor is equal to, or up to no greater than about 100 times the Kd for binding of a control antibody to said Fc receptor, wherein the control antibody is the antibody construct; and wherein the Kd for binding of said immune- stimulatory compound of the conjugate to said protein active site is equal to, or up to no greater than 100 times the Kd for binding of a control compound to said protein active site or wherein the IC50 of said immune- stimulatory compound of the conjugate is no greater than 300- fold the IC50 of a control compound, wherein the control compound is the unbound immune- stimulatory compound.
4. The immune- stimulatory conjugate of any one of claims 1 to 3, wherein the EC50 or IC50 of said immune- stimulatory compound of the conjugate is no greater than 100-fold the EC50 or IC50 of a control compound, wherein the control compound is the unbound immune- stimulatory compound.
5. The immune- stimulatory conjugate of any one of claims 1 to 3, wherein the EC50 or IC50 of said immune- stimulatory compound of the conjugate is no greater than 10-fold the EC50 or IC50 of a control compound, wherein the control compound is the unbound immune- stimulatory compound.
6. The immune- stimulatory conjugate of any one of claims 1 to 3, wherein the EC50 or IC50 of- said immune- stimulatory compound of the conjugate is equivalent to or less than the EC50 or IC50 of a control compound, wherein the control compound is the unbound immune- stimulatory compound.
7. An immune- stimulatory conjugate of any one of claims 1 to 3, wherein the EC50 or IC50 on an antigen bearing cell is equivalent to or less than the EC50 or IC50 of a control compound but EC50 or IC50 of the immune- stimulatory conjugate is 5-fold greater or more than the EC50 or IC50 of the control compound for a non-antigen bearing cell. 8. The immune- stimulatory conjugate of any one of claims 1 to 3, wherein the Kd for binding of said immune- stimulatory compound of the conjugate to said protein active site is no greater than 50 times the Kd for binding of a control compound to said protein active site, wherein the control compound is the unbound immune- stimulatory compound.
9. The immune- stimulatory conjugate of any one of claims 1 to 3, wherein the Kd for binding of said immune- stimulatory compound of the conjugate to said protein active site is no greater than 10 times the Kd for binding of a control compound to said protein active site, wherein the control compound is the unbound immune- stimulatory compound.
10. The immune- stimulatory conjugate of any one of claims 1 to 3, wherein the Kd for binding of said immune- stimulatory compound of the conjugate to said protein active site is equivalent to or less than the Kd for binding of a control compound to said protein active site, wherein the control compound is the unbound immune- stimulatory compound.
11. An immune- stimulatory conjugate comprising:
(a) an immune- stimulatory compound that optionally binds to a protein active site to stimulate an immune response;
(b) an antibody construct comprising an antigen binding domain and an Fc domain, wherein said antigen binding domain binds to a first antigen and wherein said Fc domain binds to an Fc receptor; and
(c) a linker, wherein said linker is covalently bound to said antibody construct and said linker is covalently bound to said immune- stimulatory compound; and
wherein the dissociation constant (Kd) for binding of said Fc domain of the conjugate to an Fc receptor is equal to, or up to no greater than about 100 times the Kd for binding of a control antibody to said Fc receptor, wherein the control antibody is the antibody construct; and wherein the Kd for binding of said immune- stimulatory compound when bound to a 5- 100 atom linker to said protein active site is no greater than 100 times the Kd for binding of a control compound to said protein active site or wherein the EC50 or IC50 of said immune- stimulatory compound of the conjugate is no greater than 300-fold the EC50 or IC50 of a control compound, wherein the conrol compound is the unbound immune- stimulatory compound.
12. The immune- stimulatory conjugate of claim 11, wherein the the EC50 or IC50 of said immune- stimulatory compound when bound to a 5- 100 atom linker is no greater than 100-fold the EC50 or IC50 of a control compound, wherein the control compound is the unbound immune- stimulatory compound. 13. The immune- stimulatory conjugate of claim 11, wherein the EC50 or IC50 of said immune- stimulatory compound when bound to a 5- 100 atom linker is no greater than 10-fold the EC50 or IC50 of a control compound, wherein the control compound is the unbound immune- stimulatory compound.
14. The immune- stimulatory conjugate of claim 11, wherein the EC50 or IC50 of said immune- stimulatory compound when bound to a 5- 100 atom linker is equivalent to or less than the EC50 or IC50 of a control compound, wherein the control compound is the unbound immune- stimulatory compound.
15. The immune- stimulatory conjugate of any one of claims 11 to 14, wherein the Kd for binding of said immune- stimulatory compound when bound to a 5- 100 atom linker to said protein active site is no greater than 50 times the Kd for binding of a control compound to said protein active site, wherein the control compound is the unbound immune- stimulatory compound.
16. The immune- stimulatory conjugate of any one of claims 11 to 14, wherein the Kd for binding of said of said immune- stimulatory compound when bound to a 5- 100 atom linker to said protein active site is no greater than 10 times the Kd for binding of a control compound to said protein active site, wherein the control compound is the unbound immune- stimulatory compound.
17. The immune- stimulatory conjugate of any one of claims 11 to 14, wherein the Kd for binding of said immune- stimulatory compound when bound to a 5- 100 atom linker to said protein active site is equivalent to or less than the Kd for binding of a control compound to said protein active site wherein the control compound is the unbound immune- stimulatory compound.
18. The immune- stimulatory conjugate of any one of claims 1 to 17, wherein the conjugate further comprises an E3 ubiquitin ligase binding moiety.
19. The immune- stimulatory conjugate of claim 18, wherein the E3 ubiquitin ligase binding moiety binds to VHL, cereblon, or MDM2
20. An immune- stimulatory conjugate of claims 18 or 19, wherein the E3 ubiquitin ligase binding moiety is selected from compounds 1- 1, 1-2, 1-3, 1-4, 1-5, 1-6, and 2- 1.
21. The immune stimulatory conjugate of any one of claims 18 to 20, wherein said E3 ubiquitin ligase binding moiety is attached to said linker or is part of said linker. 22. The immune- stimulatory conjugate of claim 21, wherein said E3 ubiquitin ligase binding moiety is part of said linker, wherein said E3 ubiquitin ligase binding moiety is bound through a first 5-100 atom linker to said immune- stimulatory compound and said E3 ubiquitin ligase binding moiety is bound through a second 5-100 atom linker to said antibody construct.
23. The immune- stimulatory conjugate of any one of claims 1 to 22, wherein the immune- stimulatory compound is a kinase inhibitor and the protein active site is a kinase active site.
24. The immune- stimulatory conjugate of claim 23, wherein said linker is covalently bound to said kinase inhibitor at a position on said kinase inhibitor that is at or near the solvent interface of said kinase active site as determined by modeling of said kinase inhibitor in said kinase active site.
25. The immune- stimulatory conjugate of claim 23, wherein said linker is covalently bound to said kinase inhibitor at a position on the kinase inhibitor such that when the kinase inhibitor is bound to said active site, said linker extends out from the kinase active site into the solvent, as determined by modeling of said kinase inhibitor in said kinase active site.
26. The immune- stimulatory conjugate of any one of claims 23 to 25, wherein the kinase inhibitor is selected from an inhibitor of ALK, Bcr-Abl, BRAF, BTK, c-KIT, EGFR, ErbB2, JAK, MEK, MET, PDGFRB, RET, ROS 1, Syk, SRC, TGFpR2, TNIK, TNKS, TrkA, TrkB, TrkC, VEGF, VEGFR1, VEGFR2, VEGFR3, FGFR1, FGFR2, FGFR3, FGFR4, CSF1R, RON/MS T1R, TYR03, MERTK, AXL, ΡΙ3Κδ, ΡΒΚγ, MAP4K1, PERK, and combinations thereof.
27. The immune- stimulatory conjugate of any one of claims 1 to 22, wherein said immune- stimulatory compound is a toll- like receptor agonist, STING agonist, or RIG-I agonist.
28. The immune- stimulatory conjugate of claim 27, wherein the immune- stimulatory compound is a toll-like receptor agonist selected from a TLR1 agonist, a TLR2 agonist, a TLR3 agonist, a TLR4 agonist, a TLR5 agonist, a TLR6 agonist, a TLR7 agonist, a TLR8 agonist, a TLR9 agonist, or a TLR10 agonist.
29. The immune- stimulatory conjugate of any one of claims 1 to 22, wherein said immune- stimulatory compound is selected from a pyrimidine, a purine, a guanine nucleoside, an 8- oxoadenine, an imidazoquinoline, a thiazoquinoline, a 2-amino imidazole, a furo[2,3-c]pyridine, a furo[2,3-c]quinoline, a 2-aminobenzimidazole, a 2-aminoquinoline, and a 2-aminobenzazepine. 30. The immune- stimulatory conjugate of any one of claims 1 to 22, wherein the target of the immune- stimulatory compound is a GCPR, an ion channel, a membrane transporter, a
phosphatase, or an ER protein.
31. The immune- stimulatory conjugate of any one of claims 1 to 22, wherein said immune- stimulatory compound is an antagonist of the GPCR A2aR, the sphingosine 1 -phosphate receptor 1, prostaglandin receptor EP3, prostanglandin receptor E2, Frizzled, CXCR4, or an LPA receptor.
32. The immune- stimulatory conjugate of any one of claims 1 to 22 wherein said immune- stimulatory compound is an ion channel agonist for CRAC, Kvl.3, or KCa3.1.
33. The immune- stimulatory conjugate of any one of claims 1 to 22, wherein said immune- stimulatory compound is an inhibitor of HSP90 or AAA-ATPase p97.
34. The immune- stimulatory conjugate of any one of claims 1 to 33, wherein said conjugate has immune- stimulatory activity with no or minimal cell processing.
35. The immune- stimulatory conjugate of any one of claims 1 to 34, wherein the linker is a non-cleavable linker.
36. The immune- stimulatory conjugate of any one of claims 1 to 34, wherein the linker is a Fleximer linker.
37. The immune- stimulatory conjugate of any one of claims 1 to 34, wherein the linker comprises a carbamate and one or more amide linkages.
38. The immune- stimulatory conjugate of any one of claims 1 to 34, wherein the linker is
, wherein Rx is a reactive moiety and wherein R is hydrogen, alkyl, sulfonate and methyl sulfonate. 39. The immune- stimulatory conjugate of any one of claims 1 to 34, wherein said linker is attached to the antibody construct at a cysteine or lysine residue of said antibody construct.
40. The immune- stimulatory conjugate of any one of claims 1 to 39, wherein said first antigen is a tumor antigen.
41. The immune- stimulatory conjugate of any one of claims 1 to 39, wherein said first antigen is at least 80% homologous to CD5, CD19, CD20, CD25, CD37, CD30, CD33, CD45, CAMPATH-1, BCMA, CS-1, PD-L1, B7-H3, B7-DC, HLD-DR, carcinoembryonic antigen, TAG-72, EpCAM, MUC1, fo late-binding protein, A33, G250, pro state- specific membrane antigen, ferritin, GD2, GD3, GM2, Ley, CA-125, CA19-9, epidermal growth factor, pl85HER2, IL-2 receptor, de2-7 EGFR, fibroblast activation protein, tenascin, metalloproteinases, endosialin, vascular endothelial growth factor, ανβ3, WT1, LMP2, HPV E6 E7, EGFRvIII, Her-2/neu, idiotype, MAGE A3, p53 nonmutant, NY-ESO-1, PMSA, GD2, CEA, MelanA/M ART 1 , Ras mutant, gplOO, p53 mutant, PR1, bcr-abl, tyronsinase, survivin, PSA, hTERT, Sarcoma translocation breakpoints, EphA2, PAP, ML-IAP, AFP, ERG, NA17, PAX3, ALK, androgen receptor, cyclin Bl, polysialic acid, MYCN, RhoC, TRP-2, fucosyl GM1, mesothelin, PSCA, MAGE Al, sLe(animal), CYPIBI, PLAVl, GM3, BORIS, Tn, GloboH, ETV6-AML, NY-BR-1, RGS5, SART3, STn, Carbonic anhydrase IX, PAX5, OY-TES 1, Sperm protein 17, LCK,
HMWMAA, AKAP-4, SSX2, XAGE 1, B7H3, Legumain, Tie 3, Page4, VEGFR2, MAD-CT-1, PDGFR-B, MAD-CT-2, ROR2, TRAIL 1, MUC16, MAGE A4, MAGE C2, GAGE, EGFR, CMET, HER3, MUC15, MSLN, CA6, NAPI2B, TROP2, CLDN18.2, RON, LY6E, FRA, DLL3, PTK7, LIV1, ROR1, or Fos-related antigen 1..
42. The immune- stimulatory conjugate of any one of claims 1 to 41, wherein said antibody construct further comprises a second targeting binding domain.
43. The immune- stimulatory conjugate of claim 42, wherein said targeting binding domain specifically binds an immune cell.
44. The immune- stimulatory conjugate of claim 42 or 43, wherein said targeting binding domain is conjugated to said antibody construct at a C-terminal end of said Fc domain.
45. The immune- stimulatory conjugate of any one of claims 1 to 44, wherein said antigen binding domain is from an antibody or non-antibody scaffold. 46. The immune- stimulatory conjugate of any one of claims 1 to 45, wherein said antigen binding domain is at least 97% homologous to an antigen binding domain from an antibody or non-antibody scaffold.
47. The immune- stimulatory conjugate of any one of claims 1 to 46, wherein said antibody construct is a human antibody or a humanized antibody.
48. The immune- stimulatory conjugate of any one of claims 1 to 47, wherein said Fc domain is an Fc domain variant comprising at least one amino acid residue change as compared to a wild type sequence of said Fc domain.
49. The immune- stimulatory conjugate of any one of claims 1 to 48, wherein said Fc domain is at least about 80% homologous to an Fc domain from an antibody, wherein the Fc domain from an antibody comprises amino acid residues 216 to 447 of SEQ ID NO: 898, amino acid residues 216 to 443 of SEQ ID NO: 899, or amino acid residues 216 to 444 of SEQ ID NO: 900.
