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WO2014040555A1 - Nitrogen-containing heteroaromatic ring derivative as tyrosine kinase inhibitor - Google Patents

Nitrogen-containing heteroaromatic ring derivative as tyrosine kinase inhibitor Download PDF

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Publication number
WO2014040555A1
WO2014040555A1 PCT/CN2013/083429 CN2013083429W WO2014040555A1 WO 2014040555 A1 WO2014040555 A1 WO 2014040555A1 CN 2013083429 W CN2013083429 W CN 2013083429W WO 2014040555 A1 WO2014040555 A1 WO 2014040555A1
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group
alkyl
hydrogen atom
compound
methyl
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PCT/CN2013/083429
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French (fr)
Chinese (zh)
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罗浩贤
张艳
张倩
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山东亨利医药科技有限责任公司
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Priority to CN201380041846.3A priority Critical patent/CN104583195B/en
Publication of WO2014040555A1 publication Critical patent/WO2014040555A1/en

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/56—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids
    • A61K31/58—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids containing heterocyclic rings, e.g. danazol, stanozolol, pancuronium or digitogenin
    • 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
    • C07D401/02—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 two hetero rings
    • C07D401/04—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 two hetero rings directly linked by a ring-member-to-ring-member bond
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
    • A61K31/337—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having four-membered rings, e.g. taxol
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/40—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil
    • A61K31/4025—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil not condensed and containing further heterocyclic rings, e.g. cromakalim
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D239/00—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings
    • C07D239/02—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings
    • C07D239/24—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings having three or more double bonds between ring members or between ring members and non-ring members
    • C07D239/28—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings having three or more double bonds between ring members or between ring members and non-ring members with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, directly attached to ring carbon atoms
    • C07D239/46—Two or more oxygen, sulphur or nitrogen atoms
    • C07D239/48—Two nitrogen atoms
    • 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
    • C07D401/02—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 two hetero rings
    • C07D401/12—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 two hetero rings linked by a chain containing hetero atoms as chain links
    • 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
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D403/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00
    • C07D403/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings
    • C07D403/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings linked by a chain containing hetero atoms as chain links
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D405/00—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom
    • C07D405/02—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings
    • C07D405/12—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings linked by a chain containing hetero atoms as chain links
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D413/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms
    • C07D413/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing two hetero rings
    • C07D413/12—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing two hetero rings linked by a chain containing hetero atoms as chain links

Definitions

  • the present invention relates to the field of medical technology, and particularly relates to a nitrogen-containing heteroaromatic ring derivative as a tyrosine kinase inhibitor, a pharmaceutically acceptable salt thereof or a stereoisomer thereof, a preparation method of these compounds, a drug containing the same Compositions for the prevention and/or treatment of B cell-associated blood cancers in an individual (eg, B-cell chronic lymphocytic carcinoma, non-Hodgkin's lymphoma), inflammatory and autoimmune diseases (eg, rheumatoid arthritis, Methods for systemic lupus erythematosus, etc., and the preparation of these compounds for the prevention and/or treatment of B cell-associated blood cancer (eg, B-cell chronic lymphocytic carcinoma, non-Hodgkin's lymphoma), inflammatory and autoimmune diseases Use in drugs such as rheumatoid arthritis, systemic lupus erythematosus, etc.
  • Protein Shield Kinases constitute one of the largest families of human enzymes and regulate many different signaling processes by the addition of phosphate groups to protein shields (T. Hunter, Cell 1987 50: 823-829).
  • the tyrosine kinase phosphorylation protein is shielded in the hydroxyl portion of the tyrosine residue.
  • the tyrosine kinase family includes members that control cell growth, migration, and differentiation.
  • Abnormal kinase activity has been implicated in many human diseases, including cancer, autoimmune diseases, and inflammatory diseases.
  • protein shield kinases are key regulators of cell signaling, they provide the goal of regulating cell function with small molecule kinase inhibitors and are therefore a good drug design target.
  • selective and potent inhibitors of kinase activity can be used to study cellular signaling processes and to identify other therapeutically significant cell targets.
  • B cells play a key role in the pathogenesis of autoimmune and/or inflammatory diseases.
  • Protein shield-based therapeutics that deplete B cells, such as Rituxan are effective against autoantibody-induced inflammatory diseases such as rheumatoid arthritis (Rastetter et al, Annu Rev Med 2004 55: 477). Therefore, inhibitors of protein shield kinase that play a role in B cell activation should be useful therapeutic agents for B cell mediated pathology such as autoantibody production.
  • BCR B cell receptor
  • Btk is a member of the Tec family of tyrosine kinases and has been shown to be a key regulator of early B cell formation and activation and survival of mature B cells (Khan et al, Immunity 1995 3: 283; Ellmeier et al, J. Exp. Med. 2000 192 : 1611).
  • Human Btk mutations lead to the condition X-linked gamma globulin deficiency (XLA) (Lindvall et al Immunol. Rev. 2005 203: 200). These patients are immunocompromised and show impaired B cell maturation, reduced immunoglobulin and peripheral B cell levels, reduced T cell-independent immune response, and reduced calcium use following BCR stimulation.
  • XLA X-linked gamma globulin deficiency
  • Btk-deficient mice showed a significant improvement in disease progression in a preclinical mouse model of systemic lupus erythematosus (SLE). Furthermore, Btk-deficient mice are resistant to collagen-induced arthritis (Jansson and Holmdahl Clin. Exp. Immunol. 1993 94: 459). The dose-dependent efficacy of selective Btk inhibitors in the mouse arthritis model has been demonstrated (Z. Pan et al, Chem. Med Chem. 2007 2: 58).
  • Btk also has cellular expression that may be involved in disease processes in addition to B cells.
  • Btk is expressed by mast cells and Btk-deficient bone marrow-derived mast cells show impaired antigen-induced degranulation (Iwaki J. Biol. Chem. 2005 280: 40261). This shows that Btk can be used to treat pathological mast cell responses such as allergies and asthma.
  • monocytes from XLA patients lacking Btk activity show reduced TNFa production following stimulation (Horwood et al J Exp Med 2003 197: 1603).
  • TNFa-mediated inflammation can be modulated by small molecule Btk inhibitors.
  • Btk has been reported to play a role in apoptosis (Islam and Smith Immunol. Rev. 2000 178: 49), and thus Btk inhibitors will be effective in the treatment of certain B cell lymphomas and leukemias (Feldhahn et al. Exp. Med. 2005 201 : 1837).
  • Dasatinib which was launched in 2006, is a multi-inhibitor inhibitor, which has a strong inhibitory effect on Btk and is used to treat chronic myelogenous leukemia.
  • PCI-32765 which is in clinical phase III study, is also a multi-defect inhibitor, and its inhibitory effect on Btk is Irreversible, used to treat lymphoma, leukemia and autoimmune diseases.
  • CC-292 also known as AVL-292
  • Its irreversible selective inhibition of Btk is used to treat leukemia and
  • the present invention provides nitrogen-containing heteroaryl ring derivatives useful as tyrosine kinase inhibitors, which are excellent Btk inhibitors, and which can be used for the prevention and/or treatment of B cell-associated blood cancer, inflammatory and/or Or autoimmune disease.
  • the present invention provides a compound represented by the following formula (I), a pharmaceutically acceptable salt thereof or
  • ring A and ring B each independently represent a phenyl group, a 3- 7 membered cycloalkyl group, a 3- 7 membered heterocycloalkyl group having a N, 0, S hetero atom, a 4- 7 membered heteroaryl group or 6- 10-membered two-ring structure;
  • Li and L 2 each independently represent a covalent bond, -NH-, -N(C 1-3 alkyl) -, -0- , - S(0) m - , - N(C 1-3 alkyl) C(0)- , - C(0)N(C 1-3 alkyl) -, - N(C 1 -3 alkyl)S(0) 2 - or - S(0) 2 N(C 1- 3 alkyl) -;
  • A represents a covalent bond, a substituted or unsubstituted alkylene d_ 4 alkyl substituted amino
  • b represents -CO- or -S0 2 -;
  • c represents 1,3-propenylene, 1,1- or 1,2-vinylidene, ethynylene, or unsubstituted or unsubstituted or substituted by one or two methyl or trifluoromethyl groups a 1,3-butadiene-1,4-subunit substituted with one to four methyl or trifluoromethyl groups;
  • d represents a covalent bond or an alkylene group
  • e represents a hydrogen atom, an alkoxy group, an amino group, a 3- 7 membered cycloalkyl group, a 6- to 10-membered bicyclic structure, an alkylamino group or a bis-(Cw alkyl)amino group, wherein the alkyl moieties may be the same or different
  • L 3 represents a covalent bond, -NH-, -N(C 1-3 alkyl) -, -0- , - 0- C 1-3 alkylene-, -S-Ci -3 alkylene, - S(0) m - , - C(O)- , -NHC(O)- , - N(C 1-3 alkyl) C(O)- , - C(0)NH- , -C(0) N(Ci -3 alkyl) -, - NHS(0) 2 - , -N(Ci -3 alkyl) S(0) 2 - , -S(0) 2 NH- , -S(0) 2 N (Ci -3 alkyl) -, - OC(O)- or - C(0)0- ,
  • R4 represents a hydrogen atom, an alkyl group, -N(Cw alkyl) 2 , -NHC(0)0-(Cw alkyl), -OH, - 0(C 1-4 alkyl), - S(0) 2 (C 1-3 alkyl), 3- 7 membered cycloalkyl, phenyl or 5- 6-membered heteroaryl;
  • R 2 represents - L 4 - R 5 ,
  • L 4 represents a covalent bond, -NH-, -N(C 1-3 alkyl)-, - 0-C 1-3 alkylene-, -S-C 1-3 alkylene- or -S ( 0) m - ,
  • R 5 represents a hydrogen atom, C 2-4 alkenyl, C 2-4 alkynyl, C 1-4 alkyl, amino, -NH(C 1-3 alkyl), -N(Cw alkyl) 2 , - OH , - 0 (Cw alkyl), - C(0) (Cw alkyl) or 3- 7 membered cycloalkyl,
  • R 5 cannot be C 1-4 alkyl, - OH , - 0 (C 1-3 alkyl), - C(0) (C 1-3 alkyl),
  • L 4 is - 0- d_ 3 alkylene - when, R 5 is not a hydrogen atom;
  • alkyl moiety, cycloalkyl group, heteroaryl group may be further substituted by 1 to 4 ( ⁇ ,
  • the carbon atom of the cycloalkyl or bicyclic structure may be replaced by 1-4 identical or different N, NH, N(C 1-3 alkyl), 0, S(0) m , C(O);
  • the heteroaryl group has 1-4 hetero atoms, and each of them is independently selected from N, 0 or S;
  • n 0, 1 or 2;
  • p and q are independent representations of 0, 1, 2, 3 or 4.
  • the present invention provides the above-described formula (I) a compound, a pharmaceutically acceptable salt thereof or a stereoisomer thereof, wherein:
  • Ring A and Ring B each independently represent a phenyl group, a 3- 7 membered cycloalkyl group, a 3-7 membered heterocycloalkyl group having a N, 0, S hetero atom, a 4-7 membered heteroaryl group or a 6-10 member.
  • Li and L 2 each independently represent a covalent bond, -NH-, -N(C 1-3 alkyl) -, -0- , - S(0) m - , - N(C 1-3 alkyl) C(0)- , - C(0)N(C 1-3 alkyl) -, - N(C 1 -3 alkyl)S(0) 2 - or - S(0) 2 N(C 1- 3 alkyl) -;
  • A represents a covalent bond, a substituted or unsubstituted alkylene d_ 4 alkyl substituted amino
  • b represents -CO- or -S0 2 -;
  • c represents 1, 3-propenylene, 1,1- or 1,2-vinylidene, ethynylene, or unsubstituted, which is unsubstituted or substituted by one or two methyl or trifluoromethyl groups. Or a 1,3-butadiene-1,4-subunit substituted by one to four methyl groups or by a trifluoromethyl group;
  • d represents a covalent bond or an alkylene group
  • e represents a hydrogen atom, an alkoxy group, an amino group, a 3- 7 membered cycloalkyl group, a 6- to 10-membered bicyclic structure, an alkylamino group or a bis-(Cw alkyl)amino group, wherein the alkyl moieties may be the same or different;
  • L 3 represents a covalent bond, -NH-, -N(C 1-3 alkyl) -, -0- , - 0- C 1-3 alkylene-, -S-Ci -3 alkylene, - S(0) m - , - C(O)- , -NHC(O)- , - N(C 1-3 alkyl) C(O)- , - C(0)NH- , -C(0) N(Ci -3 alkyl) -, - NHS(0) 2 - , -N(Ci -3 alkyl) S(0) 2 - , -S(0) 2 NH- , - S(0) 2 N (C 1-3 alkyl) -, - OC(O)- or - C(0)0- ,
  • R4 represents a hydrogen atom, an alkyl group, -N(Cw alkyl) 2 , -NHC(0)0-(Cw alkyl), -OH, - 0(C 1-4 alkyl), - S(0) 2 (C 1-3 alkyl), 3- 7 membered cycloalkyl, phenyl or 5- 6-membered heteroaryl;
  • R 2 represents - L 4 - R 5 ,
  • L 4 represents -S-C 1-3 alkylene- or -S(0) m - ,
  • R 5 represents a hydrogen atom, C 2-4 alkenyl, C 2-4 alkynyl, C 1-4 alkyl, amino, -NH(C 1-3 alkyl), -N(Cw alkyl) 2 , - OH , - 0 (Cw alkyl), - C(0) (Cw alkyl) or 3- 7 membered cycloalkyl,
  • n 0, 1 or 2.
  • alkyl moiety, cycloalkyl group, heteroaryl group may be further substituted by 1 to 4 ( ⁇ ,
  • the carbon atom of the cycloalkyl or bicyclic structure may be replaced by 1-4 identical or different N, NH, N(C 1-3 alkyl), 0, S(0) m , C(O);
  • the heteroaryl group contains 1-4 heteroatoms, each independently selected N, 0 or S;
  • n 0, 1 or 2;
  • p and q are independent representations of 0, 1, 2, 3 or 4.
  • the present invention provides a compound represented by the above formula (I), a pharmaceutically acceptable salt thereof or a stereoisomer thereof, wherein:
  • Ring A and Ring B each independently represent a phenyl group, a 5- to 6-membered cycloalkyl group, a 5-6 membered heterocycloalkyl group having a N, 0, S hetero atom, a 5-6 membered heteroaryl group or 8-10 members.
  • Li and L 2 each independently represent a covalent bond, -NH-, -N(CH 3 )-, - ⁇ -, - S(0) m - , -N(CH 3 )C(0)-, -C (0)N(CH 3 ) -, - N(CH 3 )S(0) 2 - or - S(0) 2 N(CH 3 ) -;
  • a represents a covalent bond, an imino group that is unsubstituted or substituted with CH 3 ;
  • b represents -CO- or -S0 2 -;
  • c represents a 1,2-vinylidene or ethynylene group which is unsubstituted or substituted by one or two methyl groups;
  • d represents a covalent bond or a methylene group;
  • e represents a hydrogen atom, a methoxy group, an amino group, a piperidinyl group, a morpholinyl group, a pyrrolidinyl group, a piperazinyl group, a TV-methyl piperazinyl group, a 6- to 9-membered spiro ring structure, a 6- to 8-membered ring structure. , 6- to 8-membered bridged ring structure, methylamino or bis-(methyl)amino;
  • L 3 represents a covalent bond, -NH-, -N(C 1-3 alkyl)-, -0-, - 0-C 1-3 alkylene-, -S-Ci -3 alkylene, - S(0) m - , - C(O)- , -NHC(O)-, - C(0)NH- , - NHS(0) 2 - , - S(0) 2 NH- , - oc(o )- or - c(o)o- ,
  • R4 represents a hydrogen atom, an alkyl group, -N(Cw alkyl)2, -NHC(0)0-(Cw alkyl), -OH, - 0(C 1-4 alkyl), - S(0) 2 (C 1-3 alkyl), 5- 6-membered cycloalkyl, phenyl or 5- 6-membered heteroaryl;
  • R 2 represents - L 4 - R 5 ,
  • L 4 represents -S-C 1-3 alkylene- or -S(0) m - ,
  • R 5 represents a hydrogen atom, a C 1-4 alkyl group, an amino group, -NH(C 1-3 alkyl), -N(C 1-3 alkyl) 2 ,
  • alkyl moiety, cycloalkyl group, phenyl group, heteroaryl group, spiro ring structure, concentric ring structure, bridged ring structure may be further substituted by 1 to 4 ( ⁇ ,
  • the carbon atom of the cycloalkyl or bicyclic structure may be replaced by 1-4 identical or different N, NH, N(C 1-3 alkyl), 0, S(0) m , C(O);
  • heteroaryl group, the spiro ring structure, the concentric ring structure, and the bridged ring structure contain 1-4 hetero atoms, and each of them is independently selected from N, 0 or S;
  • n means 0 , 1 or 2 ;
  • p and q are independent representations of 0, 1, 2, 3 or 4.
  • the present invention provides a compound represented by the above formula (I), a pharmaceutically acceptable salt thereof or a stereoisomer thereof, wherein:
  • Ring A and ring B each independently represent a phenyl group, a 5-6 membered heterocycloalkyl group having a N hetero atom or a 5-6 membered heteroaryl group;
  • Li and L 2 are independently represented by -NH- , - 0- or - S(0) m -;
  • a represents a covalent bond or an imino group
  • c 1,2-vinylidene
  • d represents a covalent bond or a methylene group
  • e represents a hydrogen atom, piperidinyl, morpholinyl, pyrrolidinyl, piperazinyl or bis-(methyl)amino;
  • Ri represents a hydrogen atom, a gas atom, a methyl group, a methoxy group, a methylamino group or a di-(methyl)amino group which is unsubstituted or substituted with one to three atomic atoms;
  • R 3 represents a hydrogen atom, a halogen atom or -L 3 - R 4 ,
  • L 3 represents a covalent bond, -NH-, -N(C 1-3 alkyl)-, -0- , - 0- C 1-3 alkylene-, -S-Ci -3 alkylene-, - S(0) m - , - C(O)- , -NHC(O)- , - C(0)NH- , - OC(O)- or -C(0)0-
  • R4 represents a hydrogen atom, Methyl, ethyl, -N(C 1-3 alkyl) 2 , - NHC(0)0-CH 3 , - 0(CH 3 ) , -0(CH 2 CH 3 ) , - 0 (C(CH) 3 ) 3 ), -S(0) 2 -CH 3 , cyclopentyl, cyclohexane, pyrrolidinyl, tetrahydrofuranyl, piperidinyl, morpholinyl, piperazinyl,
  • methylthio group, ethylthio group, methylsulfonyl group, aminosulfonyl group, methylsulfinyl group, aminosulfinyl group may be further one to two of the same or different methoxy, pyrrolidinyl, tetra Hydroimidazolyl, piperidinyl, morpholinyl, piperazinyl,
  • the pyrimidinyl group may be further substituted by one to two identical or different,
  • Derivative atom methyl, amino, methylamino, bis-(methyl)amino, hydroxy, methoxy, methoxycarbonyl, carbamoyl, methylcarbamoyl or bis-(methyl)carbamoyl ;
  • n 0, 1 or 2;
  • p and q stand independently for 0, 1 or 2.
  • the present invention provides a compound represented by the above formula (I), a pharmaceutically acceptable salt thereof or a stereoisomer thereof, wherein:
  • Ring A and Ring B each independently represent phenyl, pyridyl, piperidinyl, imidazolyl, pyrazolyl, thiazolyl, thiazyl, 1,2,3-triazolyl or 1,2,4-tri Azolyl
  • Li and L 2 are independently represented by -NH- or - 0-;
  • a represents a covalent bond or an imino group
  • c 1,2-vinylidene
  • d represents a covalent bond or a methylene group
  • e represents a hydrogen atom, piperidinyl, pyrrolidinyl, morpholinyl, piperazinyl or bis-(methyl)amino;
  • R 3 represents a hydrogen atom, a fluorine atom, a chlorine atom or -L 3 - R 4 ,
  • L 3 represents a covalent bond, -NH-, - N (Ci -3 alkyl) -, - 0- , - 0- CH 2 CH 2 - , -S-CH 2 CH 2 - or - s(o) m -,
  • R4 represents a hydrogen atom, methyl, ethyl, -N(C 1-3 alkyl) 2 , -NHC(0)0-CH 3 , - 0(CH 3 ),
  • phenyl group, pyrrolyl group, imidazolyl group, thiazolyl group, oxazolyl group, thiadiazolyl group, pyridyl group Can be further replaced by one or two identical or different,
  • R 2 represents an ethylthio group which is unsubstituted or substituted by a methoxy group, a piperidinyl group or a morpholinyl group, a methylthio group, a methylsulfonyl group, an aminosulfonyl group, a methylsulfinyl group, an aminosulfinyl group.
  • m means 0, 1 or 2;
  • the present invention provides a compound represented by the above formula (I), a pharmaceutically acceptable salt thereof or a stereoisomer thereof, wherein:
  • Ring A and ring B each independently represent a phenyl group, a pyridyl group, a pyrazolyl group or a piperidinyl group;
  • Li and L 2 are independently represented by -NH-;
  • a represents an imino group
  • c 1,2-vinylidene
  • d represents a covalent bond or a methylene group
  • e represents a hydrogen atom, a pyrrolidinyl group, a morpholinyl group, a piperazinyl group or a bis-(methyl)amino group
  • Ri represents a hydrogen atom
  • R 3 represents - L 3 - R4 , L 3 represents a covalent bond, - 0- or - 0-CH 2 CH 2 -, R4 represents -N(CH 3 ) 2 , -0(CH 3 ), tetrahydrofuranyl, Phenyl, phenyl or pyridyl,
  • the phenyl group and the pyridyl group may be further substituted by one to two identical or different, and represent a carbamoyl group, a methylcarbamoyl group or a bis-(methyl)carbamoyl group;
  • R 2 represents an ethylthio group which is unsubstituted or substituted with a methoxy group or a morpholinyl group, a methylthio group, a methylsulfonyl group, an aminosulfonyl group, a methylsulfinyl group, an aminosulfinyl group;
  • p 1 ;
  • the present invention provides a compound represented by the above formula (I), a pharmaceutically acceptable salt thereof or a stereoisomer thereof, wherein:
  • Ring A represents phenyl
  • ring B represents phenyl, pyrazolyl or pyridyl
  • L 2 independently represents -NH-
  • a represents an imino group
  • b represents -CO-
  • c represents 1,2-vinylidene
  • d represents a covalent bond
  • e represents a hydrogen atom
  • R 3 represents - L 3 - R4 , L 3 represents a covalent bond, -NH-, - N(C 1-3 alkyl) -, -0-, - 0- C 1-3 alkylene-, - S - C 1-3 alkylene-, or - C(0)NH- , R4 represents a hydrogen atom, methyl, ethyl, - 0(CH 3 ), - 0(C(CH 3 ) 3 ) or morpholinyl; R 2 represents -L 4 - R 5 , and L 4 represents -S-C 1-3 alkylene- or -S(0) m - , R 5 represents a hydrogen atom or p represents 1; q represents 1.
  • the "d- 6 alkyl group” as used in the present invention means a linear or branched alkyl group having 1 to 6 carbon atoms, and includes “alkyl", “Cw alkyl” and the like, and examples thereof include, but are not limited to, For example, methyl, ethyl, n-propyl, isopropyl, n-butyl, 2-methylpropyl, 1-methylpropyl, 1,1-dimethylethyl and the like.
  • the terms “Cw alkyl” and “Cw alkyl” refer to specific examples containing 1 to 4, 1 to 3 carbon atoms in the above examples.
  • the "d- 6 alkylene group” as used in the present invention means a structure in which a straight or branched alkyl group having 1 to 6 carbon atoms is removed by one hydrogen atom, and means a divalent alkyl group.
  • the "alkylene chain” is a polymethylene group, that is, -(CH 2 )x- , wherein X is a positive integer, preferably 1 to 6, 1 to 4, 1 to 3, 1 to 2 or 2 to 3.
  • the substituted alkylene chain is a polymethylene group in which one or more methylene hydrogen atoms are replaced by a substituent.
  • C 1-4 alkylene “C 1-3 alkylene” and the like, examples of which include, but are not limited to, for example, methylene (-CH 2 - ), ethylene ( - CH 2 CH 2 - ), propylene (-CH 2 CH 2 CH 2 - ), butylene (-CH 2 CH 2 CH 2 CH 2 - ), and the like.
  • C 1-4 alkylene “C 1-3 alkylene” refers to a specific example containing 1 to 4, 1 to 3 carbon atoms in the above examples.
  • C 2 _ 4 alkenyl group as used in the present invention means a linear or branched alkenyl group having 2 to 4 carbon atoms; examples thereof include, but are not limited to, for example, a vinyl group, a 1-propenyl group, 2-propenyl, 1-methylvinyl, 1-butenyl, 2-butenyl, 3-butenyl, 1-methyl-1-propenyl, 2-methyl-i-propenyl, 1-methyl-2-propenyl, 2-methyl-2-propenyl.
  • C 2 _ 4 alkynyl group in the present invention means the number of carbon atoms containing a triple bond to 2-4 straight or branched alkynyl group; examples thereof include, but are not limited to, ethynyl, 2-propynyl Base, 2-butynyl, 3-butynyl, 1-methyl-2-propynyl, and the like.
  • the "d- 6 alkoxy group” of the present invention means a group in which the term “d- 6 alkyl group” is bonded to another structure through an oxygen atom, such as a methoxy group, an ethoxy group, a propoxy group, an isopropoxy group, or a butyl group. Oxyl, isobutoxy, tert-butoxy, sec-butoxy, pentyloxy, neopentyloxy, hexyloxy and the like. Alkoxy groups are preferred, and d- 3 alkoxy groups are more preferred.
  • the terms “d- 4 alkoxy", “d- 3 alkoxy” refer to the group “alkyl", "d- 3 alkyl” attached to the other structure through an oxygen atom.
  • the "d- 6 alkylamido" of the present invention means a group in which "d- 6 alkyl" is bonded to other structures via an acylamino group.
  • the "prime” as used in the present invention means a fluorine atom, a chlorine atom, a bromine atom or an iodine atom.
  • the "3- 7-membered cycloalkyl group” as used in the present invention means that the ring atoms are all carbon atoms, and a hydrogen atom-derived cyclic alkyl group is removed, including, for example, "3- 6-membered cycloalkyl group", "4".
  • cycloalkyl ""5- 7-membered cycloalkyl", "5-6-membered cycloalkyl”, examples of which include, but are not limited to, cyclopropyl, cyclobutane, cyclopentyl, cyclohexyl Alkyl, cycloheptyl, cyclooctyl, cyclopentanone, cyclohexanone, and the like.
  • the "3- 7-membered heterocycloalkyl group” as used in the present invention means a 3-7 membered cyclic group having one or more hetero atoms, and the "hetero atom” means a nitrogen atom, an oxygen atom, a sulfur atom, or the like. .
  • a 3- to 6-membered heterocyclic group is preferred, and a 5- to 6-membered heterocyclic group is more preferred.
  • "4-7-membered heteroaryl” means an aromatic group consisting of 4 to 7 ring atoms (having at least one hetero atom), including "5- to 7-membered heteroaryl""5- to 6-membered heteroaryl” Specific examples include, but are not limited to, furyl, thiyl, pyrrolyl, thiazolyl, thiadiazolyl, oxazolyl, oxadiazolyl, imidazolyl, pyrazolyl, pyridyl, pyrimidinyl, 1,4-dioxadienyl, 2//- 1,2-oxazinyl, 4//- 1,2-oxazinyl, 6//- 1,2-oxazinyl, 4 //- 1,3-oxazinyl, 6//-1,3-oxazinyl, 4//- 1,4-oxazinyl, pyridazinyl, pyridyl 11 -methyl, 1,
  • 6-10 membered bicyclic structure means a bicyclic group consisting of 6 to 10 ring atoms (may not contain or contain one or more heteroatoms:), including "7-10 membered bicyclic structure”,”8-10 “Secondary ring structure”, “6-9 element spiral ring structure”, “6-8 yuan parallel ring structure”, “6-8 yuan bridge ring structure”, etc.
  • aromatic bicyclic structures including, but not limited to, benzofuranyl, benzisofuranyl, benzothianyl, fluorenyl, benzoxazolyl, benzo Imidazolyl, carbazolyl, benzotriazolyl, quinolyl, isoquinolyl, acridinyl, phenanthryl, benzoxazinyl, pyridazinyl, quinazolinyl, quinoxalinyl, Phenazinyl, acridinyl, fluorenyl, naphthyridyl, 1 ,3-dihydrobenzofuranyl, benzo[[1.3]dioxolyl, isoindolyl, chromanyl, 1,2,3,4-tetrahydropyrrolo[3,4-c]pyrrolyl, 5,6-dihydroimidazole[1.2- ⁇ ]pyrazine-7(8
  • the "6-9-membered spirocyclic group" of the present invention means that at least two rings of a class share an atomic shape.
  • a 6- to 9-membered fused ring structure include, but are not limited to, spiro[3.3]heptyl, spiro[3.4]octyl, spiro[3.5]decyl, spiro[4.4]decyl, spiro[3.4]oct-6-ene , snail [3.5] ⁇ -6-alkenyl, spiro[4.4] ⁇ -6-alkenyl, spiro[4.4] ⁇ -2,7-dienyl, 2-oxaspiro[3.3]heptanyl, 6-oxaspiro[2.5]octyl, 4-oxa-7-aminospiro[2.5]octyl, 2-aminospiro[3.3]heptanyl, 2-oxa-6-aminospiro[3.3 Heptylalkyl, 2-
  • the "6- to 8-membered ring structure" as used in the present invention refers to a 6-8 membered cyclic group formed by two or more ring structures sharing two adjacent atoms with each other, and specific examples thereof include But not limited to: bicyclo [3.1.0] hexane, bicyclo [4.1.0] heptyl, bicyclo [2.2.0] hexane, bicyclo [3.2.0] heptyl, bicyclo [4.2.0] Octyl, bicyclo[3.1.0]hex-2-enyl, bicyclo[4.1.0]hept-3-enyl, bicyclo[3.2.0]hept-3-yl,bicyclo[ 4.2.0] Oct-3-enyl, benzofuranyl, benzoisofuranyl, benzothianyl, decyl, benzoxazolyl, benzimidazolyl, oxazolyl, benzotrien Azyl, quinolyl, isoquinolyl, acridiny
  • the "6- to 8-membered bridged ring structure" of the present invention refers to a 6- to 8-membered cyclic group formed by two or more ring-shaped structures sharing two non-adjacent atoms with each other, and a specific embodiment thereof Including, ⁇ , ⁇ , ⁇ , ⁇ , ⁇ , ⁇ , ⁇ , , ⁇ , ⁇ , 0,
  • i JSr (3, ⁇ , ⁇ , The above group may be further substituted by 1 to 4, ( ⁇ represents a halogen atom, a C alkyl group, an amino group, an alkylamino group, a bis-(C alkyl)amino group, a hydroxyl group, an alkoxy group, an alkoxycarbonyl group, an amino group.
  • ⁇ represents a halogen atom, a C alkyl group, an amino group, an alkylamino group, a bis-(C alkyl)amino group, a hydroxyl group, an alkoxy group, an alkoxycarbonyl group, an amino group.
  • Formyl, alkylcarbamoyl, bis-(Cw alkyl)carbamoyl or 3- 6 membered cycloalkyl which may be the same or different.
  • Particularly preferred compounds of the formula (I) according to the invention include:
  • Particularly preferred compounds of the formula (I) according to the invention further comprise:
  • the invention provides a process for the preparation of a compound of the invention described above. In one embodiment, the invention provides a pharmaceutical group comprising a compound of the invention described above Compound.
  • the invention provides the use of a compound of the invention described above for the prevention and/or treatment of B cell associated blood cancer in an individual (eg, B cell chronic lymphocytic carcinoma, non-Hodgkin's lymphoma), inflammatory and autoimmune A method of disease (eg, rheumatoid arthritis, systemic lupus erythematosus, etc.).
  • a compound of the invention described above for the prevention and/or treatment of B cell associated blood cancer in an individual (eg, B cell chronic lymphocytic carcinoma, non-Hodgkin's lymphoma), inflammatory and autoimmune A method of disease (eg, rheumatoid arthritis, systemic lupus erythematosus, etc.).
  • the invention provides a compound of the invention described above for use in the prevention and/or treatment of B cell associated blood cancer (eg, B cell chronic lymphocytic carcinoma, non-Hodgkin's lymphoma), inflammatory and autoimmune Use in drugs for sexual diseases such as rheumatoid arthritis, systemic lupus erythematosus, etc.
  • B cell associated blood cancer eg, B cell chronic lymphocytic carcinoma, non-Hodgkin's lymphoma
  • inflammatory and autoimmune Use in drugs for sexual diseases such as rheumatoid arthritis, systemic lupus erythematosus, etc.
  • DMF dimethylformamide
  • THF tetrahydrofuran
  • DIEA V, V-diisopropylethylamine
  • BINAP 2,2,-bisdiphenylphosphino-1,1,-binaphthyl
  • Xantphos 4,5-bisdiphenylphosphine-9,9-dimethyloxaxime.
  • Intermediate 1 (1 equivalent) is mixed with a solvent amount of phosphorus oxychloride (at least 10 equivalents), about two equivalents of dimethylaniline are added, and the reaction is stirred by heating (90-110 ° C) for several hours until the intermediate 1 disappears. After cooling, pour into ice water, precipitate a solid, filter, and dry to give Intermediate 2. Alternatively, the reaction system is concentrated, and column chromatography is carried out to obtain Intermediate 2.
  • Method 1 Mix intermediate 2 (1 eq.) with starting material 2 (1.1-1.3 eq.), add a suitable solvent (such as tetrahydrofuran, n-butanol, etc.), and heat (80-120 ° C) to stir the reaction for several hours to the intermediate. 2 disappeared. Cooling, concentrating the reaction system, and column chromatography gave Intermediate 3.
  • a suitable solvent such as tetrahydrofuran, n-butanol, etc.
  • Method 2 Buch Buchwald-Hartwig coupling. Mix intermediate 2 (1 eq.) with starting material 2 (1-1.3 eq.), add the appropriate solvent (usually toluene, dioxane, etc.), catalytic amount of palladium catalyst (usually Pd 2 (dba) 3 , Pd (OAc) 2, etc., catalytic amount of ligand (usually Xantphos, BINAP, etc.), base (Cs 2 C0 3 , NaO Bu, etc.) (1 ⁇ 1 - 1.5 equivalents), fully substituted inert gas (such as nitrogen, argon) Gas, etc., heating (70-120 ° C) to stir the reaction for several hours until the intermediate 2 disappears. The mixture was cooled, concentrated, and subjected to column chromatography to give Intermediate 3.
  • solvent usually toluene, dioxane, etc.
  • catalytic amount of palladium catalyst usually Pd 2 (dba) 3 , Pd (OAc) 2, etc.
  • Method 1 Intermediate 3 (1 eq.) And starting material 3 (1 1.3 equivalents) were mixed, and a suitable solvent (e.g. tert-butanol, tert-amyl alcohol, etc.), addition of a base (e.g., DIEA and the like, 1- 1.5 eq. ), or a catalytic amount of acid (such as acetic acid, trifluoroacetic acid, etc.), heated (80-120 ° C), or heated under microwave (80-120 ° C) to stir the reaction for several hours until the intermediate 3 disappears. Cooling, concentrating the reaction system, and column chromatography gave Intermediate 4.
  • a suitable solvent e.g. tert-butanol, tert-amyl alcohol, etc.
  • a base e.g., DIEA and the like, 1- 1.5 eq.
  • a catalytic amount of acid such as acetic acid, trifluoroacetic acid, etc.
  • Method 2 Buchwald-Hartwig coupling. In the same way as in step 2 of step 3, only raw material 2 needs to be replaced with raw material 3.
  • R 2 , R 3 , Li, L 2 , a, b, c, d, e, p, q, A and B in the reaction scheme are as described above.
  • Method 1 The substrate (1 equivalent) is dissolved in methanol and water (or a mixed solvent of dichloromethane, methanol, and water), and an oxidizing agent (such as potassium persulfate complex salt, m-chloroperoxybenzoic acid, etc.) is added ( 1 equivalent), react to LCMS at room temperature to monitor the complete disappearance of the substrate, spin the solution, prepare the liquid phase for purification, or purify the column with silica gel to obtain the product.
  • an oxidizing agent such as potassium persulfate complex salt, m-chloroperoxybenzoic acid, etc.
  • Method 2 The substrate (1 equivalent) is dissolved in methanol, and opened to reflux in air for several hours. The product is purified by spin-drying or purified by silica gel column.
  • the substrate (1 equivalent) is dissolved in methanol and water (or a mixed solvent of dichloromethane, methanol, water), and an oxidizing agent (such as potassium persulfate complex salt, m-chloroperoxybenzoic acid, etc.) (greater than 2 equivalents) ), reacting at room temperature overnight or heating for several hours, until the substrate is completely disappeared by LCMS, the solution is spin-dried, purified by liquid phase preparation, or purified by silica gel column to obtain a product.
  • an oxidizing agent such as potassium persulfate complex salt, m-chloroperoxybenzoic acid, etc.
  • R 2 , R 3 , R 5 , Li, L 2 , a, b, c, d, e, p, q, A and B in the reaction scheme are as described above.
  • the compound of the formula (I) of the present invention, a stereoisomer thereof, can be used in the form of a free form or a pharmaceutically acceptable salt thereof.
  • the compound of the formula (I) of the present invention is basic and can form an acid salt with an inorganic acid or an organic acid.
  • the compound of the formula (I) of the present invention or a pharmaceutically acceptable salt thereof may exist in the form of an optical isomer due to the presence of an asymmetric carbon atom, and therefore, the present invention also includes these optical isomers and mixtures thereof.
  • the structures described herein are also intended to include all isomeric (e.g., enantiomeric, diastereomeric, and geometric (or conformational) forms) of the structure; for example, R and about each asymmetric center S configuration, Z and E double bond isomers, and Z and E conformers.
  • single stereochemical isomers as well as mixtures of enantiomers, diastereomers and geometric isomers (or conformational isomers) of the compounds of the invention are within the scope of the invention.
  • the compound of the formula (I), a pharmaceutically acceptable salt thereof or a stereoisomer thereof of the present invention may be combined with one or more pharmaceutically acceptable carriers to constitute a pharmaceutical composition.
  • the pharmaceutical composition can be formulated into a conventional preparation for clinical use, and can be administered to a patient in need of such treatment by oral or parenteral administration. Such as tablets, granules, capsules, powders, injections, inhalants, sublingual preparations, syrups, gels, ointments, suppositories, lotions, eye drops, nasal drops, sprays, transdermal formulations Wait.
  • These preparations can be prepared by a conventional method, by adding a pharmaceutically acceptable carrier such as an excipient, a binder, a moisturizer, a disintegrating agent, a thickener or the like.
  • the compound of the formula (I) of the present invention a pharmaceutically acceptable salt thereof or a stereoisomer thereof has a preferable BTK kinase inhibitory action, and is a drug which preferably has an excellent antitumor effect and a therapeutic effect of an autoimmune disease.
  • the compound of the formula (I), a pharmaceutically acceptable salt thereof or a stereoisomer thereof thereof is used for the preparation of a B cell-associated blood cancer (for example, B cell chronic lymphocytic carcinoma, non-Hodgkin's lymphoma), and autoimmunity Sexual diseases (such as rheumatoid arthritis, systemic lupus erythematosus, etc.) play an important role.
  • a B cell-associated blood cancer for example, B cell chronic lymphocytic carcinoma, non-Hodgkin's lymphoma
  • autoimmunity Sexual diseases such as rheumatoid arthritis, systemic lupus erythematosus, etc.
  • the compound of the formula (I), a pharmaceutically acceptable salt thereof or a stereoisomer thereof of the present invention is a kinase inhibitor, particularly a Btk inhibitor.
  • These inhibitors can be used to treat one or more diseases that respond to kinase inhibition in a mammal, including response to Btk inhibition and/or B cells. A disease that inhibits proliferation.
  • Btk inhibition a disease that inhibits proliferation.
  • the present invention includes a method for treating a mammal, such as a human, having a disease responsive to inhibition of Btk activity and/or inhibiting proliferation of B cells, the method comprising: administering to a mammal having such a disease an effective amount of at least A chemical entity provided herein.
  • the effective concentration can be determined experimentally, for example, by measuring the blood concentration of the compound, or theoretically by calculating the bioavailability.
  • Other kinases that may be affected in addition to Btk include, but are not limited to, other tyrosine kinases and serine/threonine kinases.
  • kinases play a significant role in controlling the signaling pathways of essential cellular processes such as proliferation, differentiation and death (apoptosis). Abnormal kinase activity has been implicated in various diseases including a variety of cancers, autoimmune and/or inflammatory diseases, and acute inflammatory responses. The versatile role of kinases in key cell signaling pathways provides a significant opportunity to identify new drugs that target the kinase and signal transduction pathways.
  • One embodiment includes a method of treating a patient having an autoimmune and/or inflammatory disease or an acute inflammatory response in response to inhibition of Btk activity and/or B cell proliferation.
  • Autoimmune and/or inflammatory diseases that can be affected by the use of the compounds and compositions according to the invention include, but are not limited to, psoriasis, allergies, localized enteritis, irritable bowel syndrome, Sjogren's disease, tissue grafts Rejection and hyperacute rejection of transplanted organs, asthma, systemic lupus erythematosus (and associated glomerulonephritis), dermatomyositis, multiple sclerosis, scleroderma, vasculitis (ANCA-related and other blood vessels) Inflammation, autoimmune hemolytic and thrombocytopenic symptoms, Goodpas syndrome (and associated glomerulonephritis and pulmonary hemorrhage), atherosclerosis, rheumatoid arthritis, chronic idiopathic thrombocytopenia Purpura (ITP), Edison's disease, Parkinson's disease, Alzheimer's disease, diabetes, septic shock and myasthenia gravis.
  • Anti-inflammatory drugs include, but are not limited to: NSAID, non-specific and COX-2 specific cyclooxygenase inhibitors, gold compounds, dermal steroids, methotrexate, tumor necrosis factor (TNF) receptor antagonists, immunization Inhibitor and methotrexate.
  • NSAIDs include, but are not limited to, ibuprofen, flurbiprofen, naproxen and naproxen sodium, diclofenac, a combination of diclofenac sodium and misoprostol, sulindac, benzopyrene, diflunis Willow, piroxicam, indomethacin, etodolac, fenoprofen calcium, ketoprofen, nabumetone sodium, sulfasalazine, tolbutin sodium and hydroxychloroquine.
  • NSAIDs also include COX-2 Specific inhibitors such as celecoxib, valdecoxib, remiclox and/or etoricoxib.
  • the anti-inflammatory agent is a salicylate or a salt.
  • Salicylates or salts include, but are not limited to, acetylsalicylic acid or aspirin, sodium salicylate, and choline salicylate and magnesium salicylate.
  • Anti-inflammatory drugs can also be Shield Steroids.
  • the skin shield steroid may be cortisone, dexamethasone, methylprednisolone, prednisolone, prednisolone sodium phosphate, or prednisone.
  • the anti-inflammatory agent is a gold compound such as gold thioudate or auranofin.
  • the invention also includes embodiments in which the anti-inflammatory agent is a metabolic inhibitor such as a dihydrofolate reductase inhibitor such as methotrexate or a dihydroorotate dehydrogenase inhibitor such as leflunomide.
  • a further embodiment of the invention relates to wherein the at least one anti-inflammatory compound is an anti-monoclonal antibody (such as eculizumab or pegizumab), a TNF antagonist such as entanercept or infliximab
  • the infliximab is an anti-TNFa monoclonal antibody.
  • a further embodiment of the invention relates to wherein at least one active agent is an immunosuppressive compound such as selected from the group consisting of methotrexate, leflunomide, cyclosporine, tacrolimus, azathioprine and mycophenolate
  • an immunosuppressive compound such as selected from the group consisting of methotrexate, leflunomide, cyclosporine, tacrolimus, azathioprine and mycophenolate
  • B-B and B-precursor expression of Btk have been implicated in the pathology of B-package malignancy, B cell malignancy including but not limited to B-cell lymphoma, lymphoma (including Hodgkin and non-Hodgkin's lymphoma) ), hair cell lymphoma, multiple myeloma, chronic and acute myeloid leukemia and chronic and acute lymphocytic leukemia.
  • Btk has been shown to be an inhibitor of Fas/APO-1 (CD-95) death-inducing signaling complex (DISC) in B-lineage lymphoid cells.
  • the fate of leukemia/lymphoma cells may lie in the balance between the reverse pre-apoptotic effect of caspase-activated caspase and the upstream anti-apoptotic regulatory mechanism including Btk and/or its substrate (Vassilev et al. , J. Biol. Chem. 1998, 274, 1646-1656).
  • Btk inhibitors can be used as chemical sensitizers, and thus can be used in combination with other chemotherapeutic drugs, particularly drugs which induce apoptosis, such as antitumor agents, immunosuppressive agents and the like.