50. The immune- stimulatory conjugate of any one of claims 1 to 49, wherein said Fc domain has at least one amino acid residue change as compared to wildtype, wherein said Fc domain is at comprises at least 80% homologous to SEQ ID NO: 296, and wherein said at least one amino acid residue change is:
a) F243L, R292P, Y300L, L235V, and P396L, wherein numbering of amino acid residues in said Fc domain is relative to SEQ ID NO: 898;
b) S239D and I332E, wherein numbering of amino acid residues in said Fc domain is relative to SEQ ID NO: 898; or
c) S298A, E333A, and K334A, wherein numbering of amino acid residues in said Fc domain is relative to SEQ ID NO: 898.
51. The immune- stimulatory conjugate of any one of claims 1 to 50, wherein said Fc domain has at least one amino acid residue change as compared to wildtype, wherein said Fc domain is at comprises at least 80% homologous to SEQ ID NO: 898, and wherein said at least one amino acid residue change is:
a) N297A, N297G, N297Q, N297D as in Kabat numbering and relative to SEQ ID NO: 898; or
b) K322A/L234A/L235A N296A as in Kabat numbering and relative to SEQ ID NO: 898; or
c) L234F/L235E/P331S N296A as in Kabat numbering and relative to SEQ ID NO:
898; or d) P329G/L234A/L235A as in Kabat numbering and relative to SEQ ID NO: 898.
52. The immune- stimulatory conjugate of any one of claims 1 to 51, wherein said Kd for binding of said antigen binding domain to said first antigen in the presence of said immune- stimulatory compound is less than about ΙΟΟηΜ and is equal to, or up to no greater than about 10 times the Kd of the binding of the antigen binding domain to said first antigen in the absence of the immune- stimulatory compound; and said Kd for binding of said Fc domain to said Fc receptor in the presence of said immune- stimulatory compound is equal to, or up to no greater than about 10 times said Kd for the binding of said Fc domain to said Fc receptor in the absence of said immune- stimulatory compound.
53. The immune- stimulatory conjugate of any one of claims 1 to 52, wherein said molar ratio of immune- stimulatory compound to antibody is less than 5.
54. The immune- stimulatory conjugate of any one of claims 1 to 53, wherein said linker is bound to said antibody construct at an amino acid residue that does not interfere with said Fc domain binding to said Fc receptor.
55. The immune- stimulatory conjugate of any one of claims 1 to 54, wherein said linker is not attached to an amino acid residue of said Fc domain selected from a group consisting of: 221, 224, 227, 228, 230, 231, 223, 233, 234, 235, 236, 237, 238, 239, 240, 241, 243, 244, 245, 247, 249, 250, 258, 262, 263, 264, 265, 266, 267, 268, 269, 270, 271, 272, 273, 275, 276, 278, 280, 281, 283, 285, 286, 291, 292, 293, 294, 295, 296, 297, 298, 299, 300, 302, 305, 313, 318, 323, 324, 325, 327, 328, 329, 330, 331, 332, 333, 335, 336, 396, or 428, wherein numbering of amino acid residues in said Fc domain is according to the EU index as in Kabat.
56. The immune- stimulatory conjugate of any one of claims 1 to 55, wherein said Fc domain is selected from a group consisting of a human IgGl Fc domain, a human IgG2 Fc domain, a human IgG3 Fc domain, and a human IgG4 Fc domain.
57. The immune- stimulatory conjugate of any one of claims 1 to 56, wherein said conjugate induces the secretion of cytokines by an antigen presenting cell.
58. A pharmaceutical composition comprising an immune- stimulatory conjugate of any one of claims 1 to 57 and a pharmaceutically acceptable excipient.
59. A method of treating cancer, comprising administering to a subject in need thereof a pharmaceutical composition of claim 58. General Schemes
Synthesis of Immune-Stimulatory Compound-Linkers and Immune-Modulatory Compound- Linker Constructs
[0551] An construct of a linker and an immune- stimulatory compound or an immune-modulatory compound (denominated ISC) can be synthesized by various methods. For example, ISC-linker constructs can be synthesized as shown in Scheme Bl.
Sche
R = NHS, pentafluorophenyl
ISC: immune-stimulatory compound
[0552] A PEGylated carboxylic acid (i) that has been activated for amide bond formation can be reacted with an appropriately substituted amine containing immune- stimulatory compound to afford an intermediate amide. Formation of an activated ester (ii) can be achieved by reaction the intermediate amide-containing carboxylic using a reagent such as N-hydroxysuccinimide or pentafluorophenol in the presence of a coupling agent such as diisopropylcarbodiimide (DIC) to provide compounds (ii).
[0553] An ISC-linker construct can be synthesized as shown in Scheme B2.
Scheme B2:
[0554] An activated carbonate such as (i) can be reacted with an appropriately substituted amine containing immune- stimulatory compound to afford carbamates (ii) which can be deprotected using standard methods based on the nature of the R3 ester group. The resulting carboxylic acid (iii) can then by coupled with an activating agent such as N-hydroxysuccinimide or
pentafluorophenol to provide compounds (iv).
[0555] A ISC-linker construct can be synthesized as shown in Scheme B3.
Scheme B3:
i-a; X = NHS ϋ
i-b; X = H
ISC: immune-stimulatory compound
[0556] An activated carboxylic ester such as (i-a) can be reacted with an appropriately
substituted amine containing immune- stimulatory compound to afford amides (ii). Alternatively, carboxylic acids of type (i-b) can be coupled to an appropriately substituted amine containing immune- stimulatory compound in the presence of an amide bond forming agent such as
dicyclohexycarbodiimde (DCC) to provide the desired ISC.
[0557] An ISC-linker construct can be synthesized by various methods such as that shown in Scheme B4.
Scheme B4:
ϋ
ISC: immune-stimulatory compound
[0558] An activated carbonate such as (i) can be reacted with an appropriately substituted amine containing immune- stimulatory compound to afford carbamates (ii) as the target ISC. [0559] An ISC-linker construct can also be synthesized as shown in Scheme B5.
Scheme B5:
[0560] An activated carboxylic acid such as (i-a, i-b, i-c) can be reacted with an appropriately substituted amine containing immune- stimulatory compound to afford amides (ii-a, ii-b, ii-c) as the target linkered immune- stimulatory compounds.
General Scheme for the Synthesis of Immune-Modulatory Conjugates Containing a PROTAC
[0561] An immune-modulatory conjugate containing a PROTAC (or PTM) as described herein can comprise an antibody construct Ab (such as an antibody) convalently attached via a linker (L) to a PROTAC, wherein the PROTAC comprises a ubiquitin E3 ligase binding group (E; also referred to as ULM), a spacer (S) and an immune-modulatory compound (K; also referred to as an IMC) (such as a kinase inhibitor). The general formula is: Ab— (L— (Ci-S-C2))n, wherein Ab is the antibody construct, C1-S-C2 is PROTAC or PTM, wherein, C2 is an E3 ubiquitin ligase binding group (E or ULM) covalently bound to a spacer group (s) that is covalently bound to Ci, an immune-modulatory compound (E or IMC), and L is a linker covalently bonded to the antibody construct and to the PROTAC; and n has a value from about 1 to about 8.
[0562] In the following exemplary scheme, the immune-modulatory compound (E in this scheme) is a kinase inhibitor. Scheme 1: deprotect
couple
deprotect
0-NH HN-© conjugate
[0563] A kinase inhibitor containing a free amine functional group can be acylated with a multifunctional amino acid derivative such as aspartate or glutamate using standard amide bond coupling reactions such as HATU in DMF containing and amine base to provide intermediates (ii). Deprotection of compounds (ii) using known methods for the conversion of carboxylic esters to carboxylic acids, such as hydrogenation when R = Bn can provide compounds (iii) which can be coupled to an E3 ubiquitin ligase such as a group that binds VHL or cereblon to provide PROTACs (iv). Compounds that bind VHL may be hydroxyproline compounds such as those disclosed in WO 2013/106643, and other compounds described in US 2016/0045607, WO 2014187777, US 20140356322, and US 9,249,153. Compounds that bind to cereblon include thalidomide, lenalidomide, pomalidomide and analogs thereof. Other small molecule compounds that bind to cereblon are also known, e.g., the compounds disclosed as an in US2016/0058872 and US2015/0291562. The amine protecting group can be converted to intermediates (v) using appropriate reagents such as TFA when PG = Boc. Acylation of amines (v) by activated linker reagents (X* = NHS) or by direct amide bond coupling can provide linked-PROTAC (L-C) compounds (vi) which can subsequently be conjugated to an antibody using known methods as described herein.
Schem
[0564] Alternatively, a kinase inhibitor containing a free amine functional group can be acylated with a multi-functional amino acid derivative such as lysine using standard amide bond coupling reactions such as HATU in DMF containing and amine base to provide intermediates (vii). Deprotection of compounds (vii) using known methods, such as hydrogenation when R = Cbz can provide compounds (ix) which can be coupled to an E3 ubiquitin ligase to provide
PROTACs (x). The second amine protecting group (PG2) can be converted to intermediates (v) using appropriate reagents such as TFA when PG = Boc. Acylation of amines (xi) by activated linker reagents (X* = NHS) or by direct amide bond coupling can provide linked-PROTAC compounds (xii) which can subsequently be conjugated to an antibody using known methods as described herein. FIGURE DESCRIPTIONS
[0565] FIGURE 1 illustrates a schematic of a conjugate comprising an antibody and a second binding domain. The antibody contains two heavy chains as shown in dark gray and two light chains as shown in light gray. A portion of the heavy chains contain Fc domains (705 and 720). The antibody comprises a binding domain comprising two antigen binding sites (710 and 715). The second binding domain is attached to the antibody (780 and 785; black spheres), for example, at the C-terminus of the heavy chains.
[0566] FIGURE 2 illustrates a schematic of an exemplary conjugate. The conjugate comprises an antibody, which contains two heavy chains as shown in dark gray and two light chains as shown in light gray. The antibody comprises a binding domain comprising two antigen binding sites (910 and 915), and a portion of the heavy chains contain Fc domains (905 and 920). The immune- stimulatory compounds (930 and 940; black stars) are attached to the antibody by linkers (960 and 970; black line). A second binding domain is attached to the antibody (980 and 985; black spheres), for example, at the C-terminus of the heavy chains.
[0567] FIGURE 3 illustrates a schematic of an exemplary conjugate. The conjugate comprises the Fc region of an antibody with the heavy chains shown in dark gray, and two scaffolds as shown in light gray. The conjugate comprises a first binding domain comprising two antigen binding sites (1110 and 1115) in the scaffolds, and a portion of the heavy chains contain Fc domains (1105 and 1120). The immune- stimulatory compounds (1130 and 1140; black starts) are conjugated to the Fc domains by linkers (1160 and 1170; black lines). A second binding domain is attached to the conjugate (1180 and 1185; black spheres), for example, at the C-terminus of the heavy chains.
[0568] FIGURE 4 illustrates a schematic of an exemplary conjugate. The conjugate comprises the F(ab')2 region of an antibody with the Fab portions of heavy chains shown in dark gray and light chains shown in light gray, and two scaffolds as shown in the darkest gray. The conjugate comprises a first binding domain comprising two antigen binding sites (1310 and 1315), and a portion of two scaffolds contain Fc domains (1340 and 1345). The immune- stimulatory compounds (1330 and 1340; black stars) are attached to the conjugate by linkers (1360 and 1370; black lines). A second binding domain is attached to the Fc domains (1380 and 1385; black spheres).
[0569] FIGURE 5 illustrates a schematic of an exemplary conjugate. The conjugate contains two scaffolds as shown in light gray and two scaffolds as shown in dark gray. The conjugate comprises a first binding domain comprising two antigen binding sites (1510 and 1515), and a portion of the two dark gray scaffolds contain Fc domains (1540 and 1545). The immune- stimulatory compounds (1530 and 1540; black stars) are conjugated to the conjugate by linkers (1560 and 1570; black lines). A second binding domain is attached to the conjugate (1580 and 1585; black spheres).
[0570] FIGURE 6 illustrates a schematic of a conjugate comprising an antibody and a second binding domain. The antibody contains two heavy chains as shown in dark gray and two light chains as shown in light gray. A portion of the heavy chains contain Fc domains (1705 and 1720). The antibody comprises a binding domain comprising two antigen binding sites (1710 and 1715). The second binding domain is attached to the antibody (1780 and 1785; black spheres), for example, at the C-terminus of the light chains.
[0571] FIGURE 7 illustrates a schematic of an exemplary conjugate. The conjugate comprises an antibody, which contains two heavy chains as shown in dark gray and two light chains as shown in light gray. The antibody comprises a binding domain comprising two antigen binding sites (1910 and 1915), and a portion of the heavy chains contain Fc domains (1905 and 1920). The immune- stimulatory compounds (1930 and 1940; black stars) are conjugated to the antibody by linkers (1960 and 1970; black lines). A second binding domain is attached to the antibody (1980 and 1985; black spheres), for example, at the C-terminus of the light chains.
[0572] FIGURE 8 illustrates a schematic of an exemplary conjugate. The conjugate comprises an Fc region of an antibody shown indark gray, and two scaffolds as shown in light gray. The conjugate comprises a first binding domain comprising two antigen binding sites (2110 and 2115) in the scaffolds, and a portion containing Fc domains (2105 and 2120). The immune- stimulatory compounds (2130 and 2140; black stars) are conjugated to the antibody construct by linkers (2160 and 2170; black lines). A second binding domain is attached to the antibody (2180 and 2185; black spheres).
[0573] FIGURE 9 illustrates a schematic of an exemplary conjugate. The conjugate comprises the F(ab')2 region of an antibody with heavy chains shown in dark gray and light chains shown in light gray, and two scaffolds as shown in darkest gray. The conjugate comprises a first binding domain comprising two antigen binding sites (2310 and 2315), and a portion of two scaffolds contain Fc domains (2340 and 2345). The immune- stimulatory compounds (2330 and 2340; black stars) are conjugated to the antibody by linkers (2360 and 2370; black lines). A second binding domain is attached to the antibody (2380 and 2385; black spheres), for example, at the C- terminus of the light chains.
[0574] FIGURE 10 illustrates a schematic of an exemplary conjugate. The conjugate contains two scaffolds as shown in light gray and two scaffolds as shown in dark gray. The conjugate comprises a first binding domain comprising two antigen binding sites (2510 and 2515), and a portion of the two dark gray scaffolds contain Fc domains (2540 and 2545). The immune- stimulatory compounds (2530 and 2540; black stars) are conjugated to the antibody construct by linkers (2560 and 2570; black lines). A second binding domain is attached to the conjugate (2580 and 2585; black spheres).
[0575] FIGURE 11 illustrates a schematic of an antibody construct comprising an antibody. The antibody contains two heavy chains and two light chains. A portion of the heavy chains contain Fc domains (2705 and 2720). The antibody comprises a binding domain comprising two antigen binding sites shown in black (2710 and 2715).
[0576] FIGURE 12 illustrates a schematic of an antibody construct comprising an antibody. The antibody contains two heavy chains and two light chains. A portion of the heavy chains contain Fc domains (2925 and 2930). The antibody comprises a first binding domain comprising two antigen binding sites shown in black (2910 and 2915). The antibody comprises a second binding domain comprising two single chain variable fragments (2905 and 2920) attached to a C- terminus of the light chains. A single chain variable fragment can be attached to a light chain chain at a heavy chain variable domain of the single chain variable fragment. A single chain variable fragment can be attached to a light chain at a light chain variable domain of the single chain variable fragment.
[0577] FIGURE 13 illustrates a schematic of an antibody construct comprising an antibody. The antibody contains two heavy chains and two light chains. A portion of the heavy chains contain Fc domains (3120 and 3125). The antibody comprises a first binding domain comprising two antigen binding sites shown in black (3110 and 3115). The antibody comprises a second binding domain comprising two single chain variable fragments (3130 and 3135) attached to a C- terminus of the heavy chains. A single chain variable fragment can be attached to a heavy chain chain at a heavy chain variable domain of the single chain variable fragment. A single chain variable fragment can be attached to a heavy chain at a light chain variable domain of the single chain variable fragment.
[0578] FIGURE 14 illustrates a schematic of an antibody construct comprising an antibody. The antibody contains two heavy chains and two light chains. A portion of the heavy chains contain
Fc domains (3330 and 3335). The antibody comprises a first binding domain comprising two antigen binding sites shown in black (3310 and 3315). The antibody comprises a second binding domain comprising two single chain variable fragments (3320 and 3325) attached to a C- terminus of the light chains. A single chain variable fragment can be attached to a light chain chain at a heavy chain variable domain of the single chain variable fragment. A single chain variable fragment can be attached to a light chain at a light chain variable domain of the single chain variable fragment. The antibody comprises a third binding domain comprising two single chain variable fragments (3340 and 3345) attached to a C-terminus of the heavy chains. A single chain variable fragment can be attached to a heavy chain chain at a heavy chain variable domain of the single chain variable fragment. A single chain variable fragment can be attached to a heavy chain at a light chain variable domain of the single chain variable fragment.
[0579] FIGURE 15A sets forth a structure of an immune- stimulatory compound (left) and an immune- stimulatory compound with an amine handle (right top).
[0580] FIGURE 15B shows the x-ray crystal structure and binding orientation (pdb code 5D7A) of the immune- stimulatory compound as a ball- and- stick model as described in FIGURE 15A in a TNIK (TRAF2 and NCK- interacting protein kinase) active site. Cysl08 (bottom left) and Glnl57 (top right) of the TNIK protein are labeled. The dashed lines represent non-covalent bonds between the immune- stimulatory compound and the active site of TNIK. FIGURE 15B indicates that the terminal oxygen on the immune- stimulatory compound interacts with Gin 157 of the TNIK active site with a non-covalent bond length of about 2.9 A. Further, FIGURE 15B indicates that one of the nitrogen atoms of the compound interacts with the Cysl08 at the carboxyl group of the cysteine residue with a non-covalent bond length of about 3.1 A, and another nitrogem atom of the compound interacts with the amine group of the cysteine residue with a non-covalent bond length of about 3.2 A.
[0581] FIGURE 15C shows a close up of the binding orientation of the immune- stimulatory compound as a ball-and-stick model as described in FIGURE 15A in a TNIK active site, where the linker and antibody portions are pointing away and sitting outside of the active site. The modeling indicates that positioning of a linker on the 4-position of the benzimidazole of the left- hand compound of FIGURE 15A would extend from the active site and not interfere with the active site interactions of the compound as depicted in FIGURE 15B and FIGURE 15C.
[0582] FIGURE 16 sets forth a structure of an immune- stimulatory compound (left) and an immune- stimulatory compound with an amine handle and linker surrogate (right). The structure on the right illustrates that the immune- stimulatory compound is predicted to sit in the enzyme active site, whereas the amine handle and linker surrogate are predicted to sit outside of the enzyme active site, in the solvent.
[0583] FIGURE 17A sets forth a structure of an immune- stimulatory compound (left) and an immune- stimulatory compound with a linker surrogate (left side of molecule on right). FIGURE 17A indicates that the TGFpRl EC50 of the compound on the left is 14 nM, and that the TGFPR2 EC50 is 5 nM. The R group on the right figure is a hydrogen atom.
[0584] FIGURE 17B shows the x-ray crystal structure and binding orientation (pdb code 5E91) of the immune- stimulatory compound as a ball-and-stick model as described in FIGURE 17A in a TGFPR2 (transforming growth factor, beta receptor II) active site. Cys396 of the TGFPR2 is depicted in the upper right corner of the figure. [0585] FIGURE 17C shows a close up of the binding orientation of the immune- stimulatory compound as a ball-and-stick model as described in FIGURE 17 A in a TGFPR2 active site, where the linker and antibody construct portions are pointing away and sitting outside of the active site. Asn332 (upper right) and His328 (bottom right) of the TGFPR2 active site are also labeled. A bond length of 2.9 A is shown between an oxygen atom of the compound and the active site. Other bond lengths labeled include 2.8 A.
[0586] FIGURE 18A sets forth a structure of an immune- stimulatory compound (left) and an immune- stimulatory compound with an amine handle (upper left in molecule on right). FIGURE 18A indicates that the TNKS 1 IC50 of the compound on the left is 13 nM, and that the TNKS2 IC50 is 5 nM.
[0587] FIGURE 18B shows the x-ray crystal structure and binding orientation (pdb code 3KR8) of the immune- stimulatory compound as a ball-and-stick model as described in FIGURE 18A in a TNKS (tankyrase) active site. Glyl l85 and Serl221 are labeled on the active site of TNKS. The figure indicates that an oxygen atom of the compound interacts with Glyl 185 and Serl221 with a non-covalent bond length of about 2.9 A.
[0588] FIGURE 18C shows a close up of the binding orientation of the immune- stimulatory compound as a ball-and-stick model as described in FIGURE 18A in a TNKS active site, where the linker and antibody construct portions are pointing away and sitting outside of the active site, including the trifluoromethyl group of the compound.
[0589] FIGURE 19A shows a structure of an immune- stimulatory compound (left) and an immune- stimulatory compound with two amine handles (on right side of molecule on the right). FIGURE 19A indicates that the TNKS 1 IC50 of the compound on the left is 8 nM, and that the TNKS 2 ICso is 2 nM.
[0590] FIGURE 19B shows the x-ray crystal structure and binding orientation (pdb code 4191) of the immune- stimulatory compound as a ball-and-stick model as described in FIGURE 19A with dual site binding in the TNKS (tankyrase) active site. Aspl 198, Glyl 185, and Tyrl213 are labeled for the active site of TNKS. The figure indicates that an oxygen atom of the compound interacts with Aspl 198 with a non-covalent bond length of about 3.2 A. Another oxygen atom of the compound interacts with Tyrl213 with a non-covalent bond length of about 2.9 A. Both a nitrogen atom and oxygen atom of the compound interact with Glyl 185 with a non-covalent bond length of about 2.8 A and 2.7 A, respectively.
[0591] FIGURE 19C shows a close up of the binding orientation of the immune- stimulatory compound as a ball-and-stick model as described in FIGURE 19A in a TNKS active site, where the linker and antibody portions are pointing away and sitting outside of the active site. The figure indicates that the compound interacts with Aspl 198 at a non-covalent bond length of about 3.2 A (middle). The figure indicates that the compound interacts with Tyrl213 at a non-covalent bond length of about 2.9 A (left).
[0592] FIGURE 20A illustrates a schematic of an exemplary conjugate and its molecular target. The conjugate comprises an antibody (3405) attached to a linker (3410) that is attached to a drug (3415) at the opposite end of the antibody (3405). The molecular target (3420) has an active site (3425) that is complementary to the drug (3415).
[0593] FIGURE 20B illustrates a schematic of an active exemplary conjugate that is bound to the the molecular target's active site. The drug (3415) sits within the active site of the molecular target (3420). The linker (3410) and antibody (3405) sit outside of the active site (3430, represented by the dashed line).
[0594] FIGURE 20C illustrates a schematic of an active drug (3415) and linker (3410) that is bound to the molecular target (3420). The linker (3410) sits outside of the active site (3430, the dashed line is the boundary of the active site).
[0595] FIGURE 20D illustrates a schematic of an active drug (3415) that is bound to the molecular target (3420). The active drug sits inside the active site (3430, the dashed line is the boundary of the active site).
[0596] FIGURE 21A shows the results of an assay for degradation of TFGpR2 by an anti-HER2 antibody-TGFpR2-VHL PROTAC conjugate. Plasmid expressing HER2 was transfected into HEK293 cells, and the cells were treated with DMSO, PROTAC T-20, HER2 antibody (IgGl), or Her2 Antibody-Protac conjugate (050-T11020). Whole cell lysates were prepared from cells after 2 (left blot), 24 (middle blot), or 48 (right blot) hours incubation and quantitated with a BCA assay. Equal amounts of lysates were run on protein gels, transferred to PVDF, and TGFPR2 (top), TGFpRl (middle), or control actin (bottom) was detected using commercially available reagents. At both tested concentrations of the conjugate, the level of target TGFPR2 was diminished at 24 and 48 hours of treatment as demonstrated by the diminished signal of TGFPR2 in the lanes containing 050-T11020. For the 2 hour blot, from left to right, the lanes represent DMSO; T-20 5 μΜ; 050 IgG 1 μΜ; 050-T11020 1 μΜ; and 050-T11020 0.5 μΜ. For the 24 hour blot, from left to right, the lanes represent DMSO; T-20 5 μΜ; 050 IgG 0.5 μΜ; 050 IgG 1 μΜ; 050-T11020 0.5 μΜ; and 050-T11020 1 μΜ. For the 48 hour blot, from left to right, the lanes represent PBS; 050 IgG 1 μΜ; 050-T11020 1 μΜ; and 050-T11020 0.5 μΜ.
[0597] FIGURE 21B provides a quantification of the western blot data for TGFPR2 shown in FIG 21 A. To quantitate the amount of protein degradation, the signals on the Western blot were normalized to actin loading control and data was presented as a percent of matched control on the y-axis, which is labeled from 0 to 140 in intervals of 20. A thick black line denotes 100 precent. The medium-gray bars at the left of each data set represent the data obtained at 2 hours of treatment. The darkest gray bars in the middle of each data set represent the data obtained at 24 hours. The lightest gray bars at the right of each data set represent the data obtained at 48 hours. On the x-axis, from the left to right, the data sets are T20 5 μΜ; 050-11020 0.5 μΜ; and 050- 11020 1 μΜ.
[0598] FIGURE 21C provides a quantification of the western blot data for TGFpRl shown in FIGURE 21 A. To quantitate the amount of protein degradation, the signals on the Western blot were adjusted to actin loading control and data was presented as a percent of matched control on the y-axis, which is labeled as 0 to 200 in intervals of 20. The medium-gray bars at the left of each data set represent the data obtained at 2 hours of treatment. The darkest gray bars in the middle of each data set represent the data obtained at 24 hours. The lightest gray bars on the right of each data set represent the data obtained at 48 hours. On the x-axis, from the left to right, the data sets are T20 5 μΜ; 050-11020 0.5 μΜ; and 050-11020 1 μΜ. Consistent with the western blot data, the amount of TGFpRl protein remained fairly constant throughout the treatment period.
[0599] FIGURE 22A and FIGURE 22B show the results of an assay for antigen targeted degradation of TGFPR2 by an antibody conjugate with PROTACs having VHL or Cereblon E3 binding moieties. BT474 cells were plated and treated the following day with either a T-15 PROTAC or a T-20 PROTAC, a conjugate of a HER2 antibody TGFpR2-VHL binding
PROTAC (050-T05020), a conjugate of a HER2 antibody TGFpR2-Cereblon binding PROTAC (050-T05015), or a conjugate of a TROP2 antibody TGFpR2-VHL binding PROTAC (130- T05020). Whole cell lysates were prepared 24 hours after treatment and quantitated with a BCA assay. Equal amounts of lysates were run on protein gels, transferred to PVDF, and TGFPR2 and actin were detected using commercially available reagents. FIGUGRE 22A shows that HER2- antigen specific degradation was found with both the HER2 binding PROTAC conjugates, but not with the control TROP2-binding PROTAC conjugate, as indicated by the retained signal of the TGFPR2 protein (top blot; actin control is bottom blot). The lanes, from left to right, represent DMSO; T-15 300 nM; T-20 300 nM; PBS; unlabeled; 050-T05015 0.5 mM; 050- T05020 0.5 μΜ; and 130-T05020 0.5 μΜ.
[0600] FIGURE 22B provides a quantitation of FIGURE 22A, and was determined by normalizing the TGFPR2 signals to actin loading control. The data are presented as a percent of vehicle control on the y-axis, which is labeled as 0 to 120 in intervals of 20. The x-axis, from left to right, represents T-15 300 nM; T-20 300 nM; 050-T05015; 050-T0520; and 130-T05020. The thick black line is at 100 of the y-axis.
[0601] FIGURE 23A and FIGURE 23B show the results of an assay for cellular levels of TGFpR2 and TGFpRl in the presence of a TGFpR2/TGFpRl-VHL PROTAC (T-20) with or without the addition of a proteasome inhibitor. Normal human lung fibroblasts were treated with or without proteasome inhibitor MG-132 followed by the addition of DMSO or T-20. Whole cell lysates were prepared and then quantitated with a BCA assay. Equal amounts of lysates were run on protein gels and transferred to PVDF membrane. TGFpRl, TGFPR2, and actin were detected using commercially available reagents. FIGURE 23A provides the western blot results of the assay. The results demonstrate that the addition of the proteasome inhibitor protected TGFpRl and TGFPR2 against degradation induced by T-20, as indicated by rescue of the TGFPR2 and TGFpR2 signals by addition of MG-132 in the presence of T-20. TGFpR2 is the top row, TGFpRl is the middle row, and actin is the bottom row. The left blots lanes represent, from left to right, MG132 concentrations of 0 (shown as -); 10; and 50 followed by addition of DMSO. The right blots represent, from left to right, MG132 concentrations of 0 (shown as -); 10; and 50 μΜ followed by addition of 5 μΜ T-20.