  • chemotherapeutic agents that can be used in combination with chemical sensitizers include, but are not limited to, topoisomerase I inhibitors (such as camptothecin or topotecan), topoisomerase II inhibitors (such as daunorubicin) And etoposide:), alkylating agents (such as cyclophosphamide, melphalan and BCNU), tubulin-directed agents (such as Taxol and Vinblastine) and biological agents (such as antibodies such as anti-CD20 antibody, IDEC8, Immunotoxins and cytokines).
  • Btk activity has been associated with some leukemias that express the bcr-abl fusion gene resulting from partial translocation of chromosomes 9 and 22.
  • Btk is shielded by bcr-abl kinase, which triggers a downstream survival signal that prevents apoptosis in bcr- abl cells (N. Feldhahn et al., J. Exp. Med. 2005, 201(11), 1837-1852 ).
  • the compound of the formula (I) of the present invention or a pharmaceutically acceptable salt thereof has a good BTK kinase inhibitory action, has high safety, a large safety window and a small side effect;
  • control compound CC-292, was prepared in accordance with WO2009158571A1.
  • the compound of the present invention its chemical name, structural formula and preparation method are shown in the preparation examples of the respective compounds.
  • ATP adenosine triphosphate
  • BTK Bruton's tyrosine kinase
  • mg mg
  • mL ml
  • g microgram
  • ⁇ microliter
  • mM millimoles per liter
  • EDTA ethylenediaminetetraacetic acid
  • DMSO dimethyl Ketosulfone.
  • HTRFR KinEASETM -TK purchased from Cisbio, lot number 62TK0PEB; BTK: purchased from Carna, Cat.No.08-080; ATP: purchased from Sigma, Cat. No. A7699, CAS
  • MgCl 2 purchased from Sigma, CAS No. 7786-30-3, Lot. No. 101M8701V; DMSO: purchased from Sigma, CAS No. 67-68-5, Lot. No. STBC0365V ;
  • Test reagent preparation available from Thermo, Cat. No. 249944, Lot. No. 1057825; 384-well plate: available from Greiner, Cat. No. 784075, Lot. No. ⁇ 1112 ⁇ 6 ⁇ . 2. Test reagent preparation
  • test sample was separately detected at 615 nm by a microplate reader.
  • the inhibitory activity of the compounds of the present invention against BTK kinase was significantly stronger than that of the control compound CC-292.
  • the compound of the present invention its chemical name and structural formula and preparation method are shown in the preparation examples.
  • mice Male, 6-8 week Balb/c mice.
  • CTG CellTiter-Glo (Cat. No. G7572, Promega), storing CTG buffer and CTG substrate at -20.
  • C It is recommended to prepare CTG reagents in the following ways:
  • CTG buffer before use and equilibrate to room temperature.
  • CTG buffer can be used. Melt well before use and store at room temperature for more than 48 hours.
  • the lyophilized CTG substrate was equilibrated to room temperature and 100 ml of buffer was taken into the amber bottle containing the substrate to obtain the CTG reagent. Gently mix until a uniform solution is obtained.
  • the substrate should be completely melted in one minute, dispensed and stored in a - 20 ° C refrigerator for a long period of time.
  • test compound gradient dilution solution First, 10 mM of the test compound stock solution was diluted 3 times in DMSO with a total of 10 concentrations. Then add ⁇ DMSO diluted compound to 90 ⁇ compound dilution buffer, and then take lO L
  • the 10% DMSO diluted compound was added to 90 ⁇ of the compound dilution buffer at a maximum concentration of 10 ⁇ M and a DMSO concentration of 0.1% for a total of 10 concentration gradients.
  • mice * The spleens of Balb/c mice were smashed, mashed in MACS buffer, and filtered through a 40 ⁇ nylon cell sieve to obtain a single cell suspension.
  • the obtained cell suspension was centrifuged at 400 g for five minutes, the supernatant was removed, and 1 ml of the red blood cell lysate was added at room temperature and the cell pellet was gently resuspended. Two minutes later, pre-chilled MACS buffer was added. The cell suspension was filtered through a 40 ⁇ cell sieve into a new centrifuge tube.
  • a biotinylated antibody mixture of ⁇ was added to every 10 7 cells. After mixing, incubate on ice for 20 minutes, add 30 ⁇ l of MACS buffer and 20 ⁇ l of avidin-containing magnetic beads per 10 7 cells and incubate on ice for 20 minutes. After centrifugation, the cells were resuspended in 500 ⁇ l of MACS buffer. The pre-cooled MACS sorting column was placed in a MACS sorter and the cell suspension was applied to a MACS sorting column. The cells under the flow of unbound antibody are collected. • Detection of cells before and after sorting by flow cytometry using PE anti-biotin antibody and CD45R (B220) antibody detection.
  • the data obtained will be analyzed using Excel 2007 and GraphPad Prism 5.0 software in order to calculate IC 5 .
  • the nonlinear S-curve regression will be used to fit the data to obtain a dose-response curve, and the GraphPad Prism 5.0 software will automatically give the IC 50 value.
  • V sample / V2 dissolved 3 ⁇ 4 pairs, X 100%
  • V sample is the reading of the compound treated well
  • V2 » 3 ⁇ 4 vs. 3 ⁇ 4 is the average of the solvent control well (V2) readings.
  • the compound of the present invention (Compound 1-106) has an IC 5 Q ⁇ 0.5 ⁇ for the inhibitory activity of Balb/c mouse B cells in vitro.
  • the IC 5 Q of some compounds is as follows: In vitro cytological inhibitory activity of some compounds of the invention against Balb/c mouse B cells
  • an ELISA protocol is used that evaluates compounds at different concentrations in spleen cells using a biotinylated probe compound that binds only to BTK that is not occupied by the compound. For the occupancy rate of BTK enzyme, calculate the %81 ⁇ occupancy rate (BTK Occupancy).
  • Mouse anti-BTK antibody (Becton Dickinson); goat anti-mouse HRP antibody (Becton Dickinson); cell lysate (Cell Signaling); Bruton tyrosine kinase (BTK) (Carna); streptavidin coated 96-well plate (Thermo); rat lymphocyte isolation kit (LTS 1083PK, Tianjin Haoyang Biological Products Technology Co., Ltd.); microplate reader (victor4, PE); centrifuge (5804R, Eppendorf); microscope ( CX31RTSF, Olympus); MACS sorter (MACS).
  • BTK Bruton tyrosine kinase
  • Probe compound solution (Probe): Weigh 1 mg of sample compound and prepare a concentration of 1 mM. Stock solution, diluted with sample diluent; Sample diluents: PBS containing 1% bovine serum albumin and 0.1% Tween-20; Washing solution: 0.05% Tween-20 PBS.
  • the cell concentration was diluted to 3 x 107 Cells/ml, 90 ⁇ l/well with PBS. Add compound ⁇ /well and incubate for 1 h at 37 °C. Centrifuge at 20 °C, 400g for 20min, discard the supernatant.
  • the protease inhibitor PMSF was added to the cell lysis buffer (note that PMSF was added before the lysate was used).
  • the lysate was added to each tube-enriched cell and mixed. According to the lysate instructions, the ice was lysed for 5 min and centrifuged at 14,000 g for 10 min. Add the supernatant ⁇ to the 96-well plate.
  • the OD value at 450 nm was detected. According to the OD value, use the microplate reader
  • the A 4 parameter logic curve calculates the amount of BTK in each sample.
  • the frozen plasma was pre-incubated in a 37 ° C constant temperature water bath and thawed for use.
  • the stability of the compound in plasma was evaluated by the retention percentage of the compound after incubation at each time point, and the concentration of the sample was expressed by the ratio of the peak area of the test compound to the internal standard peak area. Calculated as follows:
  • the concentration of the compound is determined as the final solution at the initial incubation.
  • the compound 23 of the present invention is more stable in plasma than the control drug CC-292.
  • the V patch clamp method detects the inhibition of the hERG potassium channel by the compound of the present invention.
  • the compound of the present invention (Compound 1-106): Its chemical name, structural formula and preparation method are shown in the preparation examples of the respective compounds.
  • Control compound CC-292, synthesized according to patent WO2009158571A1.
  • experimental method
  • Extracellular fluid mM: 7V ⁇ 2 hydroxyethylpiperazine-2-ethylethanesulfonic acid (HEPES) 10.
  • HEPES hydroxyethylpiperazine-2-ethylethanesulfonic acid
  • In-electrode solution (in mM): KC1 120, OH 31.25, CaC3 ⁇ 4 5.374, MgCI 2 L75, ethylene glycol-bis( ⁇ -aminoethyl ether) ⁇ ', ⁇ '"-tetraacetic acid (EGTA) 10, HEPES 10, Na. 2 -ATP 4, pH was adjusted to 7.2 with IN potassium hydroxide; osmotic pressure was adjusted to 280 290 mOsm; after filtration - 20 ° C storage.
  • the hERG current was recorded using whole cell patch clamp technique.
  • the cell suspension was applied to a 35 mm culture and placed on an inverted microscope stage. After the cells were attached, the cells were perfused with extracellular fluid at a flow rate of 12 mL/min.
  • the glass microelectrode is drawn in two steps by a microelectrode drawing device, and its water resistance is 2 - 5 ⁇ .
  • the clamping potential was maintained at -80 mV.
  • the voltage is stimulated to depolarize to +60 mV and then repolarized to -50 mV to induce the hERG tail current. All records are performed after the current has stabilized. Extracellular perfusion administration starts from a low concentration, each concentration is 5 ⁇ until the current is stable, and the next concentration is given.
  • Stable cell line CHO hERG was purchased from AVIV A.
  • the minimum sealing resistance is not less than 1GQ
  • the hERG current is not less than 0.4nA.
  • the test coverage includes the following aspects:
  • the hERG current was recorded on the CHO-K1 cell line stably expressing the hERG channel by manual patch clamp technique; the inhibition rate of each concentration was calculated from the hERG tail current; each compound was tested 5 for each concentration test 2 cells; one positive control drug.
  • n H slope; IC 5 . : The maximum half of the test substance is inhibited.
  • the positive control drug Amitriptyline is one of the most widely used drugs for blocking hERG currents, IC 5 for inhibition of hERG current in this study. For 3.15 ⁇ , this result is consistent with the results reported in the literature. This shows that the results of this trial are credible.
  • test compound Compound 1-106
  • SD rats To observe the toxicity of the test compound (Compound 1-106) administered to SD rats for 14 days, to understand the toxicity of the test substance and its sputum organs, to determine the dose of non-toxic reaction under the test conditions; Whether the test substance has delayed toxicity and whether the toxic reaction is reversible.
  • the main experimental group was the vehicle control group, the test substance (Compound 1-106), the low dose group, the middle dose group, the high dose group, and 10 animals each, male and female.
  • the middle dose group and the vehicle control group were additionally restored. There are 10 animals each, half male and half female.
  • the compound of the present invention (Compound 1-106) has good tolerance and high safety.
  • 5-Bromouracil (10.00 g, 52.4 mmol) was added to the above aqueous solution, and refluxed for 16 h. The solution was cooled to -5 ° C, and the pH was adjusted to 7 with concentrated hydrochloric acid to precipitate a solid, which was filtered and dried to give a pale yellow solid, 3.50 g, yield 33%
  • V-(2-chloroethyl)morpholine hydrochloride (9.3 g, 0.05 mol), thiourea (3.81 g, 0.05 mol) in 95% ethanol (50 mL), heated to reflux for 24 hours, cooled Concentrated to give a white solid which was taken directly to next.
  • tert-Butyl 3-(2-chloro-5-(methylthio)pyrimidin-4-ylamino)phenylcarbamate (0.25 g, 0.68 mmol) was dissolved in 5 mL of tert-amyl alcohol, then acetic acid 2 was added 6-Methoxypyridine-3-amine (0.088 g, 0.71 mmol) was added dropwise, and the mixture was stirred at 85 ° C for 1 h. The reaction was cooled to room temperature, then EtOAc (EtOAc) was evaporated.
  • N"-(3-Aminophenyl)-W 2 -(4-(2-methoxyethoxy)phenyl)-5-(methylsulfinyl)pyrimidine-2,4-diamine (1.8 g, 4.35 mmol) was dissolved in 20 mL of dry tetrahydrofuran, 4 mL of DIPEA was added to bring the pH to 10, then 4-bromobut-2-enoyl chloride (crude, about 4.35 mmol) was dissolved in 10 mL of dichloromethane. Slowly drip into the solution, and the reaction was stirred at 0 ° C for 2 h and placed for later use.
  • Methyl 4-bromocrotonate ( 1.775 g, 9.93 mmol) was dissolved in 20 mL of dichloromethane, then pyrrolidine ( 1.408 g, 19.86 mmol) was added to the solution at 0 ° C, the reaction was stirred for 2 h, and dried. , used directly in the next step.
  • Methyl 4-(pyrrolidin-1-yl)but-2-enoate ( 1.67 g, 9.9 mmol) was dissolved in 20 mL of methanol, then NaOH (0.792 g, 19.8 mmol) was added and the resulting solution was at 0°.
  • Stir for 2 h at C then add 50 mL of ethyl acetate and 50 mL of water, extract, and dilute the aqueous phase with 2 N Hydrochloric acid was adjusted to pH 6.0, then ethyl acetate (100 mL) was added and extracted three times. The obtained organic phase was dried and dried to give a yield of 0.52 g of pale red oil.
  • the crude product in the previous step was dissolved in 15 mL of methanol and 5 mL of water, then slowly added potassium hydride (oxone) (0.301 g, 0.49 mmol), reacted at room temperature for 25 min, the solution was spun dry, and added to 30 The organic layer was dried over anhydrous sodium sulfate (MgSO4), filtered, filtered, filtered,jjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjj
  • 0.21 g of the previous step was dissolved in 20 mL of methanol and 2 mL of water, then slowly added potassium hydride (oxone) (0.301 g, 0.49 mmol) dissolved in 2 mL of water, and reacted at room temperature for 25 min. After quenching with 10 mL of a saturated aqueous solution of sodium thiosulfate, 50 mL of dichloromethane and 30 mL of water were added, and the organic phase was dried over anhydrous sodium sulfate, filtered, filtered and evaporated to dryness. 20:1) 0.096 g of a white solid was obtained.
  • 4-Bromocrotonic acid (0.106 g, 0.64 mmol) was dissolved in 10 mL of dichloromethane, and 2 drops of DMF were added, then 2 mL of oxalyl chloride was added at 0 ° C, stirred for 2 h, and dried, and was taken.
  • N 4 -(3-Aminophenyl)-TV 2 -(1-methyl-p-pyrazol-4-yl)-5-(methylthio)pyrimidine-2,4-diamine (0.21 g, 0.64 Ment) was dissolved in 20 mL of dry tetrahydrofuran, DIPEA (0.248 g, 1.92 mmol) was added to bring the pH to 10, then 4-bromo crotonyl chloride was dissolved in 5 mL of dichloromethane and slowly added dropwise to the system. The reaction was stirred at 0 ° C for 2 h and placed for later use.
  • V-(3-(2-(1-methyl-p-pyrazol-4-ylamino)-5-(methylthio)pyrimidin-4-ylamino)phenyl)-4-(pyrrolidin-1) -yl)but-2-enamide (0.10 g, 0.215 mmol) dissolved in 15 mL of methanol, then slowly added potassium peroxodisulfate complex (oxone) (0.146 g, 0.237 mmol) dissolved in 2 mL of water.

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Abstract

The present invention relates to compounds as represented by general formula (I) as tyrosine kinase inhibitors, preparation method thereof, pharmaceutical compositions containing the compounds, and uses thereof for preventing and/or treating instances of B-cell related leukemia, inflammatory diseases and autoimmune diseases, wherein A, ring B, L1, L2, R1, R2, R3, a, b, c, d, e, p and q are as defined in the specification.

Description

作为酪氨酸激酶抑制剂的含氮杂芳环衍生物  Nitrogen-containing heteroaromatic ring derivatives as tyrosine kinase inhibitors
1、 技术领域 1. Technical field
本发明属于医药技术领域， 具体涉及作为酪氨酸激酶抑制剂的含氮 杂芳环衍生物、 其药学上可接受的盐或其立体异构体， 这些化合物的制 备方法， 含有这些化合物的药物组合物，使用这些化合物预防和 /或治疗 个体中 B细胞相关的血癌（例如 B细胞慢性淋巴细胞癌、 非霍奇金淋巴 瘤）， 炎性以及自身免疫性疾病（例如类风湿性关节炎、 系统性红斑狼疮 等）的方法， 以及这些化合物在制备用于预防和 /或治疗 B细胞相关的血 癌（例如 B细胞慢性淋巴细胞癌、 非霍奇金淋巴瘤）， 炎性以及自身免疫 性疾病（例如类风湿性关节炎、 系统性红斑狼疮等）的药物中的应用。  The present invention relates to the field of medical technology, and particularly relates to a nitrogen-containing heteroaromatic ring derivative as a tyrosine kinase inhibitor, a pharmaceutically acceptable salt thereof or a stereoisomer thereof, a preparation method of these compounds, a drug containing the same Compositions for the prevention and/or treatment of B cell-associated blood cancers in an individual (eg, B-cell chronic lymphocytic carcinoma, non-Hodgkin's lymphoma), inflammatory and autoimmune diseases (eg, rheumatoid arthritis, Methods for systemic lupus erythematosus, etc., and the preparation of these compounds for the prevention and/or treatment of B cell-associated blood cancer (eg, B-cell chronic lymphocytic carcinoma, non-Hodgkin's lymphoma), inflammatory and autoimmune diseases Use in drugs such as rheumatoid arthritis, systemic lupus erythematosus, etc.
2、 背景技术  2. Background technology
蛋白盾激酶组成人类酶的最大家族之一， 并且通过添加磷酸基团到 蛋白盾上来调节许多不同的信号传导过程 (T. Hunter, Cell 1987 50: 823-829)。 特别地， 酪氨酸激酶磷酸化蛋白盾在酪氨酸残基的羟基部分。 酪氨酸激酶家族包括控制细胞生长、 迁移和分化的成员。 异常的激酶活 性已经涉及许多人类疾病， 包括癌症、 自身免疫疾病和炎性疾病。 由于 蛋白盾激酶属于细胞信号传导的关键调节剂， 它们提供用小分子激酶抑 制剂来调节细胞功能的目标， 并且因此成为了良好的药物设计耙标。 除 了激酶介导的疾病过程的治疗， 激酶活性的选择性和有效抑制剂还可用 于研究细胞信号传导过程和鉴定其它具有治疗意义的细胞耙标。  Protein Shield Kinases constitute one of the largest families of human enzymes and regulate many different signaling processes by the addition of phosphate groups to protein shields (T. Hunter, Cell 1987 50: 823-829). In particular, the tyrosine kinase phosphorylation protein is shielded in the hydroxyl portion of the tyrosine residue. The tyrosine kinase family includes members that control cell growth, migration, and differentiation. Abnormal kinase activity has been implicated in many human diseases, including cancer, autoimmune diseases, and inflammatory diseases. Since protein shield kinases are key regulators of cell signaling, they provide the goal of regulating cell function with small molecule kinase inhibitors and are therefore a good drug design target. In addition to the treatment of kinase-mediated disease processes, selective and potent inhibitors of kinase activity can be used to study cellular signaling processes and to identify other therapeutically significant cell targets.
关于 B 细胞在自身免疫和 /或炎性疾病的发病机制中的关键作用存 在良好的证据。 消耗 B细胞的基于蛋白盾的治疗剂如 Rituxan针对自身 抗体导致的炎性疾病如类风湿性关节炎是有效的（Rastetter等， Annu Rev Med 2004 55: 477)。 因此， 在 B细胞活化中发挥作用的蛋白盾激酶的抑 制剂应该是对于 B细胞介导的疾病病理如自身抗体生成有用的治疗剂。  There is good evidence that B cells play a key role in the pathogenesis of autoimmune and/or inflammatory diseases. Protein shield-based therapeutics that deplete B cells, such as Rituxan, are effective against autoantibody-induced inflammatory diseases such as rheumatoid arthritis (Rastetter et al, Annu Rev Med 2004 55: 477). Therefore, inhibitors of protein shield kinase that play a role in B cell activation should be useful therapeutic agents for B cell mediated pathology such as autoantibody production.
通过 B细胞受体 (BCR)的信号传导控制一系列 B细胞应答， 包括增 殖和分化到成熟的抗体生成细胞。 BCR是 B 细胞活性的关键调节点并 且异常的信号传导可以导致失调的 B 细胞增殖和病原性自身抗体的形 成， 其导致多种自身免疫疾病和 /或炎性疾病。 布鲁顿 (Bmton' s)酪氨酸 蛋白激酶 (Btk)是在 BCR的膜近端和紧接下游的非 BCR相关的激酶。 Btk 的缺乏已经显示阻断 BCR信号传导，并且因此 Btk的抑制可以是阻断 B 细胞介导的疾病过程的有效治疗方法。 Signaling through the B cell receptor (BCR) controls a range of B cell responses, including proliferation and differentiation into mature antibody-producing cells. BCR is a key regulatory point for B cell activity and abnormal signaling can lead to dysregulated B cell proliferation and the formation of pathogenic autoantibodies that result in a variety of autoimmune and/or inflammatory diseases. Bmton's tyrosine protein kinase (Btk) is a non-BCR-associated kinase proximal to and downstream of the membrane of BCR. The lack of Btk has been shown to block BCR signaling, and thus inhibition of Btk can be blocking B An effective treatment for cell-mediated disease processes.
Btk是酪氨酸激酶 Tec家族的成员 , 并且显示是早期 B细胞形成以 及成熟 B细胞活化和存活的关键调节剂（Khan等， Immunity 1995 3: 283; Ellmeier等， J. Exp. Med. 2000 192: 1611)。 人的 Btk突变导致病症 X连 锁丙球蛋白缺乏血症 (XLA)(Lindvall等 Immunol. Rev. 2005 203: 200)。 这些患者是免疫受损的， 并且显示受损的 B细胞成熟， 降低的免疫球蛋 白和外周 B细胞水平， 减少的不依赖 T细胞的免疫应答以及在 BCR刺 激后的减弱的钙动用。  Btk is a member of the Tec family of tyrosine kinases and has been shown to be a key regulator of early B cell formation and activation and survival of mature B cells (Khan et al, Immunity 1995 3: 283; Ellmeier et al, J. Exp. Med. 2000 192 : 1611). Human Btk mutations lead to the condition X-linked gamma globulin deficiency (XLA) (Lindvall et al Immunol. Rev. 2005 203: 200). These patients are immunocompromised and show impaired B cell maturation, reduced immunoglobulin and peripheral B cell levels, reduced T cell-independent immune response, and reduced calcium use following BCR stimulation.
关于 Btk在自身免疫疾病和炎性疾病中的作用的证据已经由 Btk-缺 陷型小鼠模型提供。 在系统性红斑狼疮 (SLE)的临床前鼠模型中， Btk缺 陷型小鼠显示疾病进展的显著改善。 此外， Btk-缺陷型小鼠对胶原诱导 的关节炎具有抗性 (Jansson和 Holmdahl Clin. Exp. Immunol. 1993 94: 459)。 已经证明选择性 Btk抑制剂在小鼠关节炎模型中的剂量依赖性功 效 (Z. Pan等， Chem. Med Chem. 2007 2: 58) 。  Evidence for the role of Btk in autoimmune diseases and inflammatory diseases has been provided by the Btk-deficient mouse model. Btk-deficient mice showed a significant improvement in disease progression in a preclinical mouse model of systemic lupus erythematosus (SLE). Furthermore, Btk-deficient mice are resistant to collagen-induced arthritis (Jansson and Holmdahl Clin. Exp. Immunol. 1993 94: 459). The dose-dependent efficacy of selective Btk inhibitors in the mouse arthritis model has been demonstrated (Z. Pan et al, Chem. Med Chem. 2007 2: 58).
Btk还有除了 B细胞之外可能涉及疾病过程的细胞表达。 例如 Btk 由肥大细胞表达并且 Btk缺陷型骨髓来源的肥大细胞显示受损的抗原诱 导的脱粒 (Iwaki J. Biol. Chem. 2005 280: 40261) 。 这显示 Btk可以用 于治疗病理性肥大细胞反应如变态反应和哮喘。 此外， 其中缺乏 Btk活 性的来自 XLA 患者的单核细胞显示在刺激后减少的 TNFa 生成 (Horwood 等 J Exp Med 2003 197: 1603)。 因此， TNFa介导的炎症可以 由小分子 Btk抑制剂调节。 此外， 已经报道的 Btk在细胞凋亡中发挥作 用（Islam和 Smith Immunol. Rev. 2000 178: 49) ,并且因此 Btk抑制剂对 于治疗某些 B 细胞淋巴瘤和白血病将是有效的（Feldhahn 等 J. Exp. Med. 2005 201 : 1837) 。  Btk also has cellular expression that may be involved in disease processes in addition to B cells. For example, Btk is expressed by mast cells and Btk-deficient bone marrow-derived mast cells show impaired antigen-induced degranulation (Iwaki J. Biol. Chem. 2005 280: 40261). This shows that Btk can be used to treat pathological mast cell responses such as allergies and asthma. In addition, monocytes from XLA patients lacking Btk activity show reduced TNFa production following stimulation (Horwood et al J Exp Med 2003 197: 1603). Thus, TNFa-mediated inflammation can be modulated by small molecule Btk inhibitors. Furthermore, Btk has been reported to play a role in apoptosis (Islam and Smith Immunol. Rev. 2000 178: 49), and thus Btk inhibitors will be effective in the treatment of certain B cell lymphomas and leukemias (Feldhahn et al. Exp. Med. 2005 201 : 1837).
2006年上市的 Dasatinib是多耙点抑制剂， 对 Btk具有较强抑制作 用， 用于治疗慢性骨髓性白血病； 此外处于临床 III期研究的 PCI- 32765 也是多耙点抑制剂， 对 Btk抑制作用为不可逆性， 用于治疗淋巴瘤、 白 血病及自身免疫疾病。
Figure imgf000004_0001
目前尚未有选择性的 Btk抑制剂上市， 研究最快的药物是 CC- 292 (又称 AVL- 292 ) , 处于临床 I期研究， 其不可逆的选择性抑制 Btk, 用于治疗白血病及
Figure imgf000004_0002
Dasatinib, which was launched in 2006, is a multi-inhibitor inhibitor, which has a strong inhibitory effect on Btk and is used to treat chronic myelogenous leukemia. In addition, PCI-32765, which is in clinical phase III study, is also a multi-defect inhibitor, and its inhibitory effect on Btk is Irreversible, used to treat lymphoma, leukemia and autoimmune diseases.
Figure imgf000004_0001
At present, there is no selective Btk inhibitor listed. The fastest drug to be studied is CC-292 (also known as AVL-292), which is in clinical phase I study. Its irreversible selective inhibition of Btk is used to treat leukemia and
Figure imgf000004_0002
本发明的目的是提供优良的高选择性 Btk抑制剂， 其能够用于预防 和 /或治疗 B细胞相关的血癌、 炎性和 /或自身免疫性疾病。  It is an object of the present invention to provide an excellent highly selective Btk inhibitor which can be used for the prevention and/or treatment of B cell-associated blood cancer, inflammatory and/or autoimmune diseases.
3、 发明内容  3, the content of the invention
本发明提供了用作酪氨酸激酶抑制剂的含氮杂芳环衍生物， 所述化 合物是优良的 Btk抑制剂， 并且能够用于预防和 /或治疗 B细胞相关的 血癌、 炎性和 /或自身免疫性疾病。  The present invention provides nitrogen-containing heteroaryl ring derivatives useful as tyrosine kinase inhibitors, which are excellent Btk inhibitors, and which can be used for the prevention and/or treatment of B cell-associated blood cancer, inflammatory and/or Or autoimmune disease.
具体而言， 本发明提供了下述通式 （I ) 所示的化合物、 其药学上 可接受的盐或
Figure imgf000004_0003
Specifically, the present invention provides a compound represented by the following formula (I), a pharmaceutically acceptable salt thereof or
Figure imgf000004_0003
其中， 环 A和环 B分别独立的表示苯基， 3- 7元环烷基， 含有 N、 0、 S杂原子的 3- 7元杂环烷基， 4- 7元杂芳基或 6-10元二环结构；  Wherein ring A and ring B each independently represent a phenyl group, a 3- 7 membered cycloalkyl group, a 3- 7 membered heterocycloalkyl group having a N, 0, S hetero atom, a 4- 7 membered heteroaryl group or 6- 10-membered two-ring structure;
Li和 L2分别独立的表示共价键， -NH- , - N(C1-3烷基) -， -0- , - S(0)m- , - N(C1-3 烷基 )C(0)- , - C(0)N(C1-3 烷基） -， - N(C1 -3 烷基 )S(0)2-或― S(0)2N(C1-3烷基) -； Li and L 2 each independently represent a covalent bond, -NH-, -N(C 1-3 alkyl) -, -0- , - S(0) m - , - N(C 1-3 alkyl) C(0)- , - C(0)N(C 1-3 alkyl) -, - N(C 1 -3 alkyl)S(0) 2 - or - S(0) 2 N(C 1- 3 alkyl) -;
a表示共价键， 未被取代或被 d_4烷基取代的亚氨基； A represents a covalent bond, a substituted or unsubstituted alkylene d_ 4 alkyl substituted amino;
b表示- CO-或- S02-； b represents -CO- or -S0 2 -;
c表示未被取代或被一或两个甲基或三氟甲基取代的 1,3-亚丙烯基、 1,1-或 1,2-亚乙烯基， 亚乙炔基， 或者未被取代或被一至四个甲基或三 氟甲基取代的 1,3-丁二烯 -1,4-亚基；  c represents 1,3-propenylene, 1,1- or 1,2-vinylidene, ethynylene, or unsubstituted or unsubstituted or substituted by one or two methyl or trifluoromethyl groups a 1,3-butadiene-1,4-subunit substituted with one to four methyl or trifluoromethyl groups;
d表示共价键或 亚烷基； e表示氢原子， 烷氧基，氨基， 3- 7元环烷基， 6- 10元二环结构， 烷基氨基或二- ( Cw烷基）氨基， 其中烷基部分可相同或不同； 和 分别独立的表示氢原子， 卤素原子，氰基，硝基， CM烯基， C2-4炔基或 - L3- R4, d represents a covalent bond or an alkylene group; e represents a hydrogen atom, an alkoxy group, an amino group, a 3- 7 membered cycloalkyl group, a 6- to 10-membered bicyclic structure, an alkylamino group or a bis-(Cw alkyl)amino group, wherein the alkyl moieties may be the same or different; Respectively represent a hydrogen atom, a halogen atom, a cyano group, a nitro group, a CM alkenyl group, a C 2-4 alkynyl group or a - L 3 - R 4 group , respectively.
L3表示共价键， -NH- , - N(C1-3烷基) -， -0- , - 0- C1-3亚烷基-， -S-Ci-3 亚烷基， - S(0)m- , - C(O)- , -NHC(O)- , - N(C1-3烷基) C(O)- , - C(0)NH- , -C(0)N(Ci-3 烷基） -， - NHS(0)2- , -N(Ci-3 烷基） S(0)2- , -S(0)2NH- , -S(0)2N(Ci-3烷基) -， - OC(O)-或- C(0)0- , L 3 represents a covalent bond, -NH-, -N(C 1-3 alkyl) -, -0- , - 0- C 1-3 alkylene-, -S-Ci -3 alkylene, - S(0) m - , - C(O)- , -NHC(O)- , - N(C 1-3 alkyl) C(O)- , - C(0)NH- , -C(0) N(Ci -3 alkyl) -, - NHS(0) 2 - , -N(Ci -3 alkyl) S(0) 2 - , -S(0) 2 NH- , -S(0) 2 N (Ci -3 alkyl) -, - OC(O)- or - C(0)0- ,
R4表示氢原子， 烷基， - N(Cw烷基 )2 , - NHC(0)0- (Cw烷基)， -OH , - 0(C1-4烷基)， - S(0)2(C1-3烷基）， 3- 7元环烷基， 苯基或 5- 6元杂 芳基； R4 represents a hydrogen atom, an alkyl group, -N(Cw alkyl) 2 , -NHC(0)0-(Cw alkyl), -OH, - 0(C 1-4 alkyl), - S(0) 2 (C 1-3 alkyl), 3- 7 membered cycloalkyl, phenyl or 5- 6-membered heteroaryl;
R2表示- L4- R5 , R 2 represents - L 4 - R 5 ,
L4表示共价键， -NH- , - N(C1-3烷基) -， - 0- C1-3亚烷基-， - S- C1-3亚 烷基-或 - S(0)m- , L 4 represents a covalent bond, -NH-, -N(C 1-3 alkyl)-, - 0-C 1-3 alkylene-, -S-C 1-3 alkylene- or -S ( 0) m - ,
R5表示氢原子， C2-4烯基， C2-4炔基， C1-4烷基， 氨基， - NH(C1-3 烷基）， - N(Cw烷基 )2 , -OH , - 0(Cw烷基)， - C(0)(Cw烷基)或 3- 7元环 烷基， R 5 represents a hydrogen atom, C 2-4 alkenyl, C 2-4 alkynyl, C 1-4 alkyl, amino, -NH(C 1-3 alkyl), -N(Cw alkyl) 2 , - OH , - 0 (Cw alkyl), - C(0) (Cw alkyl) or 3- 7 membered cycloalkyl,
其中 L4为共价键时， R5不能为 C1-4烷基， - OH , - 0(C1-3烷基）， - C(0)(C1-3烷基)， Where L 4 is a covalent bond, R 5 cannot be C 1-4 alkyl, - OH , - 0 (C 1-3 alkyl), - C(0) (C 1-3 alkyl),
L4为- 0- d_3亚烷基 -时， R5不能为氢原子； L 4 is - 0- d_ 3 alkylene - when, R 5 is not a hydrogen atom;
所述 烷基部分、 环烷基、 杂芳基可以进一步被 1至 4个（^取 代，  The alkyl moiety, cycloalkyl group, heteroaryl group may be further substituted by 1 to 4 (^,
表示 素原子、 烷基、 氨基、 烷基氨基、 二-（Cw烷基） 氨基、 羟基、 烷氧基、 烷氧羰基、 氨基甲酰基、 烷基氨基甲 酰基、 二- ( C 烷基）氨基甲酰基或 3-6元环烷基， 其中 （^可以相同 或不同；  Derivative atom, alkyl, amino, alkylamino, bis-(Cw alkyl)amino, hydroxy, alkoxy, alkoxycarbonyl, carbamoyl, alkylcarbamoyl, di-(C alkyl)amino Formyl or 3-6 membered cycloalkyl, wherein (^ may be the same or different;
所述环烷基、二环结构上碳原子可以被 1 -4个相同或不同的 N、 NH、 N(C1-3烷基)、 0、 S(0)m、 C(O)替换； The carbon atom of the cycloalkyl or bicyclic structure may be replaced by 1-4 identical or different N, NH, N(C 1-3 alkyl), 0, S(0) m , C(O);
所述杂芳基含有 1-4个杂原子， 分别独立的选 N、 0或 S ;  The heteroaryl group has 1-4 hetero atoms, and each of them is independently selected from N, 0 or S;
m表示 0 , 1或 2;  m means 0, 1 or 2;
p和 q分别独立的表示 0 , 1 , 2 , 3或 4。  p and q are independent representations of 0, 1, 2, 3 or 4.
在一个优选的实施方案中， 本发明提供了上述通式（I )所示的化 合物、 其药学上可接受的盐或其立体异构体， 其中： In a preferred embodiment, the present invention provides the above-described formula (I) a compound, a pharmaceutically acceptable salt thereof or a stereoisomer thereof, wherein:
环 A和环 B分别独立的表示苯基， 3- 7元环烷基， 含有 N、 0、 S 杂原子的 3-7元杂环烷基， 4-7元杂芳基或 6-10元二环结构；  Ring A and Ring B each independently represent a phenyl group, a 3- 7 membered cycloalkyl group, a 3-7 membered heterocycloalkyl group having a N, 0, S hetero atom, a 4-7 membered heteroaryl group or a 6-10 member. Two-ring structure;
Li和 L2分别独立的表示共价键， -NH- , - N(C1-3烷基) -， -0- , - S(0)m- , - N(C1-3 烷基 )C(0)- , - C(0)N(C1-3 烷基） -， - N(C1 -3 烷基 )S(0)2-或― S(0)2N(C1-3烷基) -； Li and L 2 each independently represent a covalent bond, -NH-, -N(C 1-3 alkyl) -, -0- , - S(0) m - , - N(C 1-3 alkyl) C(0)- , - C(0)N(C 1-3 alkyl) -, - N(C 1 -3 alkyl)S(0) 2 - or - S(0) 2 N(C 1- 3 alkyl) -;
a表示共价键， 未被取代或被 d_4烷基取代的亚氨基； A represents a covalent bond, a substituted or unsubstituted alkylene d_ 4 alkyl substituted amino;
b表示- CO-或- S02-； b represents -CO- or -S0 2 -;
c表示未被取代或被一或两个甲基或经三氟甲基取代的 1 ,3-亚丙烯 基、 1,1-或 1,2-亚乙烯基， 亚乙炔基， 或者未被取代或被一至四个甲基 或经三氟甲基取代的 1,3-丁二烯 -1,4-亚基；  c represents 1, 3-propenylene, 1,1- or 1,2-vinylidene, ethynylene, or unsubstituted, which is unsubstituted or substituted by one or two methyl or trifluoromethyl groups. Or a 1,3-butadiene-1,4-subunit substituted by one to four methyl groups or by a trifluoromethyl group;
d表示共价键或 亚烷基；  d represents a covalent bond or an alkylene group;
e表示氢原子， 烷氧基，氨基， 3- 7元环烷基， 6- 10元二环结构， 烷基氨基或二- ( Cw烷基）氨基， 其中烷基部分可相同或不同； 和 分别独立的表示氢原子， 卤素原子，氰基，硝基， CM烯基， e represents a hydrogen atom, an alkoxy group, an amino group, a 3- 7 membered cycloalkyl group, a 6- to 10-membered bicyclic structure, an alkylamino group or a bis-(Cw alkyl)amino group, wherein the alkyl moieties may be the same or different; Respectively represent a hydrogen atom, a halogen atom, a cyano group, a nitro group, a CM alkenyl group,
C2-4炔基或 - L3- R4, C 2-4 alkynyl or - L 3 - R 4 ,
L3表示共价键， -NH- , - N(C1-3烷基) -， -0- , - 0- C1-3亚烷基-， -S-Ci-3 亚烷基， - S(0)m- , - C(O)- , -NHC(O)- , - N(C1-3烷基) C(O)- , - C(0)NH- , -C(0)N(Ci-3 烷基） -， - NHS(0)2- , -N(Ci-3 烷基） S(0)2- , -S(0)2NH- , - S(0)2N(C1-3烷基) -， - OC(O)-或- C(0)0- , L 3 represents a covalent bond, -NH-, -N(C 1-3 alkyl) -, -0- , - 0- C 1-3 alkylene-, -S-Ci -3 alkylene, - S(0) m - , - C(O)- , -NHC(O)- , - N(C 1-3 alkyl) C(O)- , - C(0)NH- , -C(0) N(Ci -3 alkyl) -, - NHS(0) 2 - , -N(Ci -3 alkyl) S(0) 2 - , -S(0) 2 NH- , - S(0) 2 N (C 1-3 alkyl) -, - OC(O)- or - C(0)0- ,
R4表示氢原子， 烷基， - N(Cw烷基 )2 , - NHC(0)0- (Cw烷基)， -OH , - 0(C1-4烷基)， - S(0)2(C1-3烷基）， 3- 7元环烷基， 苯基或 5- 6元杂 芳基； R4 represents a hydrogen atom, an alkyl group, -N(Cw alkyl) 2 , -NHC(0)0-(Cw alkyl), -OH, - 0(C 1-4 alkyl), - S(0) 2 (C 1-3 alkyl), 3- 7 membered cycloalkyl, phenyl or 5- 6-membered heteroaryl;
R2表示- L4- R5 , R 2 represents - L 4 - R 5 ,
L4表示- S- C1-3亚烷基 -或- S(0)m- , L 4 represents -S-C 1-3 alkylene- or -S(0) m - ,
R5表示氢原子， C2-4烯基， C2-4炔基， C1-4烷基， 氨基， - NH(C1-3 烷基）， - N(Cw烷基 )2 , -OH , - 0(Cw烷基)， - C(0)(Cw烷基)或 3- 7元环 烷基， R 5 represents a hydrogen atom, C 2-4 alkenyl, C 2-4 alkynyl, C 1-4 alkyl, amino, -NH(C 1-3 alkyl), -N(Cw alkyl) 2 , - OH , - 0 (Cw alkyl), - C(0) (Cw alkyl) or 3- 7 membered cycloalkyl,
m表示 0 , 1或 2。  m means 0, 1 or 2.
所述 烷基部分、 环烷基、 杂芳基可以进一步被 1至 4个（^取 代，  The alkyl moiety, cycloalkyl group, heteroaryl group may be further substituted by 1 to 4 (^,
表示 素原子、 烷基、 氨基、 烷基氨基、 二-（Cw烷基） 氨基、 羟基、 烷氧基、 烷氧羰基、 氨基甲酰基、 烷基氨基甲 酰基、 二- ( C 烷基）氨基甲酰基或 3-6元环烷基， 其中 （^可以相同 或不同； Expressive atom, alkyl, amino, alkylamino, di-(Cw alkyl) An amino group, a hydroxyl group, an alkoxy group, an alkoxycarbonyl group, a carbamoyl group, an alkylcarbamoyl group, a bis-(C alkyl)carbamoyl group or a 3-6 membered cycloalkyl group, wherein (^ may be the same or different;
所述环烷基、二环结构上碳原子可以被 1 -4个相同或不同的 N、 NH、 N(C1-3烷基)、 0、 S(0)m、 C(O)替换； The carbon atom of the cycloalkyl or bicyclic structure may be replaced by 1-4 identical or different N, NH, N(C 1-3 alkyl), 0, S(0) m , C(O);
所述杂芳基含有 1-4个杂原子， 分别独立的选 N、 0或 S;  The heteroaryl group contains 1-4 heteroatoms, each independently selected N, 0 or S;
m表示 0, 1或 2;  m means 0, 1 or 2;
p和 q分别独立的表示 0, 1 , 2 , 3或 4。  p and q are independent representations of 0, 1, 2, 3 or 4.