[0602] FIGURE 23B provides quantification of the results of the FIGURE 23A, and was obtained by adjusting the western signal to the actin loading control. The data are presented as a percent of the matched vehicle control on the y-axis, which is labeled from 0 to 100 at intervals of 10. The light gray bars represent the data for TGFPR2 and the dark gray bars represent the data for TGFpRl. The x-axis, from left to right, is labeled as T-20; T-20 + 10 μΜ MG132; and T-20 + 50 μΜ MG132.
EXAMPLES
[0603] The following examples illustrate the various methods of making and assaying compounds and conjugates described herein. It is understood that one skilled in the art may be able to make these compounds and conjugates by similar methods or by combining other methods known to one skilled in the art. It is also understood that one skilled in the art would be able to make compounds and conjugates in a similar manner as described below by using the appropriate starting materials and modifying the synthetic route as needed. In general, starting materials and reagents can be obtained from commercial vendors or synthesized according to sources known to those skilled in the art or prepared as described herein.
EXAMPLE 1
[0604] Synthesis of (S)-Nl-(4-(5-amino-6-((4-morpholinopyridin-3-yl)carbamoyl)pyrazin-2- yl)benzyl)-2-(6-(4-((2,5-dioxo-2,5-dihydro-lH-pyrrol-l-yl)methyl)-cyclohexane-l- carboxamido)hexanamido)-N5-(2-(3-(((S)-l-((2S,4R)-4-hydroxy-2-((4-(4-methylthiazol-5- yl)benzyl)carbamoyl)pyrrolidin-l-yl)-3,3-dimethyl-l-oxobutan-2-yl)amino)-3- oxopropoxy)ethyl)pentanediamide (Compound 1-1)
Step A: Preparation oflnt lB-1
Int 1 B-1
[0605] HATU (3.54 g, 9.36 mmol) was added to a solution containing 1.64 g (7.5 mmol) of 3- amino-6-bromopyrazine-2-carboxylic acid in 25 mL of DMF. The reaction was stirred for 5 minutes before adding 2.5 mL (22.5 mmol) of N-methylmorpholine and 1.68 g (9.36 mmol) of 4- morpholinopyridin-3-amine. The reaction mixture was stirred for 16 h then quenched with 10 mL of saturated NH4C1 solution and then 10 mL of water. The mixture was extracted with EtOAc three times; the combined organics were washed with brine and then dried over a2S04. The solvent was then evaporated and the residue was chromatographed (0% to 20% CH3OH / dichloromethane) to afford compound Int lB-1 as a yellow solid. Step B: Preparation of Int 1B-2
Int 1 B-1 Int 1 B-2
[0606] A solution containing 1.5 g (4.0 mmol) of 3-amino-6-bromo-N-(4-morpholinopyridin-3- yl)pyrazine-2-carboxamide and 1.1 g (4.4 mmol) of (4-(2-(((tert-butoxy)carbonyl)- amino)methyl)phenyl)boronic acid in 25 mL of dioxane and 4.0 mL of 2N a2C03 (8.0 mmol) was degassed and back filled with nitrogen three times. 295 mg (0.4 mmol) of PdCl2 (dppf) was added and the reaction vessel was degassed with nitrogen twice. The reaction mixture was then heated at 90 °C for 3 h then cooled and stirred overnight then filtered through a plug of Celite®. The filtrate was diluted with EtOAc, washed with water and then brine, and dried over Na2S04. The solvent was then evaporated and the residue was chromatographed (0% to 20% MeOH / dichloromethane) to afford 1.3 g of compound Int 1B-2 as a white solid. LCMS (M+H) = 506.
Step C: Preparation of Int 1B-3
2 HCl
Int 1 B-2 Int 1 B-3
[0607] A solution containing 1.2 g (2.4 mmol) of Int 1B-2 in 25 mL of EtOAc was added 10 mL of 4N HCl in dioxane at room temperature. The reaction was stirred for 3h and the solvent was evaporated. The resulting solid was triturated three times with toluene to provide the desired amine salt which was used without purification. LCMS (M+H) = 406. Step D: Preparation of Int 1B-4
[0608] To a solution containing 112 mg (0.276 mmol) of Int 1B-3 and 93 mg (0.276 mmol) of Boc-L-glutamic acid 5-benzyl ester in 2 mL of DMF was added 105 mg (0.276 mmol) of HATU and 0.06 mL (0.55 mmol) of N-methylmorpholine. The reaction mixture was stirred for 16 h then quenched with 1 mL of saturated NH4C1 solution and 1 mL of water. The mixture was extracted with EtOAc three times; the combined organic s were washed with brine and then dried over a2S04. The solvent was then evaporated and the residue was chromatographed (0% to 20% CH3OH / dichloro methane) to afford 160 mg of compound Int 1B-4 as a yellow solid. LCMS (M+H) = 725.
Step E: Preparation of Int 1B-5
[0609] A solution containing 100 mg (0.14 mmol) of Int 1B-4 in 20 mL of 1: 1 THF - EtOH was degassed and back filled with nitrogen three times. 100 mg of 20% Pd(OH)2 was added and the mixture was degassed two additional times. The reaction mixture was stirred for 16 h then filtered through Celite with EtOAc. Removal of the solvent and trituration with toluene afforded 75 mg of Int 1B-5 which was used directly in the next step. LCMS (M+H) = 635. Step F: Preparation oflnt 1B-6
[0610] To a solution containing 75 mg (0.12 mmol) of lnt 1B-5 and 82 mg (0.15 mmol) of (2S,4R)-l-((S)-2-(3-(2-aminoethoxy)propanamido)-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4- methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide in 1.0 mL of DMF was added 66 mg (0.15 mmol) of BOP reagent and 0.026 mL (0.24 mmol) of diisopropylethylamine. The reaction mixture was stirred for 16 h then quenched with 1 mL of saturated NaHC03 solution and 1 mL of water. The mixture was extracted with EtOAc three times; the combined organic extracts were washed with brine and then dried over a2S04. The solvent was then evaporated and the residue was chromatographed (0% to 20% CH3OH / dichloromethane) to afford 58 mg of the desired compound as a yellow solid which was immediately dissolved in 5 mL of EtOAc then treated with 1 mL of 4 N HC1 in dioxane at room temperature and the reaction was stirred for 3h. The solvent was removed under reduced pressure and the residue was azeotroped three times with toluene then stirred with ether and filtered to afford 43 mg of (S)-2-amino- N1-(4-(5 -amino-6-((4- morpholinopyridin-3-yl)carbamoyl)pyrazin-2-yl)benzyl)-N5-(2-(3-(((S)-l-((2S,4R)-4-hydroxy-2- ((4-(4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidin-l-yl)-3,3-dimethyl-l-oxobutan-2- yl)amino)-3-oxopropoxy)-ethyl)pentanediamide trihydrochloride as bright yellow crystalline solid. LCMS (M+H) = 1062.
[0611] Int 1B-6 is PROTAC T-015 Step G: Preparation of Compound 1-1
Compound 1-
[0612] A solution containing 43 mg (0.037 mmol) of (S)-2-amino-N1-(4-(5-amino-6-((4- morpholinopyridin-3-yl)carbamoyl)pyrazin-2-yl)benzyl)-N5-(2-(3-(((S)-l-((2S,4R)-4-hydroxy-2- ((4-(4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidin-l-yl)-3,3-dimethyl-l-oxobutan-2- yl)amino)-3-oxopropoxy)ethyl)pentanediamide trihydrochloride was combined with (16 mg, 0.037 mmol) of LC-smcc (succinimidyl 4-(N-maleimidomethyl)cyclohexane-l-carboxy-(6- amidocaproate)) in 1.5 mL of DCM and DIPEA (0.064 mL, 0.36 mmol). After stirring overnight, the reaction became cloudy and LCMS indicated the presence of product. The reaction was concentrated then taken up in a minimum amount of THF and water. The mixture was neutralized with saturated NaHC03 and the mixture was chromatographed (30 g, CI 8, H20 to CH3CN, liquid load) to provide Compound 1-1 (31.8, mg) as a yellow solid after lyophilization from CH3CN/H20. 1H NMR (CD3OD) δ 9.46 (s, IH), 8.84 (s, IH), 8.78 (s, IH), 8.26 (d, J=8.5Hz, IH), 8.03 (d, J=8.5Hz, 2H), 7.44 (d, J=8.5Hz, 2H), 7.41 (d, J=8.4Hz, 2H), 7.35 (d, J=8.4Hz, 2H), 7.26 (d, J=5.5Hz, IH), 6.78 (s, 2H), 4.66 (s, IH), 4.59 (m, 2H), 4.46 (t, J=7.0Hz, 4H), 4.37 (m, 2H), 3.88 (d, J=11.5Hz, IH), 3.81-3.70 (m, 5H), 3.69 (t, J=5.5Hz, 2H), 3.55-3.49 (m, 3H), 3.11 (t, J=11.5Hz, 2H), 3.10-3.01 (m, 5H), 2.50 (t, J=15.0Hz, 2H), 2.33 (s, 3H), 2.35- 2.22 (m, 6H), 2.11-2.01 (m, 4H), 1.94 (m, 1H), 1.76-1.58 (m, 8H), 1.50-1.25 (m, 8H), 1.11 (s, 9H), 1.05-0.95 (m, 4H). LCMS (M+H) = 1395.6.
[0613] The following compounds were prepared in an analogous manner to that described for the synthesis of Compound 1-1 by substituting the appropriate aryl boronic acid in step B and E3 ligase ligand / spacer group in step E.
TABLE 12. Exemplary Compounds
TABLE 13. Exemplary Compounds
y l)carbamo y l)pyrazin- 2- yl)phenethy 1) -
2- (6-(4-((2,5-dioxo-2,5-diliydro-lH- pyrrol- 1 -yl)methyl)cyclohexane- 1 - carboxamido)hexanamido)-N5-(2-(3- (((S)-l-((2S,4R)-4-hydroxy-2-((4-(4- methylthiazol-5- yl)benzyl)carbamoyl)pyrrolidin-l-yl)- 3 , 3 -dimethyl- 1 -oxobutan- 2- yl) amino ) -
3- oxopropoxy)ethyl)pentanediamide
ArB(OH)2 ^ - B(OH)2
E3 Ligand
M+l 1408
EXAMPLE 2
[0614] Synthesis of 3-amino-6-(4-(2-((2S)-2-(6-(4-((2,5-dioxo-2,5-diliydro-lH-pyrrol-l- yl)methyl)cyclohexane-l-carboxamido)hexanamido)-6-(2-((2-(2,6-dioxopiperidin-3-yl)-l,3- dioxoisoindolin-4-yl)oxy)acetamido)hexanamido)ethyl)phenyl)-N-(4-morpholinopyridin-3- yl)pyrazine-2-carboxamide (Compound 2-1)
Step A: Preparation oflnt 7B-1
Int 1 B-1
Int 7B-1
[0615] A solution containing 3.0 g (8.0 mmol) of 3-amino-6-bromo-N-(4-morpholinopyridin-3- yl)pyrazine-2-carboxamide and 2.6 g (8.8 mmol) of (4-(2-(((tert-butoxy)carbonyl)- amino)ethyl)phenyl)boronic acid in 50 mL of dioxane and 8 mL of 2N a2C03 (16.0 mmol) was degassed and back filled with nitrogen three times. 600 mg (0.8 mmol) of PdCl2 (dppf) was added and the reaction vessel was degassed with nitrogen twice. The reaction mixture was then heated at 90 °C for 3 h then cooled and stirred overnight then filtered through a plug of Celite®. The filtrate was diluted with EtOAc, washed with water and then brine, and dried over Na2S04. The solvent was then evaporated and the residue was chromatographed (0% to 20% MeOH / dichloromethane) to afford 2.5 g of compound Int 1.2a as a brown solid. The material was dissolved in 100 mL of 1: 1 THF : EtOH was degassed and back filled with nitrogen three times. 500 mg of 20% Pd(OH)2 was added and the mixture was degassed two additional times. The reaction mixture was stirred for 16 h then filtered through Celite with EtOAc. Removal of the solvent afforded 2.0g of lnt 7B-1 which was used directly in the next step. LCMS (M+H) = 420.
Step B: Preparation oflnt 7B-2
Int 7A-1 Int 7B-2
[0616] To a solution containing 228 mg (0.60 mmol) of Boc-L-Lys(Z)-OH in 5 mL of DMF was added 228 mg (0.60 mmol) of HATU and the reaction was stirred for 5 minutes before the addition of 210 mg (0.50 mmol) of lnt 7B-1 and 121 mg (1.2 mmol) of N-methylmorpholine. The reaction mixture was stirred for 3h then quenched with 5 mL of saturated NaHC03 solution and 2 mL of water. The mixture was extracted with EtOAc three times; the combined organics were washed with brine and then dried over a2S04. The solvent was then evaporated and the residue was chromatographed (0% to 20% CH3OH / dichloromethane) to afford 190 mg of compound Int 7B-2 as a yellow solid. LCMS (M+H) = 782.
Step C: Preparation of Int 7B-3
Int 7B-3
[0617] A solution containing 164 mg (0.21 mmol) of Int 7B-2 in 10 mL of methanol was degassed three times while back filling with nitrogen before the addition of 50 mg of 5% Pd on carbon. A balloon of hydrogen was added and the reaction was stirred for 3h then filtered through Celite with EtOAc. Removal of the solvent afforded 40 mg of Int 7B-3 as a yellow solid. 1H NMR (CD3OD) δ 9.48 (s, 1H), 8.77 (s, 1H), 8.28 (d, J=5.6Hz, 1H), 8.02 (d, J=8.0Hz, 2H), 7.40 (d, J=8.0Hz, 2H), 7.28 (d, J=5.2Hz, 1H), 3.98-3.79 (m, 5H), 3.51 (m, 2H), 3.04 (t, J=4.8Hz, 4H), 2.90 (t, J=5.1Hz, 2H), 2.62 (t, J=7.2Hz, 2H), 1.68 (m, 1H), 1.51 (s, 9H), 1.44-1.22 (m, 5H). LCMS (M+H) = 648.3.