在另一个优选的实施方案中， 本发明提供了上述通式（I )所示的 化合物、 其药学上可接受的盐或其立体异构体， 其中：  In another preferred embodiment, the present invention provides a compound represented by the above formula (I), a pharmaceutically acceptable salt thereof or a stereoisomer thereof, wherein:
环 A和环 B分别独立的表示苯基， 5- 6元环烷基， 含有 N、 0、 S 杂原子的 5-6元杂环烷基， 5-6元杂芳基或 8-10元二环结构；  Ring A and Ring B each independently represent a phenyl group, a 5- to 6-membered cycloalkyl group, a 5-6 membered heterocycloalkyl group having a N, 0, S hetero atom, a 5-6 membered heteroaryl group or 8-10 members. Two-ring structure;
Li和 L2分别独立的表示共价键， -NH-, -N(CH3)-, -Ο-, - S(0)m- , -N(CH3)C(0)-, -C(0)N(CH3) -, - N(CH3)S(0)2-或- S(0)2N(CH3) -； Li and L 2 each independently represent a covalent bond, -NH-, -N(CH 3 )-, -Ο-, - S(0) m - , -N(CH 3 )C(0)-, -C (0)N(CH 3 ) -, - N(CH 3 )S(0) 2 - or - S(0) 2 N(CH 3 ) -;
a表示共价键， 未被取代或被 CH3取代的亚氨基； a represents a covalent bond, an imino group that is unsubstituted or substituted with CH 3 ;
b表示- CO-或- S02-； b represents -CO- or -S0 2 -;
c表示未被取代或被一或两个甲基取代的 1,2-亚乙烯基或亚乙炔基； d表示共价键或亚甲基；  c represents a 1,2-vinylidene or ethynylene group which is unsubstituted or substituted by one or two methyl groups; d represents a covalent bond or a methylene group;
e表示氢原子， 甲氧基， 氨基， 哌啶基， 吗啉基， 吡咯烷基， 哌嗪 基， TV-甲基哌嗪基， 6- 9元螺环结构， 6- 8元并环结构， 6- 8元桥环结构， 甲基氨基或二- (甲基）氨基；  e represents a hydrogen atom, a methoxy group, an amino group, a piperidinyl group, a morpholinyl group, a pyrrolidinyl group, a piperazinyl group, a TV-methyl piperazinyl group, a 6- to 9-membered spiro ring structure, a 6- to 8-membered ring structure. , 6- to 8-membered bridged ring structure, methylamino or bis-(methyl)amino;
和 分别独立的表示氢原子， 卤素原子， 硝基或 - L3- R4 , And independently represent a hydrogen atom, a halogen atom, a nitro group or a - L 3 - R4 group,
L3表示共价键， -NH-, - N(C1-3烷基) -， -0-, - 0- C1-3亚烷基-， -S-Ci-3 亚烷基， - S(0)m- , - C(O)- , -NHC(O)-, - C(0)NH- , - NHS(0)2- , - S(0)2NH- , - oc(o)-或- c(o)o- , L 3 represents a covalent bond, -NH-, -N(C 1-3 alkyl)-, -0-, - 0-C 1-3 alkylene-, -S-Ci -3 alkylene, - S(0) m - , - C(O)- , -NHC(O)-, - C(0)NH- , - NHS(0) 2 - , - S(0) 2 NH- , - oc(o )- or - c(o)o- ,
R4表示氢原子， 烷基， - N(Cw烷基 )2, - NHC(0)0- (Cw烷基)， -OH, - 0(C1-4烷基)， - S(0)2(C1-3烷基）， 5- 6元环烷基， 苯基或 5- 6元杂 芳基； R4 represents a hydrogen atom, an alkyl group, -N(Cw alkyl)2, -NHC(0)0-(Cw alkyl), -OH, - 0(C 1-4 alkyl), - S(0) 2 (C 1-3 alkyl), 5- 6-membered cycloalkyl, phenyl or 5- 6-membered heteroaryl;
R2表示- L4- R5 , R 2 represents - L 4 - R 5 ,
L4表示- S- C1-3亚烷基 -或- S(0)m- , L 4 represents -S-C 1-3 alkylene- or -S(0) m - ,
R5表示氢原子， C1-4烷基， 氨基， - NH(C1-3烷基)， - N(C1-3烷基 )2,R 5 represents a hydrogen atom, a C 1-4 alkyl group, an amino group, -NH(C 1-3 alkyl), -N(C 1-3 alkyl) 2 ,
-OH, - 0(Cw烷基)， - C(0)(Cw烷基)或 4-7元环烷基， 所述 烷基部分、 环烷基、 苯基、 杂芳基、 螺环结构、 并环结构、 桥环结构可以进一步被 1至 4个（^取代， -OH, - 0 (Cw alkyl), - C(0) (Cw alkyl) or 4-7 membered cycloalkyl, The alkyl moiety, cycloalkyl group, phenyl group, heteroaryl group, spiro ring structure, concentric ring structure, bridged ring structure may be further substituted by 1 to 4 (^,
表示 素原子、 烷基、 氨基、 烷基氨基、 二-（Cw烷基） 氨基、 羟基、 烷氧基、 烷氧羰基、 氨基甲酰基、 烷基氨基甲 酰基、 二- ( C 烷基）氨基甲酰基或 5-6元环烷基， 其中 （^可以相同 或不同；  Derivative atom, alkyl, amino, alkylamino, bis-(Cw alkyl)amino, hydroxy, alkoxy, alkoxycarbonyl, carbamoyl, alkylcarbamoyl, di-(C alkyl)amino Formyl or 5-6 membered cycloalkyl, wherein (^ may be the same or different;
所述环烷基、二环结构上碳原子可以被 1 -4个相同或不同的 N、 NH、 N(C1-3烷基)、 0、 S(0)m、 C(O)替换； The carbon atom of the cycloalkyl or bicyclic structure may be replaced by 1-4 identical or different N, NH, N(C 1-3 alkyl), 0, S(0) m , C(O);
所述杂芳基、 螺环结构、 并环结构、 桥环结构含有 1-4个杂原子， 分别独立的选 N、 0或 S ;  The heteroaryl group, the spiro ring structure, the concentric ring structure, and the bridged ring structure contain 1-4 hetero atoms, and each of them is independently selected from N, 0 or S;
m表示 0 , 1或 2 ;  m means 0 , 1 or 2 ;
p和 q分别独立的表示 0 , 1 , 2 , 3或 4。  p and q are independent representations of 0, 1, 2, 3 or 4.
在另一个优选的实施方案中， 本发明提供了上述通式（I )所示的 化合物、 其药学上可接受的盐或其立体异构体， 其中：  In another preferred embodiment, the present invention provides a compound represented by the above formula (I), a pharmaceutically acceptable salt thereof or a stereoisomer thereof, wherein:
环 A和环 B分别独立的表示苯基， 含有 N杂原子的 5-6元杂环烷 基或 5-6元杂芳基；  Ring A and ring B each independently represent a phenyl group, a 5-6 membered heterocycloalkyl group having a N hetero atom or a 5-6 membered heteroaryl group;
Li和 L2分别独立的表示- NH- , — 0-或- S(0)m-； Li and L 2 are independently represented by -NH- , - 0- or - S(0) m -;
a表示共价键或亚氨基；  a represents a covalent bond or an imino group;
b表示- CO-；  b means - CO-;
c表示 1,2-亚乙烯基；  c represents 1,2-vinylidene;
d表示共价键或亚甲基；  d represents a covalent bond or a methylene group;
e表示氢原子， 哌啶基， 吗啉基， 吡咯烷基， 哌嗪基或二-（甲基） 氨基；  e represents a hydrogen atom, piperidinyl, morpholinyl, pyrrolidinyl, piperazinyl or bis-(methyl)amino;
Ri表示氢原子， 素原子， 未被取代或被一至三个 素原子取代的 甲基、 甲氧基， 甲基氨基或二 -（甲基）氨基；  Ri represents a hydrogen atom, a gas atom, a methyl group, a methoxy group, a methylamino group or a di-(methyl)amino group which is unsubstituted or substituted with one to three atomic atoms;
R3表示氢原子， 卤素原子或- L3- R4, R 3 represents a hydrogen atom, a halogen atom or -L 3 - R 4 ,
L3表示共价键， -NH- , - N(C1-3烷基) -， -0- , - 0- C1-3亚烷基-， -S-Ci-3 亚烷基-， - S(0)m- , - C(O)- , -NHC(O)- , - C(0)NH- , - OC(O)-或- C(0)0- , R4表示氢原子，甲基，乙基， - N(C1-3烷基 )2 , - NHC(0)0- CH3，- 0(CH3) , -0(CH2CH3) , - 0(C(CH3)3) , -S(0)2-CH3 , 环戊烷基， 环己烷基， 吡咯烷 基， 四氢呋喃基， 哌啶基， 吗啉基， 哌嗪基， 苯基， 吡咯基， 咪唑基， 噻唑基， 噁唑基， 噻二唑基， 吡啶基或嘧啶基； R2表示甲基硫基， 乙基硫基， 甲基磺酰基， 氨基磺酰基， 甲基亚磺 酰基或氨基亚磺酰基， L 3 represents a covalent bond, -NH-, -N(C 1-3 alkyl)-, -0- , - 0- C 1-3 alkylene-, -S-Ci -3 alkylene-, - S(0) m - , - C(O)- , -NHC(O)- , - C(0)NH- , - OC(O)- or -C(0)0- , R4 represents a hydrogen atom, Methyl, ethyl, -N(C 1-3 alkyl) 2 , - NHC(0)0-CH 3 , - 0(CH 3 ) , -0(CH 2 CH 3 ) , - 0 (C(CH) 3 ) 3 ), -S(0) 2 -CH 3 , cyclopentyl, cyclohexane, pyrrolidinyl, tetrahydrofuranyl, piperidinyl, morpholinyl, piperazinyl, phenyl, pyrrolyl, Imidazolyl, thiazolyl, oxazolyl, thiadiazolyl, pyridyl or pyrimidinyl; R 2 represents a methylthio group, an ethylthio group, a methylsulfonyl group, an aminosulfonyl group, a methylsulfinyl group or an aminosulfinyl group.
所述甲基硫基、 乙基硫基、 甲基磺酰基、 氨基磺酰基、 甲基亚磺酰 基、 氨基亚磺酰基可以进一步被一至两个相同或不同的甲氧基、 吡咯烷 基、 四氢咪唑基、 哌啶基、 吗啉基、 哌嗪基取代，  The methylthio group, ethylthio group, methylsulfonyl group, aminosulfonyl group, methylsulfinyl group, aminosulfinyl group may be further one to two of the same or different methoxy, pyrrolidinyl, tetra Hydroimidazolyl, piperidinyl, morpholinyl, piperazinyl,
所述吡咯烷基、 四氢咪唑基、 哌啶基、 吡啶基、 吗啉基、 哌嗪基、 苯基、 吡咯基、 咪唑基、 噻唑基、 噁唑基、 噻二唑基、 吡啶基或嘧啶基 可以进一步被一至两个相同或不同的 取代，  The pyrrolidinyl group, tetrahydroimidazolyl group, piperidinyl group, pyridyl group, morpholinyl group, piperazinyl group, phenyl group, pyrrolyl group, imidazolyl group, thiazolyl group, oxazolyl group, thiadiazolyl group, pyridyl group or The pyrimidinyl group may be further substituted by one to two identical or different,
表示 素原子、 甲基、 氨基、 甲基氨基、 二- (甲基）氨基、 羟 基、 甲氧基、 甲氧羰基、 氨基甲酰基、 甲基氨基甲酰基或二— （甲基） 氨基甲酰基；  Derivative atom, methyl, amino, methylamino, bis-(methyl)amino, hydroxy, methoxy, methoxycarbonyl, carbamoyl, methylcarbamoyl or bis-(methyl)carbamoyl ;
m表示 0, 1或 2;  m means 0, 1 or 2;
p和 q分别独立的表示 0, 1或 2。  p and q stand independently for 0, 1 or 2.
在另一个优选的实施方案中， 本发明提供了上述通式（I )所示的 化合物、 其药学上可接受的盐或其立体异构体， 其中：  In another preferred embodiment, the present invention provides a compound represented by the above formula (I), a pharmaceutically acceptable salt thereof or a stereoisomer thereof, wherein:
环 A和环 B分别独立的表示苯基， 吡啶基， 哌啶基， 咪唑基， 吡 唑基， 噻唑基， 噻哈基， 1,2,3-三唑基或 1,2,4-三唑基；  Ring A and Ring B each independently represent phenyl, pyridyl, piperidinyl, imidazolyl, pyrazolyl, thiazolyl, thiazyl, 1,2,3-triazolyl or 1,2,4-tri Azolyl
Li和 L2分别独立的表示- NH-或- 0-； Li and L 2 are independently represented by -NH- or - 0-;
a表示共价键或亚氨基；  a represents a covalent bond or an imino group;
b表示- CO-；  b means - CO-;
c表示 1,2-亚乙烯基；  c represents 1,2-vinylidene;
d表示共价键或亚甲基；  d represents a covalent bond or a methylene group;
e表示氢原子， 哌啶基， 吡咯烷基， 吗啉基， 哌嗪基或二-（甲基） 氨基；  e represents a hydrogen atom, piperidinyl, pyrrolidinyl, morpholinyl, piperazinyl or bis-(methyl)amino;
表示氢原子， 氟原子， 氯原子，三氟甲基， 甲氧基或三氟甲氧基； R3表示氢原子， 氟原子， 氯原子或- L3- R4, Represents a hydrogen atom, a fluorine atom, a chlorine atom, a trifluoromethyl group, a methoxy group or a trifluoromethoxy group; R 3 represents a hydrogen atom, a fluorine atom, a chlorine atom or -L 3 - R 4 ,
L3表示共价键， -NH-, - N (Ci-3烷基） -， - 0- , - 0- CH2CH2- , -S-CH2CH2- 或- s(o)m -, L 3 represents a covalent bond, -NH-, - N (Ci -3 alkyl) -, - 0- , - 0- CH 2 CH 2 - , -S-CH 2 CH 2 - or - s(o) m -,
R4表示氢原子，甲基，乙基， - N(C1-3烷基 )2, - NHC(0)0- CH3，- 0(CH3),R4 represents a hydrogen atom, methyl, ethyl, -N(C 1-3 alkyl) 2 , -NHC(0)0-CH 3 , - 0(CH 3 ),
- 0(C(CH3)3), -S(0)2-CH3 , 吡咯烷基， 四氢呋喃基， 哌啶基， 吗啉基， 苯基， 吡咯基， 咪唑基， 噻唑基， 噁唑基， 噻二唑基或吡啶基， - 0(C(CH 3 ) 3 ), -S(0) 2 -CH 3 , pyrrolidinyl, tetrahydrofuranyl, piperidinyl, morpholinyl, phenyl, pyrrolyl, imidazolyl, thiazolyl, oxazole Base, thiadiazolyl or pyridyl,
所述苯基、 吡咯基、 咪唑基、 噻唑基、 噁唑基、 噻二唑基、 吡啶基 可以进一步被一至两个相同或不同的 取代， The phenyl group, pyrrolyl group, imidazolyl group, thiazolyl group, oxazolyl group, thiadiazolyl group, pyridyl group Can be further replaced by one or two identical or different,
表示氨基甲酰基、 甲基氨基甲酰基或二- (甲基）氨基甲酰基； Represents a carbamoyl group, a methylcarbamoyl group or a bis-(methyl)carbamoyl group;
R2表示未被取代或被甲氧基、 哌啶基、 吗啉基取代的乙基硫基， 甲 基硫基， 甲基磺酰基， 氨基磺酰基， 甲基亚磺酰基， 氨基亚磺酰基； m表示 0, 1或 2; R 2 represents an ethylthio group which is unsubstituted or substituted by a methoxy group, a piperidinyl group or a morpholinyl group, a methylthio group, a methylsulfonyl group, an aminosulfonyl group, a methylsulfinyl group, an aminosulfinyl group. ; m means 0, 1 or 2;
p和 q分别独立的表示 0或 1。  p and q stand independently for 0 or 1.
在另一个优选的实施方案中， 本发明提供了上述通式（I )所示的 化合物、 其药学上可接受的盐或其立体异构体， 其中：  In another preferred embodiment, the present invention provides a compound represented by the above formula (I), a pharmaceutically acceptable salt thereof or a stereoisomer thereof, wherein:
环 A和环 B分别独立的表示苯基， 吡啶基， 吡唑基或哌啶基； Ring A and ring B each independently represent a phenyl group, a pyridyl group, a pyrazolyl group or a piperidinyl group;
Li和 L2分别独立的表示- NH-； Li and L 2 are independently represented by -NH-;
a表示亚氨基；  a represents an imino group;
b表示- CO-；  b means - CO-;
c表示 1,2-亚乙烯基；  c represents 1,2-vinylidene;
d表示共价键或亚甲基；  d represents a covalent bond or a methylene group;
e表示氢原子， 吡咯烷基， 吗啉基， 哌嗪基或二-（甲基）氨基； Ri表示氢原子；  e represents a hydrogen atom, a pyrrolidinyl group, a morpholinyl group, a piperazinyl group or a bis-(methyl)amino group; and Ri represents a hydrogen atom;
R3表示- L3- R4 , L3表示共价键， - 0-或- 0- CH2CH2- , R4表示- N(CH3)2, -0(CH3), 四氢呋喃基， 吗啉基， 苯基或吡啶基， R 3 represents - L 3 - R4 , L 3 represents a covalent bond, - 0- or - 0-CH 2 CH 2 -, R4 represents -N(CH 3 ) 2 , -0(CH 3 ), tetrahydrofuranyl, Phenyl, phenyl or pyridyl,
所述苯基、 吡啶基可以进一步被一至两个相同或不同的 取代， 表示氨基甲酰基、 甲基氨基甲酰基或二- (甲基）氨基甲酰基；  The phenyl group and the pyridyl group may be further substituted by one to two identical or different, and represent a carbamoyl group, a methylcarbamoyl group or a bis-(methyl)carbamoyl group;
R2表示未被取代或被甲氧基、 吗啉基取代的乙基硫基， 甲基硫基， 甲基磺酰基， 氨基磺酰基， 甲基亚磺酰基， 氨基亚磺酰基； R 2 represents an ethylthio group which is unsubstituted or substituted with a methoxy group or a morpholinyl group, a methylthio group, a methylsulfonyl group, an aminosulfonyl group, a methylsulfinyl group, an aminosulfinyl group;
p表示 1 ;  p represents 1 ;
q表示 1。  q means 1.
在另一个优选的实施方案中， 本发明提供了上述通式（I )所示的 化合物、 其药学上可接受的盐或其立体异构体， 其中：  In another preferred embodiment, the present invention provides a compound represented by the above formula (I), a pharmaceutically acceptable salt thereof or a stereoisomer thereof, wherein:
环 A表示苯基； 环 B表示苯基， 吡唑基或吡啶基； 和 L2分别 独立的表示 -NH-; Ring A represents phenyl; ring B represents phenyl, pyrazolyl or pyridyl; and L 2 independently represents -NH-;
a表示亚氨基； b表示- CO-； c表示 1,2-亚乙烯基； d表示共价键; e表示氢原子； 表示氢原子；  a represents an imino group; b represents -CO-; c represents 1,2-vinylidene; d represents a covalent bond; e represents a hydrogen atom; represents a hydrogen atom;
R3表示- L3- R4 , L3表示共价键， -NH-, - N(C1-3烷基) -， -0-, - 0- C1-3 亚烷基-， - S- C1-3亚烷基-， 或- C(0)NH- , R4表示氢原子， 甲基， 乙基， - 0(CH3), - 0(C(CH3)3)或吗啉基； R2表示- L4- R5 , L4表示- S- C1-3亚烷基 -或- S(0)m- , R5表示氢原子或 p表示 1； q表示 1。 R 3 represents - L 3 - R4 , L 3 represents a covalent bond, -NH-, - N(C 1-3 alkyl) -, -0-, - 0- C 1-3 alkylene-, - S - C 1-3 alkylene-, or - C(0)NH- , R4 represents a hydrogen atom, methyl, ethyl, - 0(CH 3 ), - 0(C(CH 3 ) 3 ) or morpholinyl; R 2 represents -L 4 - R 5 , and L 4 represents -S-C 1-3 alkylene- or -S(0) m - , R 5 represents a hydrogen atom or p represents 1; q represents 1.
发明详述  Detailed description of the invention
本发明所述的 "d_6烷基"是指含有 1-6个碳原子的直链或支链的烷 基，其中包括" 烷基"、 "Cw烷基"等，其实例包括但不限于例如甲基、 乙基、 正丙基、 异丙基、 正丁基、 2-甲基丙基、 1-甲基丙基、 1,1-二甲基 乙基等。 术语 "Cw烷基"、 "Cw烷基"指上述实例中的含有 1至 4个、 1 至 3个碳原子的具体实例。 The "d- 6 alkyl group" as used in the present invention means a linear or branched alkyl group having 1 to 6 carbon atoms, and includes "alkyl", "Cw alkyl" and the like, and examples thereof include, but are not limited to, For example, methyl, ethyl, n-propyl, isopropyl, n-butyl, 2-methylpropyl, 1-methylpropyl, 1,1-dimethylethyl and the like. The terms "Cw alkyl" and "Cw alkyl" refer to specific examples containing 1 to 4, 1 to 3 carbon atoms in the above examples.
本发明所述的 "d_6亚烷基 "是指含有 1-6个碳原子的直链或支链的 烷基去掉一个氢原子后的结构，是指二价烷基。 "亚烷基链"为聚亚甲基， 即- (CH2)x- , 其中 X为正整数， 优选为 1 到 6、 1 到 4、 1 到 3、 1 到 2 或 2 到 3。 经取代亚烷基链是一个或一个以上亚甲基氢原子经取代基置 换的聚亚甲基。 其中包括" C1-4亚烷基"、 "C1-3亚烷基 "等， 其实例包括但 不限于例如亚甲基（ - CH2- ),亚乙基（ - CH2CH2- ),亚丙基（ - CH2CH2CH2- ), 亚丁基（- CH2CH2CH2CH2- )等。 术语" C1-4亚烷基"、 "C1-3亚烷基"指上 述实例中的含有 1至 4个、 1至 3个碳原子的具体实例。 The "d- 6 alkylene group" as used in the present invention means a structure in which a straight or branched alkyl group having 1 to 6 carbon atoms is removed by one hydrogen atom, and means a divalent alkyl group. The "alkylene chain" is a polymethylene group, that is, -(CH 2 )x- , wherein X is a positive integer, preferably 1 to 6, 1 to 4, 1 to 3, 1 to 2 or 2 to 3. The substituted alkylene chain is a polymethylene group in which one or more methylene hydrogen atoms are replaced by a substituent. These include "C 1-4 alkylene", "C 1-3 alkylene" and the like, examples of which include, but are not limited to, for example, methylene (-CH 2 - ), ethylene ( - CH 2 CH 2 - ), propylene (-CH 2 CH 2 CH 2 - ), butylene (-CH 2 CH 2 CH 2 CH 2 - ), and the like. The term "C 1-4 alkylene", "C 1-3 alkylene" refers to a specific example containing 1 to 4, 1 to 3 carbon atoms in the above examples.
本发明所述的 "C2_4烯基"是指含有双键的碳原子数为 2-4 的直链或 支链烯基； 其实例包括但不限于例如乙烯基、 1-丙烯基、 2-丙烯基、 1- 甲基乙烯基、 1-丁烯基、 2-丁烯基、 3-丁烯基、 1—甲基— 1—丙烯基、 2-甲 基— i—丙烯基、 1—甲基— 2-丙烯基、 2-甲基- 2-丙烯基。 The "C 2 _ 4 alkenyl group" as used in the present invention means a linear or branched alkenyl group having 2 to 4 carbon atoms; examples thereof include, but are not limited to, for example, a vinyl group, a 1-propenyl group, 2-propenyl, 1-methylvinyl, 1-butenyl, 2-butenyl, 3-butenyl, 1-methyl-1-propenyl, 2-methyl-i-propenyl, 1-methyl-2-propenyl, 2-methyl-2-propenyl.
本发明所述的 "C2_4炔基"是指含有叁键的碳原子数为 2-4 的直链或 支链的炔基； 其实例包括但不限于例如乙炔基、 2-丙炔基、 2-丁炔基、 3-丁炔基、 1-甲基- 2-丙炔基等。 "C 2 _ 4 alkynyl group" in the present invention means the number of carbon atoms containing a triple bond to 2-4 straight or branched alkynyl group; examples thereof include, but are not limited to, ethynyl, 2-propynyl Base, 2-butynyl, 3-butynyl, 1-methyl-2-propynyl, and the like.
本发明所述的 "C w烷氧基"、 "Ci-4烷氨基"、 "二 (Cw烷基)氨基"、 "C1-4烷氧基羰基"、 "C1-4烷基石 基，，、 "C1-4烷基磺酰基"、 "C1-4烷基亚 磺酰基"、 "d_4烷基氨基磺酰基"、 "d_4烷基氨基亚磺酰基"， 分别是指 "Ci-4烷基- 0- "基团、 "C1-4烷基- NH- "基团、 "(C1-4烷基 )2N- "基团、 "C1-4 烷基- 0- C(O)- "基团、 "C1-4烷基- S- "基团、 "c1-4烷基- so2- "基团、 "c1-4 烷基- SO- "基团、 "C1-4烷基- S02- NH -"、 "C1-4烷基- SO- NH- "基团， 其中"C w alkoxy", "Ci -4 alkylamino", "di(Cw alkyl)amino", "C 1-4 alkoxycarbonyl", "C 1-4 alkyl" ,,, "C 1-4 alkylsulfonyl", "C 1-4 alkylsulfinyl", "d- 4 alkylaminosulfonyl", "d- 4 alkylaminosulfinyl", respectively "Ci -4 alkyl- 0-" group, "C 1-4 alkyl-NH-" group, "(C 1-4 alkyl) 2 N- " group, "C 1-4 alkyl group" - 0-C(O)-" group, "C 1-4 alkyl-S-" group, "c 1-4 alkyl-so 2 - " group, "c 1-4 alkyl-SO - "Group, "C 1-4 alkyl-S0 2 - NH -", "C 1-4 alkyl-SO-NH-" group, wherein
"Cw烷基"如前文所定义。 本发明所述 "d_6烷氧基 "指术语" d_6烷基"通过氧原子与其他结构 相连接的基团， 如甲氧基、 乙氧基、 丙氧基、 异丙氧基、 丁氧基、 异丁 氧基、 叔丁氧基、 仲丁氧基、 戊氧基、 新戊氧基、 己氧基等。 优选 烷氧基，更优选 d_3烷氧基。术语" d_4烷氧基"、 "d_3烷氧基 "指术语" 烷基"、 "d_3烷基"通过氧原子与其他结构相连接的基团。 "Cw alkyl" is as defined above. The "d- 6 alkoxy group" of the present invention means a group in which the term "d- 6 alkyl group" is bonded to another structure through an oxygen atom, such as a methoxy group, an ethoxy group, a propoxy group, an isopropoxy group, or a butyl group. Oxyl, isobutoxy, tert-butoxy, sec-butoxy, pentyloxy, neopentyloxy, hexyloxy and the like. Alkoxy groups are preferred, and d- 3 alkoxy groups are more preferred. The terms "d- 4 alkoxy", "d- 3 alkoxy" refer to the group "alkyl", "d- 3 alkyl" attached to the other structure through an oxygen atom.
本发明所述" d_6烷基酰氨基"是指" d_6烷基"通过酰氨基与其他结 构相连接的基团。 The "d- 6 alkylamido" of the present invention means a group in which "d- 6 alkyl" is bonded to other structures via an acylamino group.
本发明所述的" 素"是指氟原子、 氯原子、 溴原子或碘原子等。 本发明所述的 "3- 7 元环烷基"是指环原子全部为碳原子， 去除一个 氢原子衍生的环状烷基基团， 其中包括例如 "3- 6元环烷基"、 "4- 6元环 烷基" "5- 7元环烷基"、 "5-6元环烷基"， 其实例包括但不限于： 环丙烷 基、 环丁烷基、 环戊烷基、 环己烷基、 环庚烷基、 环辛烷基、 环戊酮基、 环己酮基等。  The "prime" as used in the present invention means a fluorine atom, a chlorine atom, a bromine atom or an iodine atom. The "3- 7-membered cycloalkyl group" as used in the present invention means that the ring atoms are all carbon atoms, and a hydrogen atom-derived cyclic alkyl group is removed, including, for example, "3- 6-membered cycloalkyl group", "4". - 6-membered cycloalkyl ""5- 7-membered cycloalkyl", "5-6-membered cycloalkyl", examples of which include, but are not limited to, cyclopropyl, cyclobutane, cyclopentyl, cyclohexyl Alkyl, cycloheptyl, cyclooctyl, cyclopentanone, cyclohexanone, and the like.
本发明所述的 "3- 7元杂环烷基"是指含有一至多个杂原子的 3-7元 环状基团， 所述"杂原子"是指氮原子、 氧原子、 硫原子等。 优选 3- 6元 杂环基， 更优选 5- 6元杂环基。 具体实例包括但不仅限于 2,5-二氢噻吩 基、 4,5-二氢吡唑基、 3,4-二氢- 2//-吡喃基、 5,6-二氢- 4//- 1,3-噁嗪基、 氮 杂环丙烷基、 氮杂环丁烷基、 氮杂环戊烷基、 哌啶基、 硫杂环丁烷基、 四氢呋喃基、 四氢吡咯基、 咪唑烷基、 吡唑烷基、 四氢呋喃基、 1 ,4-二 The "3- 7-membered heterocycloalkyl group" as used in the present invention means a 3-7 membered cyclic group having one or more hetero atoms, and the "hetero atom" means a nitrogen atom, an oxygen atom, a sulfur atom, or the like. . A 3- to 6-membered heterocyclic group is preferred, and a 5- to 6-membered heterocyclic group is more preferred. Specific examples include, but are not limited to, 2,5-dihydrothiophenyl, 4,5-dihydropyrazolyl, 3,4-dihydro-2//-pyranyl, 5,6-dihydro-4// - 1,3-oxazinyl, aziridine, azetidinyl, azetidinyl, piperidinyl, thietane, tetrahydrofuranyl, tetrahydropyrrolyl, imidazolidine Base, pyrazolidinyl, tetrahydrofuranyl, 1 ,4-di
-二氧杂环己烷基、 1,3-二硫杂环己烷基、 吗啉基、 哌
Figure imgf000012_0001
-dioxanyl, 1,3-dithiane, morpholinyl, piperidine
Figure imgf000012_0001
"4-7元杂芳基"是指含有 4-7个环原子 （其中至少含有一个杂原子） 构成的芳香性基团， 包括" 5- 7元杂芳基" " 5- 6元杂芳基"， 其具体实例包 括但不仅限于呋喃基、 噻哈基、 吡咯基、 噻唑基、 噻二唑基、 噁唑基、 噁二唑基、 咪唑基、 吡唑基、 吡啶基、 嘧啶基、 1,4-二氧杂环己二烯基、 2//- 1,2-噁嗪基、 4//- 1,2-噁嗪基、 6//- 1,2-噁嗪基、 4//- 1,3-噁嗪基、 6//- 1,3- 噁嗪基、 4//- 1,4-噁嗪基、 哒嗪基、 吡11秦基、 1,2,3-三嗪基、 1,2,4-三嗪基、 1,3,5-三嗪基、 1 ,2,4,5-四嗪基、 氧杂环庚三烯基、 硫杂环庚三烯基、 氮 杂环庚三烯基、 1,3-二氮杂环庚三烯基、 氮杂环辛四烯基等。 "4-7-membered heteroaryl" means an aromatic group consisting of 4 to 7 ring atoms (having at least one hetero atom), including "5- to 7-membered heteroaryl""5- to 6-membered heteroaryl" Specific examples include, but are not limited to, furyl, thiyl, pyrrolyl, thiazolyl, thiadiazolyl, oxazolyl, oxadiazolyl, imidazolyl, pyrazolyl, pyridyl, pyrimidinyl, 1,4-dioxadienyl, 2//- 1,2-oxazinyl, 4//- 1,2-oxazinyl, 6//- 1,2-oxazinyl, 4 //- 1,3-oxazinyl, 6//-1,3-oxazinyl, 4//- 1,4-oxazinyl, pyridazinyl, pyridyl 11 -methyl, 1,2,3- Triazinyl, 1,2,4-triazinyl, 1,3,5-triazinyl, 1,2,4,5-tetraazinyl, oxetanyl, thietane a group, azacycloheptatrienyl, 1,3-diazaheptatrienyl, azacyclotetradecenyl, and the like.
"6-10元二环结构"是指含有 6-10个环原子构成的二环基团（可以不 含有或者含有一个及一个以上的杂原子：)，包括" 7- 10元二环结构"、"8- 10 元二环结构"、 "6- 9元螺环结构"、 "6- 8元并环结构"、 "6- 8元桥环结构" 等。 其具体实例包括但不限于二环 [3丄 0]己烷基、 二环 [4.1.0]庚烷基、 二环 [2.2.0]己烷基、 二环 [3.2.0]庚烷基、 二环 [4.2.0]辛烷基、 八氢并环戊 二烯基、 八氢— 茚基、 十氢化萘基、 十四氢菲基、 双环 [3.1.0]己- 2-烯 基、 双环 [4.1.0]庚- 3-烯基、 双环 [3.2.0]庚- 3-烯基、 双环 [4.2.0]辛- 3-烯基、 1,2,3,3a-四氢并环戊二烯基、 2,3,3a,4,7,7a-六氢 - 茚基、 1,2,3,4,4α,5,6,8α-八 氢化萘基 、 1 ,2,4α,5,6,8α- 六 氢化萘基 、 1,2,3,4,5,6,7,8,9, 10-十氢菲基、 环丁烷并四氢吡咯基、 环戊烷并四氢吡咯 基、 氮杂环丁烷并咪唑烷基、 3-氧杂双环并 [3.1.0]己烷基、 六氢呋喃 "6-10 membered bicyclic structure" means a bicyclic group consisting of 6 to 10 ring atoms (may not contain or contain one or more heteroatoms:), including "7-10 membered bicyclic structure","8-10 "Secondary ring structure", "6-9 element spiral ring structure", "6-8 yuan parallel ring structure", "6-8 yuan bridge ring structure", etc. Specific examples include but not limited to two rings [3丄0 Hexyl, bicyclo[4.1.0]heptyl, bicyclo[2.2.0]hexane, bicyclo[3.2.0]heptyl, bicyclo[4.2.0]octyl, VIII Hydrogencyclopentadienyl, octahydro-indenyl, decalinyl, tetrahydrophenanthyl, bicyclo[3.1.0]hex-2-enyl, bicyclo[4.1.0]hept-3-enyl Bicyclo[3.2.0]hept-3-enyl, bicyclo[4.2.0]oct-3-enyl, 1,2,3,3a-tetrahydrocyclopentadienyl, 2,3,3a, 4,7,7a-hexahydro-indenyl, 1,2,3,4,4α,5,6,8α-octahydronaphthyl, 1,2,4α,5,6,8α-hexahydronaphthyl, 1,2,3,4,5,6,7,8,9, 10-decahydrophenanthyl, cyclobutanetetrahydropyrrolyl, cyclopentahydrotetrahydropyrrolyl, azetidine Alkyl, 3-oxabicyclo[3.1.0]hexane, hexahydrofuran
[3,4- 6][1,4]二噁英基、六氢- 2//-环戊烷并 [6] [1,4]二噁英基、 [3,4- 6][1,4]dioxin, hexahydro-2//-cyclopenta[6][1,4]dioxin,
,ΝΗ HN. HN I 冊  ,ΝΗ HN. HN I
00 OC 0 oo OCr "00、 o OCX 00 OCT 0ONH oO 0 00 00 HO0 0O 00 OC 0 oo OCr "00, o OCX 00 OCT 0O NH oO 0 00 00 H O0 0O
OC 、 Η φΗ
Figure imgf000013_0001
Θ 、 、 、 <
Figure imgf000013_0002
、 Η [$ι、 J$r OC, Η φ Η
Figure imgf000013_0001
Θ , , , , <
Figure imgf000013_0002
, Η [$ι, J$r
Θ Θ Θ φΗ c & 3、 以及芳香性的 二环结构， 包括但不限于苯并呋喃基、 苯并异呋喃基、 苯并噻哈基、 吲 哚基、 苯并噁唑基、 苯并咪唑基、 吲唑基、 苯并三唑基、 喹啉基、 异喹 啉基、 吖啶基、 菲啶基、 苯并哒嗪基、 酞嗪基、 喹唑啉基、 喹喔啉基、 酚嗪基、 喋啶基、 嘌呤基、 萘啶基、 1 ,3-二氢苯并呋喃基、 苯并 [ [1.3] 二氧杂环戊烯基、 异吲哚啉基、 色满基、 1,2,3,4-四氢吡咯并 [3,4- c]吡咯 基、 5,6-二氢咪唑 [1.2- β]吡嗪- 7(8/ )-基、 5,6-二氢- 1,7-萘啶- 7(8/ )-基、 5Η- 吡咯 [3.4- 6]吡啶- 6(7/ )-基、 7,8-二氢吡啶 [4.3- ί ]嘧啶- 6(5/ )-基、 2,3,6,7- 四氢- 吡唑 [4.3- c]吡啶- 5(4//)-基、 6,7-二氢噻唑 [5.4- c]吡啶- 5(4/ )-基 等。 Θ Θ φ φ Η c & 3, and aromatic bicyclic structures including, but not limited to, benzofuranyl, benzisofuranyl, benzothianyl, fluorenyl, benzoxazolyl, benzo Imidazolyl, carbazolyl, benzotriazolyl, quinolyl, isoquinolyl, acridinyl, phenanthryl, benzoxazinyl, pyridazinyl, quinazolinyl, quinoxalinyl, Phenazinyl, acridinyl, fluorenyl, naphthyridyl, 1 ,3-dihydrobenzofuranyl, benzo[[1.3]dioxolyl, isoindolyl, chromanyl, 1,2,3,4-tetrahydropyrrolo[3,4-c]pyrrolyl, 5,6-dihydroimidazole[1.2-β]pyrazine-7(8/)-yl, 5,6-di Hydrogen-1,7-naphthyridine-7(8/)-yl, 5Η-pyrrole[3.4-6]pyridine-6(7/)-yl, 7,8-dihydropyridine [4.3- ί]pyrimidine-6 (5/)-yl, 2,3,6,7-tetrahydro-pyrazole [4.3- c]pyridine-5(4//)-yl, 6,7-dihydrothiazole [5.4- c]pyridine- 5(4/)-based, etc.
本发明所述" 6- 9 元螺环基"是指一类至少有两个环共享一个原子形 成的 6- 9元稠环结构。 其具体实施例包括但不仅限于： 螺 [3.3]庚烷基、 螺 [3.4]辛烷基、螺 [3.5]壬烷基、螺 [4.4]壬烷基、螺 [3.4]辛- 6-烯基、螺 [3.5] 壬— 6—烯基、 螺 [4.4]壬- 6-烯基、 螺 [4.4]壬- 2,7-二烯基、 2-氧杂螺 [3.3]庚烷 基、 6-氧杂螺 [2.5]辛烷基、 4-氧杂- 7-氨基螺 [2.5]辛烷基、 2-氨基螺 [3.3] 庚烷基、 2-氧杂- 6-氨基螺 [3.3]庚烷基、 2-氨基螺 [3.4]辛烷基、 6-氧杂- 2- 氨基螺 [3.4]辛烷基、 2-氧杂 -6-氨基螺 [3.4]辛烷基、 2-氧杂螺 [3.4]辛烷基、 5-氧杂螺 [3.5]壬烷基、 7-氨基螺 [3.5]壬烷基、 2-氨基螺 [4.4]壬烷基、 2- 氧杂- 7-氨基螺 [4.4]壬烷基、 2-氧杂螺 [4.4]壬烷基、 1,7-二氧杂螺 [4.4]壬 烷基、 1,4,7-三氧杂螺 [4.4]壬烷基、 6-氨基螺 [3.4]辛- 7-烯基、 2-氧杂- 6- 氨基螺 [3.4]辛- 7-烯基、 7-氨基螺 [3.5]壬- 5-烯基、 2-氨基螺 [4.4]壬- 7-烯基 等。 The "6-9-membered spirocyclic group" of the present invention means that at least two rings of a class share an atomic shape. A 6- to 9-membered fused ring structure. Specific examples thereof include, but are not limited to, spiro[3.3]heptyl, spiro[3.4]octyl, spiro[3.5]decyl, spiro[4.4]decyl, spiro[3.4]oct-6-ene , snail [3.5] 壬-6-alkenyl, spiro[4.4]壬-6-alkenyl, spiro[4.4]壬-2,7-dienyl, 2-oxaspiro[3.3]heptanyl, 6-oxaspiro[2.5]octyl, 4-oxa-7-aminospiro[2.5]octyl, 2-aminospiro[3.3]heptanyl, 2-oxa-6-aminospiro[3.3 Heptylalkyl, 2-aminospiro[3.4]octyl, 6-oxa-2-aminospiro[3.4]octyl, 2-oxa-6-aminospiro[3.4]octyl, 2- Oxanspiro[3.4]octyl, 5-oxaspiro[3.5]decyl, 7-aminospiro[3.5]decyl, 2-aminospiro[4.4]decyl, 2-oxa--7 -aminospiro[4.4]decyl, 2-oxaspiro[4.4]decyl, 1,7-dioxaspiro[4.4]decyl, 1,4,7-trioxaspiro[4.4]壬alkyl, 6-aminospiro[3.4]oct-7-alkenyl, 2-oxa-6-aminospiro[3.4]oct-7-alkenyl, 7-aminospiro[3.5]indole-5-alkenyl , 2-Aminospiro[4.4]壬-7-alkenyl, and the like.
本发明所述" 6- 8 元并环结构"是指由两个或两个以上环状结构彼此 共用两个相邻的原子所形成的 6-8元环状基团， 其具体实施例包括但不 仅限于： 二环 [3.1.0]己烷基、 二环 [4.1.0]庚烷基、 二环 [2.2.0]己烷基、 二 环 [3.2.0]庚烷基、 二环 [4.2.0]辛烷基、 双环 [3.1.0]己- 2-烯基、 双环 [4.1.0] 庚— 3-烯基、 双环 [3.2.0]庚- 3-婦基、 双环 [4.2.0]辛- 3-烯基、 苯并呋喃基、 苯并异呋喃基、 苯并噻哈基、 吲哚基、 苯并噁唑基、 苯并咪唑基、 吲唑 基、 苯并三唑基、 喹啉基、 异喹啉基、 吖啶基、 菲啶基、 苯并哒嗪基、 酞嗪基、 喹唑啉基、 喹喔啉基、 噻吩并 [2,3- 6]噻哈基、 噻吩并 [3,2- 6]噻 哈基、 苯并 [6]噻哈基、 苯并 [6]噻唑基、 环丁烷并四氢吡咯基、 环戊烷 并四氢吡咯基、 氮杂环丁烷并咪唑烷基、 3-氧杂双环并 [3.1.0]己烷基、 六氢呋喃 [3,4- 6] [1,4]二噁英基、 六氢- 2//-环戊烷并 [6][1,4]二噁英基等。  The "6- to 8-membered ring structure" as used in the present invention refers to a 6-8 membered cyclic group formed by two or more ring structures sharing two adjacent atoms with each other, and specific examples thereof include But not limited to: bicyclo [3.1.0] hexane, bicyclo [4.1.0] heptyl, bicyclo [2.2.0] hexane, bicyclo [3.2.0] heptyl, bicyclo [4.2.0] Octyl, bicyclo[3.1.0]hex-2-enyl, bicyclo[4.1.0]hept-3-enyl, bicyclo[3.2.0]hept-3-yl,bicyclo[ 4.2.0] Oct-3-enyl, benzofuranyl, benzoisofuranyl, benzothianyl, decyl, benzoxazolyl, benzimidazolyl, oxazolyl, benzotrien Azyl, quinolyl, isoquinolyl, acridinyl, phenanthryl, benzoxazinyl, pyridazinyl, quinazolinyl, quinoxalinyl, thieno[2,3- 6]thiazide Haki, thieno[3,2- 6]thiraki, benzo[6]thiraki, benzo[6]thiazolyl, cyclobutanetetrahydropyrrolyl, cyclopentahydrotetrapyrrolyl , azetidinazolidinyl, 3-oxabicyclo[3.1.0]hexane, hexahydrofuran [3,4- 6] [1,4] dioxins , Hexahydro - 2 // - cyclopenta [6] [1,4] dioxin-yl and the like.
本发明所述" 6- 8 元桥环结构 "是指由两个或两个以上环状结构彼此 共用两个不相邻的原子所形成的 6- 8元环状基团， 其具体实施例包括但 、 Θ、 、 Φ、 Η、 φΗ、 Θ、 Θ、
Figure imgf000014_0001
、 Θ、 、 Φ、 0 、The "6- to 8-membered bridged ring structure" of the present invention refers to a 6- to 8-membered cyclic group formed by two or more ring-shaped structures sharing two non-adjacent atoms with each other, and a specific embodiment thereof Including, Θ, 、, Φ, Η , φ Η , Θ, Θ,
Figure imgf000014_0001
, Θ, Φ, 0,
i JSr、 (3、 Θ、 Θ、
Figure imgf000014_0002
以上所述基团可以进一步被 1至 4个 取代， （^表示卤素原子、 C 烷基、 氨基、 烷基氨基、 二 - （C 烷基）氨基、 羟基、 烷氧 基、 烷氧羰基、 氨基甲酰基、 烷基氨基甲酰基、 二 - （Cw烷基） 氨基甲酰基或 3- 6元环烷基， 其中 可以相同或不同。 i JSr, (3, Θ, Θ,
Figure imgf000014_0002
The above group may be further substituted by 1 to 4, (^ represents a halogen atom, a C alkyl group, an amino group, an alkylamino group, a bis-(C alkyl)amino group, a hydroxyl group, an alkoxy group, an alkoxycarbonyl group, an amino group. Formyl, alkylcarbamoyl, bis-(Cw alkyl)carbamoyl or 3- 6 membered cycloalkyl, which may be the same or different.
本发明特别优选的通式（I )所示的化合物包括:  Particularly preferred compounds of the formula (I) according to the invention include:
Figure imgf000015_0001
Figure imgf000016_0001
Figure imgf000015_0001
Figure imgf000016_0001
本发明特别优选的通式（I)所示的化合物还包括:  Particularly preferred compounds of the formula (I) according to the invention further comprise:
Figure imgf000016_0002
Figure imgf000017_0001
Figure imgf000016_0002
Figure imgf000017_0001
Figure imgf000018_0001
Figure imgf000018_0001
在一个实施方案中， 本发明提供了上述本发明化合物的制备方法。 在一个实施方案中， 本发明提供了含有上述本发明化合物的药物组 合物。 In one embodiment, the invention provides a process for the preparation of a compound of the invention described above. In one embodiment, the invention provides a pharmaceutical group comprising a compound of the invention described above Compound.
在一个实施方案中， 本发明提供了使用上述本发明化合物预防和 / 或治疗个体中 B细胞相关的血癌（例如 B细胞慢性淋巴细胞癌、 非霍奇 金淋巴瘤）， 炎性以及自身免疫性疾病（例如类风湿性关节炎、 系统性红 斑狼疮等）的方法。  In one embodiment, the invention provides the use of a compound of the invention described above for the prevention and/or treatment of B cell associated blood cancer in an individual (eg, B cell chronic lymphocytic carcinoma, non-Hodgkin's lymphoma), inflammatory and autoimmune A method of disease (eg, rheumatoid arthritis, systemic lupus erythematosus, etc.).