[0618] To a solution containing 58 mg (0.09 mmol) of Int 7B-3 and 30 mg (0.09 mmol) of 2-((2- (2,6-dioxopiperidin-3-yl)-l,3-dioxoisoindolin-4-yl)oxy)acetic acid in 1 mL of DMF was added 48 mg (0.11 mmol) of BOP reagent and 0.047 mL (0.27 mmol) of diisopropylethylamine. The reaction mixture was stirred for 16 h then quenched with 1 mL of saturated NaHC03 solution and 1 mL of water. The mixture was extracted with EtOAc three times and the combined organic extracts were washed with brine and then dried over a2S04. The solvent was then evaporated and the residue was chromatographed (0% to 20% CH3OH / dichloro methane) to afford 60 mg of the desired compound which was immediately dissolved in 4 mL of EtOAc and 1 mL of methanol then treated with 2 mL of 4 N HCl in dioxane at room temperature and the reaction was stirred for 2h. The solvent was removed under reduced pressure and the yellow solid was evaporated three times from diethyl ether to afford 49 mg of Int 7B-4 as bright yellow crystalline solid. LCMS (M+H) = 862.
[0619] Int 7B-4 is PROTAC T-20. Ste E: Preparation of Compound 2-1
[0620] To a solution containing 43 mg (0.05 mmol) of (S)-2-amino-N1-(4-(5-amino-6-((4- morpholinopyridin-3-yl)carbamoyl)pyrazin-2-yl)benzyl)-N5-(2-(3-(((S)-l-((2S,4R)-4-hydroxy-2-
((4-(4-methylthiazol-5-yl)benzyl)carbam
yl)amino)-3-oxopropoxy)ethyl)pentanediamide trihydrochloride as bright yellow crystalline solid which was combined with (32 mg, 0.07 mmol) of LC-smcc (succinimidyl 4-(N- maleimidomethyl)cyclohexane-l-carboxy-(6-amidocaproate)) in 3 mL of DCM and DIPEA (0.13 mL, 0.7 mmol). After stirring overnight, the reaction became cloudy and LCMS indicated the presence of product. The reaction was concentrated then taken up in a minimum amount of THF and water. The mixture was neutralized with saturated NaHC03 and the mixture was
chromatographed (30 g, CI 8, H20 to CH3CN, liquid load) to provide Compound 2-1 as a yellow solid after lyophilization from CH3CN/H20. LCMS (M+H) = 1194.3.
EXAMPLE 3
TRAF2 And NCK Interacting Kinase (TNIK) Inhibitors
[0621] Synthesis of 4-((S)-2-((S)-2-(6-(2,5-dioxo-2,5-dihydro-lH-pyrrol-l-yl)hexanamido)-3- methylbutanamido)-5-ureidopentanamido)benzyl((6-((8-(((ls,4s)-4-hydroxy- cyclohexyl)oxy)quinazolin-2-yl)amino)-lH-benzo[d]imidazol-4-yl)methyl)carbamate (Compound 1C)
Step A: Preparation oflnt lC-1
Int 1C-1
[0622] 4-bromo-lH-benzo[d]imidazol-6-amine (903 mg, 4.26 mmol) was dissolved in a mixture of 15 mL THF, 8 mL of H20 and 20 mL of MeOH. Solid NaHC03 (716 mg, 8.52 mmol) was added and the mixture was stirred for 15 min before adding 1.4 g (6.39 mmol) of Boc20. The reaction mixture was concentrated and covered with MeOH to give a fine dark suspension. Silica gel was then added and the mix was concentrated to dryness. Silica gel column chromatography (ISCO 125 g, DCM to 20% MeOH/DCM) provided the desired material (1.1 g) as a yellow solid. 1H NMR (CDC13) δ 8.48 (s, 1H), 8.41 (s, 1H), 8.14 (s, 1H), 7.54 (s, 1H), 1.69 (s, 9H).
Step B: Preparation oflnt lC-2
Int 1C-1 Int 1C-2
[0623] To a mixture containing Int lC-1 (0.979 g, 2.37 mmol) in 25 mL of DMF was added 4,5- bis(diphenylphosphino)-9,9-dimethylxanthene (0.137 g, 0.237 mmol) and Zn(CN)2 (0.418 g, 3.56 mmol). The mixture was purged with N2 for 10 minutes. Palladium(II) acetate (0.053 g, 0.237 mmol) was then added and the mixture purged with N2 for 10 min then heated to 80°C. After 5h the reaction was cooled and diluted with EtOAc and filtered through Celite. Chromatography (24 g Gold silica, DCM to 20% MeOH/DCM) gave tert-butyl (4-cyano-lH-benzo[d]imidazol-6- yl)carbamate gave the desired product as a pale pink solid which was used directly in the next step.
Step C: Preparation oflnt lC-3
[0624] H Cube: 22 mL of 30% concentrated NH4OH was diluted to 200 mL with MeOH. Half of this solution was used to prime and wash the H-cube lines and 2N NH4OH in MeOH (88 ml) was used to dissolved the sample. Used 70 x 4 mm Ra-Ni column, 60°C, 10 psi, 1 ml/min, 0.026 molar in NH4OH/MeOH for 4h (recirculate) on the H-cube instrument when LCMS showed product with some SM remained. The sample was concentrated and placed under high vacuum for 16h. Chromatography (40 g silica, Gold, DCM to 80: 18:2 DCM: MeOH :NH4OH) gave a partial separation and 440 mg of the desired product as a white solid which was used directly in the next step without additional purification. LCMS (M+H) = 263.
Step D: Preparation oflnt lC-4
H2N
Int 1 C-3 Int 1 C-4
[0625] To an ice-cold mixture of lnt lC-3 and 3 mL saturated NaHC03 in 17 mL of THF was added benzyl chloroformate (0.29 mL, 2.0 mmol) dropwise. The reaction mixture was stirred for 3h, then concentrated, covered with EtOAc and filtered through Na2S04, concentrated with silica gel and dry loaded onto a 24 g silica Gold cartridge. Elution with 100% heptanes to 100% EtOAc gave 450 mg of Int lC-4 as a white solid. 1H NMR (CDC13) d 8.52 (s, 1H), 8.29 (s, 1H), 7.6-7.3 (m, 11H), 6.60 (bs, 1H), 5.55 (s, 2H), 5.25 (s, 2H), 4.61 (s, 2H), 1.56 (s, 9H).
Step E: Preparation of Int lC-5
CbzHN
Int 1 C-4 Int 1 C-5
[0626] To a suspension of benzyl 4-((((benzyloxy)carbonyl)amino)methyl)-6-((tert- butoxycarbonyl)amino)-lH-benzo[d]imidazole-l-carboxylate (161 mg, 0.303 mmol) in 10 mL of MeOH was added K2CO3 (84 mg, 0.606 mmol). The reaction was stirred at room temperature for lh when TLC showed the reaction to be complete. Chromatography (4 g silica, Gold, DCM to 20% MeOH/DCM) gave the mon-deprotected compound (141.8, mg) as a white solid. This material was dissolved in 9 mL of DCM and treated with 1 mL of TFA. The reaction was stirred at room temperature for lh then concentrated. The residue taken up in DCM and treated with 1 mL of Et3N. The reaction was concentrated and chromatographed (4 g silica gel, Gold, DCM to
80: 18:2 DCM:MeOH:NH40H) to give 115 mg of Int lC-5 as a pale yellow semi-solid. 1H NMR (CDCI3) δ 7.70 (s, 1H), 7.25 (m, 5H), 6.72 (s, 1H), 6.42 (s, 1H), 5.87 (bs, 1H), 5.05 (s, 2H), 4.44
(s, 2H).
Step F: Preparation of Int lC-6
Int 1C-5
Int 1C-6
[0627] A mixture containing 56 mg (0.188 mmol) of Int lC-5 and 96 mg (0.244 mmol) of 8- (((ls,4s)-4-((tert-butyldimethylsilyl)oxy)cyclohexyl)oxy)-2-chloroquinazoline in 2 mL of isopropanol was heated in a microwave tube for 2h at 150°C. The reaction was cooled and 0.5 mL of water was added. Tetrabutylammonium fluoride (564 μΐ, 0.564 mmol) was added and the mixture was stirred for 2h. The solvent was concentrated then partitioned between EtOAc and NaHC03. The EtOAc was washed with water then brine and dried ( a2S04), filtered and concentrated. The material was adsorbed onto silica gel using DCM then concentrated.
Chromatography (24 g silica gel, Gold, DCM to 10% MeOH/DCM) to give 41 mg of Int lC-6 as a yellow solid. 1H NMR (CD3OD) δ 9.12 (s, IH), 9.05 (bs, IH), 8.06 (s, IH), 7.41-7.06 (m, 9H), 5.13 (s, 2H), 4.89 (s, IH), 4.71 (bs, 2H), 3.86 (bs, IH), 2.20-2.07 (m, 4H), 1.89-1.80 (m, 2H), 1.73 (t, J=12Hz, 2H). LCMS (M+H) = 539.6.
Step G: Preparation of Compound lC-7
[0628] A mixture containing benzyl ((6-((8-(((ls,4s)-4-hydroxycyclohexyl)oxy)quinazolin-2- yl)amino)-lH-benzo[d]imidazole-4-yl)methyl)carbamate (181 mg, 0.336 mmol) was combined with water (2 ml) and 4 N HCl in dioxane (2 ml) in a microwave tube then heated in a microwave for 2h at 100°C. The solvents were removed under reduced pressure and saturated NaHC03 was added to make the free-base. This mixture was loaded onto a 100 g C18 column using a minimum of MeOH to finish the loading. Elution with H20 to CH3CN (TFA modifier) gave 121 mg of Int lC-7 as yellow solid after co-evaporation with DCM and heptane. 1H NMR (D20) δ 9.12 (s, IH), 8.97 (s, IH), 8.53 (s, IH), 7.48 (d, J=2.0Hz, IH), 7.42 (dd, J=2.0, 8.0Hz, IH), 7.34 (d, J=8.0Hz, IH), 7.30 (m, 3H), 4.70 (s, IH), 4.44 (s, 2H), 3.80 (m, IH), 1.94 (m, 2H), 1.67 (m, 6H). LCMS (M+H) = 405.3. Step H: Preparation of Compound 1 C
[0629] To a solution containing 4-((S)-2-((S)-2-(6-(2,5-dioxo-2,5-dihydro-lH-pyrrol-l- yl)hexanamido)-3-methylbutanamido)-5-ureidopentanamido)benzyl (4-nitrophenyl) carbonate (45.6 mg, 0.062 mmol) in 1 mL of DMF was added 618 uL of a 0.1 M solution of (ls,4s)-4-((2- ((4-(aminomethyl)-lH-benzo[d]imidazol-6-yl)amino)quinazolin-8-yl)oxy)cyclohexan-l-ol (618 0.062 mmol) and N,N-diisopropylethylamine (21.53 μΐ, 0.124 mmol). The reaction was stirred for 16h then concentrated. The residue was chromatographed (30 g C18, H20 to CH3CN, liquid loaded using a mixture of THF, saturated NaHC03 (aq) and H20 to give 25 mg of
Compound 1C as a yellow solid. 1H NMR (CD3OD) δ 9.12 (s, IH), 9.07 (bs, IH), 8.06 (s, IH), 7.56 (d, J=8.0Hz, 2H), 7.40 (d, J=8.0Hz, IH), 7.33-7.22 (m, 4H), 7.12 (bs, IH), 6.76 (s, 2H), 5.08 (s, 2H), 4.89 (m, IH), 4.82 (s, 2H), 4.72 (m, 2H), 4.49 (m, IH), 4.15 (d, J=7.5Hz, IH), 3.86 (bs, IH), 3.44 (m, 3H), 3.22-3.08 (m, 4H), 2.26 (t, J=7.5Hz, 2H), 2.18-2.03 (m, 6H), 1.89 (m, 3H), 1.73 (m, 3H), 1.65-1.51 (m, 7H), 1.30 (3H), 0.96 (m, 6H). LCMS (M+H) = 1003.9.
EXAMPLE 4
Generation of Antibody- TGFb Inhibitor Conjugates through Partial Reduction of Native
Intrachain Disulfide Bonds of Non-Engineered Antibodies
[0630] The mAb (3-8 mg/mL in PBS) was exchanged into HEPES (100 mM, pH 7.0, 1 mM DTP A) via molecular weight cut-off centrifugal filtration (Millipore, 30 kDa). The resultant mAb solution was transferred to a tared 50 mL conical tube. The mAb concentration was determined to be 3-8 mg/mL by A28o. To the mAb solution was added TCEP (2.0-4.0 equivalents, 1 mM stock) at room temperature and the resultant mixture was incubated at 37 °C for 30-90 minutes, with gentle shaking. Upon being cooled to room temperature, a stir bar was added to the reaction tube. With stirring, the linker-pay load from Examples 1 and 2 (5-10 equivalents, 10 mM DMSO) was added dropwise. The resultant reaction mixture was allowed to stir at ambient temperature for 30-60 minutes, at which point N-ethyl maleimide (3.0 equivalents, 100 mM DMA) was added. After an additional 15 minutes of stirring, N-acetylcysteine (6.0-11.0 equivalents, 50 mM HEPES) was added. The crude ADC was then exchanged into PBS and purified by preparative SEC (e.g. HiLoad 26/600, Superdex 200pg) using PBS as the mobile phase. The pure fractions were concentrated via molecular weight cut-off centrifugal filtration (Millipore, 30 kDa), sterile filtered, and transferred to 15 mL conical tubes. Drug-antibody construct ratios (molar ratios) were determined by methods described in Example 4 below.
EXAMPLE 5
General Procedure for the Determination of the Drug- Antibody-Ratios [0631] Hydrophobic Interaction Chromatography
[0632] 10 μL· oΐ a 6 mg/mL solution of a conjugate is injected into an HPLC system set-up with a TOSOH TSKgel Butyl-NPR™ hydrophobic interaction chromatography (HIC) column (2.5 μΜ particle size, 4.6 mm x 35 mm) attached. Then, over the course of 18 minutes, a method is run in which the mobile phase gradient is run from 100% mobile phase A to 100% mobile phase B over the course of 12 minutes, followed by a six-minute re-equilibration at 100% mobile phase A. The flow rate is 0.8 mL/min and the detector is set at 280 nM. Mobile phase A is 1.5 M ammonium sulfate, 25 mM sodium phosphate (pH 7). Mobile phase B is 25% isopropanol in 25 mM sodium phosphate (pH 7). Post-run, the chromatogram is integrated and the molar ratio is determined by summing the weighted peak area.