在一个实施方案中， 本发明提供了上述本发明化合物在制备用于预 防和 /或治疗 B细胞相关的血癌（例如 B细胞慢性淋巴细胞癌、 非霍奇金 淋巴瘤）， 炎性以及自身免疫性疾病（例如类风湿性关节炎、 系统性红斑 狼疮等）的药物中的应用。  In one embodiment, the invention provides a compound of the invention described above for use in the prevention and/or treatment of B cell associated blood cancer (eg, B cell chronic lymphocytic carcinoma, non-Hodgkin's lymphoma), inflammatory and autoimmune Use in drugs for sexual diseases such as rheumatoid arthritis, systemic lupus erythematosus, etc.
本发明上述化合物可以采用下述流程中描述的方法和 /或本领域普 通技术人员已知的其它技术来合成， 但不仅限于以下方法。  The above compounds of the present invention can be synthesized by the methods described in the following schemes and/or other techniques known to those skilled in the art, but are not limited to the following methods.
为方便起见， 本发明使用众所周知的缩写代表多种化学化合物， 包 括但不限于  For convenience, the invention uses well-known abbreviations to represent a variety of chemical compounds, including but not limited to
DMF: 二甲基甲酰胺； THF: 四氢呋喃； DIEA: V， V-二异丙基 乙胺；  DMF: dimethylformamide; THF: tetrahydrofuran; DIEA: V, V-diisopropylethylamine;
BINAP: 2,2，-双二苯膦基- 1,1，-联萘； Xantphos: 4,5-双二苯膦- 9,9- 二甲基氧杂蒽等。  BINAP: 2,2,-bisdiphenylphosphino-1,1,-binaphthyl; Xantphos: 4,5-bisdiphenylphosphine-9,9-dimethyloxaxime.
反应路线：  Reaction route:
Figure imgf000019_0001
原料 4
Figure imgf000019_0001
Raw material 4
式 化合物  Compound
反应步骤：  Reaction steps:
步骤 i 中间体 1的制备  Step i Preparation of intermediate 1
将 5-溴尿嘧啶 (1 当量)与原料 1(至少 5当量)混合， 无溶剂， 或者加 入少量极性溶剂 （水， 乙醇， 正丁醇等）加热（100- 150°C)搅拌反应数小 时至 5-溴尿嘧啶消失。 冷却， 析出固体， 过滤， 得中间体 1。 当原料 1 为硫醇时， 需在反应时加入等当量碱 (如氢氧化钠）。 Mix 5-bromouracil (1 eq.) with starting material 1 (at least 5 equivalents), without solvent, or add a small amount of polar solvent (water, ethanol, n-butanol, etc.) to heat (100-150 ° C) to stir the reaction small When the 5-bromouracil disappeared. After cooling, the solid was precipitated and filtered to give Intermediate 1. When the starting material 1 is a mercaptan, an equivalent amount of a base such as sodium hydroxide is added during the reaction.
步骤 2 中间体 2的制备  Step 2 Preparation of intermediate 2
将中间体 1(1 当量)与溶剂量三氯氧磷 (至少 10 当量)混合， 加入约 两当量 二甲基苯胺， 加热（90- 110°C)搅拌反应数小时至中间体 1消 失。 冷却， 倒入冰水中， 析出固体， 过滤， 干燥， 得中间体 2。 或者浓 缩反应体系， 柱层析， 得中间体 2。  Intermediate 1 (1 equivalent) is mixed with a solvent amount of phosphorus oxychloride (at least 10 equivalents), about two equivalents of dimethylaniline are added, and the reaction is stirred by heating (90-110 ° C) for several hours until the intermediate 1 disappears. After cooling, pour into ice water, precipitate a solid, filter, and dry to give Intermediate 2. Alternatively, the reaction system is concentrated, and column chromatography is carried out to obtain Intermediate 2.
步骤 3 中间体 3的制备  Step 3 Preparation of intermediate 3
方法 1 : 将中间体 2(1 当量)与原料 2(1.1- 1.3 当量)混合， 加入适当 溶剂（如四氢呋喃， 正丁醇等）， 加热（80- 120°C)搅拌反应数小时至中间体 2消失。 冷却， 浓缩反应体系， 柱层析， 得中间体 3。  Method 1 : Mix intermediate 2 (1 eq.) with starting material 2 (1.1-1.3 eq.), add a suitable solvent (such as tetrahydrofuran, n-butanol, etc.), and heat (80-120 ° C) to stir the reaction for several hours to the intermediate. 2 disappeared. Cooling, concentrating the reaction system, and column chromatography gave Intermediate 3.
方法 2: 釆用 Buchwald-Hartwig偶联。 将中间体 2(1 当量）与原料 2(1-1.3当量)混合， 加入适当溶剂（通常为甲苯， 二氧六环等）， 催化量的 钯催化剂（通常为 Pd2(dba)3, Pd(OAc)2 等）， 催化量的配体（通常为 Xantphos, BINAP等）， 碱 (Cs2C03, NaO Bu等）（1· 1- 1.5当量）， 充分置换 惰性气体 (如氮气， 氩气等）， 加热 (70- 120°C)搅拌反应数小时至中间体 2 消失。 冷却， 浓缩反应体系， 柱层析， 得中间体 3。 Method 2: Buch Buchwald-Hartwig coupling. Mix intermediate 2 (1 eq.) with starting material 2 (1-1.3 eq.), add the appropriate solvent (usually toluene, dioxane, etc.), catalytic amount of palladium catalyst (usually Pd 2 (dba) 3 , Pd (OAc) 2, etc., catalytic amount of ligand (usually Xantphos, BINAP, etc.), base (Cs 2 C0 3 , NaO Bu, etc.) (1·1 - 1.5 equivalents), fully substituted inert gas (such as nitrogen, argon) Gas, etc., heating (70-120 ° C) to stir the reaction for several hours until the intermediate 2 disappears. The mixture was cooled, concentrated, and subjected to column chromatography to give Intermediate 3.
步骤 4 中间体 4的制备  Step 4 Preparation of intermediate 4
方法 1 : 将中间体 3(1 当量)与原料 3(1- 1.3 当量)混合， 加入适当溶 剂（如叔丁醇， 叔戊醇等）， 加入碱 (如 DIEA等， 1- 1·5 当量）， 或者催化 量的酸（如乙酸， 三氟乙酸等）， 加热（80- 120°C) , 或者微波下加热 (80- 120°C)搅拌反应数小时至中间体 3 消失。 冷却， 浓缩反应体系， 柱 层析， 得中间体 4。 Method 1: Intermediate 3 (1 eq.) And starting material 3 (1 1.3 equivalents) were mixed, and a suitable solvent (e.g. tert-butanol, tert-amyl alcohol, etc.), addition of a base (e.g., DIEA and the like, 1- 1.5 eq. ), or a catalytic amount of acid (such as acetic acid, trifluoroacetic acid, etc.), heated (80-120 ° C), or heated under microwave (80-120 ° C) to stir the reaction for several hours until the intermediate 3 disappears. Cooling, concentrating the reaction system, and column chromatography gave Intermediate 4.
方法 2: 釆用 Buchwald-Hartwig偶联。 操作同步骤 3的方法 2 , 只 需将原料 2换成原料 3。  Method 2: Buchwald-Hartwig coupling. In the same way as in step 2 of step 3, only raw material 2 needs to be replaced with raw material 3.
步骤 5 中间体 5的制备  Step 5 Preparation of intermediate 5
将中间体 4 (1 当量)溶于二氯甲烷， 加入溶剂量三氟乙酸， 或者通入 盐酸气， 室温或冷却下搅拌反应至中间体 4消失。 浓缩， 得中间体 5 , 直接用于下步反应。  Intermediate 4 (1 equivalent) was dissolved in dichloromethane, and a solvent amount of trifluoroacetic acid was added thereto, or hydrochloric acid gas was passed through, and the reaction was stirred at room temperature or under cooling to the intermediate 4 to disappear. Concentration, intermediate 5 was obtained and used directly in the next step.
步骤 6 式（ I )化合物的制备  Step 6 Preparation of the compound of formula (I)
将中间体 5 (1 当量)溶于适当的溶剂（如 THF ,二氯甲烷，丙酮， DMF , 或者为混合溶剂）， 加入 2-3 当量的碱 (如 DIEA), 冷却下 (- 20°C到- 10度 °C), 緩慢滴入原料 4 (0.9-1.1 当 量)搅拌反应至中间体 5 消失。 淬灭反 应， 浓缩， 柱层析或者中低压制备液相纯化得式 I化合物。 Dissolve intermediate 5 (1 eq.) in a suitable solvent (eg THF, dichloromethane, acetone, DMF, or as a solvent mixture), add 2-3 equivalents of base (eg DIEA), cool down (-20 ° C) To - 10 degrees °C), slowly instill the raw material 4 (0.9-1.1 eq.) and stir the reaction until the intermediate 5 disappears. The compound of formula I is purified by quenching, concentration, column chromatography or medium to low pressure preparative liquid phase purification.
反应路线中的 、 R2、 R3、 Li、 L2、 a、 b、 c、 d、 e、 p、 q、 A和 B 如前文所述。 R 2 , R 3 , Li, L 2 , a, b, c, d, e, p, q, A and B in the reaction scheme are as described above.
另外，本发明上述化合物或者中间体还可以采用下述流程中描述的 方法
Figure imgf000021_0001
In addition, the above compounds or intermediates of the present invention may also adopt the methods described in the following schemes.
Figure imgf000021_0001
方法 1 : 将底物 （ 1 当量）溶于甲醇和水中 （或者二氯甲烷、 甲醇、 水的混合溶剂中） ， 加氧化剂 （如过硫酸氢钾复合盐、 间氯过氧苯甲酸 等） （1 当量） , 在常温下反应至 LCMS监测底物完全消失， 将溶液旋 干， 制备液相纯化， 或硅胶柱纯化得到产品。  Method 1: The substrate (1 equivalent) is dissolved in methanol and water (or a mixed solvent of dichloromethane, methanol, and water), and an oxidizing agent (such as potassium persulfate complex salt, m-chloroperoxybenzoic acid, etc.) is added ( 1 equivalent), react to LCMS at room temperature to monitor the complete disappearance of the substrate, spin the solution, prepare the liquid phase for purification, or purify the column with silica gel to obtain the product.
方法 2: 将底物 （ 1 当量）溶于甲醇中， 敞口于空气中回流数小时， 旋干 制备液相纯化或硅胶柱纯化得到产品。
Figure imgf000021_0002
Method 2: The substrate (1 equivalent) is dissolved in methanol, and opened to reflux in air for several hours. The product is purified by spin-drying or purified by silica gel column.
Figure imgf000021_0002
将底物 （ 1 当量） 溶于甲醇和水中 （或者二氯甲烷、 甲醇、 水的混 合溶剂中）， 加氧化剂（如过硫酸氢钾复合盐、 间氯过氧苯甲酸等） （大 于 2当量） , 在常温下反应过夜或者加热数小时， 至 LCMS监测底物完 全消失， 将溶液旋干， 制备液相纯化， 或硅胶柱纯化得到产品。  The substrate (1 equivalent) is dissolved in methanol and water (or a mixed solvent of dichloromethane, methanol, water), and an oxidizing agent (such as potassium persulfate complex salt, m-chloroperoxybenzoic acid, etc.) (greater than 2 equivalents) ), reacting at room temperature overnight or heating for several hours, until the substrate is completely disappeared by LCMS, the solution is spin-dried, purified by liquid phase preparation, or purified by silica gel column to obtain a product.
反应路线中的 、 R2、 R3、 R5、 Li、 L2、 a、 b、 c、 d、 e、 p、 q、 A 和 B如前文所述。 本发明式 （I ) 化合物、 其立体异构体可以以游离的形式或其药学 上可接受的盐的形式使用。 本发明式 （I ) 化合物显碱性， 可以与无机 酸或有机酸形成酸式盐。 如盐酸盐、 氢氟酸盐、 氢溴酸盐、 氢碘酸盐、 硫酸盐、 三氟乙酸盐、 苯磺酸盐、 甲磺酸盐、 三氟甲磺酸盐、 乙磺酸盐、 碳酸盐、 硝酸盐、 磷酸盐、 亚磷酸盐、 马来酸盐、 酒石酸盐、 柠檬酸盐、 醋酸盐、 苯甲酸盐、 富马酸盐、 草酸盐、 葡萄糖酸盐、 羟基乙酸盐、 羟 乙磺酸盐、 乳酸盐、 乳糖酸盐、 苹果酸盐、 琥珀酸盐、 对甲苯磺酸盐、 甘氨酸盐、 三甲基甘氨酸盐、 精氨酸盐、 鸟氨酸盐、 谷氨酸盐、 天冬氨 酸盐等。 R 2 , R 3 , R 5 , Li, L 2 , a, b, c, d, e, p, q, A and B in the reaction scheme are as described above. The compound of the formula (I) of the present invention, a stereoisomer thereof, can be used in the form of a free form or a pharmaceutically acceptable salt thereof. The compound of the formula (I) of the present invention is basic and can form an acid salt with an inorganic acid or an organic acid. Such as hydrochloride, hydrofluoride, hydrobromide, hydroiodide, sulfate, trifluoroacetate, besylate, methanesulfonate, triflate, ethanesulfonate , carbonate, nitrate, phosphate, phosphite, maleate, tartrate, citrate, acetate, benzoate, fumarate, oxalate, gluconate, hydroxyl Acetate, isethionate, lactate, lactobionate, malate, succinate, p-toluenesulfonate, Glycinate, trimethylglycine, arginine, ornithine, glutamate, aspartate, and the like.
本发明式 （I ) 化合物或其药学上可接受的盐由于存在不对称碳原 子， 可以以一种旋光异构体形式存在， 因此， 本发明还包括这些旋光异 构体及其混合物。 本文描述的结构也拟包括所述结构的所有异构（例如 对映异构、 非对映异构和几何异构（或构象异构)） 形式； 例如， 关于每 一不对称中心的 R和 S构型、 Z和 E双键异构体以及 Z和 E构象异构 体。 因此， 本发明化合物的单一立体化学异构体以及对映异构体、 非对 映异构体和几何异构体 (或构象异构体） 的混合物都在本发明的范围内。 除非另作规定， 否则本发明化合物的所有互变异构形式都在本发明的范 围内。 此外， 除非另作规定， 否则本文所述的结构也拟包括不同之处仅 在于存在一个或一个以上同位素富集的原子的化合物。 举例来说， 具有 本发明的结构但包括氢经氘或氚置换或碳经富集 13C或 14C 的碳置换 的化合物在本发明的范围内。 此类化合物可用作例如分析工具、 生物分 析中的探针或本发明的治疗剂。 在一些实施例中， 式 I中的包含一个或 一个以上氘原子。  The compound of the formula (I) of the present invention or a pharmaceutically acceptable salt thereof may exist in the form of an optical isomer due to the presence of an asymmetric carbon atom, and therefore, the present invention also includes these optical isomers and mixtures thereof. The structures described herein are also intended to include all isomeric (e.g., enantiomeric, diastereomeric, and geometric (or conformational) forms) of the structure; for example, R and about each asymmetric center S configuration, Z and E double bond isomers, and Z and E conformers. Thus, single stereochemical isomers as well as mixtures of enantiomers, diastereomers and geometric isomers (or conformational isomers) of the compounds of the invention are within the scope of the invention. Unless otherwise specified, all tautomeric forms of the compounds of the invention are within the scope of the invention. Moreover, unless otherwise specified, the structures described herein are intended to include compounds that differ only in the presence of one or more isotopically enriched atoms. For example, compounds having the structure of the present invention but including hydrogen replaced by ruthenium or osmium or carbon substituted by carbon enriched 13C or 14C are within the scope of the invention. Such compounds are useful, for example, as analytical tools, as probes in biological assays or as therapeutic agents in the present invention. In some embodiments, Formula I contains one or more deuterium atoms.
本发明式 （I ) 化合物、 其药学上可接受的盐或其立体异构体可以 与一种或多种药用载体组成药物组合物。 所述药物组合物可以制成临床 上使用的常规制剂， 可以口服或肠胃外给药等方式施用于需要这种治疗 的患者。 如片剂、 颗粒、 胶嚢、 粉末、 注射剂、 吸入剂、 舌下给药制剂、 糖浆、 凝胶、 油膏、 栓剂、 洗剂、 眼部滴剂、 鼻腔滴剂、 喷雾剂、 透皮 制剂等。这些制剂可以通过常规方法， 添加药用载体如赋形剂、黏合剂、 增湿剂、 崩解剂、 增稠剂等制备而成。  The compound of the formula (I), a pharmaceutically acceptable salt thereof or a stereoisomer thereof of the present invention may be combined with one or more pharmaceutically acceptable carriers to constitute a pharmaceutical composition. The pharmaceutical composition can be formulated into a conventional preparation for clinical use, and can be administered to a patient in need of such treatment by oral or parenteral administration. Such as tablets, granules, capsules, powders, injections, inhalants, sublingual preparations, syrups, gels, ointments, suppositories, lotions, eye drops, nasal drops, sprays, transdermal formulations Wait. These preparations can be prepared by a conventional method, by adding a pharmaceutically acceptable carrier such as an excipient, a binder, a moisturizer, a disintegrating agent, a thickener or the like.
本发明式 （I ) 化合物、 其药学上可接受的盐或其立体异构体具有 较好的 BTK激酶抑制作用， 是较好具有优良的抗肿瘤作用以及自身免 疫疾病治疗作用的药物 。 同时本发明式（I )化合物、 其药学上可接受 的盐或其立体异构体在制备治疗 B细胞相关的血癌 （例如 B细胞慢性淋 巴细胞癌、 非霍奇金淋巴瘤）， 以及自身免疫性疾病 （例如类风湿性关节 炎、 系统性红斑狼疮等）中起着重要作用。  The compound of the formula (I) of the present invention, a pharmaceutically acceptable salt thereof or a stereoisomer thereof has a preferable BTK kinase inhibitory action, and is a drug which preferably has an excellent antitumor effect and a therapeutic effect of an autoimmune disease. At the same time, the compound of the formula (I), a pharmaceutically acceptable salt thereof or a stereoisomer thereof thereof is used for the preparation of a B cell-associated blood cancer (for example, B cell chronic lymphocytic carcinoma, non-Hodgkin's lymphoma), and autoimmunity Sexual diseases (such as rheumatoid arthritis, systemic lupus erythematosus, etc.) play an important role.
本发明式（I )化合物、 其药学上可接受的盐或其立体异构体是一 种激酶抑制剂， 特别是 Btk抑制剂。 这些抑制剂可以用于治疗哺乳动物 中的一种或多种响应激酶抑制的疾病， 包括响应 Btk抑制和 /或 B细胞 增殖的抑制的疾病。 不希望束縛于任何特定的理论， 相信本发明化合物 与 Btk的相互作用导致 Btk活性的抑制， 并因此得到这些化合物药学应 用。 因此， 本发明包括用于治疗具有响应 Btk 活性的抑制和 /或抑制 B 细胞增殖的疾病的哺乳动物， 例如人的方法， 该方法包括： 向具有这样 的疾病的哺乳动物给药有效量的至少一种在本文中提供的化学实体。 可 以在实验上例如通过测定化合物的血液浓度， 或理论上通过计算生物利 用度， 确定有效浓度。 除了 Btk之外， 还可能受到影响的其它激酶包括 但不限于， 其它酪氨酸激酶和丝氨酸 /苏氨酸激酶。 The compound of the formula (I), a pharmaceutically acceptable salt thereof or a stereoisomer thereof of the present invention is a kinase inhibitor, particularly a Btk inhibitor. These inhibitors can be used to treat one or more diseases that respond to kinase inhibition in a mammal, including response to Btk inhibition and/or B cells. A disease that inhibits proliferation. Without wishing to be bound by any particular theory, it is believed that the interaction of the compounds of the invention with Btk results in inhibition of Btk activity, and thus the pharmaceutical use of these compounds. Accordingly, the present invention includes a method for treating a mammal, such as a human, having a disease responsive to inhibition of Btk activity and/or inhibiting proliferation of B cells, the method comprising: administering to a mammal having such a disease an effective amount of at least A chemical entity provided herein. The effective concentration can be determined experimentally, for example, by measuring the blood concentration of the compound, or theoretically by calculating the bioavailability. Other kinases that may be affected in addition to Btk include, but are not limited to, other tyrosine kinases and serine/threonine kinases.
激酶在控制基本细胞过程如增殖、 分化和死亡（细胞凋亡） 的信号 传导路径方面起着显著的作用。 异常的激酶活性已经暗示于各种疾病 中，所述的疾病包括多种癌症、自身免疫和 /或炎性疾病和急性炎性反应。 激酶在关键细胞信号传导路径中的多面性作用提供识别耙向激酶和信 号传导路径的新药物的显著机会。  Kinases play a significant role in controlling the signaling pathways of essential cellular processes such as proliferation, differentiation and death (apoptosis). Abnormal kinase activity has been implicated in various diseases including a variety of cancers, autoimmune and/or inflammatory diseases, and acute inflammatory responses. The versatile role of kinases in key cell signaling pathways provides a significant opportunity to identify new drugs that target the kinase and signal transduction pathways.
一个实施方案包括治疗具有自身免疫和 /或炎性疾病或响应 Btk 活 性和 /或 B细胞增殖的抑制的急性炎性反应的患者的方法。  One embodiment includes a method of treating a patient having an autoimmune and/or inflammatory disease or an acute inflammatory response in response to inhibition of Btk activity and/or B cell proliferation.
使用根据本发明的化合物和组合物可以影响的自身免疫和 /或炎性 疾病包括但不限于： 银屑病， 变态反应， 局限性肠炎， 肠易激综合征， 舍格伦病， 组织移植物排斥反应和移植器官的超急性排斥反应， 哮喘， 系统性红斑狼疮（和相关的肾小球肾炎）， 皮肌炎， 多发性硬化， 硬皮 病， 血管炎 （ANCA-相关的和其它的血管炎）， 自身免疫溶血性和血小 板减少性症状， 古德帕斯综合征 （和相关的肾小球肾炎和肺出血）， 动 脉粥样硬化， 类风湿性关节炎，慢性的特发性血小板减少性紫癜（ITP ), 艾迪生病， 帕金森病， 阿尔茨海默病， 糖尿病， 脓毒性休克和重症肌无 力。  Autoimmune and/or inflammatory diseases that can be affected by the use of the compounds and compositions according to the invention include, but are not limited to, psoriasis, allergies, localized enteritis, irritable bowel syndrome, Sjogren's disease, tissue grafts Rejection and hyperacute rejection of transplanted organs, asthma, systemic lupus erythematosus (and associated glomerulonephritis), dermatomyositis, multiple sclerosis, scleroderma, vasculitis (ANCA-related and other blood vessels) Inflammation, autoimmune hemolytic and thrombocytopenic symptoms, Goodpas syndrome (and associated glomerulonephritis and pulmonary hemorrhage), atherosclerosis, rheumatoid arthritis, chronic idiopathic thrombocytopenia Purpura (ITP), Edison's disease, Parkinson's disease, Alzheimer's disease, diabetes, septic shock and myasthenia gravis.
本文中包括的是治疗方法， 其中将本文中提供的至少一种化学实 体与抗炎药组合给药。抗炎药包括但不限于： NSAID ,非特异性和 COX- 2 特异性环氧合酶抑制剂， 金化合物， 皮盾类固醇类， 甲氨蝶呤， 肿瘤坏 死因子 （TNF ) 受体拮抗剂， 免疫抑制剂和甲氨蝶呤。  Included herein are methods of treatment wherein at least one of the chemical entities provided herein is administered in combination with an anti-inflammatory agent. Anti-inflammatory drugs include, but are not limited to: NSAID, non-specific and COX-2 specific cyclooxygenase inhibitors, gold compounds, dermal steroids, methotrexate, tumor necrosis factor (TNF) receptor antagonists, immunization Inhibitor and methotrexate.
NSAID的实例包括但不限于， 布洛芬， 氟比洛芬， 萘普生和萘普 生钠， 双氯芬酸， 双氯芬酸钠和米索前列醇的组合， 舒林酸， 苯曙丙酸， 二氟尼柳， 吡罗昔康， 吲哚美辛， 依托度酸， 非诺洛芬钙， 酮洛芬， 萘 丁美酮钠，柳氮磺吡啶，托美丁钠和羟氯喹。 NSAID的实例还包括 COX- 2 特异性抑制剂如塞来考昔， 伐地考昔， 芦米考昔和 /或艾托考昔。 Examples of NSAIDs include, but are not limited to, ibuprofen, flurbiprofen, naproxen and naproxen sodium, diclofenac, a combination of diclofenac sodium and misoprostol, sulindac, benzopyrene, diflunis Willow, piroxicam, indomethacin, etodolac, fenoprofen calcium, ketoprofen, nabumetone sodium, sulfasalazine, tolbutin sodium and hydroxychloroquine. Examples of NSAIDs also include COX-2 Specific inhibitors such as celecoxib, valdecoxib, remiclox and/or etoricoxib.
在一些实施方案中， 抗炎药是水杨酸酯或盐。 水杨酸酯或盐包括 但不限于乙酰基水杨酸或阿斯匹林， 水杨酸钠以及水杨酸胆碱和水杨酸 镁。  In some embodiments, the anti-inflammatory agent is a salicylate or a salt. Salicylates or salts include, but are not limited to, acetylsalicylic acid or aspirin, sodium salicylate, and choline salicylate and magnesium salicylate.
抗炎药还可以是皮盾类固醇类。例如，皮盾类固醇类可以是可的松， 地塞米松， 甲泼尼龙， 泼尼松龙， 泼尼松龙磷酸钠， 或泼尼松。  Anti-inflammatory drugs can also be Shield Steroids. For example, the skin shield steroid may be cortisone, dexamethasone, methylprednisolone, prednisolone, prednisolone sodium phosphate, or prednisone.
在另外的实施方案中，抗炎药是金化合物，如金硫丁二钠或金诺芬。 本发明还包括其中抗炎药是代谢抑制剂如二氢叶酸还原酶抑制剂， 如甲氨蝶呤或二氢乳清酸盐脱氢酶抑制剂如来氟米特的实施方案。  In other embodiments, the anti-inflammatory agent is a gold compound such as gold thioudate or auranofin. The invention also includes embodiments in which the anti-inflammatory agent is a metabolic inhibitor such as a dihydrofolate reductase inhibitor such as methotrexate or a dihydroorotate dehydrogenase inhibitor such as leflunomide.
本发明的其它实施方案涉及其中至少一种抗炎化合物是抗单克隆 抗体 （如依库珠单抗或培克珠单抗）， TNF 拮抗剂如依那西普 ( entanercept ) 或英利昔单抗的组合， 所述英利昔单抗是一种抗 TNFa 单克隆抗体。  A further embodiment of the invention relates to wherein the at least one anti-inflammatory compound is an anti-monoclonal antibody (such as eculizumab or pegizumab), a TNF antagonist such as entanercept or infliximab In combination, the infliximab is an anti-TNFa monoclonal antibody.
本发明的其它的实施方案涉及其中至少一种活性药是免疫抑制剂 化合物如选自甲氨蝶呤， 来氟米特， 环胞素， 他克莫司， 硫唑嘌呤和吗 替麦考酚酯中的免疫抑制剂化合物的组合。  A further embodiment of the invention relates to wherein at least one active agent is an immunosuppressive compound such as selected from the group consisting of methotrexate, leflunomide, cyclosporine, tacrolimus, azathioprine and mycophenolate A combination of immunosuppressant compounds in an ester.
表达 Btk的 B细包和 B细包前体已经暗示于 B细包恶性的病理学 中， B细胞恶性包括但不限于 B细胞淋巴瘤， 淋巴瘤（包括霍奇金和非 霍奇金淋巴瘤）， 毛细胞淋巴瘤， 多发性骨髓瘤， 慢性的和急性的髓细 胞源性白血病和慢性的和急性的淋巴细胞白血病。  B-B and B-precursor expression of Btk have been implicated in the pathology of B-package malignancy, B cell malignancy including but not limited to B-cell lymphoma, lymphoma (including Hodgkin and non-Hodgkin's lymphoma) ), hair cell lymphoma, multiple myeloma, chronic and acute myeloid leukemia and chronic and acute lymphocytic leukemia.
已经表明 Btk是在 B-系淋巴样细胞中 Fas/APO- 1(CD- 95)死亡诱导 信号传导复合物 (DISC)的抑制剂。白血病 /淋巴瘤细胞的命运可能在于由 DISC活化的半胱天冬蛋白酶的反向前细胞凋亡作用和包括 Btk和 /或其 底物的上游抗细胞凋亡调节机理之间的平衡 (Vassilev等， J. Biol. Chem. 1998, 274, 1646-1656) 。  Btk has been shown to be an inhibitor of Fas/APO-1 (CD-95) death-inducing signaling complex (DISC) in B-lineage lymphoid cells. The fate of leukemia/lymphoma cells may lie in the balance between the reverse pre-apoptotic effect of caspase-activated caspase and the upstream anti-apoptotic regulatory mechanism including Btk and/or its substrate (Vassilev et al. , J. Biol. Chem. 1998, 274, 1646-1656).
还发现 Btk抑制剂可以用作化学敏化剂 , 因此可以用于与其它化 学治疗药组合， 所述的化学治疗药特别是诱导细胞凋亡的药， 如抗肿瘤 剂、 免疫抑制剂等。 可以与化学敏化抑制剂组合使用的其它化学治疗药 的实例包括但不仅限于拓朴异构酶 I抑制剂 （如喜树碱或托泊替康）， 拓朴异构酶 II抑制剂（如道诺霉素和依托泊苷：)， 烷化剂（如环磷酰胺， 美法仑和 BCNU ), 微管蛋白导向的药剂 （如泰素和长春碱） 和生物制 剂 （例如抗体如抗 CD20抗体， IDEC8 , 免疫毒素和细胞因子）。 Btk 活性已经与一些表达由部分染色体 9 和 22 的易位导致的 bcr-abl融合基因的白血病相关。这种异常通常在慢性髓细胞源性白血病 中观察到。 Btk本盾上由 bcr- abl激酶磷酸化， 这引发在 bcr- abl细胞中 防止细胞凋亡的下游生存信号（ N. Feldhahn等 , J.Exp.Med.2005, 201(11), 1837-1852 ) 。 It has also been found that Btk inhibitors can be used as chemical sensitizers, and thus can be used in combination with other chemotherapeutic drugs, particularly drugs which induce apoptosis, such as antitumor agents, immunosuppressive agents and the like. Examples of other chemotherapeutic agents that can be used in combination with chemical sensitizers include, but are not limited to, topoisomerase I inhibitors (such as camptothecin or topotecan), topoisomerase II inhibitors (such as daunorubicin) And etoposide:), alkylating agents (such as cyclophosphamide, melphalan and BCNU), tubulin-directed agents (such as Taxol and Vinblastine) and biological agents (such as antibodies such as anti-CD20 antibody, IDEC8, Immunotoxins and cytokines). Btk activity has been associated with some leukemias that express the bcr-abl fusion gene resulting from partial translocation of chromosomes 9 and 22. This abnormality is usually observed in chronic myeloid leukemia. Btk is shielded by bcr-abl kinase, which triggers a downstream survival signal that prevents apoptosis in bcr- abl cells (N. Feldhahn et al., J. Exp. Med. 2005, 201(11), 1837-1852 ).
本发明化合物与最接近的现有技术相比， 具有以下优点：  The compounds of the invention have the following advantages over the closest prior art:
( 1 )本发明式（I )化合物或其药学上可接受的盐具有较好的 BTK 激酶抑制作用， 安全性高， 安全窗口大并且副作用小；  (1) The compound of the formula (I) of the present invention or a pharmaceutically acceptable salt thereof has a good BTK kinase inhibitory action, has high safety, a large safety window and a small side effect;
( 2 )本发明式（I )化合物或其药学上可接受的盐显示出良好的生 物稳定性， 药效时程长， 作用持久， 生物利用度高；  (2) The compound of the formula (I) of the present invention or a pharmaceutically acceptable salt thereof exhibits good biostability, long pot life, long lasting effect, and high bioavailability;
( 3 ) 本发明化合物制备工艺简单， 药品纯度高、 盾量稳定， 易于 进行大规模工业生产。  (3) The preparation process of the compound of the invention is simple, the drug has high purity, the shield volume is stable, and it is easy to carry out large-scale industrial production.
以下通过药理实验进一步阐述本发明化合物有益效果，但不应将此 理解为本发明化合物仅具有下列有益效果。  The beneficial effects of the compounds of the present invention are further illustrated by pharmacological experiments below, but it should not be construed that the compounds of the present invention have only the following beneficial effects.
试验例 本发明化合物的药理活性试验  Test Example Pharmacological activity test of the compound of the present invention
I 本发明化合物的体外抗布鲁顿酪氨酸激酶 (BTK)活性测定  I In vitro anti-brown tyrosine kinase (BTK) activity assay of compounds of the invention
供试品：  testing sample:
对照化合物： CC- 292 , 按照 WO2009158571A1制得。  The control compound: CC-292, was prepared in accordance with WO2009158571A1.
本发明化合物：其化学名称和结构式与制备方法见各化合物的制备实 施例。  The compound of the present invention: its chemical name, structural formula and preparation method are shown in the preparation examples of the respective compounds.
实验方法：  experimental method:
下述实验中缩写所代表的含义如下：  The meanings of the abbreviations in the following experiments are as follows:
ATP: 三磷酸腺苷； BTK: 布鲁顿酪氨酸激酶； mg: 毫克； mL: 毫升； g: 微克； μί: 微升； mM: 毫摩尔每升； EDTA: 乙二胺四乙 酸； DMSO: 二甲基亚砜。  ATP: adenosine triphosphate; BTK: Bruton's tyrosine kinase; mg: mg; mL: ml; g: microgram; μί: microliter; mM: millimoles per liter; EDTA: ethylenediaminetetraacetic acid; DMSO: dimethyl Ketosulfone.
1. 试验材料  Test material
HTRFR KinEASE™ -TK: 购自 Cisbio, 批号 62TK0PEB; BTK: 购 自 Carna, Cat.No.08- 080； ATP： 购 自 Sigma, Cat. No. A7699, CAS HTRFR KinEASETM -TK: purchased from Cisbio, lot number 62TK0PEB; BTK: purchased from Carna, Cat.No.08-080; ATP: purchased from Sigma, Cat. No. A7699, CAS
No.34369-07-8； MgCl2： 购自 Sigma , CAS No. 7786-30-3, Lot. No. 101M8701V; DMSO:购自 Sigma, CAS No.67-68-5, Lot. No. STBC0365V;No. 34369-07-8; MgCl 2 : purchased from Sigma, CAS No. 7786-30-3, Lot. No. 101M8701V; DMSO: purchased from Sigma, CAS No. 67-68-5, Lot. No. STBC0365V ;
96孔板： 购自 Thermo, Cat. No. 249944, Lot. No. 1057825 ; 384孔板： 购 自 Greiner, Cat. No. 784075, Lot. No. Ε1112Φ6Υ。 2. 试验用试剂配制 96-well plate: available from Thermo, Cat. No. 249944, Lot. No. 1057825; 384-well plate: available from Greiner, Cat. No. 784075, Lot. No. Ε1112Φ6Υ. 2. Test reagent preparation
① 1 X激酶緩沖液（ Kinase buffer ) ( 5 mM MgCl2, IMm DTT, 50nM SEB ) ； ② DTT 将 DTT原液用灭菌注射用水稀释到 lOOmM作为储备 液备用； ③ ATP 用灭菌注射用水配制 5mM的储备液备用； ④ 10mM化 合物溶液： 釆用 100% DMSO将化合物溶解配制成 10mM的化合物储备 液备用。 1 1 X Kinase buffer ( 5 mM MgCl 2 , IMm DTT, 50 nM SEB ) ; 2 DTT DDT stock solution was diluted with sterile water for injection to 100 mM as a stock solution; 3 ATP was prepared with sterile water for injection 5 mM Stock solution spare; 4 10 mM compound solution: 化合物 Dissolve the compound in 100% DMSO to make a 10 mM compound stock solution.
3. 酶反应阶段  3. Enzymatic reaction stage
①将 10 mM的化合物溶液用 100% DMSO稀释 20倍， 再稀释 2倍后进 行一系列的 3倍稀释， 共 10个浓度梯度， 再用 l x kinase buffer将每个浓度 的溶液稀释 100倍作为试验用化合物浓度， 4μΙ7孔。 ②配制 5x酶溶液： 将酶加入 lxkinase buffer, 2μΙ7孔。③ 25 °C条件下孵育 30min。④配制 5xTK Substrate-biotin: 将 TK Substrate-biotin加入 l xkinase buffer中 , 2mL/孔； ⑤配制 5χΑΤΡ: 将 ATP加入 l xkinase base buffer中， 2mL/孔； ⑥在 25 °C 条件下孵育 40min。  1 Dilute 10 mM compound solution 20 times with 100% DMSO, dilute 2 times, then perform a series of 3-fold dilutions, a total of 10 concentration gradients, and then dilute each concentration of the solution by 100 times with lx kinase buffer. With compound concentration, 4 μΙ 7 wells. 2 Preparation 5x enzyme solution: Add enzyme to lxkinase buffer, 2μΙ7 well. Incubate at 25 °C for 30 min. 4 Preparation 5xTK Substrate-biotin: Add TK Substrate-biotin to l xkinase buffer, 2mL/well; 5 Preparation 5χΑΤΡ: Add ATP to l xkinase base buffer, 2mL/well; 6 incubate at 25 °C for 40min.
4. 检测反应阶段  4. Detection reaction stage
①配制 4xStreptavidin- XL665： 将 Streptavidin- XL665加入 HTRF® Detection buffer中， 5μΙ7孔。 ②每孔再加入 5mL TK Antibody- Cryptate。 ③ 25°C条件下孵育 60 min。  1 Preparation 4xStreptavidin- XL665: Add Streptavidin- XL665 to HTRF® Detection buffer, 5μΙ7 well. 2 Add 5 mL TK Antibody- Cryptate to each well. Incubate at 25 °C for 60 min.
5. 数据读取  5. Data reading
检测反应阶段完成后， 用酶标仪分别检测检测样品在 615nm 和 After the completion of the reaction reaction phase, the test sample was separately detected at 615 nm by a microplate reader.
665nm处的荧光值。 Fluorescence value at 665 nm.
6. 曲线拟合得出 IC50 6. Curve fitting yields IC 50
釆用 GraphPad 5.0 软件进行曲线拟合， 拟合方程为 Y=Bottom + (Top-Bottom)/(l+10A((LogIC50-X)*HillSlope)) , 得出 IC50值。 曲线 Curve fitting with GraphPad 5.0 software, the fitting equation is Y=Bottom + (Top-Bottom)/(l+10 A ((LogIC50-X)*HillSlope)), and the IC 50 value is obtained.
实验结果： 在上述测定中， 本发明化合物和对照化合物 CC- 292对 布鲁顿酪氨酸激酶 (BTK)的体外细胞抑制活性的 IC5。值列在表 1 中： 表 1 Experimental Results: IC 5 of the in vitro cytostatic activity of the compound of the present invention and the control compound CC-292 against Bruton's tyrosine kinase (BTK) in the above assay. The values are listed in Table 1: Table 1
化合物 BTK酶学抑制活性 IC5。 ( nM ) Compound BTK enzymatic inhibitory activity IC 5 . ( nM )
CC-292 16.0  CC-292 16.0
化合物 16 1.08  Compound 16 1.08
化合物 17 1.66  Compound 17 1.66
化合物 19 0.90  Compound 19 0.90
化合物 21 1.48  Compound 21 1.48
化合物 23 0.84  Compound 23 0.84
化合物 24 0.60  Compound 24 0.60
化合物 103 1.20  Compound 103 1.20
化合物 104 3.19  Compound 104 3.19
实验结论：  Experimental results:
综上所述， 本发明化合物对 BTK激酶的抑制活性显著强于对照化 合物 CC- 292。  In summary, the inhibitory activity of the compounds of the present invention against BTK kinase was significantly stronger than that of the control compound CC-292.
II 本发明化合物的体外抗细胞学活性测定  II Determination of in vitro anti-cytotoxic activity of the compounds of the invention
本发明化合物： 其化学名称和结构式与制备方法见制备实施例。 The compound of the present invention: its chemical name and structural formula and preparation method are shown in the preparation examples.
1. 实验仪器 Experimental instrument
Envision 2104 读板仪， PerkinElmer, (美国）； C02培养箱， SA YO.(日 本)； 倒置显微镜， XDS-1B, 重庆广电. （重庆， 中国）； PH 计， Mettler Toledo Five easy. (中国）； MACS 分离器 (Miltenyi, 美国）； FACSCalibur (BD, 美国）。 Envision 2104 Plate Reader, PerkinElmer, (USA); C0 2 Incubator, SA YO. (Japan); Inverted Microscope, XDS-1B, Chongqing Broadcasting. (Chongqing, China); PH Meter, Mettler Toledo Five easy. (China ); MACS Separator (Miltenyi, USA); FACSCalibur (BD, USA).
2. 细胞  2. Cell
在 37°C, 5% C02培养箱中， 用含有 10%的超低免疫球蛋白的胎牛 血清， 青霉素 /链霉素， 5mM HEPES, 50μΜ β-巯基乙醇的 RPMI1640培 养基培养 Balb/c小鼠原代脾脏 B细胞。 培养基购自美国 GIBCO。  Breeding Balb/c in RPMI1640 medium containing 10% ultra-low immunoglobulin fetal bovine serum, penicillin/streptomycin, 5 mM HEPES, 50 μΜ β-mercaptoethanol at 37 ° C, 5% CO 2 incubator Rat primary spleen B cells. The medium was purchased from GIBCO, USA.
3. 小鼠  3. Mouse
雄性， 6- 8周的 Balb/c 小鼠。  Male, 6-8 week Balb/c mice.
4. 试剂与化合物配制  4. Reagents and compound preparation
( 1 ) CellTiter-Glo (CTG) (货号: G7572, Promega), 将 CTG緩沖液 和 CTG底物保存于- 20。C, 推荐以以下的方法准备 CTG 试剂：  (1) CellTiter-Glo (CTG) (Cat. No. G7572, Promega), storing CTG buffer and CTG substrate at -20. C, It is recommended to prepare CTG reagents in the following ways:
使用前融化 CTG緩沖液并平衡至室温， 方便起见， CTG緩沖液可 在使用前融化好并在室温保存到 48小时以上。 将冻干的 CTG底物平衡 到室温，取 100ml的緩沖液到装有底物的琥珀色瓶子里， 就得到了 CTG 试剂。 轻轻混勾直到得到均一的溶液， 底物应在一分钟内完全融解， 分 装并在 - 20°C冰箱中长期保存 CTG试剂。 Melt CTG buffer before use and equilibrate to room temperature. For convenience, CTG buffer can be used. Melt well before use and store at room temperature for more than 48 hours. The lyophilized CTG substrate was equilibrated to room temperature and 100 ml of buffer was taken into the amber bottle containing the substrate to obtain the CTG reagent. Gently mix until a uniform solution is obtained. The substrate should be completely melted in one minute, dispensed and stored in a - 20 ° C refrigerator for a long period of time.
( 2 ) MACS® B 细胞分离试剂盒 （定购号： 130- 090- 862, Miltenyi) ( 3 ) PE 抗生物素 ( anti-biotin ) 抗体 （货号： 409003, Biolegend) ( 4 ) PE/cy7 免抗小鼠 CD45R/B220抗体 (货号： 103221, Biolegend) ( 5 ) 细胞株 （货号： 352340, BD Falcon)  ( 2 ) MACS® B Cell Isolation Kit (Order No. 130- 090- 862, Miltenyi) ( 3 ) PE anti-biotin antibody (Cat. No. 409003, Biolegend) ( 4 ) PE/cy7 Mouse CD45R/B220 Antibody (Cat. No.: 103221, Biolegend) ( 5 ) Cell line (Cat. No.: 352340, BD Falcon)
( 6 ) AffiniPure F(ab')2 fragment goat anti mouse IgM, μ chain specific (货号： 115-006-020, Jackson)  (6) AffiniPure F(ab')2 fragment goat anti mouse IgM, μ chain specific (Cat. No. 115-006-020, Jackson)
( 7 ) 配制测试化合物  (7) Formulating test compounds
• 配制测试化合物储液:将化合物粉末溶解于 DMSO中，到 10 mM 浓度。  • Prepare a test compound stock solution: Dissolve the compound powder in DMSO to a concentration of 10 mM.
• 配制测试化合物梯度稀释溶液: 首先， 取 10 mM的测试化合物 储液用 DMSO 3倍连续梯度稀释，共 10个浓度。再分别取 ΙΟμί的 DMSO 稀释的化合物加到 90μί 化合物稀释緩沖液中， 再分别取 lO L 的 含 • Prepare a test compound gradient dilution solution: First, 10 mM of the test compound stock solution was diluted 3 times in DMSO with a total of 10 concentrations. Then add ΙΟμί DMSO diluted compound to 90μί compound dilution buffer, and then take lO L
10%DMSO 稀释的化合物加到 90μί化合物稀释緩沖液中，化合物最高 浓度为 10μΜ, DMSO 浓度为 0.1%, 共 10个浓度梯度。 The 10% DMSO diluted compound was added to 90 μί of the compound dilution buffer at a maximum concentration of 10 μM and a DMSO concentration of 0.1% for a total of 10 concentration gradients.