Mass Spectrometry
One microgram of immune- stimulatory conjugate (antibody construct immune- stimulatory compound conjugate) is injected into an LC/MS such as an Agilent 6550 iFunnel Q-TOF equipped with an Agilent Dual Jet Stream ESI source coupled with Agilent 1290 Infinity UHPLC system. Raw data is obtained and is deconvoluted with software such as Agilent MassHunter Qualitative Analysis Software with BioConfirm using the Maximum Entropy deconvolution algorithm. The average mass of the conjugates is calculated by the software, which used top peak height at 25% for the calculation. This data is then imported into another program to calculate the molar ratio of the conjugates such as Agilent molar ratio calculator
EXAMPLE 6
Determination of Kd Values for antigens and FcgRs
[0633] Kd is measured using surface plasmon resonance assays using a BIACORE®-2000 or a BIACORE®-3000 (BIAcore, Inc., Piscataway, N.J.) at 25 °C with immobilized antigen CM5 chips at "10 response units (RU). Briefly, carboxymethylated dextran biosensor chips (CM5, BIACORE, Inc.) are activated with N-ethyl-N'-(3-dimethylaminopropyl)-carbodiimide hydrochloride (EDC) and N-hydroxysuccinimide (NHS) according to the supplier's instructions. Antigen is diluted with 10 mM sodium acetate, pH 4.8, to 5 μg/mL (~0.2 μΜ) before injection at a flow rate of 5 μΕ/ητίηυίε to achieve approximately 10 response units (RU) of coupled protein. Following the injection of antigen, 1 M ethanolamine is injected to block unreacted groups. For kinetics measurements, two-fold serial dilutions of Fab (0.78 nM to 500 nM) are injected in PBS with 0.05% polysorbate 20 (TWEEN-20™) surfactant (PBST) at 25 °C at a flow rate of approximately 25 μΕ/ηιίη. Association rates (kon) and dissociation rates (koff) are calculated using a simple one-to-one Langmuir binding model (BIACORE® Evaluation Software version 3.2) by simultaneously fitting the association and dissociation sensorgrams. The equilibrium dissociation constant (IQ) is calculated as the ratio k0ff/kon. See, e.g., Chen et al., J. Mol. Biol. 293:865-881 (1999). If the on-rate exceeds 106 M- l s- 1 by the surface plasmon resonance assay above, then the on-rate can be determined by using a fluorescent quenching technique that measures the increase or decrease in fluorescence emission intensity (excitation=295 nm; emission=340 nm, 16 nm band-pass) at 25 °C of a 20 nM anti-antigen antibody (Fab form) in PBS, pH 7.2, in the presence of increasing concentrations of antigen as measured in a spectrometer, such as a stop- flow equipped spectrophometer (Aviv Instruments) or a 8000-series SLM-AMINCO™
spectrophotometer (ThermoSpectronic) with a stirred cuvette.
EXAMPLE 7
PBMC Screening Assay
[0634] Materials and general procedures. Human peripheral blood mononuclear cells (PBMC) were obtained from BenTek, frozen at 25 x 106 cell/mL in 10 % DMSO (Sigma) prepared in fetal bovine serum (Gibco) and stored in liquid nitrogen. For the culture, PBMC were thawed quickly in a 37 °C water bath and diluted into pre-warmed RPMI 1640 (Lonza) supplemented with 10% fetal bovine serum, 2 mM glutamine, 50 μg/mL penicillin, 50 U/mL streptomycin (all from Gibco) and centrifuged for 5 minutes at 500 x g. PBMC were suspended into the growth media described above and cultured at a concentration of 1 x 106 cells per mL at 37 °C in a 5% C02 incubator.
[0635] General procedure for in vitro small molecule screening. PBMC were thawed, suspended at a concentration of 1 x 106 cell/mL in growth media and 200 was aliquoted into each well of a 96-well plate for a total of 0.2 x 106 cells per well. PBMC were incubated for approximately 16-18 hours at 37 °C in a 5% C02 humidified incubator. PBMC plates were centrifuged at 500 x g for 5 minutes and the growth media was removed. 150 of twelve concentrations ranging from 1000 to 0.000238 nM of small molecules prepared in growth media were added to PBMC in duplicate and incubated for 24 hours at 37 °C in a 5% C02 incubator. Prior to supernatant harvest, cells were spun at 500 x g for 5 minutes to remove cell debris. TNF-a activity was assessed in the supernatant by ELISA (eBioscience) or HTRF (CisBio) per the manufacturer's instructions. The optical density at 450 nm and 570 nm (ELISA) or luminescence (HTRF) was analyzed using an Envision (Perkin Elmer) plate reader.
EXAMPLE 8
General Procedure for measuring the Kd of kinase inhibitor by BLI
[0636] Measurement of the Kd for a kinase target is determined using the following reactants: (1) immune- stimulatory conjugate, (2) non-conjugated immune- stimulatory compound-linker constructs, and (3) non-conjugated immune- stimulatory control compound is performed using an Octet Red 96 instrument (ForteBio) with a biotin coated biosensor chip. As a negative control and baseline setting for the conjugate, the unconjugated control antibody is also included as a binding reactant (4). Purified, active, recombinant strep-tagged protein kinase domain is immobilized onto the bio-sensor surface at several concentrations. The analysis for each reactant is performed in the following steps: (1) baseline acquisition (60 s); (2) loading of the kinase domain onto the biotin coated biosensor (120 s); (3) second baseline acquisition (60 s); (4) association of the reactant (120 s); and (5) dissociation of reactant (480-900 s). The control reactant (1) is used at 6 concentrations of 3-fold concentration series diluted from a DMSO stock into PBS buffer for incubations. Data are analyzed using Octet Data Analysis Software 9.0 (ForteBio) and fitted to the 1: 1 binding model. Equilibrium dissociation constants (KD) are calculated by the ratio of kon to kDff. Reactants (2-4) are then used at 6 concentrations of 3-fold concentration series bracketing the observed Kd for reactant (1). Reactant (3) is diluted from a DMSO stock into PBS buffer for incubations. The antibody reactants (2) and (4) are diluted for incubations into PBS/0. l%BSA/0.02%Tween 20. Data for reactants (2-4) are analyzed using Octet Data Analysis Software 9.0 (ForteBio) and fitted to the 1: 1 binding model. Equilibrium dissociation constants (KD) are calculated by the ratio of kon to kDff.
EXAMPLE 9
CEA-TNIK Immune-stimulatory Compounds Retain Inhibitor Affinity for TNIK
[0637] The Kd for CEA-TNIK immune- stimulatory conjugates with TNIK inhibitors and the inhibitors as free, control compounds are determined using Bio-layer Inter ferometry with an Octet Red instrument (ForteBio) as follows. Dual N-terminal tagged (strep, GST) active kinase TNIK kinase (aa 1-367) is expressed as a recombinant protein by baculo virus in sf9 insect cells and purified by a GST-affinity column. The TNIK kinase is captured onto the surface of biotin- coated biosensor wells at several different loading concentrations. The analysis for each reactant is performed in the following steps: (1) baseline acquisition (60 s); (2) loading of the kinase domain onto the biotin coated biosensor (120 s); (3) second baseline acquisition (60 s); (4) association of the reactant (120 s); and (5) dissociation of reactant (480-900 s). First, the control compound is used at 6 concentrations of 3-fold concentration series diluted from a DMSO stock into PBS buffer for incubations. Data are analyzed using Octet Data Analysis Software 9.0 (ForteBio) and fitted to the 1: 1 binding model. Equilibrium dissociation constants (KD) are calculated by the ratio of kon to kDff. Next, the CEA-TNIK conjugates are then used at 6 concentrations of 3-fold concentration series bracketing the observed Kd for reactant (1).
Reactant (3) is diluted from a DMSO stock into PBS buffer for incubations. The antibody reactants (2) and (4) are diluted for incubations into PBS/0. l%BSA/0.02%Tween 20. Data for reactants (2-4) are analyzed using Octet Data Analysis Software 9.0 (ForteBio) and fitted to the 1: 1 binding model. Equilibrium dissociation constants (KD) are calculated by the ratio of kon to k0ff. The results show that the Kd of the tested CEA-TNIK immune- stimulatory conjugates is measured at 1-10 times the Kd of its control compound.
EXAMPLE 10
Immune-Stimulatory Conjugates Inhibit Signaling by A2aR without Processing
[0638] To demonstrate immune- stimulatory conjugates do not require extensive processing for activity, immune- stimulatory conjugates with A2aR inhibitors are tested in a short-term cell- based assay for antagonism of signaling by a cell surface GPCR. CHO transfectants expressing human A2aR are seeded into 96-well white bottom plates at 2-3xl04 cells/well in the absence of G418 20 hours prior to assay. Equimolar titrations of immune- stimulatory compounds and immune- stimulatory conjugates (conjugates containing the immune- stimulatory compounds at DARs 2 to 4) are added to the cells, incubated for 15-30 minutes, followed by addition of 10 nM adenine and 10 further minutes of incubation. Irrelevant immune- stimulatory compounds and conjugates are used as negative controls. (In some experiments, adenine deaminase (3U/ml) and adenosine 5' [ ,β-methylene] diphosphate (50uM) are added along with test items to lower cAMP baseline for the assay.) After the 10-minute incubation, cAMP levels are measured by an HTRF assay using a GS cAMP Assay Kit (CisBio) and an Envision multi-modal plate reader (Perkin-Elmer). The results show that the immune- stimulatory conjugates inhibit the cAMP increase generated by A2aR activation within minutes indicating that little to no processing is needed for immune- stimulatory activity and do so with IC50s close to those of the non- conjugated immuno stimulatory compound.
EXAMPLE 11
General Procedure for Determining Protein Degradation by Immune-Modulatory
Conjugates Containing Proteolysis targeting modules
[0639] Proteolysis targeting modules (PTMs) and immune-modulatory compounds are prepared as described above. Conjugates of PTMs and antibody constructs are prepared as described in Example 4. The average DAR is about 4.
[0640] Cells are plated, allowed to adhere, and treated with vehicle, an inhibitor, a PROTAC or an immune-modulatory conjugate in the presence or absence of proteasome inhibitor, such as
MG-132. After treatment, media is aspirated and cells are rinsed with ice cold PBS. Ice cold lysis buffer (20 mM TrisHCl pH 7.5, 150 mM NaCl, 1% Triton X-100, 2 mM EDTA and 10% glycerol) containing phosphatase and protease inhibitors is added to wells and cells are removed from the plate using a cell scraper. Lysates are transferred to a 1.5 ml tube and rocked for one hour at 4°C with vortexing every -15 minutes. Tubes are spun at 8500xg for 10 minutes and supernatants are drawn into an insulin needle twice. Cell lysates are frozen at -80°C. Protein concentration is determined using a BCA assay and equal amounts of samples are boiled with reducing loading buffer. The samples are then subjected to electrophoresis on 4-20%
polyacrylamide gels which are then transferred to PVDF membranes. Blocking and staining are done in 5% w/v soy milk PBS with 0.05% Tween 20 and washing using PBS with 0.05% Tween.
For PROTACs and immune-modulatory conjugates targeting TGFPR2, blots are incubated overnight with rocking at 4°C with 1:200 primary anti-TGFpR2 antibody (Santa Cruz, sc-17791).
For PROTACs and immune-modulatory conjugates targeting TGFpRl, blots are incubated overnight with rocking at 4°C with 1:3000 primary anti-TGFpRl antibody (R&D, MAB5871).
Loading controls are detected with 1: 15000 diluted primary antibody incubation at room temperature for 1 hour with rocking (Tubulin - Abeam, ab7291 ; Actin - Abeam, ab8224).
Secondary antibodies are diluted 1: 10000 and blots are incubated for 1 hour at room temperature with rocking (Jackson ImmunoResearch, 115-035-003 or 112-035-003). ECL reagent is used to detect the signal and blots are imaged using the ChemiDoc MP (Biorad). Analysis of densitometry is done using the ImageLab software and signals are adjusted based on loading control.
EXAMPLE 12
Degradation of TFGPR2 by a TGFpR2-VHL PROTAC Conjugated to an Anti-HER2
Antibody.
[0641] Protac T-20 was prepared as described in Example 2. Pertuzumab was used as the HER2 antibody. Her2 Antibody-Protac conjugate (050-T11020; Compound 2.1 (Example 2)) was prepared by attachment of maleimidomethylcyclohexane-l-carboxylate linker to T-20 to form a linker-T-20 construct (T11020) followed by conjugation of Tl 1020) to the Her2 antibody generally following the protocol in Example 4 for interchain cysteine conjugation. The average drug loading was about 2 to 4.
[0642] Plasmid expressing HER2 was transfected into HEK293 cells using commercially available materials and conditions. Twenty four hours after transfection, cells were treated with DMSO, PROTAC T-20, HER2 antibody (IgGl), or Her2 Antibody-Protac conjugate (050- Tl 1020). Whole cell lysates were prepared from cells after 2, 24, or 48 hours incubation and quantitated with a BCA assay. Equal amounts of lysates were run on protein gels, transferred to PVDF, and TGFPR2 was detected using commercially available reagents. To quantitate the amount of protein degradation, the signals on the Western blot were adjusted to actin loading control and data is presented as a percent of matched control. Referring to FIGURE 21A, FIGURE 21B, and FIGURE 21C, at both tested concentrations, 0.5uM and luM of conjugate, the level of target TGFPR2 was diminished at 24 and 48 hours of treatment, while TGFPR2.
EXAMPLE 13
Antigen Targeted Degradation of TGFpR2 by an Antibody Conjugate using VHL and
Cereblon E3 Binding Moieties
[0643] A HER2 antigen positive, TROP2 antigen negative cell line BT474 was used to demonstrate antigen specific delivery of PROTAC conjugates.