5. 实验方法  5. Experimental methods
( 1 ) 分离 Balb/c小鼠 B细胞：  (1) Isolation of Balb/c mouse B cells:
*取 Balb/c小鼠脾脏， 在 MACS緩沖液中捣碎， 用 40μι 的尼龙细 胞筛网过滤得到单细胞悬液。  * The spleens of Balb/c mice were smashed, mashed in MACS buffer, and filtered through a 40 μιη nylon cell sieve to obtain a single cell suspension.
•4。C, 将得到的细胞悬液在 400g 离心五分钟， 去上清， 室温下加 入 lml的红细胞裂解液并轻轻地重悬起细胞团。 两分钟后， 加入预冷的 MACS緩沖液。用 40μι 的细胞筛网过滤细胞悬液到一个新的离心管中。 • 4. C, the obtained cell suspension was centrifuged at 400 g for five minutes, the supernatant was removed, and 1 ml of the red blood cell lysate was added at room temperature and the cell pellet was gently resuspended. Two minutes later, pre-chilled MACS buffer was added. The cell suspension was filtered through a 40 μιη cell sieve into a new centrifuge tube.
4。C, 400g离心 5分钟来收集细胞。 4. C, 400 g was centrifuged for 5 minutes to collect the cells.
*在加入磁珠前， 用 MACS 緩沖液将细胞密度调整到 107个细胞* Adjust the cell density to 10 7 cells with MACS buffer before adding the beads
/40μ1。 每 107个细胞中加入 ΙΟμΙ的生物素化的抗体混合物。 混匀后在冰 上孵育 20分钟，每 107个细胞中加入 30μ1的 MACS緩沖液和 20μ1的抗 生物素的磁珠并在冰上孵育 20分钟。 离心后用 500μ1的 MACS緩沖液 重悬细胞。 将预冷的 MACS分选柱置于 MACS分选器中， 将细胞悬液 加到 MACS分选柱中。 收集流下的未结合抗体的细胞。 •通过流式细胞术用 PE anti- biotin抗体和 CD45R(B220)抗体检测检 测分选前和分选后的细胞。 /40μ1. A biotinylated antibody mixture of ΙΟμΙ was added to every 10 7 cells. After mixing, incubate on ice for 20 minutes, add 30 μl of MACS buffer and 20 μl of avidin-containing magnetic beads per 10 7 cells and incubate on ice for 20 minutes. After centrifugation, the cells were resuspended in 500 μl of MACS buffer. The pre-cooled MACS sorting column was placed in a MACS sorter and the cell suspension was applied to a MACS sorting column. The cells under the flow of unbound antibody are collected. • Detection of cells before and after sorting by flow cytometry using PE anti-biotin antibody and CD45R (B220) antibody detection.
( 2 ) IC5o测定 (2) IC 5 o determination
•用血球计数板计数新鲜分离的小鼠 B 细胞， 通过眙盼蓝染色法检 测细胞活率应在 98%以上。  • Freshly isolated mouse B cells were counted using a hemocytometer and the cell viability was determined to be above 98% by trypan blue staining.
•用培养基将细胞密度调整到每毫升 3.89χ 105个细胞. 用多道移液 器取 90μ1细胞悬液到 96孔板中， 得到最终细胞密度为 3.5x l04个细胞 每孔。 • Adjust the cell density to 3.89 χ 10 5 cells per ml with medium. A 90 μl cell suspension was pipetted into a 96-well plate using a multichannel pipette to give a final cell density of 3.5×10 4 cells per well.
*用 DMSO 溶解稀释被测化合物和阳性化合物形成储存液， 加入 ΙΟμΙ配制的一系列化合物溶液到 96孔板里 （每个化合物的每个浓度做 三点重复）。 在 37 °C , 5% C02培养箱中孵育 30分钟， 然后再加入 50μ1 Β细胞刺激混合液， 刺激混合液中 anti- Igm的终浓度是 10 g/ml。 * Dissolve the test compound and the positive compound in DMSO to form a stock solution, and add a series of compound solutions prepared in ΙΟμΙ to a 96-well plate (three-point repetition for each concentration of each compound). Incubate for 30 minutes at 37 ° C in a 5% CO 2 incubator, then add 50 μl of sputum cell stimulating mixture and stimulate the final concentration of anti- Igm in the mixture to be 10 g/ml.
•将细胞板在 37 °C , 5% C02培养箱中继续孵育 72小时后用 CTG的 方法进行检测。 • The cell plates were incubated for 72 hours at 37 ° C in a 5% C0 2 incubator and tested by CTG.
·融化 CTG试剂并平衡至室温， 用多道移液器转入到 96孔板中， • Thaw the CTG reagent and equilibrate to room temperature and transfer to a 96-well plate using a multi-channel pipette.
50μ1 CTG试剂 /孔， 在微孔板快速震荡器上震荡 2分钟后在黑暗中放置 10分钟， 用 Envision检测发光读值。 50 μl CTG reagent / well, vortexed on a microplate fast shaker for 2 minutes, then placed in the dark for 10 minutes, and the luminescence reading was detected with Envision.
6. 数据分析  6. Data analysis
得到的数据会用 Excel 2007和 GraphPad Prism 5.0软件进行分析， 为了计算 IC5。， 将利用非线性 S曲线回归来拟合数据得出一条剂量 -效应 曲线， GraphPad Prism 5.0软件会自动给出 IC50值。 The data obtained will be analyzed using Excel 2007 and GraphPad Prism 5.0 software in order to calculate IC 5 . The nonlinear S-curve regression will be used to fit the data to obtain a dose-response curve, and the GraphPad Prism 5.0 software will automatically give the IC 50 value.
细胞存活率用以下公式进行计算： V样品 / V2 溶 ¾对,照 X 100% , V样品是 化合物处理孔的读值， V2 »¾对¾是溶剂对照孔 (V2)读值的平均值。 Cell viability was calculated using the following formula: V sample / V2 dissolved 3⁄4 pairs, X 100%, V sample is the reading of the compound treated well, V2 » 3⁄4 vs. 3⁄4 is the average of the solvent control well (V2) readings.
7. 实验结果：  7. Experimental results:
本发明化合物 （化合物 1-106 )对 Balb/c小鼠 B细胞体外细胞的抑 制活性的 IC5Q < 0.5μΜ。 部分化合物的 IC5Q如下： 本发明部分化合物对 Balb/c小鼠 B细胞体外细胞学抑制活性 化合物 The compound of the present invention (Compound 1-106) has an IC 5 Q < 0.5 μΜ for the inhibitory activity of Balb/c mouse B cells in vitro. The IC 5 Q of some compounds is as follows: In vitro cytological inhibitory activity of some compounds of the invention against Balb/c mouse B cells
化合物 1 ++  Compound 1 ++
化合物 8 ++  Compound 8 ++
化合物 13 +++  Compound 13 +++
化合物 15 +++  Compound 15 +++
化合物 16 +++  Compound 16 +++
化合物 17 +++  Compound 17 +++
化合物 18 +++  Compound 18 +++
化合物 19 ++++  Compound 19 ++++
化合物 20 +++  Compound 20 +++
化合物 21 +++  Compound 21 +++
化合物 22 +++  Compound 22 +++
化合物 23 ++++  Compound 23 ++++
其中， ++++代表 IC50 ( μΜ ) <0.01 μΜ; +++代表 Ο.ΟΙμΜ <IC50 ( μΜ ) <0.1μΜ; ++代表 0.1 <IC50 ( μΜ )≤0·5μΜ o Where ++++ represents IC 50 ( μΜ ) <0.01 μΜ; +++ represents Ο.ΟΙμΜ <IC 50 ( μΜ ) <0.1μΜ; ++ represents 0.1 <IC 50 ( μΜ )≤0·5μΜ o
。  .
实验结论： 综上所述， 由表 2可见， 本发明化合物对 Balb/c小鼠 Β 细胞增殖均有较强的抑制作用。  Experimental conclusions: In summary, it can be seen from Table 2 that the compounds of the present invention have a strong inhibitory effect on the proliferation of Balb/c mouse sputum cells.
ΠΙ 本发明化合物对大鼠脾细胞 BTK酶占有率测定实验  测定 Determination of BTK enzyme occupancy rate in rat spleen cells by the compound of the present invention
为了测定细胞或组织溶解产物中未被化合物占用 BTK 的量， 使用 ELISA方案， 其利用一种只结合未被化合物占用 BTK的生物素化探针 化合物,评价化合物在不同浓度下，在脾细胞中对 BTK酶的占有率情况， 计算％81^占有率 （ BTK Occupancy ) 。  To determine the amount of BTK that is not occupied by a compound in a cell or tissue lysate, an ELISA protocol is used that evaluates compounds at different concentrations in spleen cells using a biotinylated probe compound that binds only to BTK that is not occupied by the compound. For the occupancy rate of BTK enzyme, calculate the %81^ occupancy rate (BTK Occupancy).
1、 实验材料  1. Experimental materials
小鼠抗 BTK抗体 (Becton Dickinson); 山羊抗小鼠 HRP抗体 (Becton Dickinson); 细胞裂解液 (Cell Signaling); 布鲁顿酪氨酸激酶 （ BTK ) (Carna); 链霉亲和素包被的 96 孔板 (Thermo); 大鼠淋巴细胞分离试剂 盒 (LTS 1083PK , 天津市灏洋生物制品科技有限责任公司）； 酶标仪 (victor4, PE) ； 离心机 (5804R, Eppendorf); 显微镜 (CX31RTSF, 奥林巴 斯）； MACS分选器 (midiMACS separation unit, MACS)。  Mouse anti-BTK antibody (Becton Dickinson); goat anti-mouse HRP antibody (Becton Dickinson); cell lysate (Cell Signaling); Bruton tyrosine kinase (BTK) (Carna); streptavidin coated 96-well plate (Thermo); rat lymphocyte isolation kit (LTS 1083PK, Tianjin Haoyang Biological Products Technology Co., Ltd.); microplate reader (victor4, PE); centrifuge (5804R, Eppendorf); microscope ( CX31RTSF, Olympus); MACS sorter (MACS).
2、 实验步骤  2, the experimental steps
(1)试剂配制  (1) reagent preparation
探针化合物溶液（Probe ): 称取样品化合物 lmg, 配制浓度为 ImM 的储备液， 使用时用样品稀释液稀释； 样品稀释液（ Sample diluents ) ： 含 1 %牛血清白蛋白和 0.1 % Tween-20 的 PBS ; 洗涤液 （Washing solution ): 含 0.05 % Tween-20的 PBS。 Probe compound solution (Probe): Weigh 1 mg of sample compound and prepare a concentration of 1 mM. Stock solution, diluted with sample diluent; Sample diluents: PBS containing 1% bovine serum albumin and 0.1% Tween-20; Washing solution: 0.05% Tween-20 PBS.
(2) 大鼠脾脏单细胞制备  (2) Preparation of rat spleen single cells
将脾脏用 lmL的 PBS緩沖液沖洗（在脾脏的一端剪个小口， 用注射 器在脾脏另一端注入 lml预冷 PBS沖洗） ， 然后转移至 200 目无菌滤 网， 用手术剪剪碎， 再用注射器研磨， 注意边研磨边加预冷的 PBS緩沖 液沖洗， 共计用 5ml的 PBS緩沖液沖洗。 4°C , 400g离心 3min, 去上 清， 力口入 20ml细胞洗涤液 PBS , 4°C , 600Rpm, 离心 lOmin, 洗涤 3 次。  Rinse the spleen with 1 mL of PBS buffer (cut a small opening at one end of the spleen and inject 1 ml of pre-cooled PBS into the other end of the spleen with a syringe), then transfer to a 200-mesh sterile filter, cut with a surgical scissors, and reuse The syringe was ground, and rinsed with pre-cooled PBS buffer while grinding, and rinsed with 5 ml of PBS buffer. After centrifugation at 400 °C for 3 min at 4 °C, remove the supernatant, and add 20 ml of cell washing solution to PBS, 4 ° C, 600 Rpm, centrifuge for 10 min, and wash 3 times.
(3) 化合物与细胞作用  (3) Compound and cell action
将细胞计数后， 用 PBS将细胞浓度稀释到 3xl07Cells/ml, 90μ1/孔。 加入化合物 ΙΟμΙ/孔， 37°C孵育 lh。 20 °C , 400g离心 20min, 弃掉上清。  After counting the cells, the cell concentration was diluted to 3 x 107 Cells/ml, 90 μl/well with PBS. Add compound ΙΟμΙ/well and incubate for 1 h at 37 °C. Centrifuge at 20 °C, 400g for 20min, discard the supernatant.
(4) 裂解细胞  (4) Lysis cells
将蛋白酶抑制剂 PMSF加入到细胞裂解液（ cell lysis buffer ) 中 （注 意 PMSF在裂解液使用前加入）。将裂解液加入每管富集的细胞中混匀 , 根据裂解液说明书操作， 冰上裂解 5min, 14000g 离心 10min。 取上清 ΙΟΟμΙ加入 96孔板中。  The protease inhibitor PMSF was added to the cell lysis buffer (note that PMSF was added before the lysate was used). The lysate was added to each tube-enriched cell and mixed. According to the lysate instructions, the ice was lysed for 5 min and centrifuged at 14,000 g for 10 min. Add the supernatant ΙμΙ to the 96-well plate.
(5) ΒΤΚ测定实验步骤  (5) ΒΤΚ measurement experiment steps
每孔加入 lOOul 标准品或样品与 ΙΟμΙ 探针化合物溶液（终浓度为 ΙμΜ ) 混合， 28 °C震荡孵育 lh。 孵育后， 取 ΙΟΟμΙ加入到链霉亲和素包 被的 ELISA板上 ( Streptavidin-coatd ELISA plate ) , 28 °C震荡孵育 lh。 用洗涤液（Washing sol ution ) 洗板 5次。 向每孔中加入 ΙΟΟμΙ 纯化的 鼠抗人 ΒΤΚ抗体 ( Purified mouse anti-human antibody ) ( 1 : 1000倍稀 释） 。 28°C震荡孵育 lh。 用洗涤液洗板 5次。 向每孔中加入 ΙΟΟμΙ ΗΙ Ρ 标记的羊抗鼠抗体 ( HRP goat anti-mouse Ig ) ( 1 : 1000稀释） 。 28 °C震 荡孵育 lh。用洗涤液洗板 5次后向每孔加入 ΙΟΟμΙ底物 ΤΜΒ溶液， 28 °C 孵育 15min。 每孔加入 50μ1 1M H2S04终止反应。  Add lOOul of standard or sample to each well and mix with ΙΟμΙ probe compound solution (final concentration ΙμΜ) and incubate for 1 h at 28 °C. After incubation, ΙΟΟμΙ was added to a streptavidin-coated ELISA plate and incubated at 28 °C for 1 h. Wash the plate 5 times with Washing sol ution. Purified mouse anti-human antibody (1: 1000-fold dilution) was added to each well. Incubate at 28 °C for 1 h. Wash the plate 5 times with washing solution. ΙΟΟμΙ ΗΙ 标记 labeled goat anti-mouse Ig (1:1000 dilution) was added to each well. Incubate at 28 °C for 1 h. After washing the plate with the washing solution 5 times, a ΙΟΟμΙ substrate ΤΜΒ solution was added to each well, and incubated at 28 ° C for 15 min. The reaction was stopped by adding 50 μl of 1 M H2S04 to each well.
3、 检测指标及检测方法  3. Detection indicators and detection methods
反应终止后， 检测 450nm处的 OD值。 根据 OD值， 使用酶标仪中 After the reaction was terminated, the OD value at 450 nm was detected. According to the OD value, use the microplate reader
A 4参数逻辑曲线计算各样品中 BTK的量。 The A 4 parameter logic curve calculates the amount of BTK in each sample.
4、 数据处理及结果 *%BTK占有率 = (对照组 BTK量-化合物组 BTK量） /对照组 BTK 量 χ100% 4, data processing and results *% BTK occupancy = (control group BTK amount - compound group BTK amount) / control group BTK amount χ 100%
**变异系数 CV=57 X 100%  **Coefficient of variation CV=57 X 100%
实验结果如表 3所示：  The experimental results are shown in Table 3:
表 3 : 化合物对大鼠脾细胞 BTK占有率实验结果  Table 3: Experimental results of BTK occupancy rate of rat spleen cells
%BTK占有率  %BTK share
化合物  Compound
50nM 200nM  50nM 200nM
化合物 24 85.05% 86.83%  Compound 24 85.05% 86.83%
化合物 103 77.96% 87.93%  Compound 103 77.96% 87.93%
实验结论： 由表 3可见， 本发明化合物在大鼠脾细胞中的 BTK 占 有率较高， 体现出较好的药效。  Experimental conclusion: It can be seen from Table 3 that the compound of the present invention has a higher BTK occupancy rate in rat spleen cells, and exhibits better pharmacological effects.
IV 本发明化合物血浆稳定性实验  IV. Plasma stability test of the compound of the present invention
实验方法  experimental method
1、 评价体系  1. Evaluation system
(1) 测试化合物及对应母液信息： 取化合物 5mg左右， 先用 DMSO 配制成 10mM的储备液, 再用 DMSO: 7j =l :l稀释成 50uM的测试化合 物工作溶液， 待用。 （反应体系中 DMSO含量≤0.5%)  (1) Test compound and corresponding mother liquor information: Take about 5mg of compound, first prepare 10mM stock solution with DMSO, then dilute to 50uM test compound working solution with DMSO: 7j = l:1, and use. (The DMSO content in the reaction system is ≤0.5%)
(2) 测试介盾  (2) Test shield
基盾 1种属 来源 抗凝剂 储藏温度（。C ) 人血浆 男性： 女性 =1: 1 ( V: V ) 肝素钠 -80 beagle犬血浆 雄性健康 beagle犬 肝素钠 -80  Base Shield 1 species Source Anticoagulant Storage temperature (.C) Human plasma Male: Female =1: 1 ( V: V ) Heparin sodium -80 beagle dog plasma Male health beagle dog Heparin sodium -80
SD大鼠血浆 雄性健康 SD大鼠 肝素钠 -80  SD rat plasma male health SD rat heparin sodium -80
2. 样品制备方法  2. Sample preparation method
(1) 将冰冻的血浆置 37°C恒温水浴锅中预孵育解冻备用。  (1) The frozen plasma was pre-incubated in a 37 ° C constant temperature water bath and thawed for use.
(2) 取 10 待测化合物 （ 50 μΜ ) 加到 490 μ 各种属血浆中， 待 测物终浓度 Ι μΜ, 两个复样。  (2) Take 10 test compounds (50 μΜ) and add them to 490 μ of various plasmas. The final concentration of the analyte is Ι μΜ, two replicates.
(3) 涡旋混匀， 置于 37°C恒温水浴锅中温育。  (3) Vortex and mix, and incubate in a constant temperature water bath at 37 °C.
(4) 在对应的时间点（ T=0 h, 2 h, 4 h )取反应液 50μί, 加入 300μί 的终止液， 混匀后置- 80°C冻存。  (4) Take 50 μί of the reaction solution at the corresponding time point (T=0 h, 2 h, 4 h), add 300 μί of the stop solution, mix and store at -80 °C.
(5) 待温育实验完成后， 解冻各时间点样品管， 混匀后至离心机中 12000rpm离心 5min。  (5) After the incubation experiment is completed, thaw the sample tubes at each time point, mix and centrifuge at 12000 rpm for 5 min in a centrifuge.
(6) 取干净 96孔样品板加入 150 μ 的水， 取上清 50 μ 至样品孔 中 , 混匀后递交样品板进行进样分析。 (6) Take a clean 96-well sample plate and add 150 μl of water. Take the supernatant 50 μ to the sample well. In the middle, mix and submit the sample plate for injection analysis.
3. LC-MS/MS 进样分析  3. LC-MS/MS injection analysis
4. 数据分析  4. Data analysis
化合物在血浆中的稳定性釆用化合物经各时间点孵育后的保留百 分率来评价， 样品的浓度用测试化合物的峰面积和内标峰面积之比表 示。 计算公式如下：  The stability of the compound in plasma was evaluated by the retention percentage of the compound after incubation at each time point, and the concentration of the sample was expressed by the ratio of the peak area of the test compound to the internal standard peak area. Calculated as follows:
剩余量 （％) = CTn/CT0 Remaining amount (%) = C Tn /C T0
其中， CTn为化合物在各孵育时间点后的终溶液测定浓度， n=2 h,Wherein, C Tn is the final solution concentration of the compound after each incubation time point, n=2 h,
4 h; 4 h;
CT。为化合物在起始孵育时的终溶液测定浓度。 C T . The concentration of the compound is determined as the final solution at the initial incubation.
实验结果  Experimental result
表 4 在不同血浆中温孵后 (T=2h)剩余量(％)比较 Table 4 Comparison of residual amount (%) after incubation in different plasmas (T=2h)
~化合物 Ajk浆 beagle犬血浆 SD大鼠血浆 ~ compound Ajk pulp beagle dog plasma SD rat plasma
~ CC-292 76.8% 89.5% 83.5% ~ CC-292 76.8% 89.5% 83.5%
化合物 23 95.4% 100.3% 104.5%  Compound 23 95.4% 100.3% 104.5%
表 5 在不同血浆中温孵后 (T=4h)剩余量(％)比较 Table 5 Comparison of residual amount (%) after incubation in different plasmas (T=4h)
~化合物 Ajk浆 beagle犬血浆 SD大鼠血浆 ~ compound Ajk pulp beagle dog plasma SD rat plasma
~ CC-292 54.8% 75.0% 63.9% ~ CC-292 54.8% 75.0% 63.9%
化合物 23 86.7% 94.3% 90.7%  Compound 23 86.7% 94.3% 90.7%
综上所述， 本发明化合物 23与对照药 CC- 292相比， 在血浆中的稳 定性更好。  In summary, the compound 23 of the present invention is more stable in plasma than the control drug CC-292.
V 膜片钳方法检测本发明化合物的对 hERG钾离子通道的抑制作 歷  The V patch clamp method detects the inhibition of the hERG potassium channel by the compound of the present invention.
实验材料： 如下所示：  Experimental material: as shown below:
材料 来源  Material source
盐酸阿米替林 * Sigma- Aldrich  Amitriptyline hydrochloride * Sigma- Aldrich
DMSO Merck  DMSO Merck
CHO-hERG AVIVA  CHO-hERG AVIVA
*盐酸阿米替林， 国际通用的标准 hERG通道阻断剂。  * Amitriptyline hydrochloride, the internationally accepted standard hERG channel blocker.
本发明化合物 （化合物 1-106 ) ： 其化学名称和结构式与制备方法 见各化合物的制备实施例。 对照化合物： CC-292 , 按照专利 WO2009158571A1合成。 实验方法 The compound of the present invention (Compound 1-106): Its chemical name, structural formula and preparation method are shown in the preparation examples of the respective compounds. Control compound: CC-292, synthesized according to patent WO2009158571A1. experimental method
(i) 溶液.及化合物的配制  (i) Preparation of solutions and compounds
细胞外液 (mM): 7V~2羟基乙基哌嗪 - - 2-乙磺酸 (HEPES) 10、 NaCI 145、 KC1 4、 CaCI2 2、 MgCI2 l、 Glucose 10, 用 IN氢氧化钠调节 pH至 7.4; 渗透压调至 290 3()() mOsm; 过滤后 保存。 Extracellular fluid (mM): 7V~2 hydroxyethylpiperazine-2-ethylethanesulfonic acid (HEPES) 10. NaCI 145, KC1 4, CaCI 2 2, MgCI 2 l, Glucose 10, adjusted with IN sodium hydroxide pH to 7.4; osmotic pressure adjusted to 290 3 () () mOsm; filtered and stored.
电极内液 (in mM): KC1 120、 OH 31.25 , CaC¾ 5.374、 MgCI2 L75 , 乙二醇—二（β—氨基乙基醚) ΛζΛ^Υ',Λ'"-四乙酸 (EGTA) 10、 HEPES 10、 Na.2-ATP 4, 用 IN氢氧化钾调节 pH至 7.2; 渗透压调至 280 290mOsm; 过 滤后- 20°C保存。 In-electrode solution (in mM): KC1 120, OH 31.25, CaC3⁄4 5.374, MgCI 2 L75, ethylene glycol-bis(β-aminoethyl ether) ΛζΛ^Υ', Λ'"-tetraacetic acid (EGTA) 10, HEPES 10, Na. 2 -ATP 4, pH was adjusted to 7.2 with IN potassium hydroxide; osmotic pressure was adjusted to 280 290 mOsm; after filtration - 20 ° C storage.
化合物的配制： 阳性对照药盐酸阿米替林和 2个样本 CC- 292和本发 明化合物（化合物 1-106 )先溶于 1()0% DMSO ( Sigma- Aldrich, 34869 ) , 配置成 30 mM的储备溶液。 实验前用 DMSO将上述储备溶液稀释为各个 试验浓度 1000倍的溶液， 然后再用细胞外液稀释 1000倍到所需浓度。 细 胞外液中 DMSO最终浓度为 0.1 %。  Preparation of the compound: The positive control drug amitriptyline hydrochloride and 2 samples CC-292 and the compound of the invention (compound 1-106) were first dissolved in 1 () 0% DMSO (Sigma-Aldrich, 34869), configured to 30 mM Stock solution. The above stock solution was diluted with DMSO to a solution of 1000 times each test concentration before the experiment, and then diluted 1000 times with the extracellular liquid to the desired concentration. The final concentration of DMSO in the extracellular fluid was 0.1%.
(2) 电生理实验。  (2) Electrophysiological experiments.
采用全细胞膜片钳技术记录 hERG电流。取细胞悬液加于 35 mm的培 养 i中， 置于倒置显微镜载物台上。 待细胞贴壁后， 用细胞外液灌流， 流速为 1 2 mL/min。 玻璃微电极由微电极拉制仪两步拉制， 其入水电阻 值为 2- 5 ΜΩ。 建立全细胞记录后， 保持钳制电位为- 80 mV。 给予电压刺 激时去极化至 +60 mV, 然后复极化至 -50 mV引出 hERG尾电流。 所有记 录均在电流稳定后进行。胞外灌流给药从低浓度开始，每个浓度 5 ΙΟηώι 至电流稳定， 再给下一个浓度。  The hERG current was recorded using whole cell patch clamp technique. The cell suspension was applied to a 35 mm culture and placed on an inverted microscope stage. After the cells were attached, the cells were perfused with extracellular fluid at a flow rate of 12 mL/min. The glass microelectrode is drawn in two steps by a microelectrode drawing device, and its water resistance is 2 - 5 Ω. After establishing a whole cell recording, the clamping potential was maintained at -80 mV. The voltage is stimulated to depolarize to +60 mV and then repolarized to -50 mV to induce the hERG tail current. All records are performed after the current has stabilized. Extracellular perfusion administration starts from a low concentration, each concentration is 5 ΙΟηώι until the current is stable, and the next concentration is given.
(3) 细  (3) Fine
稳定细胞株 CHO hERG购自 AVIV A公司。 为了盾量控制， 最小的封 接电阻不小于 1GQ, 并且 hERG电流不小于 0.4nA。  Stable cell line CHO hERG was purchased from AVIV A. For shield control, the minimum sealing resistance is not less than 1GQ, and the hERG current is not less than 0.4nA.
此次试险包括以下几个方面：  The test coverage includes the following aspects:
利用手动膜片钳技术在稳定表达 hERG通道的 CHO-K1细胞株上记 录 hERG电流； 根据 hERG尾电流计算每个浓度的抑制率； 每个化合物测 试 5 每个浓度测试 2个细胞； 一个阳性对照药物。 The hERG current was recorded on the CHO-K1 cell line stably expressing the hERG channel by manual patch clamp technique; the inhibition rate of each concentration was calculated from the hERG tail current; each compound was tested 5 for each concentration test 2 cells; one positive control drug.
Figure imgf000034_0001
Figure imgf000034_0001
通过 Digidata 1440 ( Molecular Devices ) 和 pCLAMP软件 （ 10,2.版， Through Digidata 1440 (Molecular Devices) and pCLAMP software (10, 2. version,
Molecular Devices ) A/D D/A数模转换， 进行刺激发放及信号釆集； 膜 片¾| "放大 ( Miilticlamp 700B , Molecular Devices ) 放大 4言号， 滤波为 1 ΚΉζ。 4吏用（！^111^1¾ ( 10.2¾_, Molecular Devices )和 Prism进--—步进亍数 据分析和曲线拟合。 数据均以均值 ±标准差表示。 IC5G数值由 Logistic方 程进行拟合所得： Molecular Devices ) A/DD/A digital-to-analog conversion for stimulus distribution and signal collection; 3⁄4| "Magnification (Miilticlamp 700B, Molecular Devices) Enlarge 4 words, filter to 1 ΚΉζ. 4吏 (!^111^13⁄4 ( 10.23⁄4_, Molecular Devices) and Prism---stepping data analysis and Curve fitting. Data are expressed as mean ± standard deviation. IC 5G values are fitted by Logistic equation:
max— mm  Max- mm
-mm  -mm
IC, y: 抑制百分比； max: 为 100%; min: 为 0%; [drug] : 测试  IC, y: percent inhibition; max: 100%; min: 0%; [drug] : test
nH: 斜率； IC5。： 测试物的最大半数抑制浓度。 n H : slope; IC 5 . : The maximum half of the test substance is inhibited.
实验结果  Experimental result
本试验利用全细胞膜片钳技术， 在稳定表达 hERG通道的 CHO-K1 细胞株上检测了化合物对 hERG电流的阻断作用。 每个测试化合物的半 数抑制浓度（IC5o ) 由 Logistic方程最佳拟合得出。 在本试验的检测浓度 范围内我们选择的药物浓度范围为国际药物研发公司所最常用的 hERG 通道测试范围， 因为这一浓度范围已经涵盖通常的给药剂量。 绝大多数 In this experiment, whole cell patch clamp technique was used to detect the blocking effect of compounds on hERG currents on CHO-K1 cells stably expressing hERG channels. The half-inhibitory concentration (IC 5 o ) of each test compound was derived from the best fit of the Logistic equation. The range of drug concentrations we selected within the range of concentrations tested in this test is the most commonly used hERG channel test range for international drug discovery companies, as this concentration range already covers the usual doses administered. The vast majority
〇  〇
对 hERG通道有抑制作用的样本都能够在这些浓度下对 hERG电流产生 抑制作用。 化合物对 hERG的阻断效应见表 5。 Samples that inhibited hERG channels were able to inhibit hERG currents at these concentrations. The blocking effect of compounds on hERG is shown in Table 5.
表 6 在 CHO-K1 稳定细胞株上所记录到的化合物对 hERG电流的 IC5Q数值 Table 6 IC 5Q values of hERG currents recorded by compounds on CHO-K1 stable cell lines
样品  Sample
盐酸阿米替林 3.15  Amitriptyline hydrochloride 3.15
CC-292 21.50 本发明化合物 （化合物 1-106 ) >30.00  CC-292 21.50 Compound of the invention (Compound 1-106) >30.00
阳性对照药物 Amitriptyline是使用最为广泛的阻断 hERG电流工具 药物之一， 在本次研究中对 hERG电流抑制的 IC5。为 3.15 μΜ, 这一结果 与文献报道的结果相符合。 这表明本次试验的结果是可信的。 The positive control drug Amitriptyline is one of the most widely used drugs for blocking hERG currents, IC 5 for inhibition of hERG current in this study. For 3.15 μΜ, this result is consistent with the results reported in the literature. This shows that the results of this trial are credible.
本研究所检测的化合物对 hERG电流抑制在最高测试浓度 ( 30.00 μΜ )对 hERG电流的抑制作用均远未达到 IC5。， 从而说明了本发明化合 物（化合物 1-106 )对 hERG通道没有明显的抑制作用。 而对照药 CC- 292 对 hERG电流的 IC5。值为 21.50 μΜ, 对 hERG通道有抑制作用。 因此， 本 发明化合物 （化合物 1-106 ) 与对照药相比， 安全性更高。 The inhibitory effect of the compounds tested in this study on hERG current inhibition at the highest concentration (30.00 μΜ) on hERG current was far from IC 5 . Thus, it was demonstrated that the compound of the present invention (Compound 1-106) has no significant inhibitory effect on the hERG channel. The control drug CC-292 has an IC 5 for hERG current. The value is 21.50 μΜ, which has an inhibitory effect on the hERG channel. Therefore, the compound of the present invention (Compound 1-106) is safer than the control drug.
参考文献： Blockade of the HERG human cardiac K+ channel by the antidepressant drug amitriptyline. British Journal of Pharmacology, (2000) 129: 1474-1480. References: Blockade of the HERG human cardiac K+ channel by the Antidepressant drug amitriptyline. British Journal of Pharmacology, (2000) 129: 1474-1480.
VI 本发明化合物的安全性评价实验  VI Safety Evaluation Experiment of the Compound of the Invention
本发明化合物 （化合物 1-106 ) ： 其化学名称和结构式与制备方法 见各化合物的制备实施例。  The compound of the present invention (Compound 1-106): Its chemical name, structural formula and preparation method are shown in the preparation examples of the respective compounds.
实验方法  experimental method
观察 SD大鼠灌冒给予受试化合物 （化合物 1-106 ) 重复给药 14天的 毒性， 了解受试物毒性作用及其耙器官， 确定此试验条件下无毒反应剂 量； 并通过恢复期观察受试物是否有延迟毒性以及出现的毒性反应是否 可逆。  To observe the toxicity of the test compound (Compound 1-106) administered to SD rats for 14 days, to understand the toxicity of the test substance and its sputum organs, to determine the dose of non-toxic reaction under the test conditions; Whether the test substance has delayed toxicity and whether the toxic reaction is reversible.
主试验组分别为溶媒对照组、 受试物 （化合物 1-106 ) 低剂量组、 中剂量组、 高剂量组， 动物各 10只， 雌雄各半； 其中中剂量组及溶媒对 照组另设恢复期动物各 10只， 雌雄各半。  The main experimental group was the vehicle control group, the test substance (Compound 1-106), the low dose group, the middle dose group, the high dose group, and 10 animals each, male and female. The middle dose group and the vehicle control group were additionally restored. There are 10 animals each, half male and half female.
按 10ml/kg容量灌胃给药， 每天 1次， 连续给药 14天。 试验期间每天 观察动物的外观和一般行为学变化， 并进行体重、 摄食量、 血液学、 凝 血功能、血清生化检查；给药期结束后对所有安乐死动物进行大体解剖， 主要脏器称重， 脏器进行病理组织学检查。  It was intragastrically administered at a dose of 10 ml/kg once a day for 14 days. During the test, the appearance and general behavioral changes of the animals were observed every day, and the body weight, food intake, hematology, coagulation function, and serum biochemical examination were performed. After the end of the administration period, all the euthanized animals were grossly dissected, the main organs were weighed, and the organs were dirty. The device was examined for histopathology.
实验结果  Experimental result
本发明化合物 （化合物 1-106 ) 的耐受性好， 安全性高。  The compound of the present invention (Compound 1-106) has good tolerance and high safety.
4、 具体实施方式  4, the specific implementation
以下通过实施例形式的具体实施方式，对本发明的上述内容作进一 步的详细说明。但不应将此理解为本发明上述主题的范围仅限于以下实 施例。 凡基于本发明上述内容所实现的技术均属于本发明的范围。  The above content of the present invention will be further described in detail below by way of specific embodiments in the form of embodiments. However, the scope of the above-mentioned subject matter of the present invention should not be construed as being limited to the following embodiments. Any technique implemented based on the above description of the present invention is within the scope of the present invention.
实施例 1 4-(4-(4-(3-丙烯酰氨基苯氨基) -5- (甲硫基)嘧啶 -2-基氨 基)苯氧基) -N-甲基吡啶 -2-甲酰胺 (化合物 1)的制备
Figure imgf000037_0001
Example 1 4-(4-(4-(3-acrylamidophenylamino)-5-(methylthio)pyrimidin-2-ylamino)phenoxy)-N-methylpyridine-2-carboxamide Preparation of (Compound 1)
Figure imgf000037_0001
(1) 5- (甲硫基)嘧啶- 2,4(1//,3/ )-二酮的制备
Figure imgf000037_0002
(1) Preparation of 5-(methylthio)pyrimidine-2,4(1//,3/)-dione
Figure imgf000037_0002
将 5-溴尿嘧啶 (3 g, 15.7 mmol) 和甲石 醇钠 20%的水溶液 (10 mL)混 合， 加热至 100 °C反应 5 h, 冷却至室温， 过滤，得到固体 1.5 g, 收率： 60.5%。  5-bromouracil (3 g, 15.7 mmol) and 20% aqueous solution of sodium methoxide (10 mL) were mixed, heated to 100 ° C for 5 h, cooled to room temperature, filtered to give a solid 1.5 g, yield : 60.5%.
(2) 2,4-二氯- 5- (甲硫基)嘧啶的制备
Figure imgf000037_0003
(2) Preparation of 2,4-dichloro-5-(methylthio)pyrimidine
Figure imgf000037_0003
将 5- (甲硫基)嘧啶- 2,4(1//,3/ )-二酮（1.5 g, 9.5 mmol) , 二甲基苯 胺 (2.3 g, 19 mmol)混合， 加入三氯氧磷 (15 g, 97.8 mmol) , 在 100 °C反应 4 h, 冷却， 倒入冰水中 （100 mL) , 析出固体， 过滤， 干燥， 得到固体 1.1 g, 收率： 59.4%。  Mix 5-(methylthio)pyrimidine-2,4(1//,3/)-dione (1.5 g, 9.5 mmol), dimethylaniline (2.3 g, 19 mmol), add phosphorus oxychloride (15 g, 97.8 mmol), reacted at 100 °C for 4 h, cooled, poured into ice water (100 mL), and the solid was precipitated, filtered, and dried to give a solid 1.1 g, yield: 59.4%.
(3) 3- (2-氯- 5- (甲硫基)嘧啶- 4-基氨基)苯基氨基甲酸叔丁酯的制备
Figure imgf000037_0004
(3) Preparation of tert-butyl 3-(2-chloro-5-(methylthio)pyrimidin-4-ylamino)phenylcarbamate
Figure imgf000037_0004
将 2,4-二氯- 5- (甲硫基)嘧啶 (0.5 g, 2.56 mmol) , 3-氨基苯基氨基甲酸 叔丁酯 (0.68 g , 3.26 mmol) 溶于 THF (20 mL) ,加入 DIPEA (0.42 g, 3·25 mmol) , 加热回流 7 h, 冷至室温， 浓缩， 柱层析 (PE:EA=4: 1)得到产品 0.4 g, 收率： 43%。  2,4-Dichloro-5-(methylthio)pyrimidine (0.5 g, 2.56 mmol), tert-butyl 3-aminophenylcarbamate (0.68 g, 3.26 mmol) dissolved in THF (20 mL) DIPEA (0.42 g, 3·25 mmol), heated under reflux for 7 h, cooled to room temperature, concentrated and purified by column chromatography (PE: EA = 4: 1) to give product 0.4 g, yield: 43%.
(4) 3- (2- (4- (2- (甲基氨基甲酰基) p比啶- 4-基氧基)苯氨基 )-5- (甲硫基) 嘧啶- 4-基氨基)苯基氨基甲酸叔丁酯的制备
Figure imgf000038_0001
(4) 3-(2-(4-(2-(methylcarbamoyl) p-pyridin-4-yloxy)phenylamino)-5-(methylthio)pyrimidin-4-ylamino)benzene Preparation of tert-butyl carbamate
Figure imgf000038_0001
将 3- (2-氯- 5- (甲硫基)嘧啶- 4-基氨基)苯基氨基甲酸叔丁酯 (0.3 g, 0.82 mmol), 4- (4-氨基苯氧基) -N-甲基吡啶- 2-甲酰胺 (0.24 g, 0.99 mmol) 溶于叔丁醇 (5 mL), 加入 DIPEA (0.127 g, 0.98 mmol), 加热到 90°C反应 8 h, 冷至室温， 过滤， 干燥， 得固体 0.4 g, 收率： 85%。  tert-Butyl 3-(2-chloro-5-(methylthio)pyrimidin-4-ylamino)phenylcarbamate (0.3 g, 0.82 mmol), 4-(4-aminophenoxy)-N- Methylpyridine-2-formamide (0.24 g, 0.99 mmol) was dissolved in tert-butanol (5 mL), DIPEA (0.127 g, 0.98 mmol) was added, and the mixture was heated to 90 ° C for 8 h, cooled to room temperature and filtered. Drying gave a solid of 0.4 g, yield: 85%.
(5) 4- (4- (4- (3-氨基苯氨基) -5- (甲硫基)嘧啶- 2-基氨基)苯氧基)- 甲基吡啶 -2-甲酰胺的制备  (5) Preparation of 4-(4-(4-(3-aminophenylamino)-5-(methylthio)pyrimidin-2-ylamino)phenoxy)-methylpyridine-2-carboxamide
Figure imgf000038_0002
Figure imgf000038_0002
将 3- (2- (4- (2- (甲基氨基甲酰基) p比啶- 4-基氧基)苯氨基 )-5- (甲硫基) 嘧啶- 4-基氨基)苯基氨基甲酸叔丁酯 (0.4 g , 0.697 mmol)溶于 DCM (10 mL), 加入 CF3COOH (2 mL) 室温搅拌 2 h, 旋干， 直接用于下步反应。 3-(2-(4-(2-(methylcarbamoyl) p-pyridin-4-yloxy)phenylamino)-5-(methylthio)pyrimidin-4-ylamino)phenylamino carboxylate (0.4 g, 0.697 mmol) was dissolved in DCM (10 mL), was added CF 3 COOH (2 mL) was stirred at room temperature 2 h, spin dry, was used directly in the next step.
(6) 4- (4- (4- (3-丙烯酰氨基苯氨基)- 5- (甲硫基)嘧啶- 2-基氨基)苯氧 基)—TV-甲基吡啶- 2-甲酰胺  (6) 4-(4-(4-(3-acrylamidophenylamino)-5-(methylthio)pyrimidin-2-ylamino)phenoxy)-TV-picoline-2-formamide
Figure imgf000038_0003
Figure imgf000038_0003
将上步得到的 4- (4- (4- (3-氨基苯氨基) -5- (甲硫基)嘧啶- 2-基氨基)苯 氧基)—TV-甲基吡啶- 2-甲酰胺, DIPEA (0.18 g, 1.39 mmol) 溶于 THF (5 mL),冷却至 - 20 °C ,在 20 min内緩慢向体系中滴加丙烯酰氯 (0.63 g 0.696 mmol)的 2 mL THF溶液。直接浓缩，制备色语纯化 （H20: MeOH = 100： 50)得到产品白色固体 90 mg, 两步收率 24.5%。 4-(4-(4-(3-Aminophenylamino)-5-(methylthio)pyrimidin-2-ylamino)phenoxy)-TV-methylpyridine-2-formamide obtained in the above step , DIPEA (0.18 g, 1.39 mmol) dissolved in THF (5 mL), cooled to - 20 ° C, and slowly added acryloyl chloride (0.63 g 0.696 mmol) in 2 mL THF over 20 min. Direct concentration, preparative color purification (H 2 0: MeOH = 100: 50) gave product white solid, 90 mg, mp.
分子式： C27H25N703S 分子量： 527.17 盾语 (m/z): 528·2 (M+ Molecular formula: C 27 H 25 N 7 0 3 S Molecular weight: 527.17 Shield (m/z): 528·2 (M+
Ή- NMR(i 6- DMSO, 400 MHz) 5(ppm): 10.12 (1H, s), 9.51 (1H, s) 8.80-8.72 (2H, m), 8.47 (1H, d), 8.23 (1H, s), 7.87 (1H, m), 7.75 (2H, d) 7.42 (1H, d), 7.37-7.26 (3H, m), 7.07 (1H, dd), 6.95 (2H, d), 6.34 (1H, dd) 6.14 (1H, dd), 5.59 (1H, d), 2.77 (3H, d), 2.31 (3H, s). 实施例 2 4-(4-(4-(3-丙烯酰氨基苯氨基) -5- (甲磺酰基)嘧啶 -2-基 J 苯氧 -TV-甲基吡啶 -2-甲酰胺 (化合物 8)的制备 Ή-NMR (i 6 - DMSO, 400 MHz) 5 (ppm): 10.12 (1H, s), 9.51 (1H, s) 8.80-8.72 (2H, m), 8.47 (1H, d), 8.23 (1H, s), 7.87 (1H, m), 7.75 (2H, d) 7.42 (1H, d), 7.37-7.26 (3H, m), 7.07 (1H, dd), 6.95 (2H, d), 6.34 (1H, Dd) 6.14 (1H, dd), 5.59 (1H, d), 2.77 (3H, d), 2.31 (3H, s). Example 2 4-(4-(4-(3-acrylamidophenylamino)-5-(methylsulfonyl)pyrimidin-2-yl J phenoxy-TV-methylpyridine-2-carboxamide (Compound 8 Preparation
Figure imgf000039_0001
Figure imgf000039_0001
(1) 3- (2- (4- (2- (甲基氨基甲酰基) p比啶- 4-基氧基)苯氨基 )-5- (甲磺酰 基)嘧啶- 4-基氨基)苯基氨  (1) 3-(2-(4-(2-(methylcarbamoyl) p-pyridin-4-yloxy)phenylamino)-5-(methylsulfonyl)pyrimidin-4-ylamino)benzene Base ammonia
Figure imgf000039_0002
Figure imgf000039_0002
将 3- (2- (4- (2- (甲基氨基甲酰基) p比啶- 4-基氧基)苯氨基 )-5- (甲硫基) 嘧啶- 4-基氨基)苯基氨基甲酸叔丁酯 （0.32 g, 0.56 mmol )溶于 15 mL 甲醇和 5 mL水中， 然后加过氧化 酸氢钟复合盐 (oxone) ( 0.857 g, 1.39 mmol ) , 体系在常温下搅拌过夜， 用 50 mL二氯甲烷萃取， 200 mL水 洗涤两次， 有机相旋干得到 0.34 g淡黄色油状物， 直接用于下一步。  3-(2-(4-(2-(methylcarbamoyl) p-pyridin-4-yloxy)phenylamino)-5-(methylthio)pyrimidin-4-ylamino)phenylamino Tert-butyl formate (0.32 g, 0.56 mmol) was dissolved in 15 mL of methanol and 5 mL of water, then oxone (0.857 g, 1.39 mmol) was added and the system was stirred at room temperature overnight. It was extracted with methylene chloride, washed twice with 200 mL of water, and then evaporated to dryness.