[0644] Protac T-15 and T-20 were prepared as described above in Examples 1 and 2, respectively. Pertuzumab was used as the Her2 antibody. Sacituzumab was used as the Trop2 antibody. Her2 Antibody-Protac conjugates (050-T05015 and 050-T05020) were prepared by attachment of an MC-VC-PAB linker to T-15 or T-20 for form T05015 and T05020 constructs, respectively, followed by conjugation to the Her2 antibody generally following the protocol in Example 4 for interchain cysteine conjugation. The average drug loading was about 2 to 4. Trop2-Protac conjugate (130-T05020) was similarly prepared.
[0645] BT474 cells were plated and treated the following day with either a small molecule (T-15 or T-20), a conjugate of a HER2 antibody TGFpR2-VHL binding PROTAC (050-T05020), a conjugate of a HER2 antibody TGFpR2-Cereblon binding PROTAC (050-T05015) or a conjugate of a TROP2 antibody TGFpR2-VHL binding PROTAC (130-T05020). Whole cell lysates were prepared 24 hours after treatment and quantitated with a BCA assay. Equal amounts of lysates were run on protein gels, transferred to PVDF, and TGFPR2 and actin were detected using commercially available reagents. Quantitation of protein bands was performed and Western signal was adjusted to actin loading control and data is presented as a percent of vehicle control. Referring to FIGURE 22A and FIGURE 22B, HER2-antigen specific degradation was found with both the HER2 binding PROTAC conjugates, but not with the control TROP2- binding PROTAC conjugates.
EXAMPLE 14
Lowered Cellular Level of TGFpR2 and TGFpRl by a TGFpR2/TGFpRl-VHL PROTAC is Proteasome Inhibitor Sensitive.
[0646] Normal human lung fibroblasts were treated with or without proteasome inhibitor MG- 132 followed by the addition of DMSO or T-20. Whole cell lysates were prepared and then quantitated with a BCA assay. Equal amounts of lysates were run on protein gels and transferred to PVDF membrane. TGFpRl, TGFPR2, and actin were detected using commercially available reagents. Western signal was adjusted to actin loading control and data is presented as a percent of the matched vehicle control. Referring to FIGURE 23A and FIGURE 23B, addition of the proteasome inhibitor protected TGFpRl and TGFPR2 against degradation induced by T-20.

Claims

CLAIMS WHAT IS CLAIMED IS:
1. An immune-modulatory conjugate comprising:
(a) an antibody construct comprising an antigen binding domain and an Fc domain, wherein the antigen binding domain binds to a first antigen;
(b) a proteolysis targeting module, comprising:
(i) a protein targeting moiety that binds to a target protein;
(ii) an E3 ubiquitin ligase binding moiety; and
(iii) a spacer S that is covalently bound to the protein targeting moiety and to the E3 ubiquitin ligase binding moiety, wherein the spacer is optionally has 1-25 consecutive no n- hydrogen atoms; and
(c) a linker L that is covalently attached to the antibody construct and to the proteolysis targeting module.
2. The conjugate of claim 1, wherein the conjugate is represented by one of the following formulae:
(VIII), or
(IX)
wherein:
Ab is the antibody construct;
L is the linker;
ULM is the E3 ubiquitin ligase binding moiety;
IMC is the protein targeting moiety and comprises an immune-modulatory compound;
S is the spacer;
n is selected from 1 to 20; and
z is selected from 1 to 20.
3. The conjugate of any one of claims 1 to 2, wherein the Fc domain is an Fc null.
4. The conjugate of any one of claims 1 to 3, wherein the E3 ubiquitin ligase is selected from the group consisting of E3 ubiquitin ligase, such as Von Hippel-Lindaue E3 ubiquitin ligase (VHL), cereblon, mouse double minute 2 homo log (MDM2), AMFR, APC/Cdc20, APC/Cdhl, C6orfl57, Cbl, CBLL1, CHFR, CHIP, DTL (Cdt2), E6-AP, HACE1, HECTD1, HECTD2, HECTD3, HECWl, HECW2, HERC2, HERC3, HERC4, HERC5, HUWEl, HYD, ITCH, LNXl, mahogunin, MARCH-I, MARCH-II, MARCH-III, MARCH-IV, MARCH- VI, MARCH- VII, MARCH- VIII, MARCH-X, MEKKl, MIBl, MIB2, MycBP2, NEDD4, NEDD4L, Parkin, PELIl, Pirh2, PJA1, PJA2, RFFL, RFWD2, Rictor, RNF5, RNF8, RNF19, RNF190, RNF20, RNF34, RNF40, RNF125, RNF128, RNF138, RNF168, SCF/p-TrCP, SCF/FBW7, SCF/Skp2, SHPRH, SIAH1, SIAH2, SMURF1, SMURF2, TOPORS, TRAF6, TRAF7, TRIM63, UBE3B, UBE3C, UBR1, UBR2, UHRF2, WWP1, WWP2, and ZNRF1.
5. The conjugate of claim 4, wherein the E3 ubiquitin ligase binding moiety binds to VHL, cereblon, or MDM2.
6. The conjugate of any one of claims 1 to 5, wherein the protein targeting moiety binds to a protein selected from one of the groups consisting of: a) an aryl hydrocarbon receptor, androgen receptor, estrogen receptor, FK506-binding protein 12, fibroblast growth factor receptor substrate 2, phosphatidylinositol -4,5- biphosphate 3-kinase, SMAD family member 3, bromodomain and extra-territorial family of proteins (BET), bromodomain-containing protein 4 member of the BET family, Abelson tyrosine kinase, receptor- interacting serine/threonine-protein kinase 1, estrogen-related receptor, and transforming growth factor beta (TGFP); or b) ALK, Bcr-Abl, BRAF, BTK, c-KIT, EGFR, ErbB2, JAK, MEK, MET, PDGFRB, RET, ROS 1, Syk, SRC, TNIK, TNKS, TNKS2, TrkA, TrkB, TrkC, VEGFR1, VEGFR2, VEGFR3, FGFR1, FGFR2, FGFR3, FGFR4, CSF1R, RON/MS T1R, TYR03, MERTK, AXL, PI3K, PI3K, MAP4K1, PERK, and KIT; or
c) TGFpR2, TGFpRl, SMAD2, SMAD3, SMAD4, beta-catenin, CREBB2, Beta
catenin/TCF4, beta catenin/LEF, beta catenin/CREBBP, YAP, TAZ, YAP/TAZ, TNKS 1, TNKS 2, MST1, MST2, NRAS, HRAS, KRAS, RASmutl2, RASmutl3, PERK (EIF2AK3), RON/MS T1R, STAT3, MCT1, MCT2, and MCT4; or d) CSFR1, RON/MST1, PI3Kd, PDKg, PARP1, PD-L1, PP2A, A2ar, TYR03, AXL, and MER.
7. The conjugate of any one of claims 1 to 6, wherein the first antigen is a tumor antigen.
8. The conjugate of claim 7, wherein the first antigen is selected from the group consisting of MUC16, CD5, CD19, CD20, CD25, CD37, CD30, CD33, CD45, CAMPATH-1, BCMA, CS- 1, PD-L1, B7-H3, B7-DC, HLD-DR, carcinoembryonic antigen (CEA), TAG-72, EpCAM, MUCl, folate-binding protein, A33, G250, pro state- specific membrane antigen (PSMA), ferritin, GD2, GD3, GM2, Ley, CA-125, CA19-9, epidermal growth factor, pl85HER2, IL-2 receptor, fibroblast activation protein (FAP), tenascin, a metalloproteinase, endosialin, vascular endothelial growth factor, ανβ3, WT1, LMP2, HPV E6, HPV E7, EGFRvIII, Her-2/neu, MAGE A3, p53 nonmutant, NY-ESO-1, MelanA/M ART 1 , Ras mutant, gplOO, p53 mutant, PR1, bcr-abl, tyronsinase, survivin, PSA, hTERT, a Sarcoma translocation breakpoint fusion protein, EphA2, PAP, ML-IAP, AFP, ERG, NA17, PAX3, ALK, androgen receptor, cyclin Bl, poly sialic acid, MYCN, RhoC, TRP-2, fucosyl GM1, mesothelin (MSLN), PSCA, MAGE Al, sLe(animal), CYP1B1, PLAV1, GM3, BORIS, Tn, GloboH, ETV6-AML, NY-BR-1, RGS5, SART3, STn, Carbonic anhydrase IX, PAX5, OY-TES l, Sperm protein 17, LCK, HMWMAA, AKAP-4, SSX2, XAGE 1, Legumain, Tie 3, Page4, VEGFR2, MAD-CT-1, PDGFR-B, MAD-CT-2, ROR2, TRAIL 1, MAGE A4, MAGE C2, GAGE, EGFR, CMET, HER3, MUC15, CA6, NAPI2B, TROP2, CLDN18.2, RON, LY6E, FRA, DLL3, PTK7, LIV1, ROR1, and Fos-related antigen 1.
9. The conjugate of any one of claims 1 to 6, wherein the first antigen is an immune cell antigen.
10. The conjugate of claim 9, wherein the immune cell antigen is an antigen present on the surface of an antigen presenting cell selected from a dendritic cell and a macrophage.
11. The conjugate of any one of claims 9 to 10, wherein the immune cell antigen is selected from the group consisting of CD40, DEC-205, CD36 mannose scavenger receptor 1, CLEC9A, DC-SIGN, CLEC12A, BDCA-2, OX40L, 41BBL, CD204, MARCO, CLEC5A, Dectin 1, Dectin 2, CLECIOA, CD206, CD64, CD32A, CD 16 A, HVEM, and CD32B.
12. The conjugate of any one of claims 1 to 10, wherein the protein targeting moiety is selected from a peptide and a non-proteinaceous molecule.
13. The conjugate of any one of claims 1 to 12, wherein the proteolysis targeting module is selected from compounds 1-1, 1-2, 1-3, 1-4, 1-5, 1-6, and 2-1.
14. The conjugate of any one of claims 1 to 13, wherein L is a cleavable linker.
15. The conjugate of any one of claims 1 to 13, wherein L is a non-cleavable linker.
The con ugate of any one of claims 1 to 15, wherein L is selected from:
line indicates a bond of G to the antibody construct, peptide is selected from 1 to 10 amino acids, R is selected from hydrogen, Ci-ioalkyl, sulfonate and methyl sulfonate, and the wavy line at the left side indicates a bond to the remainder of the linker.
17. The conjugate of claim 16, wherein G comprises a succinimide or a hydrolyzed succinimide.
18. The conjugate of any one of claims 1 to 17, wherein L is covalently bound to the antibody construct at a cysteine residue, an engineered cysteine residue, a lysine residue, a glutamate residue, or a glutamine residue of the antibody construct or is covalently bound to the antibody construct using a Sortase linker.
19. The conjugate of any one of claims 1 to 18, wherein the spacer S is an optionally substituted C1-25 alkylene or optionally substituted C1-25 heteroalkylene, wherein the
heteroalkylene is a C1-24 alkylene chain interspersed with one or more groups independently selected from: -0-, -S-, -NH2-, and -C(0)NH-; and optionally substituted with a reactive group, RX, that can form a functional group selected from an amide bond, an ester bond, an ether bond, a carbonate bond, a carbamate bond, or a thioether bond.
20. The conjugate of any one of claims 1 to 19, wherein the antibody construct further comprises a second binding domain.
21. The conjugate of claim 20, wherein the second binding domain specifically binds to an antigen on an immune cell.
22. The conjugate of claim 21, wherein the immune cell antigen is selected from the group consisting of CD40, PD-1, PD-L1, DEC-205, CD36 mannose scavenger receptor 1, CLEC9A, DC-SIGN, CLEC12A, BDCA-2, OX40L, 41BBL, CD204, MARCO, CLEC5A, Dectin 1, Dectin 2, CLECIOA, CD206, CD64, CD32A, CD 16 A, HVEM, and CD32B.
23. The conjugate of any one of claims 20 to 22, wherein the second binding domain is attached to the antibody construct at a C-terminal end of the Fc domain.
24. The conjugate of any one of claims 11 to 23, wherein the Fc domain is an Fc domain variant comprising at least one amino acid residue change as compared to a wild type sequence of the Fc domain.
25. The conjugate of claim 24, wherein the Fc domain is an Fc domain variant that decreases binding of the Fc domain to an Fc receptor.
26. The conjugate of claim 24, wherein the at least one amino acid residue change is:
a) F243L, R292P, Y300L, L235V, and P396L, wherein numbering of amino acid residues in the Fc domain is relative to SEQ ID NO: 898;
b) S239D and I332E, wherein numbering of amino acid residues in the Fc domain is relative to SEQ ID NO: 898; or c) S298A, E333A, and K334A, wherein numbering of amino acid residues in the Fc domain is relative to SEQ ID NO: 898.
27. The conjugate of claim 24, wherein the at least one amino acid residue change is:
a) N297A, N297G, N297Q, N297D as in Eu index of Kabat numbering and
relative to SEQ ID NO: 898; or
b) K322A/L234A/L235A N296A as in the EU index of Kabat numbering and relative to SEQ ID NO: 898; or
c) L234F/L235E/P33 IS N296A as in the EU index of Kabat numbering and
relative to SEQ ID NO: 898; or
d) P329G/L234A/L235A as in the EU index of Kabat numbering and relative to SEQ ID NO: 898.
28. A pharmaceutical composition, comprising the conjugate of any one of claims 1 to 27 and a pharmaceutically acceptable excipient.
29. The pharmaceutical composition of claim 28, wherein the average ratio of proteolysis targeting modules to antibody construct in the conjugate is from 2 to 6, from 3 to 5, or 1 to 3.
30. The pharmaceutical composition of one of claims 28 to 29, which is lyophilized.
31. A method of treating cancer, comprising administering the conjugate of any one of claims 1 to 27 or the pharmaceutical composition of any one of claims 28 to 30 to a subject in need thereof.
32. The method of claim 31, wherein the cancer is a solid tumor.
33. The method of claim 32, wherein the solid tumor is breast cancer, pancreatic cancer, colorectal cancer, renal cell cancer, gastric cancer, or lung cancer.
34. The method of any one of claims 31 to 33, wherein the pharmaceutical composition is administered parenterally.
35. The method of any one of claims 31 to 33, wherein the pharmaceutical composition is administered intravenously.
36. The use of the conjugate of any one of claims 1 to 27 in the preparation of a medicament for the treatment of cancer.