(2) 4-(4-(4-(3-氨基苯氨基) -5- (甲磺酰基)嘧啶 -2-基氨基)苯氧基)- 甲基吡啶 -2-甲酰胺的制  (2) Preparation of 4-(4-(4-(3-aminophenylamino)-5-(methylsulfonyl)pyrimidin-2-ylamino)phenoxy)-methylpyridine-2-carboxamide
Figure imgf000039_0003
Figure imgf000039_0003
将上步得到的 3- (2- (4- (2- (甲基氨基甲酰基) p比啶- 4-基氧基)苯氨 基)— 5— (甲磺酰基)嘧啶— 4-基氨基)苯基氨基甲酸叔丁酯 0.34 g溶于 10 mL 的 DCM中， 冰浴下通 HC1气体反应 1 h, 将体系旋干， 得到产物粗品 0.36 g, 直接用于下一步。  3-(2-(4-(2-(methylcarbamoyl) p-pyridin-4-yloxy)phenylamino)-5-(methylsulfonyl)pyrimidin-4-ylamino 0.34 g of tert-butyl phenylcarbamate was dissolved in 10 mL of DCM, reacted with HC1 gas for 1 h in an ice bath, and the system was dried to give a crude product (0.36 g).
(3) 4- (4- (4- (3-丙烯酰氨基苯氨基) -5- (甲磺酰基)嘧啶- 2-基氨基)苯氧 基) -TV-甲基吡啶- 2-甲酰胺的制备
Figure imgf000040_0001
(3) 4-(4-(4-(3-Acrylamidophenylamino)-5-(methylsulfonyl)pyrimidin-2-ylamino)phenoxy)-TV-picoline-2-formamide Preparation
Figure imgf000040_0001
将上步产物粗品 0.36 g溶于 10 mL的 THF中 , 滴加 DIPEA, 直至 溶液 pH值为 7, - 15 °C下滴加含丙烯酰氯（ 0,035 g, 0.39 mmol )的 THF 溶液 1.5 mL, 加毕此温度下反应 2.5 h。 滴加 5滴甲醇， 将体系旋干， 制备液相纯化（甲醇： 水 =57% )得到白色固体 13 mg。 三步收率： 4.1% 分子式： C27H25N705S 分子量： 559.16 盾语 (m/z): 559.8 (M+0.36 g of the crude product in the previous step was dissolved in 10 mL of THF, and DIPEA was added dropwise until the pH of the solution was 7, and 15 mL of THF solution containing acryloyl chloride (0,035 g, 0.39 mmol) was added dropwise at -15 °C. The reaction was carried out for 2.5 h at this temperature. 5 drops of methanol were added dropwise, and the system was spin-dried to give a liquid phase purified (methanol: water = 57%) to afford a white solid. Three-step yield: 4.1% Molecular formula: C 27 H 25 N 7 0 5 S Molecular weight: 559.16 Shield (m/z): 559.8 (M+
H)+. H) + .
NMR(i 6- DMSO, 400 MHz) 5(ppm): 10.19 (2H, s), 8.93 (IH, s), 8.76 (IH, q), 8.50-8.47 (2H, m), 7.83-7.60 (3H, m), 7.45 (IH, d), 7.38-7.30 (2H, m), 7.23-7.16 (IH, m), 7.12-6.90 (3H, m), 6.40-6.27 (IH, m), 6.22-6.08 (IH, m), 5.67-5.57 (IH, m), 3.38 (3H, s), 2.77 (3H, d). NMR (i 6 - DMSO, 400 MHz) 5 (ppm): 10.19 (2H, s), 8.93 (IH, s), 8.76 (IH, q), 8.50-8.47 (2H, m), 7.83-7.60 (3H , m), 7.45 (IH, d), 7.38-7.30 (2H, m), 7.23-7.16 (IH, m), 7.12-6.90 (3H, m), 6.40-6.27 (IH, m), 6.22-6.08 (IH, m), 5.67-5.57 (IH, m), 3.38 (3H, s), 2.77 (3H, d).
实施例 3 4-(4-(4-(3-丙烯酰 J^ jQ-5-(2-甲氧基乙硫基)嘧啶 -2-基 JQ苯氧基 V-甲基吡啶 -2-甲酰胺 (化合物 10)的制备  Example 3 4-(4-(4-(3-acryloyl)J^jQ-5-(2-methoxyethylthio)pyrimidin-2-yl JQ phenoxy V-methylpyridine-2-methyl Preparation of amide (compound 10)
Figure imgf000040_0002
Figure imgf000040_0002
( 1 ) 2-甲氧基乙基硫代乙
Figure imgf000040_0003
(1) 2-methoxyethylthioethane
Figure imgf000040_0003
将 1-氯- 2-甲氧基乙烷 ( 10.00 g, 106 mmol ) 碳酸钾 （ 17.82 g, 129 mmol ) , 硫代醋酸钾 ( 14.50 g, 127 mmol )加入到 50 mL DMA中， 室温搅拌 5 h, 过滤除去固体， 倒入 200 mL水中， 乙酸乙酯萃取， 有机 相旋干得无色油状化合物， 直接用于下一步。 Add 1-chloro-2-methoxyethane (10.00 g, 106 mmol) potassium carbonate ( 17.82 g, 129 mmol), potassium thioacetate (1450 g, 127 mmol) to 50 mL DMA and stir at room temperature 5 h, remove solids by filtration, pour into 200 mL of water, extract with ethyl acetate, organic The compound was dried in the form of a colorless oily compound and used directly in the next step.
(2) 2-甲氧基乙硫醇的制备 将上步得到的产物溶于 50 mL丙酮和 50 mL水中， 加入氢氧化钠 ( 5.08 g, 12,7 mmol ) , 室温反应搅拌 5 h后， 30°CO下减压除去丙酮， 得到的水溶液直接用于下一步。 (2) Preparation of 2-methoxyethanethiol The product obtained in the above step was dissolved in 50 mL of acetone and 50 mL of water, and sodium hydroxide (5.08 g, 12,7 mmol) was added thereto, and the mixture was stirred at room temperature for 5 hours. the acetone was removed under reduced pressure at 30 ° C O, the resulting solution was used directly in the next step.
(3) 5- (2-甲氧基乙硫基)嘧 - 2,4(1//,3/ )-二酮的制备
Figure imgf000041_0001
(3) Preparation of 5-(2-methoxyethylthio)pyrimidine-2,4(1//,3/)-dione
Figure imgf000041_0001
将 5-溴尿嘧啶（ 10.00 g, 52.4 mmol)加入到上述的水溶液中， 回 流反应 16 h。溶液冷却至- 5°C,用浓盐酸调 pH值至 7,析出固体，抽滤， 干燥得到淡黄色固体 3.50 g, 收率 33%  5-Bromouracil (10.00 g, 52.4 mmol) was added to the above aqueous solution, and refluxed for 16 h. The solution was cooled to -5 ° C, and the pH was adjusted to 7 with concentrated hydrochloric acid to precipitate a solid, which was filtered and dried to give a pale yellow solid, 3.50 g, yield 33%
(4) 2,4-二氯- 5- (2-甲氧基 制备
Figure imgf000041_0002
(4) Preparation of 2,4-dichloro-5-(2-methoxy)
Figure imgf000041_0002
将 5- (2-甲氧基乙硫基)嘧啶- 2,4(1//,3/ )-二酮 ( 3.00 g, 14.8 mmol)加 到 50 mL P0C13中，滴加 2 mL的 二甲基苯胺，加毕 110 °C反应 4 h 旋蒸除去溶剂，加入 20 mL甲苯再次旋千体系 ,得到油状物倒入冰水中 , 用 Na2C03水溶液中和到中性， 乙酸乙酯萃取， 浓缩有机相， 过柱 ( PE/EA:1:4 ) 得到棕色固体 620 mg， 收率 17,5% Add 5-(2-methoxyethylthio)pyrimidine-2,4(1//,3/)-dione (3.100 g, 14.8 mmol) to 50 mL of P0C1 3 and add 2 mL of 2 Methyl aniline was added to the reaction at 110 °C for 4 h. The solvent was removed by rotary evaporation. Toluene (20 mL) was added to the residue to give the oil. The oil was poured into ice water, neutralized with Na 2 CO 3 aqueous solution to neutral, and extracted with ethyl acetate. , concentrated organic phase, passed through a column (PE/EA: 1:4) to give a brown solid 620 mg, yield 17.5%
(5) 3- (2-氯- 5- (2-甲氧基乙硫基)嘧啶- 4-基氨基)苯基氨基甲酸叔丁酯 的制备
Figure imgf000041_0003
(5) Preparation of 3-(2-chloro-5-(2-methoxyethylthio)pyrimidin-4-ylamino)phenylcarbamic acid tert-butyl ester
Figure imgf000041_0003
将 2,4-二氯- 5- (2-甲氧基乙硫基)嘧啶（0.60 g, 2.51 mmol) , DIPEA ( 0.356 g, 2.76 mmol)加到 10 mL的正丁醇中， 再加入 3-氨基苯基氨 基甲酸叔丁酯 （ 0.523 g, 2.51 mmol) , 体系在 100°C搅拌 6 ii。 浓缩， 过柱（ PE:EA 10:1- 1:2 )得到白色固体 0.50 g， 收率 48.6%,  Add 2,4-dichloro-5-(2-methoxyethylthio)pyrimidine (0.60 g, 2.51 mmol), DIPEA (0.356 g, 2.76 mmol) to 10 mL of n-butanol, then add 3 tert-Butyl aminophenylcarbamate (0.523 g, 2.51 mmol), the system was stirred at 100 ° C for 6 ii. Concentration, passing through a column (PE: EA 10:1 - 1:2) gave a white solid, 0.50 g, yield 48.6%.
(6) 3- (5- (2-甲氧基乙硫基 )-2- (4- (2- (甲基氨基甲酰基)吡啶- 4-基氧基) 苯氨基)嘧啶- 4-基氨基)苯基氨基甲酸叔丁酯的制备
Figure imgf000042_0001
(6) 3-(5-(2-Methoxyethylthio)-2-(4-(2-(methylcarbamoyl)pyridine-4-yloxy)phenylamino)pyrimidin-4-yl Preparation of tert-butyl amino)phenylcarbamate
Figure imgf000042_0001
将 3- (2-氯- 5- (2-甲氧基乙硫基)嘧啶- 4-基氨基)苯基氨基甲酸叔丁 酯（0.50 g, 1.22 mmol )溶于 10 mL的叔戊醇中， 再加入 4- (4-氨基苯氧 基)—TV-甲基吡啶- 2-甲酰胺（ 0.296 mg, 1.22 mmol ) , 醋酸 2滴， 反应在 85 °C搅拌 1 h。 浓缩， 柱层析（ PE: EA-10: 1-—2: 1 )得到白色固体 0.25 g， 收率 33.2%,  tert-Butyl 3-(2-chloro-5-(2-methoxyethylthio)pyrimidin-4-ylamino)phenylcarbamate (0.50 g, 1.22 mmol) was dissolved in 10 mL of tert-pentanol Further, 4-(4-aminophenoxy)-TV-methylpyridine-2-formamide (0.296 mg, 1.22 mmol) and 2 ml of acetic acid were added, and the mixture was stirred at 85 ° C for 1 h. Concentration, column chromatography (PE: EA-10: 1- 2:1) gave 0.25 g of white solid, yield 33.2%.
(7) 4- (4- (4- (3-氨基苯氨基) -5- (2-甲氧基乙硫基)嘧啶- 2-基氨基)苯氧 基)—TV-甲基吡啶- 2-甲酰胺  (7) 4-(4-(4-(3-Aminophenylamino)-5-(2-methoxyethylthio)pyrimidin-2-ylamino)phenoxy)-TV-methylpyridine-2 -formamide
Figure imgf000042_0002
Figure imgf000042_0002
将 3- (5- (2-甲氧基乙硫基 )-2- (4- (2- (甲基氨基甲酰基) p比啶- 4-基氧基) 苯氨基)嘧啶- 4-基氨基)苯基氨基甲酸叔丁酯 （0.15 g, 0.24 mmol )溶于 10 mL的 DCM中， 冰浴下通 HC1气体反应 1 h, 浓缩， 得到产物粗品 160 mg, 直接用于下一步。  3-(5-(2-Methoxyethylthio)-2-(4-(2-(methylcarbamoyl) p-pyridin-4-yloxy)phenylamino)pyrimidin-4-yl tert-Butyl amino)phenylcarbamate (0.15 g, 0.24 mmol) was dissolved in 10 mL of DCM. EtOAc was evaporated.
(8) 4- (4- (4- (3-丙烯酰氨基苯氨基 )-5- (2-甲氧基乙硫基)嘧啶- 2-基氨 基)苯氧基) -TV-甲基吡啶- 2-  (8) 4-(4-(4-(3-Acrylamidophenylamino)-5-(2-methoxyethylthio)pyrimidin-2-ylamino)phenoxy)-TV-methylpyridine - 2-
Figure imgf000042_0003
Figure imgf000042_0003
将上步得到的粗品 160 mg溶于 10 mL的 THF中， 滴加 DIPEA, 直至溶液 pH值为 7 , -15 °C下滴加含丙烯酰氯（ 0,022 g, 0.24 mmol )的 THF溶液 0.5 mL , 加毕此温度下反应 15 min。 滴加 5滴甲醇， 将体系 旋干， 制备液相纯化（甲醇： 水 =62% )得到白色固体 17 mg。 两步收率： 12.5%。 分子式： C29H29N704S 分子量： 571.20 盾语 (m/z): 572.2The 160 mg of the crude product obtained in the previous step was dissolved in 10 mL of THF, and DIPEA was added dropwise until the pH of the solution was 7, and 0.5 mL of a solution of acryloyl chloride (0,022 g, 0.24 mmol) in THF was added dropwise at -15 °C. The reaction was carried out at this temperature for 15 min. 5 drops of methanol were added dropwise, the system was spun dry, and purified by liquid phase (methanol: water = 62%) to afford 17 mg of white solid. Two-step yield: 12.5%. Molecular formula: C 29 H 29 N 7 0 4 S Molecular weight: 571.20 Shield (m/z): 572.2
(M+ H)+. (M+ H) + .
NMR(CX)(¾, 400 MHz) 5(ppm): 8.40-8.36 (2H, m), 8.25 (IH, s), 8.21 (IH, br s), 8.07 (IH, d), 8.00 (IH, s), 7.69 (IH, d), 7.57 (2H, d), 7.52 (IH, s), 7.38-7.30 (IH, m), 7.29-7.21 (2H, m), 7.02-6.94 (3H, m), 6.38 (IH, d), 6.18 (IH, dd), 5.63 (IH, d), 3.53 (2H, t), 3.39 (3H, s), 3.03 (3H, d), 2.85 (2H, t).  NMR (CX) (3⁄4, 400 MHz) 5 (ppm): 8.40-8.36 (2H, m), 8.25 (IH, s), 8.21 (IH, br s), 8.07 (IH, d), 8.00 (IH, s), 7.69 (IH, d), 7.57 (2H, d), 7.52 (IH, s), 7.38-7.30 (IH, m), 7.29-7.21 (2H, m), 7.02-6.94 (3H, m) , 6.38 (IH, d), 6.18 (IH, dd), 5.63 (IH, d), 3.53 (2H, t), 3.39 (3H, s), 3.03 (3H, d), 2.85 (2H, t).
实施例 4 4-(4-(4-(3-丙烯酰 J^ J -5-i2-吗啉基乙硫基)嘧啶 - -基 JQ苯氧基) -N-甲基吡啶 -2-甲酰胺 (化合物 11)的制备  Example 4 4-(4-(4-(3-acryloyl J^J-5-i2-morpholinylethylthio)pyrimidin-yl-JQ phenoxy)-N-methylpyridine-2-methyl Preparation of amide (Compound 11)
Figure imgf000043_0001
Figure imgf000043_0001
(1) 2-吗啉- 4-基乙基亚氨 酯的制备
Figure imgf000043_0002
(1) Preparation of 2-morpholine-4-ylethyliminoimide
Figure imgf000043_0002
将 V-(2-氯乙基)吗啉盐酸盐 （9.3 g, 0.05 mol) 、 硫脲（3.81 g, 0.05 mol)溶于 95%的乙醇 （50 mL) 中， 加热回流 24小时， 冷却， 浓缩， 得到白色固体直接用于下一步。  V-(2-chloroethyl)morpholine hydrochloride (9.3 g, 0.05 mol), thiourea (3.81 g, 0.05 mol) in 95% ethanol (50 mL), heated to reflux for 24 hours, cooled Concentrated to give a white solid which was taken directly to next.
(2) 4,4'- [二硫基双（乙烷 - -二基)]二吗啉的制备
Figure imgf000043_0003
(2) Preparation of 4,4'-[dithiobis(ethane-diyl)]dimorpholine
Figure imgf000043_0003
将上步所得粗产品（约 0.05 mol)溶于氢氧化钠（ 2 g, 0.05 mol)的 水溶液（30 mL) 中， 加热回流 2 h, 冷却， 用稀盐酸调 pH到 5, 乙酸 乙酯萃取， 无水 酸钠干燥， 浓缩得产品 6.02 g, 收率 82.4%。  The crude product obtained in the previous step (about 0.05 mol) was dissolved in an aqueous solution (30 mL) of sodium hydroxide (2 g, 0.05 mol), heated under reflux for 2 h, cooled, adjusted to pH 5 with dilute hydrochloric acid and extracted with ethyl acetate. The organic sodium salt was dried and concentrated to give a product of 6.02 g, yield: 82.4%.
(3) 2-吗啉基乙硫醇的制备 将 4,4'- [二硫基双（乙烷 -2,1-二基)]二吗啉（6.02 g, 20.6 mmol) 溶于 无水乙醇 （40 mL) 中， 加入硼氢化钠 （3.8 g, 0.1 mol) 室温下反应 12 小时， 加入水， 减压浓缩去大部分溶剂， 乙酸乙酯萃取， 无水石 酸钠干 燥， 旋干， 得油状粗产品直接用于下一步。 (3) Preparation of 2-morpholinylethanethiol 4,4'-[Dithiobis(ethane-2,1-diyl)]dimorpholine (6.02 g, 20.6 mmol) was dissolved in absolute ethanol (40 mL) and sodium borohydride (3.8) was added. g, 0.1 mol) After reacting for 12 hours at room temperature, water is added, and the solvent is concentrated under reduced pressure. The solvent is evaporated, ethyl acetate is evaporated, dried over sodium sulfate, and evaporated to dryness.
(4) 5- (2-吗啉基乙硫基) - 2,4(1//,3/)-二酮的制备
Figure imgf000044_0001
(4) Preparation of 5-(2-morpholinoethylthio)-2,4(1//,3/)-dione
Figure imgf000044_0001
将上步所得粗品 （约 41.2 mol) 、 氢氧化钠 （ 1.6 g, 0.04 mol)和 5- 溴尿嘧啶（ 6.36 g, 33.3 mmol) 溶于水（ 50 mL ) 中加热回流反应 5 h, 冷却， 加稀盐酸调至中性， 析出白色固体， 抽滤， 干燥， 得到产品 3.7 g, 收率 43.2%。  The crude product (about 41.2 mol), sodium hydroxide (1.6 g, 0.04 mol) and 5-bromouracil (6.36 g, 33.3 mmol) dissolved in water (50 mL) were heated and refluxed for 5 h, and cooled. The mixture was adjusted to neutral with dilute hydrochloric acid, and a white solid was precipitated, filtered, and dried to give 3.7 g of product, yield 43.2%.
(5) 4- (2- (2,4-二氯嘧啶- -基硫基）乙基)吗啉的制备
Figure imgf000044_0002
(5) Preparation of 4-(2-(2,4-dichloropyrimidinyl-yl)ethyl)morpholine
Figure imgf000044_0002
将 5- (2-吗啉基乙硫基)嘧啶- 2,4(1//,3/)-二酮（ 3.7 g, 14.4 mol)溶于 三氯氧磷 （50mL) 中， 回流反应 8h, 冷却， 减压浓缩大部分溶剂后， 将剩余物倒入冰水中， 乙酸乙酯萃取， 无水硫酸钠干燥， 浓缩后硅胶柱 层析 （石油醚： 乙酸乙酯 =5:1 )得到淡黄色固体 1.72 g, 收率 40.6%。  5-(2-morpholinylethylthio)pyrimidine-2,4(1//,3/)-dione (3.7 g, 14.4 mol) was dissolved in phosphorus oxychloride (50 mL) and refluxed for 8 h. After cooling, the solvent was concentrated under reduced pressure. EtOAc was evaporated. The yellow solid was 1.72 g, yield 40.6%.
(6) 3- (2-氯- 5- (2-吗啉基乙硫基)嘧啶- 4-基氨基)苯基氨基甲酸叔丁酯 的制备
Figure imgf000044_0003
(6) Preparation of 3-(2-chloro-5-(2-morpholinoethylthio)pyrimidin-4-ylamino)phenylcarbamic acid tert-butyl ester
Figure imgf000044_0003
将 4- (2- (2,4-二氯嘧啶- 5-基硫基）乙基)吗啉 ( 1.72 g, 5.85 mmol) 、 3-氨基苯基氨基甲酸叔丁酯 （ 1.22 g, 5.85 mmol) 和 二异丙基乙胺 ( 908 mg, 7.03 mmol) 溶于叔戊醇中， 回流状态下反应 12 h, 冷却， 浓 缩后硅胶柱层析 （石油醚： 乙酸乙酯 =3:1)得到淡黄色固体 1.48 g, 收 率 54.4%。  4-(2-(2,4-Dichloropyrimidin-5-ylthio)ethyl)morpholine ( 1.72 g, 5.85 mmol), tert-butyl 3-aminophenylcarbamate ( 1.22 g, 5.85 mmol And diisopropylethylamine (908 mg, 7.03 mmol) dissolved in tert-amyl alcohol, reacted under reflux for 12 h, cooled, concentrated and purified by silica gel column chromatography ( petroleum ether: ethyl acetate = 3:1) Light yellow solid 1.48 g, yield 54.4%.
(7) 3- (2- (4- (2- (甲基氨基甲酰基) p比啶- 4-基氧基)苯氨基 )-5- (2-吗啉基 乙硫基)嘧啶- 4-基氨基)苯基氨基甲酸叔丁酯的制备
Figure imgf000045_0001
(7) 3-(2-(4-(2-(methylcarbamoyl) p-pyridin-4-yloxy)phenylamino)-5-(2-morpholinylethylthio)pyrimidine-4 Preparation of tert-butyl phenylamino)phenylcarbamate
Figure imgf000045_0001
将 3- (2-氯- 5- (2-吗啉基乙硫基)嘧啶- 4-基氨基)苯基氨基甲酸叔丁酯 ( 1.48 g, 3.18 mmol )、 4- (4-氨基苯氧基) -TV-甲基吡啶- 2-甲酰胺（ 774 mg, 3.18 mmol ) 和两滴冰乙酸溶于正丁醇中， 回流状态下反应 12 h, 冷却， 浓缩后硅胶柱层析（石油醚： 乙酸乙酯 =1 : 1 )得到白色固体 820 mg, 收 率 38.4%。  3-(2-Chloro-5-(2-morpholinoethylthio)pyrimidin-4-ylamino)phenylcarbamic acid tert-butyl ester ( 1.48 g, 3.18 mmol), 4-(4-aminophenoxyl) -TV-methylpyridine-2-ylformamide (774 mg, 3.18 mmol) and two drops of glacial acetic acid dissolved in n-butanol, reacted under reflux for 12 h, cooled, concentrated and purified by silica gel column chromatography : ethyl acetate = 1 : 1 ) gave 820 mg of white solid.
(8) 4- (4- (4- (3-丙烯酰氨基苯氨基) -5- (2-吗啉基乙硫基)嘧啶- 2-基氨 基)苯氧基)—TV-甲基吡啶- 2-  (8) 4-(4-(4-(3-Acrylamidophenylamino)-5-(2-morpholinylethylthio)pyrimidin-2-ylamino)phenoxy)-TV-methylpyridine - 2-
Figure imgf000045_0002
Figure imgf000045_0002
将 3- (2- (4- (2- (甲基氨基甲酰基) p比啶- 4-基氧基)苯氨基 )-5- (2-吗啉基 乙硫基)嘧啶- 4-基氨基)苯基氨基甲酸叔丁酯（ 820 g, 1.22 mmol )溶于二 氯甲烷 （60 mL ) , 慢慢滴加三氟乙酸（ 695 mg, 6. 10 mmol ) , 室温下 搅拌 6 h后， 减压浓缩后溶于无水四氢呋喃 （30 mL ) , 加入 二异 丙基乙胺（ 1.26 g, 9.75 mmol ) , - 20 °C下向其中慢慢滴加丙烯酰氯（ 1 10 mg, 1.22 mmol ) , 搅拌 2小时， 加入无水甲醇淬灭， 浓缩， 硅胶柱层析 (石油醚： 乙酸乙酯 =2: 1 ) , 得到白色固体 177 mg, 收率 23.1 %。  3-(2-(4-(2-(methylcarbamoyl) p-pyridin-4-yloxy)phenylamino)-5-(2-morpholinylethylthio)pyrimidin-4-yl tert-Butyl amino)phenylcarbamate (820 g, 1.22 mmol) was dissolved in dichloromethane (60 mL), then trifluoroacetic acid ( 695 mg, 6.10 mmol) was slowly added dropwise and stirred at room temperature for 6 h. After concentration under reduced pressure, it was dissolved in anhydrous tetrahydrofuran (30 mL), diisopropylethylamine (1.26 g, 9.75 mmol) was added, and acryloyl chloride (1 10 mg, 1.22 mmol) was slowly added dropwise at -20 °C. The mixture was stirred for 2 hr. EtOAc (EtOAc)EtOAc.
分子式： C32H34N804S 分子量： 626.24 盾语 (m/z): 627.3(M+H)+ NMR(CX)(¾, 400 MHz) 5(ppm): 8.35 (IH, d), 8.04 (IH, q), 7.77 (IH, s), 7.72-7.64 (2H, m), 7.53 (2H, d), 7.15 (IH, s), 7.05 (IH, t), 7.01-6.88 (4H, m), 6.68 (IH, d), 6.41 (IH, dd), 6.32 (IH, d), 6.24 (IH, dd), 5.72 (IH, d), 4.02 (IH, br s), 3.93-3.87 (2H, m), 3.84 (2H, t), 3.75 (2H, t), 3.65 (2H, t), 3.26 (2H, t), 3.01 (3H, d), 2.95-2.89 (2H, m). Molecular formula: C 32 H 34 N 8 0 4 S Molecular weight: 626.24 Shield (m/z): 627.3 (M+H) + NMR (CX) (3⁄4, 400 MHz) 5 (ppm): 8.35 (IH, d) , 8.04 (IH, q), 7.77 (IH, s), 7.72-7.64 (2H, m), 7.53 (2H, d), 7.15 (IH, s), 7.05 (IH, t), 7.01-6.88 (4H , m), 6.68 (IH, d), 6.41 (IH, dd), 6.32 (IH, d), 6.24 (IH, dd), 5.72 (IH, d), 4.02 (IH, br s), 3.93-3.87 (2H, m), 3.84 (2H, t), 3.75 (2H, t), 3.65 (2H, t), 3.26 (2H, t), 3.01 (3H, d), 2.95-2.89 (2H, m).
实施例 5 Λ^3-ί2-(4-(2-甲氧基乙氧基)^ jQ-5- (甲硫基)嘧啶 -4- 基氨基)苯基)丙烯酰胺 (化合物 13)的制备 ■HC
Figure imgf000046_0001
Example 5 Preparation of Λ^3-ί2-(4-(2-methoxyethoxy)^jQ-5-(methylthio)pyrimidin-4-ylamino)phenyl)acrylamide (Compound 13) ■HC
Figure imgf000046_0001
(1) 3- (2- (4- (2-甲氧基乙氧基)苯氨基 )-5- (甲硫基)嘧啶- 4-基氨基)苯 基氨基甲酸叔丁酯的制  (1) Preparation of tert-butyl 3-(2-(4-(2-methoxyethoxy)phenylamino)-5-(methylthio)pyrimidin-4-ylamino)phenylcarbamate
Figure imgf000046_0002
Figure imgf000046_0002
将 3- (2-氯- 5- (甲硫 - 0.25 g, 0.68 mmol )溶于 10 mL的叔戊醇中， 再加入醋酸 2滴， 4- (2-甲氧基乙 氧基)苯胺（0.119 g, 0.71 mmol ) , 反应在 85 Ό搅拌 1 h。 将体系旋千， 柱层析（ PE: EA-8: 1—2:1 )得到白色固体 0.15 g， 收率 44.1%。  3-(2-Chloro-5-methylsulfate-0.25 g, 0.68 mmol) was dissolved in 10 mL of tert-amyl alcohol, followed by 2 drops of acetic acid, 4-(2-methoxyethoxy)aniline ( 0.119 g, 0.71 mmol), the reaction was stirred at 85 ° C for 1 h. The system was purified by column chromatography (PE: EA-8: 1-2:1) to give a white solid 0.15 g, yield 44.1%.
(2) (3-氨基苯基)- Λ^- (4- (2-甲氧基乙氧基)苯基)—5- (甲硫基)嘧啶 -2,4-二胺的制备  (2) Preparation of (3-aminophenyl)- Λ^-(4-(2-methoxyethoxy)phenyl)-5-(methylthio)pyrimidine-2,4-diamine
Figure imgf000046_0003
Figure imgf000046_0003
将 3- (2- (4- (2-甲氧基乙氧基)苯氨基 )-5- (甲硫基)嘧啶- 4-基氨基)苯基 氨基甲酸叔丁酯 （0.14 g, 0.28 mmol )溶于 10 mL的 DCM中， 冰浴下 通 HC1气体反应 l h, 将体系旋干， 得到粗品 0.14 g, 直接用于下一步。  tert-Butyl 3-(2-(4-(2-methoxyethoxy)phenylamino)-5-(methylthio)pyrimidin-4-ylamino)phenylcarbamate (0.14 g, 0.28 mmol Dissolved in 10 mL of DCM, reacted with HC1 gas for 1 h in an ice bath, and the system was spun dry to obtain a crude product of 0.14 g, which was used directly in the next step.
(3) V-(3- (2- (4- (2-甲氧基乙氧基)苯氨基 )-5- (甲硫基)嘧啶- 4-基氨基) 苯基）丙烯酰胺的制备
Figure imgf000047_0001
(3) Preparation of V-(3-(2-(4-(2-methoxyethoxy)phenylamino)-5-(methylthio)pyrimidin-4-ylamino)phenyl)acrylamide
Figure imgf000047_0001
将上步产物粗品 0.14 g溶于 10 mL的 THF中 , 滴加 DIPEA, 直至 溶液 pH值为 7, -15 °C下滴加含丙烯酰氯（ 0.026 g, 0,29 mmol )的 THF 溶液 0.5 mL, 加毕此温度下反应 15 min。 滴加 5滴甲醇， 将体系旋干， 制备液相纯化（甲醇 /水 =65% )得到白色固体 11 mg。 两步收率： 8.7%。  0.14 g of the crude product in the previous step was dissolved in 10 mL of THF, and DIPEA was added dropwise until the pH of the solution was 7, and THF solution containing acryloyl chloride (0.026 g, 0,29 mmol) was added dropwise at -15 °C. , add the reaction at this temperature for 15 min. 5 drops of methanol were added dropwise, and the system was spin-dried to prepare a liquid phase (methanol / water = 65%) to afford a white solid 11 mg. Two-step yield: 8.7%.
分子式： C23H25N503S 分子量： 451.17 盾语 (m/z): 452.2Molecular formula: C 23 H 25 N 5 0 3 S Molecular weight: 451.17 Shield (m/z): 452.2
(M+ H)+. (M+ H) + .
NMR(0)C¾, 400 MHz) 5(ppm): 8.24 (IH, s), 8.1 1 (IH, s), 8.03 (IH, s), 7.55-7.47 (IH, m), 7.45 (2H, d), 7.32-7.27 (IH, m), 7.25-7.20 (IH, m), 7.16-7.04 (2H, m), 6.93 (2H, d), 6.46 (IH, dd), 6.26 (IH, dd), 5.80 (IH, dd), 4.12 (2H, t), 3.76 (2H, t), 3.46 (3H, s), 2.28 (3H, s).  NMR(0)C3⁄4, 400 MHz) 5(ppm): 8.24 (IH, s), 8.1 1 (IH, s), 8.03 (IH, s), 7.55-7.47 (IH, m), 7.45 (2H, d ), 7.32-7.27 (IH, m), 7.25-7.20 (IH, m), 7.16-7.04 (2H, m), 6.93 (2H, d), 6.46 (IH, dd), 6.26 (IH, dd), 5.80 (IH, dd), 4.12 (2H, t), 3.76 (2H, t), 3.46 (3H, s), 2.28 (3H, s).
实施例 6 V-i3-i2-(6-甲氧基吡啶 -3-基 J -5- (甲硫基)嘧啶 -4-基  Example 6 V-i3-i2-(6-methoxypyridin-3-yl J-5-(methylthio)pyrimidin-4-yl
Figure imgf000047_0002
Figure imgf000047_0002
(1) 3- (2- (6-甲氧基 p比啶- 3-基氨基 )-5- (甲硫基)嘧啶- 4-基氨基)苯基氨 基甲酸叔丁酯的制备  (1) Preparation of tert-butyl 3-(2-(6-methoxy p-pyridin-3-ylamino)-5-(methylthio)pyrimidin-4-ylamino)phenylcarbamate
Figure imgf000047_0003
Figure imgf000047_0003
将 3- (2-氯- 5- (甲硫基)嘧啶- 4-基氨基)苯基氨基甲酸叔丁酯 （ 0.25 g, 0.68 mmol )溶于 5 mL的叔戊醇中， 再加入醋酸 2滴， 6-甲氧基吡啶- 3- 胺（0.088 g, 0.71 mmol ) , 反应在 85°C搅拌 1 h。 反应冷却至室温， 加 入 iO mL 乙醚， 过滤， 得灰白色固体 0。21 g, 收率 67.6%,  tert-Butyl 3-(2-chloro-5-(methylthio)pyrimidin-4-ylamino)phenylcarbamate (0.25 g, 0.68 mmol) was dissolved in 5 mL of tert-amyl alcohol, then acetic acid 2 was added 6-Methoxypyridine-3-amine (0.088 g, 0.71 mmol) was added dropwise, and the mixture was stirred at 85 ° C for 1 h. The reaction was cooled to room temperature, then EtOAc (EtOAc) was evaporated.
(2) (3-氨基苯基)- N2- (6-甲氧基 p比啶- 3-基)- 5- (甲硫基)嘧啶- 2,4-二 胺的制备
Figure imgf000048_0001
(2) Preparation of (3-aminophenyl)-N 2 -(6-methoxyp-pyridin-3-yl)- 5-(methylthio)pyrimidine-2,4-diamine
Figure imgf000048_0001
将 3- (2- (6-甲氧基 p比啶- 3-基氨基 )-5- (甲硫基)嘧啶- 4-基氨基)苯基氨 基甲酸叔丁酯 （0.21 g, 0.46 mmol )溶于 10 mL的 DCM中， 冰浴下通 HC1气体反应 1 h,将体系旋干，得到产物粗品 0.22 g,直接用于下一步。  tert-Butyl 3-(2-(6-methoxyp-pyridin-3-ylamino)-5-(methylthio)pyrimidin-4-ylamino)phenylcarbamate (0.21 g, 0.46 mmol) Dissolved in 10 mL of DCM, reacted with HC1 gas for 1 h in an ice bath, and the system was spun dry to obtain 0.22 g of crude product, which was directly used for the next step.
(3) V-(3-(2-(6-甲氧基 p比啶- 3-基氨基 )-5- (甲硫基)嘧啶- 4-基氨基)苯 基）丙烯酰胺的制备  (3) Preparation of V-(3-(2-(6-methoxy p-pyridin-3-ylamino)-5-(methylthio)pyrimidin-4-ylamino)phenyl)acrylamide
Figure imgf000048_0002
Figure imgf000048_0002
将上步产物粗品 0.22 g溶于 10 mL的 THF中， 滴加 DIPEA, 直至 溶液 pH值为 7 , - 15 °C下滴加含丙烯酰氯 ( 0.042 g， 0.46 mmol )的 THF 溶液 0.5 mL , 加毕此温度下反应 15 min。 滴加 5滴甲醇， 将体系旋干， 制备液相纯化（甲醇：水 =67% )得到白色固体 27 mg。两步收率： 14.3%。  0.22 g of the crude product in the previous step was dissolved in 10 mL of THF, and DIPEA was added dropwise until the pH of the solution was 7, and 15 mL of THF solution containing acryloyl chloride (0.042 g, 0.46 mmol) was added dropwise at -15 °C. The reaction was carried out for 15 min at this temperature. 5 drops of methanol were added dropwise, and the system was spin-dried to prepare a liquid phase (methanol: water = 67%) to afford a white solid. Two-step yield: 14.3%.
分子式： C20H20N6O2S 分子量： 408.14 盾语 (m/z): 409.0Molecular formula: C 20 H 20 N 6 O 2 S Molecular weight: 408.14 Shield (m/z): 409.0
(M+ H)+. (M+ H) + .
NMR(DMSO-i 6, 400 MHz) 5(ppm): 10.1 1 (IH, s), 9.25 (IH, s), 8.69 (IH, s), 8.26 (IH, s), 8. 18 (IH, s), 7.96 (IH, dd), 7.87 (IH, s), 7.45-7.32 (2H, m), 7.26 (IH, t), 6.58 (IH, d), 6.43 (IH, dd), 6.23 (IH, dd), 5.74 (IH, dd), 3.75 (3H, s), 2.29 (3H, s).  NMR (DMSO-i 6, 400 MHz) 5 (ppm): 10.1 1 (IH, s), 9.25 (IH, s), 8.69 (IH, s), 8.26 (IH, s), 8. 18 (IH, s), 7.96 (IH, dd), 7.87 (IH, s), 7.45-7.32 (2H, m), 7.26 (IH, t), 6.58 (IH, d), 6.43 (IH, dd), 6.23 (IH , dd), 5.74 (IH, dd), 3.75 (3H, s), 2.29 (3H, s).
实施例 7 V-i3-i5-i甲硫基 )-2-(4-吗啉苯 JQ嘧啶 -4-基 ^)苯基) 丙烯酰胺 (化合物 16)的制备  Example 7 Preparation of V-i3-i5-imethylthio]-2-(4-morpholinylbenzene JQ pyrimidin-4-yl^)phenyl)acrylamide (Compound 16)
Figure imgf000048_0003
将对氟硝基苯 ( 5.00 g, 35.4 mmol )和吗啉 ( 9.25 g, 106 mmol ) 溶于甲苯溶液中， 反应体系回流 5 h, 旋干得到黄色固体 7.06 g, 收率 95.8%。
Figure imgf000048_0003
The p-fluoronitrobenzene (5.00 g, 35.4 mmol) and morpholine (9.25 g, 106 mmol) were dissolved in a toluene solution, and the reaction system was refluxed for 5 h, and then dried to give a white solid, 7.06 g, yield 95.8%.
(2) 4-吗啉苯胺的制备
Figure imgf000049_0001
(2) Preparation of 4-morpholinaniline
Figure imgf000049_0001
将 4— (4-硝基苯基)吗啉 ( 2.20 g, 10.6 mmol )溶于 12 mL甲醇中, 置换氮气， 加入 30 mg 10%的钯炭， 体系置换氢气三次， 常温下反应搅 拌 17 h, 滤去钯炭， 旋干滤液得淡黄色油状物 1.65 g, 收率： 87.4%。  Dissolve 4-(4-nitrophenyl)morpholine (2.20 g, 10.6 mmol) in 12 mL of methanol, replace nitrogen, add 30 mg of 10% palladium on carbon, replace the hydrogen three times, and stir at room temperature for 17 h. Palladium on charcoal was filtered off, and the filtrate was evaporated to give a pale yellow oil, 1.65 g, yield: 87.4%.
(3) 3- (5- (甲硫基)- 2- (4-吗啉苯氨基)嘧啶- 4-基氨基)苯基氨基甲酸叔 丁酯的制备  (3) Preparation of 3-(5-(methylthio)-2-(4-morpholinylamino)pyrimidin-4-ylamino)phenylcarbamic acid tert-butyl ester
Figure imgf000049_0002
Figure imgf000049_0002
将 3- (2-氯- 5- (甲硫基)嘧啶- 4-基氨基)苯基氨基甲酸叔丁酯 （ 0.25 g, 0.68 mmol ),溶于 5 mL的叔戊醇中，再加入醋酸 2滴， 4-吗啉苯胺（ 0.121 g, 0.68 mmol ) , 反应在 85 °C搅拌 1 h。 反应冷却至室温， 加入 10 mL 乙醚， 过滤， 得灰白色固体 0.27 g, 收率 77,9%。  tert-Butyl 3-(2-chloro-5-(methylthio)pyrimidin-4-ylamino)phenylcarbamate (0.25 g, 0.68 mmol), dissolved in 5 mL of tert-amyl alcohol, then acetic acid Two drops of 4-morpholinaniline (0.121 g, 0.68 mmol) were stirred at 85 ° C for 1 h. The reaction was cooled to room temperature, then added with 10 mL of ethyl ether and filtered to afford white crystals (yield:
(4) (3-氨基苯基)- 5- (甲硫基)- Λ^- (4-吗啉苯基)嘧啶- 2,4-二胺的制 备  (4) Preparation of (3-aminophenyl)-5-(methylthio)- Λ^-(4-morpholinylphenyl)pyrimidine-2,4-diamine
Figure imgf000049_0003
Figure imgf000049_0003
将 3- (5- (甲硫基)- 2- (4-吗啉苯氨基)嘧啶- 4-基氨基)苯基氨基甲酸叔 丁酯（ 0.27 g, 0.53 mmol )溶于 10 mL的 DCM中， 冰浴下通 HC1气体 反应 l h , 将体系旋干， 得到粗品 0.28 g,直接用于下一步。  tert-Butyl 3-(5-(methylthio)-2-(4-morpholinophenyl)pyrimidin-4-ylamino)phenylcarbamate (0.27 g, 0.53 mmol) was dissolved in 10 mL DCM Under the ice bath, the HC1 gas was reacted for 1 h, and the system was spun dry to obtain a crude product of 0.28 g, which was directly used in the next step.
(5) V-(3- (5- (甲硫基)- 2- (4-吗啉苯氨基)嘧啶- 4-基氨基)苯基）丙烯酰 胺的制备
Figure imgf000050_0001
(5) Preparation of V-(3-(5-(methylthio)-2-(4-morpholinanilinyl)pyrimidin-4-ylamino)phenyl)acrylamide
Figure imgf000050_0001
将 N4— (3-氨基苯基)- 5- (甲硫基)- TV2- (4-吗啉苯基)嘧啶- 2,4-二胺粗品 0.28 g溶于 10 mL的 THF中， 滴加 DIPEA, 直至溶液 pH值为 7, -15°C 下滴加含丙烯酰氯 （ 0.048 g, 0.53 mmol ) 的 THF溶液 0.5 mL, 加毕此 温度下反应 15 min。 滴加 5滴甲醇， 将体系旋干， 制备液相纯化（甲醇： 水 =66%)得到白色固体 50 mg。 两步收率： 20.4%。 0.28 g of crude N 4 —(3-aminophenyl)-5-(methylthio)-TV 2 -(4-morpholinylphenyl)pyrimidine-2,4-diamine was dissolved in 10 mL of THF. DIPEA was added dropwise until the pH of the solution was 7, and 0.5 mL of a solution of acryloyl chloride (0.048 g, 0.53 mmol) in THF was added dropwise at -15 ° C, and the reaction was carried out for 15 min at this temperature. 5 drops of methanol were added dropwise, the system was spun dry, and purified by liquid phase (methanol: water = 66%) to afford 50 mg of white solid. Two-step yield: 20.4%.
分子式： C24H26N602S 分子量： 462.18 盾语 (m/z): 463.3Molecular formula: C 24 H 26 N 6 0 2 S Molecular weight: 462.18 Shield (m/z): 463.3
(M+ H)+. (M+ H) + .
NMR(DMSO-i6, 400 MHz) 5(ppm): 10.14 (1H, s), 9.11 (1H, s), 8.65 (1H, s), 8.16 (1H, s), 7.84 (1H, s), 7.50-7.44 (3H, m), 7.42-7.33 (1H, m), 7.28 (1H, t), 6.72 (2H, d), 6.44 (1H, dd), 6.24 (1H, dd), 5.75 (1H, dd), 3.70 (4H, t), 2.95 (4H, t), 2.28 (3H, s).  NMR (DMSO-i6, 400 MHz) 5 (ppm): 10.14 (1H, s), 9.11 (1H, s), 8.65 (1H, s), 8.16 (1H, s), 7.84 (1H, s), 7.50 -7.44 (3H, m), 7.42-7.33 (1H, m), 7.28 (1H, t), 6.72 (2H, d), 6.44 (1H, dd), 6.24 (1H, dd), 5.75 (1H, dd ), 3.70 (4H, t), 2.95 (4H, t), 2.28 (3H, s).
实施例 8 Κ3-ί2-(6-ί2-甲氧基乙氧基)吡啶 -3-基 J -5- (甲硫基)  Example 8 Κ3-ί2-(6-ί2-methoxyethoxy)pyridine-3-yl J -5-(methylthio)
Figure imgf000050_0002
Figure imgf000050_0002
(1) 2- (2-甲氧基乙氧基 )- -硝基的制备
Figure imgf000050_0003
(1) Preparation of 2-(2-methoxyethoxy)--nitro group
Figure imgf000050_0003
将 2-氯- 5-硝基吡啶 (3.00 g, 18.9 mmol)和 2-甲氧基乙醇 (2.16 g, 28.4mmol)溶于 lOmLDMF中， 然后分批加入叔丁醇钾（ 2.55 g, 22.7 mmol) , 体系在常温下搅拌 2 h, 然后将溶液滴加到 100 mL水中， 析 出固体， 过滤， 干燥得 2.30 g棕色固体， 收率： 61.4%。  2-Chloro-5-nitropyridine (3.00 g, 18.9 mmol) and 2-methoxyethanol (2.16 g, 28.4 mmol) were dissolved in 10 mL of DMF, then potassium t-butoxide (2.55 g, 22.7 mmol) The system was stirred at room temperature for 2 h, and then the solution was added dropwise to 100 mL of water to precipitate a solid, which was filtered and dried to give 2.30 g of a brown solid. Yield: 61.4%.
(2) 6- (2-甲氧基乙氧基)吡啶- 3-胺的制备
Figure imgf000051_0001
(2) Preparation of 6-(2-methoxyethoxy)pyridine-3-amine
Figure imgf000051_0001
将 2- (2-甲氧基乙氧基 )-5-硝基 ( 1.53 g, 7.72 mmol )溶于 10 mL甲 醇中， 置换氮气， 加入 35 mg 10%的钯炭， 体系置换氢气三次， 常温下 反应搅拌 17 h,滤去钯炭，旋干滤液得棕色油状物 1.05 g,收率： 80.8%。  2-(2-methoxyethoxy)-5-nitro (1.53 g, 7.72 mmol) was dissolved in 10 mL of methanol, nitrogen was replaced, 35 mg of 10% palladium on carbon was added, and the system was replaced with hydrogen three times at room temperature. The reaction was stirred for 17 h, the palladium charcoal was filtered off, and the filtrate was evaporated to give a white oil, 1.05 g, yield: 80.8%.
(3) 3- (2- (6- (2-甲氧基乙氧基) p比啶- 3-基氨基 )-5- (甲硫基)嘧啶- 4-基氨 基)苯基氨基甲酸叔丁酯的  (3) 3-(2-(6-(2-methoxyethoxy)p-pyridyl-3-ylamino)-5-(methylthio)pyrimidin-4-ylamino)phenylcarbamic acid Butyl ester
Figure imgf000051_0002
Figure imgf000051_0002
将 3- (2-氯- 5- (甲硫基)嘧啶- 4-基氨基)苯基氨基甲酸叔丁酯 （ 0.22 g, 0.60 mmol )溶于 5 mL的叔戊醇中， 再加入醋酸 2滴， 6- (2-甲氧基乙氧 基)吡啶- 3-胺（ 0.106 g, 0.63 mmol ) , 反应在 85 °C搅拌 30 min。 反应冷 却至室温， 将体系旋干， 柱层析（PE:AE二 8:1— 2: 1 )得到白色固体 0.17 g， 收率 56.7%。  tert-Butyl 3-(2-chloro-5-(methylthio)pyrimidin-4-ylamino)phenylcarbamate (0.22 g, 0.60 mmol) was dissolved in 5 mL of tert-pentanol, then acetic acid 2 was added 6-(2-Methoxyethoxy)pyridine-3-amine (0.106 g, 0.63 mmol) was added dropwise, and the mixture was stirred at 85 ° C for 30 min. The reaction was cooled to room temperature, and the mixture was evaporated to dryness. &lt;RTI ID=0.0&gt;&gt;
(4) (3-氨基苯基) -TV2- (6- (2-甲氧基乙氧基) p比啶- 3-基)- 5- (甲硫基) 嘧啶 -2,4-二胺的制备 (4) (3-Aminophenyl)-TV 2 -(6-(2-methoxyethoxy) p-pyridyl-3-yl)- 5-(methylthio)pyrimidine-2,4-di Preparation of amine
Figure imgf000051_0003
Figure imgf000051_0003
将 3- (2- (6- (2-甲氧基乙氧基)吡啶- 3-基氨基 )-5- (甲硫基)嘧啶- 4-基氨 基)苯基氨基甲酸叔丁酯 （ 0.17 g, 0.34 mmol )溶于 10 mL的 DCM中 , 冰浴下通 HC1气体反应 l h, 将体系旋干， 得到产物粗品 0.17g, 直接用 于下一步。  tert-Butyl 3-(2-(6-(2-methoxyethoxy)pyridine-3-ylamino)-5-(methylthio)pyrimidin-4-ylamino)phenylcarbamate (0.17 g, 0.34 mmol) was dissolved in 10 mL of DCM. The mixture was reacted with HC1 gas for 1 hour in an ice bath, and the system was dried to give a crude product (0.17 g).
(5) V-(3-(2-(6-(2-甲氧基乙氧基) p比啶- 3-基氨基 )-5- (甲硫基)嘧啶- 4- 基氨基)苯基)丙烯酰胺的  (5) V-(3-(2-(6-(2-methoxyethoxy) p-pyridin-3-ylamino)-5-(methylthio)pyrimidin-4-ylamino)phenyl Acrylamide
Figure imgf000051_0004
Figure imgf000051_0004
将上步产物粗品 0.17 g溶于 10 mL的 THF中， 滴加 DIPEA, 直至 溶液 pH值为 7, - 15 °C下滴加含丙烯酰氯（0,031 g, 0.34 mmol )的 THF 溶液 1 mL, 加毕， 此温度下反应 25 min。 滴加 5滴甲醇， 将体系旋干， 制备液相纯化（甲醇： 水 =70% )得到白色固体 87 mg。两步收率： 55.9%。 0.17 g of the crude product of the above step was dissolved in 10 mL of THF, and DIPEA was added dropwise until The pH of the solution was 7, and 1 mL of a solution of acryloyl chloride (0,031 g, 0.34 mmol) in THF was added dropwise at -15 °C, and the reaction was carried out at this temperature for 25 min. 5 drops of methanol were added dropwise, and the system was spin-dried to give a liquid phase purified (methanol: water = 70%) to afford white crystals. Two-step yield: 55.9%.
分子式： C22H24N603S 分子量： 452.16 盾语 (m/z): 452.9Molecular formula: C 22 H 24 N 6 0 3 S Molecular weight: 452.16 Shield (m/z): 452.9
(M+ H)+. (M+ H) + .
NMR(DMSO-i 6, 400 MHz) 5(ppm): 10.12 (1H, s), 9.25 (1H, s), 8.71 (1H, s), 8.25 (1H, s), 8.19 (1H, s), 7.95 (1H, dd), 7.86 (1H, s), 7.44-7.32 (2H, m), 7.26 (1H, t), 6.59 (1H, d), 6.43 (1H, dd), 6.23 (1H, dd), 5.74 (1H, dd), 4.26 (2H, t), 3.60 (2H, t), 3.27 (3H, s), 2.29 (3H, s).  NMR (DMSO-i 6, 400 MHz) 5 (ppm): 10.12 (1H, s), 9.25 (1H, s), 8.71 (1H, s), 8.25 (1H, s), 8.19 (1H, s), 7.95 (1H, dd), 7.86 (1H, s), 7.44-7.32 (2H, m), 7.26 (1H, t), 6.59 (1H, d), 6.43 (1H, dd), 6.23 (1H, dd) , 5.74 (1H, dd), 4.26 (2H, t), 3.60 (2H, t), 3.27 (3H, s), 2.29 (3H, s).
实施例 9 V-i3-i2-(6- (二甲氨基)吡啶 -3-基氨基 )-5- (甲硫基)嘧啶 -4-基氨基)苯基)丙烯酰胺 (化合物 18)的制备  Example 9 Preparation of V-i3-i2-(6-(dimethylamino)pyridin-3-ylamino)-5-(methylthio)pyrimidin-4-ylamino)phenyl)acrylamide (Compound 18)
Figure imgf000052_0001
Figure imgf000052_0001
(1) 二甲基 - 5-硝基吡 2-胺的制备
Figure imgf000052_0002
(1) Preparation of dimethyl-5-nitropyridin-2-amine
Figure imgf000052_0002
将 2-氯- 5-硝基吡啶 ( 2.50 g, 15.8 mmol )和二甲胺盐酸盐 ( 1.93 g, 23.7 mmol )溶于 10 mL DMF中， 然后分批加入叔丁醇钾（ 3.78 g, 33.7 mmol ) , 体系在常温下搅拌 4 h, 然后将溶液滴加到 100 mL水中， 析 出固体， 过滤， 干燥， 得 2.10 g浅棕色固体， 收率： 79.7%。  2-Chloro-5-nitropyridine (2.50 g, 15.8 mmol) and dimethylamine hydrochloride ( 1.93 g, 23.7 mmol) were dissolved in 10 mL DMF, then potassium t-butoxide ( 3.78 g, 33.7 mmol), the system was stirred at room temperature for 4 h, then the solution was added dropwise to 100 mL of water, and the solid was precipitated, filtered, and dried to give 2.10 g of light brown solid. Yield: 79.7%.
(2) A^N2-二甲基吡啶- 2,5-
Figure imgf000052_0003
(2) A^N 2 -lutidine- 2,5-
Figure imgf000052_0003
将 二甲基 - 5-硝基吡啶- 2-胺 （1.90 g, 11.4 mmol ) 溶于 10 mL 甲醇中， 置换氮气， 加入 25 mg 10%的钯炭， 体系置换氢气三次， 常温 下反应搅拌 17 h,滤去钯炭，旋干滤液得棕色油状物 1.25 g,收率： 79.9%。  Dissolve dimethyl 5-nitropyridine-2-amine (1.90 g, 11.4 mmol) in 10 mL of methanol, replace nitrogen, add 25 mg of 10% palladium on carbon, replace the hydrogen three times, and stir at room temperature. h, palladium on charcoal was filtered off, and the filtrate was evaporated to give 1.25 g of a brown oil. Yield: 79.9%.
(3) 3- (2- (6- (二甲氨基) p比啶- 3-基氨基 )-5- (甲硫基)嘧啶- 4-基氨基)苯 基氨基甲酸叔丁酯的制备 将 3- (2-氯- 5- (甲硫基)嘧啶- 4-基氨基)苯基氨基甲酸叔丁酯 （ 0.20 g, 0.55 mmol )溶于 5 mL的叔戊醇中， 再加入醋酸 2滴， A^N2-二甲基吡 啶- 2,5-二胺（ 0.079 g, 0.58 mmol ) , 反应在 85 °C搅拌 30 min。 反应冷 却至室温， 加入 15 mL乙醚， 析出固体， 过滤， 得白色固体 0.17 g， 收 率 65.5%。 (3) Preparation of tert-butyl 3-(2-(6-(dimethylamino) p-pyridin-3-ylamino)-5-(methylthio)pyrimidin-4-ylamino)phenylcarbamate tert-Butyl 3-(2-chloro-5-(methylthio)pyrimidin-4-ylamino)phenylcarbamate (0.20 g, 0.55 mmol) was dissolved in 5 mL of tert-pentanol, then acetic acid 2 was added A^N 2 -lutidine- 2,5-diamine (0.079 g, 0.58 mmol) was added dropwise, and the mixture was stirred at 85 ° C for 30 min. The reaction was cooled to room temperature, and 15 mL of diethyl ether was added, and the solid was precipitated and filtered to give a white solid (0.17 g, yield: 65.5%).
(4) (3-氨基苯基)- Λ^- (6- (N B-二甲氨基） p比啶- 3-基)- 5- (甲硫基）嘧啶  (4) (3-Aminophenyl)- Λ^-(6-(N B-dimethylamino) p-pyridin-3-yl)- 5-(methylthio)pyrimidine
∞ H  ∞ H
-2,4-二胺的制备
Figure imgf000053_0001
-2,4-diamine preparation
Figure imgf000053_0001
将 3- (2- (6- (二甲氨基) ρ比啶- 3-基氨基 )-5- (甲硫基)嘧啶- 4-基氨基)苯 基氨基甲酸叔丁酯 （0.17 g, 0.36 mmol )溶于 10 mL的 DCM中， 冰浴 下通 HC1气体反应 1 h, 将体系旋干， 得到产物粗品 0.17 g, 直接用于 下一步。  3-(2-(6-(Dimethylamino) ρ-pyridin-3-ylamino)-5-(methylthio)pyrimidin-4-ylamino)phenylcarbamic acid tert-butyl ester (0.17 g, 0.36 Methyl acetate was dissolved in 10 mL of DCM, reacted with HC1 gas for 1 h in an ice bath, and the system was spun dry to obtain 0.17 g of crude product, which was directly used for the next step.
(5) V-(3- (2- (6- (二甲氨基) p比啶- 3-基氨基 )-5- (甲硫基)嘧啶- 4-基氨基) 苯基）丙烯酰胺的制备  (5) Preparation of V-(3-(2-(6-(dimethylamino) p-pyridin-3-ylamino)-5-(methylthio)pyrimidin-4-ylamino)phenyl)acrylamide
Figure imgf000053_0002
Figure imgf000053_0002
将上步产物粗品 0.17 g溶于 10 mL的 THF中， 滴加 DIPEA, 直至 溶液 pH值为 7, - 15 °C下滴加含丙烯酰氯 ( 0.035 g, 0.39 mmol )的 THF 溶液 0.5 mL,加毕，此温度下反应 15 min。 滴加 5滴甲醇，将体系旋干， 制备液相纯化（甲醇：水 =68% )得到白色固体 34 mg。两步收率： 22.5%。  0.17 g of the crude product in the previous step was dissolved in 10 mL of THF, and DIPEA was added dropwise until the pH of the solution was 7, and 15 mL of THF solution containing acryloyl chloride (0.035 g, 0.39 mmol) was added dropwise at -15 °C. Complete, react at this temperature for 15 min. 5 drops of methanol were added dropwise, and the system was spin-dried to prepare a liquid phase (methanol: water = 68%) to afford a white solid. Two-step yield: 22.5%.
子式： C2iH23N7OS 子量： 421.17 盾语 (m/z): 422.Sub-form: C 2 iH 23 N 7 OS Sub-amount: 421.17 Shield (m/z): 422.
2 (M+ H)十. 2 (M+ H) ten.
NMR(DMSO-i 6, 400 MHz) 5(ppm): 10.10 (IH, s), 8.99 (IH, s), 8.60 (IH, s), 8.14 (2H, s), 7.84 (IH, t), 7.79 (IH, dd), 7.40 (2H, d), 7.24 (IH t), 6.49-6.38 (2H, m), 6.24 (IH, dd), 5.74 (IH, dd), 2.92 (6H, s), 2.27 (3H, s). NMR (DMSO-i 6, 400 MHz) 5 (ppm): 10.10 (IH, s), 8.99 (IH, s), 8.60 (IH, s), 8.14 (2H, s), 7.84 (IH, t), 7.79 (IH, dd), 7.40 (2H, d), 7.24 (IH t), 6.49-6.38 (2H, m), 6.24 (IH, dd), 5.74 (IH, dd), 2.92 (6H, s), 2.27 (3H, s).
实施例 10 V-(3-i5-i甲基亚磺酰基 )-2-(4-吗啉苯氨基)嘧啶 -4-基氨 基)苯基)丙烯酰胺 ί化合物 19)的制备  Example 10 Preparation of V-(3-i5-imethylsulfinyl)-2-(4-morpholinylamino)pyrimidin-4-ylamino)phenyl)acrylamide ί Compound 19)
Figure imgf000054_0001
Figure imgf000054_0001
将 V-(3- (5- (甲硫基)- 2- (4-吗啉苯氨基)嘧啶- 4-基氨基)苯基）丙烯酰胺 ( 0.140 g, 0.303 mmol )溶于 15 mL甲醇和 5 mL水中， 然后加过石 酸氢 钾复合盐 (oxone) ( 0.186 g, 0.303 mmol ) , 在常温下反应 25 min, 将溶 液旋干， 制备液相纯化（甲醇 /水 =58% )得到白色固体 45 mg。 收率： 31%。  V-(3-(5-(methylthio)-2-(4-morpholinanilinyl)pyrimidin-4-ylamino)phenyl)acrylamide (0.140 g, 0.303 mmol) was dissolved in 15 mL of methanol and 5 mL of water, then add potassium octoate complex salt (oxone) (0.186 g, 0.303 mmol), react at room temperature for 25 min, spin the solution, prepare liquid phase purification (methanol / water = 58%) to obtain white Solid 45 mg. Yield: 31%.
分子式： C24H26N603S 分子量： 478.18 盾语 (m/z): 478.9Molecular formula: C 24 H 26 N 6 0 3 S Molecular weight: 478.18 Shield (m/z): 478.9
(M+ H)+. (M+ H) + .
NMR(DMSO-i 6, 400 MHz) 5(ppm): 10.18 (IH, s), 9.54 (2H, s), 8.20 (IH, s), 7.72 (IH, s), 7.55-7.38 (4H, m), 7.29 (IH, t), 6.79 (2H, d), 6.43 (IH, dd), 6.25 (IH, dd), 5.76 (IH, dd), 3.71 (4H, t), 2.99 (4H, t), 2.95 (3H, s).  NMR (DMSO-i 6, 400 MHz) 5 (ppm): 10.18 (IH, s), 9.54 (2H, s), 8.20 (IH, s), 7.72 (IH, s), 7.55-7.38 (4H, m ), 7.29 (IH, t), 6.79 (2H, d), 6.43 (IH, dd), 6.25 (IH, dd), 5.76 (IH, dd), 3.71 (4H, t), 2.99 (4H, t) , 2.95 (3H, s).
实施例 11 V-(3-i2-(6-甲氧基吡啶 -3-基 J -5- (甲基亚磺酰基)嘧 啶 -4-基氨基)苯基)丙烯酰胺 (化合物 20)的制备  Example 11 Preparation of V-(3-i2-(6-methoxypyridin-3-yl J-5-(methylsulfinyl)pyrimidin-4-ylamino)phenyl)acrylamide (Compound 20)
Figure imgf000054_0002
Figure imgf000054_0002
将化合物 V-(3- (2- (6-甲氧基吡啶- 3-基氨基 )-5- (甲硫基)嘧啶- 4-基氨 基)苯基)丙烯酰胺（0.150 g, 0.367 mmol )溶于 15 mL甲醇和 5 mL ? 中， 然后加过硫酸氢钟复合盐（oxone) ( 0.226 g, 0.368 mmol ) , 在常温下反 应 25 min, 将溶液旋干， 制备液相纯化（甲醇 /水 =61% )得到白色固体 25 mg„ 收率： 16%。  Compound V-(3-(2-(6-Methoxypyridine-3-ylamino)-5-(methylthio)pyrimidin-4-ylamino)phenyl)acrylamide (0.150 g, 0.367 mmol) Dissolve in 15 mL of methanol and 5 mL?, then add hydrogen peroxide oxone (0.226 g, 0.368 mmol), react at room temperature for 25 min, spin dry the solution, prepare liquid phase purification (methanol/water) =61%) Obtained a white solid 25 mg „ Yield: 16%.
分子式： C2。H2。N603S 分子量： 424.13 盾语 (m/z): 424.9 (M)+. Molecular formula: C 2 . H 2 . N 6 0 3 S Molecular Weight: 424.13 Shield (m/z): 424.9 (M) + .
NMR(DMSO-i 6, 400 MHz) 5(ppm): 10.15 (IH, s), 9.68-9.55 (2H, m), 8.33-8.25 (IH, m), 8.22 (IH, s), 7.95 (IH, d), 7.74 (IH, s), 7.41-7.23 (3H, m), 6.73-6.60 (IH, m), 6.42 (IH, dd), 6.23 (IH, dd), 5.75 (IH, dd), 3.78 (3H, s), 2.95 (3H, s). NMR (DMSO-i 6, 400 MHz) 5 (ppm): 10.15 (IH, s), 9.68-9.55 (2H, m), 8.33-8.25 (IH, m), 8.22 (IH, s), 7.95 (IH , d), 7.74 (IH, s), 7.41-7.23 (3H, m), 6.73-6.60 (IH, m), 6.42 (IH, dd), 6.23 (IH, dd), 5.75 (IH, dd), 3.78 (3H, s), 2.95 (3H, s).
实施例 12 V-(3-i2-(6- (二甲氨基)吡啶 -3-基氨基 )-5- (甲基亚磺酰 基)嘧啶 -4-基氨基)  Example 12 V-(3-i2-(6-(Dimethylamino)pyridine-3-ylamino)-5-(methylsulfinyl)pyrimidin-4-ylamino)
Figure imgf000055_0001
Figure imgf000055_0001
将 V-(3- (2- (6- (二甲氨基) p比啶- 3-基氨基 )-5- (甲硫基)嘧啶- 4-基氨基) 苯基）丙烯酰胺（0.150 g, 0.356 mmol )溶于 15 mL甲醇和 5 mL水中， 然后加过硫酸氢钟复合盐（oxone) ( 0.220 g, 0.358 mmol ) , 常温下反应 25 min,将溶液旋干，制备液相纯化（甲醇 /水 =51% )得到白色固体 35 mg。 收率： 22.5%。  V-(3-(2-(6-(Dimethylamino) p-pyridin-3-ylamino)-5-(methylthio)pyrimidin-4-ylamino)phenyl)acrylamide (0.150 g, 0.356 mmol ) dissolved in 15 mL of methanol and 5 mL of water, then added with hydrogen peroxide oxone (0.220 g, 0.358 mmol), reacted at room temperature for 25 min, and the solution was spun dry to prepare liquid phase purification (methanol/ Water = 51%) gave a white solid, 35 mg. Yield: 22.5%.
分子式： C21H23N702S 分子量： 437.16 盾语 (m/z): 437.9 (M+Molecular formula: C 21 H 23 N 7 0 2 S Molecular weight: 437.16 Shield (m/z): 437.9 (M+
H)+. H) + .
NMR(DMSO-i 6, 400 MHz) 5(ppm): 10.13 (IH, s), 9.56 (IH, s), 9.50-9.32 (IH, m), 8.17 (2H, s), 7.76 (IH, dd), 7.72-7.66 (IH, dd), 7.43-7.15 (3H, m), 6.60-6.46 (IH, m), 6.41 (IH, dd), 6.24 (IH, dd), 5.75 (IH, dd), 2.96 (6H, s), 2.94 (3H, s).  NMR (DMSO-i 6, 400 MHz) 5 (ppm): 10.13 (IH, s), 9.56 (IH, s), 9.50-9.32 (IH, m), 8.17 (2H, s), 7.76 (IH, dd ), 7.72-7.66 (IH, dd), 7.43-7.15 (3H, m), 6.60-6.46 (IH, m), 6.41 (IH, dd), 6.24 (IH, dd), 5.75 (IH, dd), 2.96 (6H, s), 2.94 (3H, s).
实施例 13 4-(4-(4-(3-丙烯酰氨基苯 J -5- (甲基亚磺酰基)嘧啶 -2-基 JQ苯氧 V-甲基吡啶 -2-甲酰胺 (化合物 22)的制备  Example 13 4-(4-(4-(3-acrylamidobenzene J-5-(methylsulfinyl)pyrimidin-2-yl JQ phenoxy V-methylpyridine-2-carboxamide (Compound 22 Preparation
Figure imgf000055_0002
Figure imgf000055_0002
将 4- (4- (4- (3-丙烯酰氨基苯氨基) -5- (甲硫基）嘧啶- 2-基氨基）苯氧 基)—TV-甲基吡啶- 2-甲酰胺（0.120 g, 0.227 mmol )溶于 15 mL甲醇和 5 mL 水中， 然后加入硫酸氢钾复合盐（oxone) ( 0.142 g, 0.231 mmol ) , 在常 温下反应 25 min, 将溶液旋干， 制备液相纯化（甲醇 /水 =58% )得到白 色固体 45 mg, 收率： 36.5%。  4-(4-(4-(3-acrylamidophenylamino)-5-(methylthio)pyrimidin-2-ylamino)phenoxy)-TV-methylpyridine-2-formamide (0.120 g, 0.227 mmol) dissolved in 15 mL of methanol and 5 mL of water, then added potassium sulphate complex salt (oxone) (0.142 g, 0.231 mmol), reacted at room temperature for 25 min, and the solution was spun dry to prepare liquid phase purification ( Methanol / water = 58%) gave a white solid, 45 mg, yield: 36.5%.
分子式： C27H25N704S 分子量： 543.17 盾语 (m/z): 543.9 (M+ Ή- NMR(i 6- DMSO, 400 MHz) 5(ppm): 10.18 (IH, s), 9.88 (IH, s), 9.58 (IH, s), 8.77 (IH, q), 8.48 (IH, d), 8.28 (IH, s), 7.82-7.72 (3H, m), 7.45-7.25 (4H, m), 7.15-6.98 (3H, m), 6.35 (IH, dd), 6.17 (IH, d), 5.63 (IH, d), 2.98 (3H, s), 2.77 (3H, d). Molecular formula: C 27 H 25 N 7 0 4 S Molecular weight: 543.17 Shield (m/z): 543.9 (M+ Ή-NMR (i 6 - DMSO, 400 MHz) 5 (ppm): 10.18 (IH, s), 9.88 (IH, s), 9.58 (IH, s), 8.77 (IH, q), 8.48 (IH, d ), 8.28 (IH, s), 7.82-7.72 (3H, m), 7.45-7.25 (4H, m), 7.15-6.98 (3H, m), 6.35 (IH, dd), 6.17 (IH, d), 5.63 (IH, d), 2.98 (3H, s), 2.77 (3H, d).
实施例 14 V-i3-i2-(4-(2-甲氧基乙氧基)^ jQ-5- (甲基亚磺酰基) 嘧啶 -4-基氨基  Example 14 V-i3-i2-(4-(2-methoxyethoxy)^jQ-5-(methylsulfinyl)pyrimidin-4-ylamino
Figure imgf000056_0001
Figure imgf000056_0001
将 TV- (3- (2- (4- (2-甲氧基乙氧基)苯氨基 )-5- (甲硫基)嘧啶- 4-基氨基) 苯基）丙烯酰胺（0.045 g, 0.1 mmol )溶于甲醇中敞开口回流 3 h, 旋干， 制备液相反相纯化（甲醇 /水 =57% )得到白色固体 9 mg。 收率： 19.3%。  TV-(3-(2-(4-(2-methoxyethoxy)phenylamino)-5-(methylthio)pyrimidin-4-ylamino)phenyl)acrylamide (0.045 g, 0.1 Methyl acetate was dissolved in methanol and refluxed for 3 h, dried, and purified by preparative liquid phase (methanol / water = 57%) to afford 9 mg of white solid. Yield: 19.3%.
分子式： C23H25N504S 分子量： 467.16 盾语 (m/z): 468.2 (M+ H)+. Molecular formula: C 23 H 25 N 5 0 4 S Molecular weight: 467.16 Shield (m/z): 468.2 (M+ H) + .
NMR(DMSO- ί 6,400 MHz)5(ppm): 10.17 (IH, s), 9.54 (2H, s), 8.20 (IH, s), 7.73 (IH, s), 7.55-7.45 (3H, m), 7.39 (IH, d), 7.29 (IH, t), 6.82-6.73 (2H, m), 6.42 (IH, dd), 6.24 (IH, dd), 5.75 (IH, dd), 4.00 (2H, t), 3.62 (2H, t), 3.29 (3H, s), 2.95 (3H, s). NMR (DMSO- ί 6 , 400 MHz) 5 (ppm): 10.17 (IH, s), 9.54 (2H, s), 8.20 (IH, s), 7.73 (IH, s), 7.55-7.45 (3H, m ), 7.39 (IH, d), 7.29 (IH, t), 6.82-6.73 (2H, m), 6.42 (IH, dd), 6.24 (IH, dd), 5.75 (IH, dd), 4.00 (2H, t), 3.62 (2H, t), 3.29 (3H, s), 2.95 (3H, s).
实施例 15 V-i3-i2-(4-(2-甲氧基乙氧基)苯氨基 )-5-ί甲亚磺酰基) -4-基氨基)苯基) -4- (派嗪 -1-基)丁 -2-烯酰胺 (化合物 2)的制备  Example 15 V-i3-i2-(4-(2-methoxyethoxy)phenylamino)-5-methylsulfinyl)-4-ylamino)phenyl)-4-(pyrazine- Preparation of 1-yl)but-2-enamide (Compound 2)
Figure imgf000056_0002
(1)3— (2— (4- (2-甲氧基乙氧基)苯氨基 )-5- (甲亚磺酰基)嘧啶- 4-基氨基) 苯基氨基甲酸叔
Figure imgf000056_0002
(1) 3-(2-(4-Methoxyethoxy)phenylamino)-5-(methylsulfinyl)pyrimidin-4-ylamino)phenyl carbamic acid
Figure imgf000057_0001
Figure imgf000057_0001
将 3- (2- (4- (2-甲氧基乙氧基)苯氨基 )-5- (甲硫基)嘧啶- 4-基氨基)苯基 氨基甲酸叔丁酯（5.7 g, 11.47 mmol )溶于 50 mL二氯甲烷与甲醇的混 合溶液中（二氯甲烷 /甲醇 =15: 1 ) , 将过硫代氢钾复合盐（7.05 g, 11.47 mmol )分批加入到该溶液中， TLC监测原料消失然后加入 100 mL二氯 甲烷和 lOO mL水， 萃取， 有机相干燥， 旋干， 过硅胶柱（二氯甲烷 /甲 醇 =15: 1 )得白色固体 5.1 g, 收率： 86.6%。  tert-Butyl 3-(2-(4-(2-methoxyethoxy)phenylamino)-5-(methylthio)pyrimidin-4-ylamino)phenylcarbamate (5.7 g, 11.47 mmol Dissolved in 50 mL of a mixed solution of dichloromethane and methanol (dichloromethane/methanol = 15:1), and added potassium peroxodihydrogenate (7.05 g, 11.47 mmol) to the solution in portions, TLC The disappearance of the starting material was followed by the addition of 100 mL of dichloromethane and 100 mL of water, and the organic phase was dried and evaporated to dryness. Toluene (methylene chloride/methanol = 15:1) gave a white solid, 5.1 g, yield: 86.6%.
(2) (3-氨基苯基)- (4- (2-甲氧基乙氧基)苯基) -5- (甲亚磺酰基) 嘧啶 -2,4-二胺的制备  (2) Preparation of (3-aminophenyl)-(4-(2-methoxyethoxy)phenyl)-5-(methylsulfinyl)pyrimidine-2,4-diamine
Figure imgf000057_0002
Figure imgf000057_0002
将 3- (2- (4- (2-甲氧基乙氧基)苯氨基 )-5- (甲亚磺酰基)嘧啶- 4-基氨基) 苯基氨基甲酸叔丁酯（2.3 g, 4.48 mmol )溶于 100 mL二氯甲烷， 然后 加入三氟醋酸（2.56 g, 22.55 mmol ) 然后搅拌 4 h, 旋干， 直接用于下 一步。  tert-Butyl 3-(2-(4-(2-methoxyethoxy)phenylamino)-5-(methylsulfinyl)pyrimidin-4-ylamino)phenylcarbamate (2.3 g, 4.48 Methyl acetate was dissolved in 100 mL of dichloromethane, then trifluoroacetic acid (2.56 g, 22.55 mmol) was added and stirred for 4 h, then dried and taken directly to the next.
(3) 4-溴丁- 2-烯酰氯制备
Figure imgf000057_0003
(3) Preparation of 4 -bromobut- 2 -enoic acid chloride
Figure imgf000057_0003
将 4-溴巴豆酸（0.718 g, 4.35 mmol )溶于 20 mL二氯甲烷中， 加入 2滴 DMF , 然后在 0°C时加入 2 mL草酰氯。 所得溶液搅拌 2 h, 旋干直 接用于下一步。  4-Bromocrotonic acid (0.718 g, 4.35 mmol) was dissolved in 20 mL of dichloromethane, 2 drops of DMF were added, and then 2 mL of oxalyl chloride was added at 0 °C. The resulting solution was stirred for 2 h and spun directly for the next step.
(4) 4-溴- V-(3- (2- (4- (2-甲氧基乙氧基)苯氨基 )-5- (甲亚磺酰基)嘧啶- 4-基 氨基)苯基)丁- 2-烯酰胺的制备
Figure imgf000058_0001
(4) 4-Bromo-V-(3-(2-(4-(2-methoxyethoxy)phenylamino)-5-(methylsulfinyl)pyrimidin-4-ylamino)phenyl) Preparation of butyl-2-enamide
Figure imgf000058_0001
将 N"- (3-氨基苯基)- W2- (4- (2-甲氧基乙氧基)苯基)—5- (甲亚磺酰基)嘧 啶- 2,4-二胺（ 1.8 g, 4.35 mmol)溶于 20 mL干燥的四氢呋喃中， 加入 4 mL DIPEA使得 pH值到 10, 然后将 4-溴丁- 2-烯酰氯（粗品， 约 4.35 mmol)溶于 10 mL二氯甲烷中， 緩慢的滴加到该溶液中， 反应在 0°C搅 拌 2 h, 放置备用。 N"-(3-Aminophenyl)-W 2 -(4-(2-methoxyethoxy)phenyl)-5-(methylsulfinyl)pyrimidine-2,4-diamine (1.8 g, 4.35 mmol) was dissolved in 20 mL of dry tetrahydrofuran, 4 mL of DIPEA was added to bring the pH to 10, then 4-bromobut-2-enoyl chloride (crude, about 4.35 mmol) was dissolved in 10 mL of dichloromethane. Slowly drip into the solution, and the reaction was stirred at 0 ° C for 2 h and placed for later use.
(5) 4- (4- (3- (2- (4- (2-甲氧基乙氧基)苯氨基 )-5- (甲亚磺酰基)嘧啶- 4- 基氨基)苯氨基 )-4-氧代丁 -2- 基)哌嗪- 1 -甲酸叔丁酯的制备  (5) 4-(4-(3-(2-(4-(2-methoxyethoxy)phenylamino)-5-(methylsulfinyl)pyrimidine-4-ylamino)phenylamino)- Preparation of 4-oxobutan-2-yl)piperazine-1-carboxylic acid tert-butyl ester
Figure imgf000058_0002
Figure imgf000058_0002
将 4-溴-V-(3- (2- (4- (2-甲氧基乙氧基)苯氨基 )-5- (甲亚磺酰基)嘧啶- 4- 基氨基)苯基)丁- 2-烯酰胺（0.812 g, 1.45 mmol) 和哌嗪 -1-甲酸叔丁酯 ( 0.323 g, 1.74 mmol)溶于 10 mL干燥的四氢呋喃中， 再加入 DIPEA ( 2 mL ) , 在 0°C时， 搅拌 0.5 h, 待原料消失， 旋干， 过硅胶柱（ DCM/ 甲醇 =10:1 )得 0.14 g白色固体， 收率： 14.5%。  4-Bromo-V-(3-(2-(4-(2-methoxyethoxy)phenylamino)-5-(methylsulfinyl)pyrimidin-4-ylamino)phenyl)- 2-enamide (0.812 g, 1.45 mmol) and piperazine-1-carboxylic acid tert-butyl ester (0.323 g, 1.74 mmol) were dissolved in 10 mL dry THF, then DIPEA (2 mL) at 0 ° C After stirring for 0.5 h, the starting material disappeared, and dried, and passed through a silica gel column (DCM / methanol = 10:1) to give 0.14 g of white solid. Yield: 14.5%.
(6) TV- (3- (2- (4- (2-甲氧基乙氧基)苯氨基 )-5- (甲亚磺酰基)嘧啶- 4-基 氨基)苯基)—4- (哌嗪- 1 -基 - 2-烯酰胺的制备  (6) TV-(3-(2-(4-(2-methoxyethoxy)phenylamino)-5-(methylsulfinyl)pyrimidin-4-ylamino)phenyl)-4-( Preparation of piperazine-1-yl-2-enamide
Figure imgf000058_0003
Figure imgf000058_0003
将 4- (4- (3- (2- (4- (2-甲氧基乙氧基)苯氨基 )-5- (甲亚磺酰基)嘧啶- 4-基 氨基)苯氨基 )-4-氧代丁 -2-烯基)哌嗪- 1-甲酸叔丁酯（0.14 g, 0.21 mmol), 溶于 10mL二氯甲烷中， 慢慢加入 0.24 g三氟醋酸， 反应搅拌 4h后， 旋干， 再加入 100 mL二氯甲烷和 50 mL饱和的碳酸氢钠溶液， 萃取， 有机相干燥， 旋干后， 加入 2 mL甲醇， 重结晶， 得白色固体 0.078 g, 收率： 65.7%。 4-(4-(3-(2-(4-(2-methoxyethoxy)phenylamino)-5-(methylsulfinyl)pyrimidin-4-ylamino)phenylamino)-4- Oxybutan-2-enyl) piperazine-1-carboxylic acid tert-butyl ester (0.14 g, 0.21 mmol), dissolved in 10 mL of dichloromethane, slowly added 0.24 g of trifluoroacetic acid, stirred for 4 h, then dried. , add 100 mL of dichloromethane and 50 mL of saturated sodium bicarbonate solution, and extract. The organic phase was dried, dried, and then added with 2 mL of methanol and then recrystallized to give white solid (0.078 g, yield: 65.7%).
分子式： C28H35N704S 分子量： 565.2 盾 f (m/z)：Molecular formula: C 28 H 35 N 7 0 4 S Molecular weight: 565.2 Shield f (m/z):
283.7(Μ/2+1)+. 283.7(Μ/2+1) + .
NMR(i 6- DMSO, 400 MHz) 5(ppm): 10.18 (IH, s), 9.58, 9.53 (2H, 两个单峰)， 8.19 (IH, s), 7.80 (IH, s), 7.65-7.33 (5H, m), 7.26 (IH, t), 6.88-6.65 (3H, m), 6.28 (IH, d), 4.05-3.95 (2H, m), 3.62 (2H, t), 3.29 (3H, s), 3.05 (2H, d), 2.95 (3H, s), 3.75-3.65 (4H, m), 2.50-2.22 (4H, m). NMR (i 6 - DMSO, 400 MHz) 5 (ppm): 10.18 (IH, s), 9.58, 9.53 (2H, two single peaks), 8.19 (IH, s), 7.80 (IH, s), 7.65- 7.33 (5H, m), 7.26 (IH, t), 6.88-6.65 (3H, m), 6.28 (IH, d), 4.05-3.95 (2H, m), 3.62 (2H, t), 3.29 (3H, s), 3.05 (2H, d), 2.95 (3H, s), 3.75-3.65 (4H, m), 2.50-2.22 (4H, m).
实施例 16 V-i3-i2-(4-(2-甲氧基乙氧基)苯氨基 )-5-ί甲亚磺酰基) 嘧啶 -4-基  Example 16 V-i3-i2-(4-(2-methoxyethoxy)phenylamino)-5-methylsulfinyl)pyrimidin-4-yl
Figure imgf000059_0001
Figure imgf000059_0001
将 4—溴—TV- (3- (2- (4- (2-甲氧基乙氧基)苯氨基 )-5- (甲亚磺酰基)嘧啶- 4- 基氨基)苯基)丁- 2-烯酰胺（0.812 g, 1.45 mmol ) , 吗啉（0.189 g, 2.17 mmol )溶于 10 mL干燥的四氢呋喃中， 再加入 2 mL DIPEA, 在 0°C时， 搅拌 0.5 h, 待原料消失， 旋干， 过硅胶柱（DCM/甲醇 =10:1 )得 0.070 g 白色固体， 收率： 8.5%。  4-Bromo-TV-(3-(2-(4-(2-methoxyethoxy)phenylamino)-5-(methylsulfinyl)pyrimidin-4-ylamino)phenyl)- 2-enamide (0.812 g, 1.45 mmol), morpholine (0.189 g, 2.17 mmol) was dissolved in 10 mL of dry tetrahydrofuran, then 2 mL of DIPEA was added and stirred at 0 ° C for 0.5 h until the starting material disappeared. The mixture was dried with EtOAc (EtOAc:EtOAc)
分子式： C28H34N605S 分子量： 566.2 盾语 (m/z): 284.2(M/2+H)+. Molecular formula: C 28 H 34 N 6 0 5 S Molecular weight: 566.2 Shield (m/z): 284.2 (M/2+H) + .
NMR(i 6- DMSO , 400 MHz) 5(ppm): 10.08 (IH, s), 9.53 (2H, s), 8.20 (IH, s), 7.74 (IH, s), 7.55-7.45 (3H, m), 7.36 (IH, d), 7.27 (IH, t), 6.82-6.65 (3H, m), 6.27 (IH, d), 4.05-3.95 (2H, m), 3.66-3.54 (6H, m), 3.29 (3H, s), 3.10 (2H, d), 2.95 (3H, s), 2.41-2.34 (4H, m). NMR (i 6 - DMSO , 400 MHz) 5 (ppm): 10.08 (IH, s), 9.53 (2H, s), 8.20 (IH, s), 7.74 (IH, s), 7.55-7.45 (3H, m ), 7.36 (IH, d), 7.27 (IH, t), 6.82-6.65 (3H, m), 6.27 (IH, d), 4.05-3.95 (2H, m), 3.66-3.54 (6H, m), 3.29 (3H, s), 3.10 (2H, d), 2.95 (3H, s), 2.41-2.34 (4H, m).
实施例 17 4- (二甲氨基 V-(3-i2-(4-(2-甲氧基乙氧基）苯氨 基) 甲 -4-基 ^)苯基 -2-烯酰胺 (化合物 4)的制备  Example 17 4-(Dimethylamino V-(3-i2-(4-(2-methoxyethoxy)phenylamino)methyl-4-yl^)phenyl-2-enoamide (Compound 4) Preparation
Figure imgf000059_0002
Figure imgf000059_0002
将 15 mL二甲胺的水溶液用 10 mL的二氯甲烷萃取，所得的有机相 干燥， 备用。 An aqueous solution of 15 mL of dimethylamine was extracted with 10 mL of dichloromethane to obtain an organic phase. Dry, spare.
将 4—溴—TV- (3- (2- (4- (2-甲氧基乙氧基)苯氨基 )-5- (甲亚磺酰基)嘧啶- 4- 基氨基)苯基)丁- 2-烯酰胺（0.812 g, 1.45 mmol )和 10 mL二甲胺的二 氯甲烷溶液溶于 10 mL干燥的四氢呋喃中， 再加入 DIPEA ( 2 mL ) , 在 0°C , 搅拌 0.5 h, 待原料消失， 旋干， 过硅胶柱（DCM/甲醇 =10:1 ) 得 0.030 g白色固体， 收率： 3.9%。  4-Bromo-TV-(3-(2-(4-(2-methoxyethoxy)phenylamino)-5-(methylsulfinyl)pyrimidin-4-ylamino)phenyl)- 2-enamide (0.812 g, 1.45 mmol) and 10 mL of dimethylamine in dichloromethane were dissolved in 10 mL of dry tetrahydrofuran, then DIPEA (2 mL) was added and stirred at 0 ° C for 0.5 h. Disappeared, spin dry, passed through a silica gel column (DCM / methanol = 10:1) to give 0.030 g of white solid, yield: 3.9%.