37. An immune- stimulatory conjugate comprising:
a) an immune-modulatory compound that binds to a protein active site of an intracellular protein target to stimulate an immune response by degradation of the protein target;
b) an antibody construct comprising an antigen binding domain and an Fc domain, wherein the antigen binding domain binds to a first antigen expressed on cells having the intracellular protein target; and
c) a linker, wherein the linker is covalently bound to the antibody construct and the linker is covalently bound to the immune-modulatory compound; wherein the linker is optionally a linker having 5- 100 consecutive atoms; wherein the Kd for binding of the immune-modulatory compound, attached to the antibody construct of the conjugate, to the protein active site is equal to, or up to no greater than 100 times the Kd for binding of a control compound to the protein active site or wherein the IC50 of the immune-modulatory compound, attached to the antibody construct of the conjugate, is no greater than 300-fold the IC50 of a control compound, wherein the control compound is the unbound immune-modulatory compound.
38. The immune- stimulatory conjugate of claim 37, wherein the Fc domain is an Fc null.
39. The immune- stimulatory conjugate of claim 37, wherein the Fc domain binds to an Fc receptor and the dissociation constant (Kd) for binding of the Fc domain of the conjugate to the Fc receptor is equal to, or up to no greater than about 100 times the Kd for binding of a control antibody construct to the Fc receptor, wherein the control antibody construct is the unconjugated antibody construct.
40. The immune- stimulatory conjugate of any one of claims 37 to 39, wherein the EC50 or IC50 of the immune-modulatory compound attached to the antibody construct of the conjugate is no greater than 10-fold the EC50 or IC50 of a control compound, wherein the control compound is the unbound immune-modulatory compound.
41. The immune- stimulatory conjugate of any one of claims 37 to 40, wherein the EC50 or IC50 of the immune-modulatory compound attached to the antibody construct of the conjugate is equivalent to or less than the EC50 or IC50 of a control compound, wherein the control compound is the unbound immune-modulatory compound.
42. The immune- stimulatory conjugate of any one of claims 37 to 41, wherein the conjugate further comprises an E3 ubiquitin ligase binding moiety.
43. The immune- stimulatory conjugate of claim 42, wherein the E3 ubiquitin ligase binding moiety binds to VHL, cereblon or MDM2
44. The immune- stimulatory conjugate of any one of claims 42 to 43, wherein the E3 ubiquitin ligase binding moiety is selected from compounds 1-1, 1-2, 1-3, 1-4, 1-5, 1-6, and 2-1.
45. The immune- stimulatory conjugate of any one of claims 42 to 44, wherein the E3 ubiquitin ligase binding moiety is attached to the immune-modulatory compound through a spacer, optionally a 1-25 atom spacer, and the spacer is bound through the 5-100 atom linker to the antibody construct.
46. The immune- stimulatory conjugate of any one of claims 37 to 45, wherein the immune- modulatory compound is a kinase inhibitor and the protein active site is a kinase active site.
47. The immune- stimulatory conjugate of claim 46, wherein the linker is covalently bound to the kinase inhibitor at a position on the kinase inhibitor that is at or near the solvent interface of the kinase active site as determined by modeling of the kinase inhibitor in the kinase active site.
48. The immune- stimulatory conjugate of claim 46, wherein the linker is covalently bound to the kinase inhibitor at a position on the kinase inhibitor such that when the kinase inhibitor is bound to the active site, the linker extends out from the kinase active site into the solvent, as determined by modeling of the kinase inhibitor in the kinase active site.
49. The immune- stimulatory conjugate of any one of claims 46 to 48, wherein the kinase inhibitor is selected from an inhibitor of ALK, Bcr-Abl, BRAF, BTK, c-KIT, EGFR, ErbB2, JAK, MEK, MET, PDGFRB, RET, ROS 1, Syk, SRC, TGFpR2, TNIK, TNKS, TNKS2, TrkA, TrkB, TrkC, VEGF, VEGFRl, VEGFR2, VEGFR3, FGFRl, FGFR2, FGFR3, FGFR4, CSFIR, RON/MS T1R, TYR03, MERTK, AXL, ΡΙ3Κδ, ΡΒΚγ, MAP4K1, PERK, and combinations thereof.
50. The immune- stimulatory conjugate of any one of claims 37 to 45, wherein the linker is covalently bound to the immune-modulatory compound at a site on the compound that does not interfere with binding of the compound to the protein active site.
51. An immune- stimulatory conjugate comprising:
(a) an immune- stimulatory compound that binds to a protein active site of a protein to stimulate an immune response, wherein the protein is present on the extracellular membrane or in the endoplasmic reticulum of a cell; (b) an antibody construct comprising an antigen binding domain and an Fc domain, wherein the antigen binding domain binds to a first antigen; and
(c) a linker having 5- 100 consecutive atoms, wherein the linker is covalently bound to the antibody construct and the linker is covalently bound to the immune- stimulatory compound; and wherein the Kd for binding of the immune- stimulatory compound, attached to the antibody construct of the conjugate, to the protein active site is no greater than 100 times the Kd for binding of a control compound to the protein active site or wherein the EC50 or IC50 of the immune- stimulatory compound, attached to the antibody construct of the conjugate, is no greater than 300-fold the EC50 or IC50 of a control compound, wherein the control compound is the unbound immune- stimulatory compound.
52. The immune- stimulatory conjugate of claim 51, wherein the Fc domain is an Fc null.
53. The immune- stimulatory conjugate of claim 51, wherein the Fc domain binds to an Fc receptor and the dissociation constant (Kd) for binding of the Fc domain of the conjugate to an Fc receptor is equal to, or up to no greater than about 100 times the Kd for binding of a control antibody construct to the Fc receptor, wherein the control antibody construct is the unconjugated antibody construct.
54. The immune- stimulatory conjugate of any one of claims 51 to 53, wherein the EC50 or IC50 of the immune- stimulatory compound, when bound to the antibody construct by the 5- 100 atom linker, is no greater than 10-fold the EC50 or IC50 of a control compound, wherein the control compound is the unbound immune- stimulatory compound.
55. The immune- stimulatory conjugate of any one of claims 51 to 54, wherein the EC50 or IC50 of the immune- stimulatory compound, when bound to the antibody construct by the 5- 100 atom linker, is equivalent to or less than the EC50 or IC50 of a control compound, wherein the control compound is the unbound immune- stimulatory compound.
56. The immune- stimulatory conjugate of any one of claims 51 to 55, wherein the immune- stimulatory compound is a toll- like receptor agonist, STING agonist, or RIG-I agonist.
57. The immune- stimulatory conjugate of claim 56, wherein the immune- stimulatory compound is a toll-like receptor agonist selected from a TLR1 agonist, a TLR2 agonist, a TLR3 agonist, a TLR4 agonist, a TLR5 agonist, a TLR6 agonist, a TLR7 agonist, a TLR8 agonist, a TLR9 agonist, or a TLR10 agonist.
58. The immune- stimulatory conjugate of any one of claims 51 to 57, wherein the immune- stimulatory compound is selected from a pyrimidine, a purine, a guanine nucleoside, an 8- oxoadenine, an imidazoquinoline, a thiazoquinoline, a 2-amino imidazole, a furo[2,3-c]pyridine, a furo[2,3-c]quinoline, a 2-aminobenzimidazole, a 2-aminoquinoline, and a 2-aminobenzazepine.
59. The immune- stimulatory conjugate of any one of claims 51 to 55, wherein the target of the immune- stimulatory compound is a GCPR, an ion channel, a membrane transporter, a phosphatase or an ER protein.
60. The immune- stimulatory conjugate of any one of claims 51 to 55, wherein the immune- stimulatory compound is an antagonist of the GPCR A2aR, the sphingosine 1 -phosphate receptor 1, prostaglandin receptor EP3, prostanglandin receptor E2, Frizzled, CXCR4 or an LPA receptor.
61. The immune- stimulatory conjugate of any one of claims 51 to 55, wherein the immune- stimulatory compound is an ion channel agonist for CRAC, Kvl.3 or KCa3.1.
62. The immune- stimulatory conjugate of any one of claims 51 to 55, wherein the immune- stimulatory compound is an inhibitor of HSP90 or AAA-ATPase p97.
63. The immune- stimulatory conjugate of any one of claims 51 to 62, wherein the conjugate has immune- stimulatory activity with no or minimal cell processing of the conjugate.
64. The immune- stimulatory conjugate of any one of claims 51 to 63, wherein the linker is a non-cleavable linker.
65. The immune- stimulatory conjugate of any one of claims 51 to 63, wherein the linker is a Fleximer linker.
66. The immune- stimulatory conjugate of any one of claims 51 to 63, wherein the linker comprises a carbamate and one or more amide linkages.
67. The immune- stimulatory conjugate of any one of claims 37 to 66, wherein the linker is
peptide— Rx
, wherein Rx is a reactive moiety and wherein R is hydrogen, alkyl, sulfonate and methyl sulfonate.
68. The immune- stimulatory conjugate of any one of claims 37 to 67, wherein the linker is attached to the antibody construct at a cysteine or lysine residue of the antibody construct.
69. The immune- stimulatory conjugate of any one of claims 37 to 68, wherein the first antigen is a tumor antigen.
70. The immune- stimulatory conjugate of any one of claims 37 to 69, wherein the first antigen is at least 80% homologous to CD5, CD19, CD20, CD25, CD37, CD30, CD33, CD45, CAMPATH-1, BCMA, CS-1, PD-L1, B7-H3, B7-DC (PD-L2), HLD-DR, carcinoembryonic antigen (CEA), TAG-72, EpCAM, MUCl, MUCl 5, MUCl 6, folate-binding protein, A33, G250, pro state- specific membrane antigen (PSMA), ferritin, GD2, GD3, GM2, Ley, CA-125, CA19-9, epidermal growth factor, pl85HER2, IL-2 receptor, EGFRvIII (de2-7 EGFR), fibroblast activation protein (FAP), tenascin, a metalloproteinase, endosialin, vascular endothelial growth factor, ανβ3, WT1, LMP2, HPV E6 E7, Her-2/neu, p53 nonmutant, NY-ESO-1,
MelanA/MARTl, Ras mutant, gplOO, p53 mutant, PR1, bcr-abl, tyrosinase, survivin, PSA, hTERT, a Sarcoma translocation breakpoint protein, EphA2, PAP, ML-IAP, AFP, ERG, NA17, PAX3, ALK, androgen receptor, cyclin Bl, polysialic acid, MYCN, RhoC, TRP-2, fucosyl GMl, mesothelin (MSLN), PSCA, MAGE Al, MAGE A3, sLe(animal), CYP1B1, PLAV1, GM3, BORIS, Tn, GloboH, ETV6-AML, NY-BR-1, RGS5, SART3, STn, Carbonic anhydrase IX, PAX5, OY-TES 1, Sperm protein 17, LCK, HMWMAA, AKAP-4, SSX2, XAGE 1, Legumain, Tie 3, VEGFR2, MAD-CT-1, PDGFR-B, MAD-CT-2, ROR2, , TRAILl, MAGE A4, MAGE C2, GAGE, EGFR, CMET, HER3, EPCAM, CA6, NAPI2B, TROP2, Claudin-6 (CLDN6), Claudin- 16 (CLDN16), CLDN18.2, RON, LY6E, FRA, DLL3, PTK7, Uroplakin-IB (UPK1B), LIV1, ROR1, STRA6, TMPRSS3, TMPRSS4, TMEM238, Clorfl86, Fos-related antigen 1, VEGFR1, endoglin, PD-L1, VTCN1 (B7-H4), VISTA, gpNMB, or any fragment thereof.
71. The immune- stimulatory conjugate of any one of claims 37 to 70, wherein the antibody construct further comprises a second target binding domain.
72. The immune- stimulatory conjugate of claim 71, wherein the target binding domain specifically binds an immune cell.
73. The immune- stimulatory conjugate of claim 71 or 72, wherein the target binding domain is attached to the antibody construct at a C-terminal end of the Fc domain.
74. The immune- stimulatory conjugate of any one of claims 37 to 73, wherein the antibody construct is an antibody.
75. The immune- stimulatory conjugate of claim 74, wherein the antibody construct is a human antibody or a humanized antibody.
76. The immune- stimulatory conjugate of any one of claims 37 to 75, wherein the Fc domain is an Fc domain variant comprising at least one amino acid residue change as compared to a wild type sequence of the Fc domain.
77. The immune- stimulatory conjugate of any one of claims 37 to 38, 39 to 52 and 54 to 76, wherein the Fc domain has at least one amino acid residue change as compared to wildtype, wherein the Fc domain is at comprises at least 80% homologous to SEQ ID NO: 296, and wherein the at least one amino acid residue change is:
a) F243L, R292P, Y300L, L235V, and P396L, wherein numbering of amino acid residues in the Fc domain is relative to SEQ ID NO: 898;
b) S239D and I332E, wherein numbering of amino acid residues in the Fc domain is relative to SEQ ID NO: 898; or
c) S298A, E333A, and K334A, wherein numbering of amino acid residues in the Fc domain is relative to SEQ ID NO: 898.
78. The immune- stimulatory conjugate of any one of claims 37 to 38, 39 to 52, and 54 to 76, wherein the Fc domain has at least one amino acid residue change as compared to wildtype, wherein the Fc domain is at comprises at least 80% homologous to SEQ ID NO: 898, and wherein the at least one amino acid residue change is:
a) N297A, N297G, N297Q, N297D as in Kabat numbering and relative to SEQ ID NO: 898; or
b) K322A/L234A/L235A N296A as in Kabat numbering and relative to SEQ ID NO: 898; or
c) L234F/L235E/P331S N296A as in Kabat numbering and relative to SEQ ID NO: 898; or d) P329G/L234A/L235A as in Kabat numbering and relative to SEQ ID NO: 898.
79. The immune- stimulatory conjugate of any one of claims 37 to 78, wherein the molar ratio of immune- stimulatory compound to antibody is less than 5.
80. A pharmaceutical composition comprising the conjugate of any one of claims 37 to 79 and a pharmaceutically acceptable excipient.
81. A method of treating cancer, comprising administering to a subject in need thereof the pharmaceutical composition of claim 80.
82. The use of the conjugate of any one of claims 37 to 79 or the pharmaceutical composition of claims 80 to 81 as a medicament for the treatment of cancer.
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