分子式： C26H32N604S 分子量： 524.2 盾 f (m/z)：Molecular formula: C 26 H 32 N 6 0 4 S Molecular weight: 524.2 Shield f (m/z):
263.1(Μ/2+Η ) +· 263.1(Μ/2+Η) +·
NMR(i 6- DMSO , 400 MHz) 5(ppm): 10.19 (1H, s), 9.65-7.50 (2H, m), 8.20 (1H, s), 7.77 (1H, s), 7.60-7.35 (4H, m), 7.28 (1H, t), 6.85-6.68 (3H, m), 6.31 (1H, d), 4.00 (2H, t), 3.62 (2H, t), 3.30-3.15 (5H, m), 2.95 (3H, s), 2.32 (6H, s). NMR (i 6 - DMSO, 400 MHz) 5 (ppm): 10.19 (1H, s), 9.65-7.50 (2H, m), 8.20 (1H, s), 7.77 (1H, s), 7.60-7.35 (4H , m), 7.28 (1H, t), 6.85-6.68 (3H, m), 6.31 (1H, d), 4.00 (2H, t), 3.62 (2H, t), 3.30-3.15 (5H, m), 2.95 (3H, s), 2.32 (6H, s).
实施例 18 V-i3-i2-(4-(2-甲氧基乙氧基)苯氨基 )-5-ί甲亚磺酰基) 嘧 -4-基 JQ苯基) -4- (吡咯烷 -1-基)丁 -2-烯酰胺 (化合物 5)的制备  Example 18 V-i3-i2-(4-(2-methoxyethoxy)phenylamino)-5-methylsulfinyl)pyrimidin-4-yl JQphenyl)-4-(pyrrolidine- Preparation of 1-yl)but-2-enamide (Compound 5)
Figure imgf000060_0001
Figure imgf000060_0001
4- (吡咯烷- 1 -基)丁- 2 制备
Figure imgf000060_0002
Preparation of 4- (pyrrolidine-1-yl)butyl- 2
Figure imgf000060_0002
将 4-溴巴豆酸甲酯（ 1.775 g, 9.93 mmol ) , 溶于 20 mL二氯甲烷, 然后在 0°C向溶液中加入吡咯烷（ 1.408 g, 19.86 mmol ) ,反应搅拌 2 h, 旋干， 直接用于下一步。  Methyl 4-bromocrotonate ( 1.775 g, 9.93 mmol) was dissolved in 20 mL of dichloromethane, then pyrrolidine ( 1.408 g, 19.86 mmol) was added to the solution at 0 ° C, the reaction was stirred for 2 h, and dried. , used directly in the next step.
(2) 4- (吡咯烷- 1 -基)丁- 2-
Figure imgf000060_0003
(2) 4- (pyrrolidin-1-yl)butyl- 2 -
Figure imgf000060_0003
将 4- (吡咯烷- 1-基)丁- 2-烯酸甲酯 ( 1.67 g, 9.9 mmol )溶于 20 mL甲 醇中， 然后加入 NaOH ( 0.792 g, 19.8 mmol ) , 所得的溶液在 0°C时搅 拌 2 h, 然后加入乙酸乙酯 50 mL和水 50mL, 萃取， 将水相用 2 N的稀 盐酸调节 pH值到 6.0, 然后加入乙酸乙酯 100 mL, 分三次萃取， 所得 有机相干燥， 旋干， 得 0.52 g淡红色油状物收率 33.9%。 Methyl 4-(pyrrolidin-1-yl)but-2-enoate ( 1.67 g, 9.9 mmol) was dissolved in 20 mL of methanol, then NaOH (0.792 g, 19.8 mmol) was added and the resulting solution was at 0°. Stir for 2 h at C, then add 50 mL of ethyl acetate and 50 mL of water, extract, and dilute the aqueous phase with 2 N Hydrochloric acid was adjusted to pH 6.0, then ethyl acetate (100 mL) was added and extracted three times. The obtained organic phase was dried and dried to give a yield of 0.52 g of pale red oil.
(3) 4- (吡咯烷- 1 -基)丁- 2- 备
Figure imgf000061_0001
(3) 4- (pyrrolidine-1-yl)butyl- 2 -
Figure imgf000061_0001
将 4- (吡咯烷- 1-基)丁- 2-烯酸 ( 0.50 g, 3.23 mmol )溶于 50 mL二氯 甲烷中， 加入 2滴 DMF , 然后在 0°C时緩慢加入草酰氯（ 1.64 g, 12.9 mmol ) , 所得溶液搅拌 2 h, 旋干， 直接用于下一步。  Dissolve 4-(pyrrolidin-1-yl)but-2-enoic acid (0.50 g, 3.23 mmol) in 50 mL of dichloromethane, add 2 drops of DMF, then slowly add oxalyl chloride at 0 ° C ( 1.64 g, 12.9 mmol), the resulting solution was stirred for 2 h, dried and applied directly to the next step.
(4) TV- (3- (2- (4- (2-甲氧基乙氧基)苯氨基 )-5- (甲亚磺酰基)嘧啶- 4-基 氨基)苯基)—4- (吡咯烷- 1  (4) TV-(3-(2-(4-(2-methoxyethoxy)phenylamino)-5-(methylsulfinyl)pyrimidin-4-ylamino)phenyl)-4-( Pyrrolidine-1
Figure imgf000061_0002
Figure imgf000061_0002
将 (3-氨基苯基)- (4- (2-甲氧基乙氧基)苯基)—5- (甲亚磺酰基)嘧 啶- 2,4-二胺（0.89 g, 2.15 mmol )溶于 10 mL干燥的四氢呋喃中， 加入 DIPEA ( 0.554 g, 4.3 mmol ) , 然后将 4- (吡咯烷- 1-基)丁- 2-烯酰氯（ 0·56 g, 3.23 mmol )溶于 10 mL二氯甲烷中， 緩慢的滴加到该溶液中， 反应 在 0°C搅拌 2 h, LC- MS监测原料消失， 然后加入 I mL甲醇， 旋干， 过 硅胶柱（DCM/甲醇 =10:1 )得 35 mg白色固体。  Dissolve (3-aminophenyl)-(4-(2-methoxyethoxy)phenyl)-5-(methylsulfinyl)pyrimidine-2,4-diamine (0.89 g, 2.15 mmol) DIPEA (0.554 g, 4.3 mmol) was added to 10 mL of dry tetrahydrofuran, then 4-(pyrrolidin-1-yl)but-2-enoyl chloride (0.56 g, 3.23 mmol) was dissolved in 10 mL. In methyl chloride, slowly added dropwise to the solution, the reaction was stirred at 0 ° C for 2 h, LC-MS was used to monitor the disappearance of the starting material, then 1 mL of methanol was added, spin-dried, and passed through a silica gel column (DCM / methanol = 10:1) Obtained 35 mg of a white solid.
分子式： C28H34N604S 分子量： 550.2 盾语 (m/z): 276.1(Μ/2+1)+. Molecular formula: C 28 H 34 N 6 0 4 S Molecular weight: 550.2 Shield (m/z): 276.1 (Μ/2+1) + .
NMR(i 6- DMSO+HC1, 400 MHz) 5(ppm): 11.52 (1H, s), 11.14 (1H, s), 10.93 (1H, s), 10.51 (1H, s), 8.42-8.25 (1H, m), 7.95-7.80 (1H, m), 7.68-7.50 (1H, m), 7.42-7.18 (4H, m), 6.95-6.73 (3H, m), 6.62 (1H, d), 4.12-3.88 (4H, m), 3.68-3.58 (2H, m), 3.50-3.37 (2H, m), 3.28 (3H, s), 3.09-2.93 (5H, m), 2.05-1.80 (4H, m). NMR (i 6 - DMSO + HC1, 400 MHz) 5 (ppm): 11.52 (1H, s), 11.14 (1H, s), 10.93 (1H, s), 10.51 (1H, s), 8.42-8.25 (1H , m), 7.95-7.80 (1H, m), 7.68-7.50 (1H, m), 7.42-7.18 (4H, m), 6.95-6.73 (3H, m), 6.62 (1H, d), 4.12-3.88 (4H, m), 3.68-3.58 (2H, m), 3.50-3.37 (2H, m), 3.28 (3H, s), 3.09-2.93 (5H, m), 2.05-1.80 (4H, m).
实施例 19 V-i3-i2-(l-甲基 -1H-吡唑 -4-基 jQ-5-i甲亚磺酰基)嘧 啶 -4-基氨基)苯基)丙烯酰胺 (化合物 24)的制备 Example 19 V-i3-i2-(l-methyl-1H-pyrazol-4-yl jQ-5-imethylsulfinyl)pyrimidin-4-ylamino)phenyl)acrylamide (Compound 24) preparation
Figure imgf000062_0001
Figure imgf000062_0001
( 1 ) 3- (2- (1-甲基 比唑- 4-基氨基 )-5- (甲硫基)嘧啶- 4-基氨基)苯基 氨基甲酸叔丁酯的制备  Preparation of (1) 3-(2-(1-methylpyrazol-4-ylamino)-5-(methylthio)pyrimidin-4-ylamino)phenylcarbamic acid tert-butyl ester
Figure imgf000062_0002
Figure imgf000062_0002
将 3- (2-氯- 5- (甲硫基)嘧啶- 4-基氨基)苯基氨基甲酸叔丁酯（ 0.60 g, 1.64 mmol) , 溶于 5 mL的叔戊醇中， 再加入醋酸 2滴， 1-甲基- 1H-吡 唑— 4—胺（0.163 g, 1.68 mmol) , 反应在 90 oC搅拌 13 h。 旋干， 过硅 胶柱（PE:EA=2:1 )得 0.52 g淡红色固体， 收率： 74.4%。  tert-Butyl 3-(2-chloro-5-(methylthio)pyrimidin-4-ylamino)phenylcarbamate (0.60 g, 1.64 mmol) was dissolved in 5 mL of tert-pentanol, then acetic acid was added 2 drops of 1-methyl-1H-pyrazole-4-amine (0.163 g, 1.68 mmol). The reaction was stirred at 90 ° C for 13 h. Spin dry, through a silica gel column (PE: EA = 2:1) gave 0.52 g of a pale red solid, yield: 74.4%.
( 2 ) (3-氨基苯基) -TV2- (1-甲基- 比唑- 4-基)- 5- (甲硫基）嘧啶 -2,4-二胺的制备 (2) Preparation of (3-aminophenyl)-TV 2 -(1-methyl-biazole-4-yl)- 5-(methylthio)pyrimidine-2,4-diamine
Figure imgf000062_0003
Figure imgf000062_0003
将 3- (2- (1-甲基 比唑- 4-基氨基 )-5- (甲硫基)嘧啶- 4-基氨基)苯基 氨基甲酸叔丁酯（ 0.52 g, 1.22 mmol)溶于 20 mL二氯甲烷中，加入 2 mL 三氟醋酸， 反应 2h , 将体系旋干， 得到粗品 0.39 g, 直接用于下一步。  Dissolve tert-butyl 3-(2-(1-methylpyrazol-4-ylamino)-5-(methylthio)pyrimidin-4-ylamino)phenylcarbamate (0.52 g, 1.22 mmol) In 20 mL of dichloromethane, 2 mL of trifluoroacetic acid was added and reacted for 2 h, and the system was spun dry to obtain a crude product (0.39 g) which was directly used for the next step.
(3 ) V-(3-(2-(l-甲基- 吡唑- 4-基氨基 )-5- (甲硫基)嘧啶- 4-基氨基) 苯基）丙烯酰胺的制备  (3) Preparation of V-(3-(2-(l-methyl-pyrazol-4-ylamino)-5-(methylthio)pyrimidin-4-ylamino)phenyl)acrylamide
Figure imgf000062_0004
Figure imgf000062_0004
将 N4- (3-氨基苯基)- TV2- (1-甲基- 吡唑- 4-基)- 5- (甲硫基)嘧啶- 2,4- 二胺粗品 0.16 g溶于 10 mL的 THF中，滴加 DIPEA( 0.126 g, 0.98 mmol ), 至溶液 pH值为 9, - 15oC下滴加含丙烯酰氯（ 0.048 g, 0.53 mmol ) 的 THF溶液 0.5 mL , 此温度下反应 15 min。 滴加 5滴甲醇， 将体系旋干， 直接用于下一步。 N 4 - (3-Aminophenyl)- TV 2 - (1-methyl-pyrazol-4-yl)-5-(methylthio)pyrimidine-2,4- 0.16 g of crude diamine was dissolved in 10 mL of THF, DIPEA (0.126 g, 0.98 mmol) was added dropwise, and the pH of the solution was 9, and the THF solution containing acryloyl chloride (0.048 g, 0.53 mmol) was added dropwise at -15 °C. mL, react at this temperature for 15 min. Five drops of methanol were added dropwise, and the system was spun dry and used directly in the next step.
( 4 ) V-(3- (2- (1-甲基- 吡唑- 4-基氨基 )- 5- (甲亚磺酰基)嘧啶- 4-基 氨基)苯基)丙烯酰胺的制  (4) Preparation of V-(3-(2-(1-methyl-pyrazol-4-ylamino)-5-(methylsulfinyl)pyrimidin-4-ylamino)phenyl)acrylamide
Figure imgf000063_0001
Figure imgf000063_0001
将上一步粗品溶于 15 mL甲醇和 5 mL水中， 然后緩慢加入过^ L酸 氢钾复合盐 (oxone) ( 0.301 g, 0.49 mmol ) , 在常温下反应 25 min, 将溶 液旋干， 加入 30 mL二氯甲烷和 30 mL水， 萃取， 有机相用无水硫酸钠 干燥， 抽滤， 滤液旋干， 过硅胶柱 (DCM: 甲醇 =20:1)得 0.040 g白色固 体。  The crude product in the previous step was dissolved in 15 mL of methanol and 5 mL of water, then slowly added potassium hydride (oxone) (0.301 g, 0.49 mmol), reacted at room temperature for 25 min, the solution was spun dry, and added to 30 The organic layer was dried over anhydrous sodium sulfate (MgSO4), filtered, filtered,jjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjj
分子式： C18H19N702S 分子量： 397.1 盾语 (m/z): 398.1 (Μ+1)+· NMR(i 6- DMSO+HC1, 400 MHz) 5(ppm): 10.2 (IH, m), 9.68-9.35 (2H, m), 8.20 (IH, s), 7.91-7.15 (6H, m), 6.42 (IH, dd), 6.24 (IH, d), 5.75 (IH, dd), 3.78, 3.59 (3H, 两个单峰)， 2.94 (3H, s). Molecular formula: C 18 H 19 N 7 0 2 S Molecular weight: 397.1 Shield (m/z): 398.1 (Μ+1)+· NMR (i 6 - DMSO+HC1, 400 MHz) 5 (ppm): 10.2 (IH , m), 9.68-9.35 (2H, m), 8.20 (IH, s), 7.91-7.15 (6H, m), 6.42 (IH, dd), 6.24 (IH, d), 5.75 (IH, dd), 3.78, 3.59 (3H, two single peaks), 2.94 (3H, s).
实施例 20 V-i3-i2-a-i2-甲氧基乙基) -1 -吡唑 -4-基 jQ-5- (甲亚 磺酰 -4-基氨基)苯基)丙烯酰胺 (化合物 103)  Example 20 V-i3-i2-a-i2-methoxyethyl)-1-pyrazol-4-yl jQ-5-(methylsulfinyl-4-ylamino)phenyl)acrylamide (compound) 103)
Figure imgf000063_0002
Figure imgf000063_0002
( 1 ) 1— (2-甲氧基乙基) -4-硝基 吡唑的制备 将 4-硝基吡唑 (2.0 g, 17.68 mmol) , 2-氯乙基甲基醚 (2.01 g, 21.26 mmol)和碳酸钾（4.88 g, 35.31 mmol)溶于 40 mL乙腈中所得 溶液回流 18 h, 旋干， 过硅胶柱（PE/EA=4:1 )得 1.5 g淡黄色固体， 收 率： 49.5%。 (1) Preparation of 1-(2-methoxyethyl)-4-nitropyrazole 4-Nitropyrazole (2.0 g, 17.68 mmol), 2-chloroethyl methyl ether (2.01 g, 21.26 mmol) and potassium carbonate (4.88 g, 35.31 mmol) were dissolved in 40 mL acetonitrile. h, spin dry, passed through a silica gel column (PE/EA = 4:1) yielding 1.5 g of pale yellow solid, yield: 49.5%.
(2) 1- (2-甲氧基乙基) - -吡唑- 4-胺的制备
Figure imgf000064_0001
(2) Preparation of 1-(2-methoxyethyl)--pyrazole-4-amine
Figure imgf000064_0001
将 1- (2-甲氧基乙基) - 4-硝基- 吡唑（ 1.2 g, 7.01 mmol)溶于 25 mL 甲醇中， 加入 0.08 g 10%的 Pd/C, 用氢气置换三次， 在氢气氛围下搅拌 过夜， 反应完全后抽滤， 甲醇洗涤滤饼， 旋干滤液得淡红色油状物 0.80 g, 收率： 80.9%。  1-(2-Methoxyethyl)-4-nitro-pyrazole (1.2 g, 7.01 mmol) was dissolved in 25 mL of methanol, and 0.08 g of 10% Pd/C was added and replaced with hydrogen three times. The mixture was stirred overnight under a hydrogen atmosphere. After the reaction was completed, the mixture was filtered, and the filter cake was washed with methanol. The filtrate was evaporated to give a pale red oil (yield: 0.80 g, yield: 80.9%).
(3) 3- (2- (1- (2-甲氧基乙基) 吡唑- 4-基氨基 )-5- (甲硫基)嘧啶- 4- 基氨基)苯基氨基甲酸  (3) 3-(2-(1-(2-methoxyethyl)pyrazole-4-ylamino)-5-(methylthio)pyrimidin-4-ylamino)phenylcarbamic acid
Figure imgf000064_0002
Figure imgf000064_0002
将 3- (2-氯- 5- (甲硫基)嘧啶- 4-基氨基)苯基氨基甲酸叔丁酯（ 0.560 g, tert-Butyl 3-(2-chloro-5-(methylthio)pyrimidin-4-ylamino)phenylcarbamate (0.560 g,
1.53 mmol) ,溶于 5 mL的叔戊醇中，加入 1- (2-甲氧基乙基) - 吡唑- 4- 胺 ( 0.234 g, 1.66 mmol)和 DIPEA ( 0.292 g, 2.26 mmol) , 在 110°C 搅拌 18 h。 旋干， 过硅胶柱（PE:EA=2:1 )得 0.42 g白色固体， 收率： 58.2%。 1.53 mmol), dissolved in 5 mL of tert-amyl alcohol, 1-(2-methoxyethyl)-pyrazole-4-amine (0.234 g, 1.66 mmol) and DIPEA (0.292 g, 2.26 mmol). Stir at 110 ° C for 18 h. It was spin-dried and passed through a silica gel column (PE: EA = 2:1) to give 0.42 g of white solid, yield: 58.2%.
(4) (3-氨基苯基 )-V2- (1- (2-甲氧基乙基) - p比唑- 4-基)- 5- (甲硫 基)嘧啶- 2,4-二胺三氟乙酸盐的制备 (4) (3-Aminophenyl)-V 2 - (1-(2-methoxyethyl) - p-pyrazol-4-yl)- 5-(methylthio)pyrimidine-2,4-di Preparation of amine trifluoroacetate
Figure imgf000064_0003
Figure imgf000064_0003
将 3- (2- (1- (2-甲氧基乙基) 比唑- 4-基氨基 )-5- (甲硫基)嘧啶- 4-基 氨基)苯基氨基甲酸叔丁酯 （0.15 g, 0.318 mmol)溶于 20 mL二氯甲烷 中， 加入 2 mL三氟醋酸， 反应 2 h , 将体系旋干， 得到粗品 0.12 g, 直接用于下一步。 tert-Butyl 3-(2-(1-(2-methoxyethyl)-benza-4-ylamino)-5-(methylthio)pyrimidin-4-ylamino)phenylcarbamate (0.15 g, 0.318 mmol) was dissolved in 20 mL of dichloromethane, 2 mL of trifluoroacetic acid was added, the reaction was carried out for 2 h, and the system was dried to give a crude product (0.12 g). Used directly in the next step.
( 5 ) V-(3-(2-(l-(2-甲氧基乙基) - 吡唑- 4-基氨基 )- 5- (甲硫基)嘧啶 -4-基氨基)苯基)丙烯  (5) V-(3-(2-(l-(2-methoxyethyl)-pyrazol-4-ylamino)-5-(methylthio)pyrimidin-4-ylamino)phenyl) Propylene
Figure imgf000065_0001
Figure imgf000065_0001
将上步粗品 0.118 g溶于 10 mL的 THF中， 滴加 DIPEA ( 0.082 g, The above crude product 0.118 g was dissolved in 10 mL of THF, and DIPEA (0.082 g,
0.635 mmol ) , 直至溶液 ρΗ值为 9, -15 °C下滴加含丙烯酰氯（ 0.029 g, 0.320 mmol )的 THF溶液 0.5 mL,加毕此温度下反应 15 min。滴加 2 mL 甲醇， 将体系旋干， 得棕黑色油状物 0.21 g, 直接用于下一步。 0.635 mmol ) , 0.5 mL of THF solution containing acryloyl chloride (0.029 g, 0.320 mmol) was added dropwise at a concentration of 9, -15 °C, and the reaction was carried out for 15 min at this temperature. 2 mL of methanol was added dropwise, and the system was dried to give a brown-brown oil (0.21 g).
( 6 ) V-(3-(2-(l-(2-甲氧基乙基)- 吡唑- 4-基氨基 )- 5- (甲亚磺酰基) 嘧啶- 4-基氨基)苯基）丙  (6) V-(3-(2-(l-(2-methoxyethyl)-pyrazol-4-ylamino)- 5-(methylsulfinyl)pyrimidin-4-ylamino)phenyl ) C
Figure imgf000065_0002
Figure imgf000065_0002
将上一步粗品 0.21 g溶于 20 mL甲醇和 2 mL水中， 然后緩慢加入 溶于 2 mL水的过 酸氢钾复合盐（oxone) ( 0.301 g, 0.49 mmol ) , 在常 温下反应 25 min, 加入 10 mL饱和的硫代硫酸钠水溶液淬灭， 再加入 50 mL二氯甲烷和 30 mL水， 萃取， 有机相用无水硫酸钠干燥， 抽滤， 滤液旋干， 过硅胶柱 (DCM: 甲醇 =20:1)得 0.096 g白色固体。  0.21 g of the previous step was dissolved in 20 mL of methanol and 2 mL of water, then slowly added potassium hydride (oxone) (0.301 g, 0.49 mmol) dissolved in 2 mL of water, and reacted at room temperature for 25 min. After quenching with 10 mL of a saturated aqueous solution of sodium thiosulfate, 50 mL of dichloromethane and 30 mL of water were added, and the organic phase was dried over anhydrous sodium sulfate, filtered, filtered and evaporated to dryness. 20:1) 0.096 g of a white solid was obtained.
分子式： C20H23N7O3S 分子量： 441.2 盾语 (m/z): 442.2 (M+l)+. Molecular formula: C 20 H 23 N 7 O 3 S Molecular weight: 441.2 Shield (m/z): 442.2 (M+l) + .
NMR(i 6- DMSO+HC1, 400 MHz) 5(ppm): 10.25-10.12 (IH, m), 9.68-9.35 (2H, m), 8.20 (IH, s), 7.95-7.17 (6H, m), 6.42 (IH, dd), 6.24 (IH, d), 5.75 (IH, dd), 4.24-3.92 (2H, m), 3.68-3.47 (2H, m), 3.21, 3.11 (3H, 两 个单峰)， 2.94 (3H, s). NMR (i 6 - DMSO + HC1, 400 MHz) 5 (ppm): 10.25-10.12 (IH, m), 9.68-9.35 (2H, m), 8.20 (IH, s), 7.95-7.17 (6H, m) , 6.42 (IH, dd), 6.24 (IH, d), 5.75 (IH, dd), 4.24-3.92 (2H, m), 3.68-3.47 (2H, m), 3.21, 3.11 (3H, two single peaks ), 2.94 (3H, s).
实施例 21 V-(3-i2-(l-甲基 -1 -吡唑 -4-基 jQ-5-i甲亚磺酰基)嘧 啶 -4-基 JQ苯基) -4- (吡咯烷 -1-基)丁 -2-烯酰胺 (化合物 104)的制备
Figure imgf000066_0001
Example 21 V-(3-i2-(l-methyl-1-pyrazol-4-yl jQ-5-imethylsulfinyl)pyrimidin-4-yl JQphenyl)-4-(pyrrolidine- Preparation of 1-yl)but-2-enamide (Compound 104)
Figure imgf000066_0001
( 1 ) 4-溴-V-(3- (2- (1-甲基- 吡唑- 4-基氨基 )- 5- (甲硫基)嘧啶- 4-基 氨基)苯基)丁- 2-烯酰胺的  (1) 4-Bromo-V-(3-(2-(1-methyl-pyrazol-4-ylamino)-5-(methylthio)pyrimidin-4-ylamino)phenyl)butane-2 -enamide
Figure imgf000066_0002
Figure imgf000066_0002
将 4-溴巴豆酸（ 0.106 g, 0.64 mmol)溶于 10 mL二氯甲烷中， 加入 2滴 DMF, 然后在 0°C时加入 2 mL草酰氯，搅拌 2 h,旋干，备用。  4-Bromocrotonic acid (0.106 g, 0.64 mmol) was dissolved in 10 mL of dichloromethane, and 2 drops of DMF were added, then 2 mL of oxalyl chloride was added at 0 ° C, stirred for 2 h, and dried, and was taken.
将 N4- (3-氨基苯基)- TV2- (1-甲基- p比唑- 4-基)- 5- (甲硫基)嘧啶- 2,4- 二胺（ 0.21 g, 0.64 mmol)溶于 20 mL干燥的四氢呋喃中， 加入 DIPEA ( 0.248 g, 1.92 mmol)使得 pH值到 10,然后将 4-溴巴豆酰氯溶于 5 mL 二氯甲烷中， 緩慢的滴加到体系中， 反应在 0°C搅拌 2 h, 放置备用。 N 4 -(3-Aminophenyl)-TV 2 -(1-methyl-p-pyrazol-4-yl)-5-(methylthio)pyrimidine-2,4-diamine (0.21 g, 0.64 Ment) was dissolved in 20 mL of dry tetrahydrofuran, DIPEA (0.248 g, 1.92 mmol) was added to bring the pH to 10, then 4-bromo crotonyl chloride was dissolved in 5 mL of dichloromethane and slowly added dropwise to the system. The reaction was stirred at 0 ° C for 2 h and placed for later use.
(2 ) V-(3- (2- (1-甲基- p比唑- 4-基氨基 )- 5- (甲硫基)嘧啶- 4-基氨基) 苯基)—4— (吡咯烷— 1—基)丁  (2) V-(3-(2-(1-methyl-p-butyr-4-ylamino)- 5-(methylthio)pyrimidin-4-ylamino)phenyl)-4-(pyrrolidine) — 1—基)丁
Figure imgf000066_0003
Figure imgf000066_0003
向上一步得到的溶液中加入 0.5 mL DIPEA, 吡咯烷（ 0.055 g, 0.77 mmol)在 0°C时， 搅拌 0.5 h, 加入 30 mL二氯甲烷和 30 mL水， 萃取， 有机相用无水硫酸钠干燥，抽滤，滤液旋干，过硅胶柱 (DCM: 甲醇 =40:1) 得 0.12 g白色固体， 两步收率： 40.2%。  Add 0.5 mL of DIPEA to the solution obtained in the previous step, pyrrolidine (0.055 g, 0.77 mmol) at 0 ° C, stir for 0.5 h, add 30 mL of dichloromethane and 30 mL of water, extract, and extract the organic phase with anhydrous sodium sulfate. Drying, suction filtration, and the filtrate was evaporated to dryness eluting with silica gel (DCM: MeOH = 40:1) to give 0.12 g of white solid.
( 3 ) V-(3- (2- (1-甲基- 吡唑- 4-基氨基 )- 5- (甲亚磺酰基)嘧啶- 4-基氨 基)苯基)—4- (吡咯烷- 1-基)丁- 2-烯酰胺的制备
Figure imgf000067_0001
(3) V-(3-(2-(1-methyl-pyrazol-4-ylamino)-5-(methylsulfinyl)pyrimidin-4-ylamino)phenyl)-4-(pyrrolidine) -1-Based Preparation of Butyl-2-Enamide
Figure imgf000067_0001
将 V-(3- (2- (1-甲基- p比唑- 4-基氨基 )- 5- (甲硫基)嘧啶- 4-基氨基)苯 基)— 4- (吡咯烷- 1-基)丁- 2-烯酰胺 ( 0.10 g, 0.215 mmol )溶于 15 mL甲醇 中， 然后緩慢加入溶于 2 mL水的过硫酸氢钾复合盐 (oxone) ( 0.146 g, 0.237 mmol ) , 在常温下反应 25 min, 然后力口入 10 mL饱和石 代石 酸納 溶液淬灭， 然后再加入 50 mL二氯甲烷和 30 mL水， 萃取， 有机相用无 水硫酸钠干燥， 抽滤， 滤液旋干， 过硅胶柱 (DCM: 甲醇 =20:1)得 0.070 g 白色固体， 收率： 67.9%。  V-(3-(2-(1-methyl-p-pyrazol-4-ylamino)-5-(methylthio)pyrimidin-4-ylamino)phenyl)-4-(pyrrolidin-1) -yl)but-2-enamide (0.10 g, 0.215 mmol) dissolved in 15 mL of methanol, then slowly added potassium peroxodisulfate complex (oxone) (0.146 g, 0.237 mmol) dissolved in 2 mL of water. The reaction was carried out at room temperature for 25 min, then quenched with 10 mL of saturated sodium sulphate solution, then 50 mL of dichloromethane and 30 mL of water were added, and the organic phase was dried over anhydrous sodium sulfate, suction filtered, filtrate The mixture was dried with EtOAc (EtOAc:EtOAc)
分子量: 480.2 盾语 (m/z): 241.2 ( M/2+1 ) +, 481.2 (M+1 )+· Molecular weight: 480.2 Shield (m/z): 241.2 ( M/2+1 ) + , 481.2 (M+1 )+·
NMR(i 6- DMSO+HC1, 400 MHz) 5(ppm): 10.58-10.40 (1H, m), 9.72-9.48 (2H, m), 8.20 (1H, s), 8.02-7.15 (6H, m), 6.85-6.38 (2H, m), 3.86 (2H, s), 3.77 (3H, s), 3.20-3.05 (4H, m), 2.93 (3H, s), 1.90 (4H, m). NMR (i 6 - DMSO + HC1, 400 MHz) 5 (ppm): 10.58-10.40 (1H, m), 9.72-9.48 (2H, m), 8.20 (1H, s), 8.02-7.15 (6H, m) , 6.85-6.38 (2H, m), 3.86 (2H, s), 3.77 (3H, s), 3.20-3.05 (4H, m), 2.93 (3H, s), 1.90 (4H, m).
实施例 22 V-i3-i2-(4-(2-甲氧基乙氧基)苯基 jQ-5-i甲亚磺酰基)  Example 22 V-i3-i2-(4-(2-methoxyethoxy)phenyl jQ-5-i methylsulfinyl)
Figure imgf000067_0002
Figure imgf000067_0002
( 1 ) 4-甲基戊 -2-烯酰氯的
Figure imgf000067_0003
(1) 4-methylpent-2-enoyl chloride
Figure imgf000067_0003
将 4-甲基- 2-戊酸（0.50 g, 4.38 mmol ) , 溶于 20 mL二氯甲烷中， 再加入 1滴 DMF ,冷却至 0°C ,然后向该溶液中緩' I"曼滴加草酰氯（ 1.11 g, 8.75 mmol ) , 滴加完毕后， 在常温下搅拌 1 h, 旋干， 得粗品 0.58 g。 ( 2 ) TV- (3- (2- (4- (2-甲氧基乙氧基)苯基氨基)- 5- (甲硫基)嘧啶- 4-基氨 基)苯基)—4-甲基戊 -2- 4-Methyl-2-pentanoic acid (0.50 g, 4.38 mmol) was dissolved in 20 mL of dichloromethane, then 1 drop of DMF was added, cooled to 0 ° C, and then the solution was slowly cooled. Add oxalyl chloride ( 1.11 g, 8.75 mmol ), after the addition was completed, stir at room temperature for 1 h, and spin dry to obtain a crude product of 0.58 g. (2) TV-(3-(2-(4-(2-methoxyethoxy)phenylamino)- 5-(methylthio)pyrimidin-4-ylamino)phenyl)-4-yl Kepentan-2-
Figure imgf000068_0001
Figure imgf000068_0001
-(3-氨基苯基)- Λ^- (4- (2-甲氧基乙氧基)苯基) -5- (甲硫基)嘧啶- 2,4- 二胺（ 0.20 g, 0.503 mmol )溶于 15 mL的 THF中 , 滴加 DIPEA ( 0.194 g, 1.50 mmol ) , 直至溶液 pH值为 9 , - 15 °C下滴加含有 4-甲基戊 -2-烯 酰氯（ 0.070 g, 0.528 mmol ) 的 THF溶液 0.5 mL , 加毕此温度下反应 15 min。 滴加 5滴甲醇， 将体系旋干， 得 0.425 g棕黑色油状粗品， 直 接用于下一步。  -(3-aminophenyl)- Λ^-(4-(2-methoxyethoxy)phenyl)-5-(methylthio)pyrimidine-2,4-diamine (0.20 g, 0.503 mmol Dissolve in 15 mL of THF, add DIPEA (0.194 g, 1.50 mmol) dropwise, until the pH of the solution is 9, at - 15 °C, add 4-methylpent-2-enoyl chloride (0.070 g, 0.528) Methanol) 0.5 mL of THF solution, and reacted at this temperature for 15 min. 5 drops of methanol were added dropwise, and the system was spun dry to give a crude product of 0.425 g of brownish brown oil, which was used directly for the next step.
( 3 ) TV- (3- (2- (4- (2-甲氧基乙氧基)苯基氨基)- 5- (甲亚磺酰基)嘧啶- 4- 基氨基)苯基) -4-甲基 -2-烯酰胺的制备  (3) TV-(3-(2-(4-(2-methoxyethoxy)phenylamino)-5-(methylsulfinyl)pyrimidin-4-ylamino)phenyl)-4- Preparation of methyl-2-enoamide
Figure imgf000068_0002
Figure imgf000068_0002
将上一步粗品 0.425 g溶于 10 mL甲醇和 2 mL水中，然后緩慢加入 溶于 2 mL水的过硫酸氢钾复合盐 (oxone) ( 0.309 g, 0.503 mmol )， 在常 温下反应 10 min, 加入 15 mL饱和的硫代硫酸钠水溶液淬灭， 再加入 50 mL二氯甲烷和 30 mL水， 萃取， 有机相用无水硫酸钠干燥， 抽滤， 滤液旋干， 过硅胶柱 (DCM: 甲醇 =20: 1)得 0.060 g白色固体， 两步收率： 23.5%。  The above crude product 0.425 g was dissolved in 10 mL of methanol and 2 mL of water, then slowly added potassium persulfate complex salt (oxone) (0.309 g, 0.503 mmol) dissolved in 2 mL of water, and reacted at room temperature for 10 min, added After quenching with 15 mL of saturated aqueous sodium thiosulfate solution, 50 mL of dichloromethane and 30 mL of water were added, and the organic phase was dried over anhydrous sodium sulfate, filtered, filtered, and evaporated to dryness. 20: 1) 0.060 g of white solid was obtained in two steps yield: 23.5%.
分子式： C26H31N504S 分子量： 509.2 盾语 (m/z): 510.1 (Μ+1)+·Molecular formula: C 26 H 31 N 5 0 4 S Molecular weight: 509.2 Shield (m/z): 510.1 (Μ+1)+·
NMR(i 6- DMSO+HC1, 400 MHz) 5(ppm): 10.02 (1H, s), 9.48 (2H, s), 8. 19 (1H, s), 7.73 (1H, s), 7.53-7.22 (5H, m), 6.84-6.72 (3H, m), 6.05 (1H, dd), 4.04-3.96 (2H, m), 3.65-3.59 (2H, m), 3.29 (3H, s), 2.95 (3H, s), 1.03 (6H, d). 烯丙基的 CH被 DMSO峰压住。 参考上述制备方法， 还制备了以下化合物: NMR (i 6 - DMSO + HC1, 400 MHz) 5 (ppm): 10.02 (1H, s), 9.48 (2H, s), 8. 19 (1H, s), 7.73 (1H, s), 7.53-7.22 (5H, m), 6.84-6.72 (3H, m), 6.05 (1H, dd), 4.04-3.96 (2H, m), 3.65-3.59 (2H, m), 3.29 (3H, s), 2.95 (3H , s), 1.03 (6H, d). The allylic CH is pressed by the DMSO peak. Referring to the above preparation method, the following compounds were also prepared:
Figure imgf000069_0001
Figure imgf000070_0001
Figure imgf000069_0001
Figure imgf000070_0001
Figure imgf000071_0001
Figure imgf000071_0001
SSSO請 ΪΟΖ OAV SSSO please ΪΟΖ OAV

Claims

权 利 要 求 Rights request
1、 通式 （ I ) 所示的化合物、 其药学上可接受的盐或其立体异构 1. Compounds represented by general formula (I), their pharmaceutically acceptable salts or their stereoisomers
Figure imgf000072_0001
Figure imgf000072_0001
其中， 环 A和环 B分别独立的表示苯基， 3- 7元环烷基， 含有 N、 0、 S杂原子的 3- 7元杂环烷基， 4- 7元杂芳基或 6-10元二环结构； Among them, Ring A and Ring B independently represent phenyl, 3-7-membered cycloalkyl, 3-7-membered heterocycloalkyl containing N, 0, S heteroatoms, 4-7-membered heteroaryl or 6- 10-membered bicyclic structure;
Li和 L2分别独立的表示共价键， -NH-, - N(C1-3烷基) -， -0-, - S(0)m- , - N(C1-3 烷基 )C(0)- , - C(0)N(C1-3 烷基） -， - N(C1-3 烷基 )S(0)2-或― S(0)2N(C1-3烷基) -； Li and L 2 independently represent covalent bonds, -NH-, - N(C 1-3 alkyl) -, -0-, - S(0) m -, - N(C 1-3 alkyl) C(0)- , - C(0)N(C 1-3 alkyl) -, - N(C 1-3 alkyl)S(0) 2 -or- S(0) 2 N(C 1- 3alkyl )-;
a表示共价键， 未被取代或被 d_4烷基取代的亚氨基； a represents a covalent bond, an imino group that is unsubstituted or substituted by d_4 alkyl;
b表示- CO-或- S02-； b means - CO- or - S0 2 -;
c表示未被取代或被一或两个甲基或三氟甲基取代的 1,3-亚丙烯基、 1,1-或 1,2-亚乙烯基， 亚乙炔基， 或者未被取代或被一至四个甲基或三 氟甲基取代的 1,3-丁二烯 -1,4-亚基； c represents 1,3-propenylene, 1,1- or 1,2-vinylidene, ethynylene, which is unsubstituted or substituted by one or two methyl or trifluoromethyl groups, or is unsubstituted or 1,3-butadiene-1,4-ylidene substituted by one to four methyl or trifluoromethyl groups;
d表示共价键或 亚烷基； d represents covalent bond or alkylene group;
e表示氢原子， 烷氧基，氨基， 3- 7元环烷基， 6- 10元二环结构， 烷基氨基或二-（Cw烷基）氨基， 其中烷基部分可相同或不同； 和 分别独立的表示氢原子， 卤素原子，氰基，硝基， CM烯基， C2-4炔基或 - L3- R4, e represents a hydrogen atom, an alkoxy group, an amino group, a 3-7-membered cycloalkyl group, a 6-10-membered bicyclic structure, an alkylamino group or a di-(Cw alkyl)amino group, in which the alkyl parts may be the same or different; and Each independently represents a hydrogen atom, a halogen atom, a cyano group, a nitro group, a CM alkenyl group, a C 2-4 alkynyl group or - L 3 - R 4 ,
L3表示共价键, -ΝΗ-, - N(C1-3烷基) -， - O- , - Ο- C1-3亚烷基-， -S-Ci-3 亚烷基-， -S(0)m -, -C(O)-, -NHC(O)-, - N(C1-3烷基) C(O)- , - C(0)NH- , -C(0)N(Ci-3 烷基） - , - NHS(0)2- , -N(Ci-3 烷基） S(0)2- , -S(0)2NH- , - S(0)2N(C1-3烷基) -, - oc(o)-或- c(o)o- , L 3 represents a covalent bond, -NH-, - N(C 1-3 alkyl) -, - O-, - O-C 1-3 alkylene-, -S-Ci -3 alkylene-, -S(0) m -, -C(O)-, -NHC(O)-, - N(C 1-3 alkyl) C(O)- , - C(0)NH- , -C(0 )N(Ci -3alkyl ) - , - NHS(0) 2 - , -N(Ci -3alkyl ) S(0) 2 - , -S(0) 2 NH- , - S(0) 2 N(C 1-3 alkyl) -, - oc(o)- or - c(o)o-,
R4表示氢原子, C1-4烷基， - N(C1-3烷基 )2, - NHC(0)0- (C1-4烷基)， -OH, - 0(C1-4烷基)， - S(0)2(C 烷基）， 3- 7元环烷基， 苯基或 5- 6元杂 芳基； R4 represents a hydrogen atom, C 1-4 alkyl, - N(C 1-3 alkyl) 2 , - NHC(0)0- (C 1-4 alkyl), -OH, - 0(C 1-4 Alkyl), -S(0) 2 (C alkyl), 3-7-membered cycloalkyl, phenyl or 5-6-membered heteroaryl;
R2表示- L4- R5 , R 2 means - L 4 - R 5 ,
表示共价键， -NH- -N(C1-3烷基) -， - 0- C1-3亚烷基-， - S- C1-3亚 烷基-或 - S(0)m- , R5表示氢原子， C2-4烯基， C2-4炔基， C1-4烷基， 氨基， - NH(C1-3 烷基）， - N(Cw烷基 )2, -OH, - 0(Cw烷基)， - C(0)(Cw烷基)或 3- 7元环 烷基， Represents a covalent bond, -NH- -N(C 1-3 alkyl) -, - 0- C 1-3 alkylene-, - S- C 1-3 alkylene - or - S(0) m - , R 5 represents a hydrogen atom, C 2-4 alkenyl, C 2-4 alkynyl, C 1-4 alkyl, amino , - NH (C 1-3 alkyl), - N (C alkyl) 2 , - OH, - 0(Cw alkyl), - C(0)(Cw alkyl) or 3-7 membered cycloalkyl,
其中 L4为共价键时， R5不能为 C1-4烷基， - OH, - 0(C1-3烷基）， - c(o)(c1-3烷基)， When L 4 is a covalent bond, R 5 cannot be C 1-4 alkyl, - OH, - 0(C 1-3 alkyl), - c(o)(c 1-3 alkyl),
L4为- 0- d_3亚烷基 -时， R5不能为氢原子； When L 4 is -0- d_3alkylene- , R 5 cannot be a hydrogen atom;
所述 烷基部分、 环烷基、 杂芳基可以进一步被 1至 4个（^取 代， The alkyl part, cycloalkyl group, and heteroaryl group may be further substituted by 1 to 4 (^,
表示 素原子、 烷基、 氨基、 烷基氨基、 二-（Cw烷基） 氨基、 羟基、 烷氧基、 烷氧羰基、 氨基甲酰基、 烷基氨基甲 酰基、 二- ( C 烷基）氨基甲酰基或 3-6元环烷基， 其中 （^可以相同 或不同； Represents element atom, alkyl group, amino group, alkylamino group, di-(C alkyl) amino group, hydroxyl group, alkoxy group, alkoxycarbonyl group, carbamoyl group, alkyl carbamoyl group, di-(C alkyl) amino group Formyl or 3-6 membered cycloalkyl, where (^ can be the same or different;
所述环烷基、二环结构上碳原子可以被 1 -4个相同或不同的 N、 NH、 N(C1-3烷基)、 0、 S(0)m、 C(O)替换； The carbon atoms on the cycloalkyl and bicyclic structures can be replaced by 1-4 identical or different N, NH, N(C 1-3 alkyl), 0, S(0) m , C(O);
所述杂芳基含有 1-4个杂原子， 分别独立的选 N、 0或 S; The heteroaryl group contains 1-4 heteroatoms, each of which is independently selected from N, 0 or S;
m表示 0, 1或 2; m represents 0, 1 or 2;
p和 q分别独立的表示 0, 1 , 2 , 3或 4。 p and q independently represent 0, 1, 2, 3 or 4.
2、 如权利要求 1 所述的化合物、 其药学上可接受的盐或其立体异 构体： 2. The compound of claim 1, its pharmaceutically acceptable salt or its stereoisomer:
其中， R2表示- L4- R5 , Where, R 2 represents - L 4 - R 5 ,
L4表示- S- C1-3亚烷基 -或- S(0)m- , L 4 represents - S- C 1-3 alkylene - or - S(0) m -,
R5表示氢原子， C2-4烯基， C2-4炔基， C1-4烷基， 氨基， - NH(C1-3 烷基）， - N(Cw烷基 )2, -OH, - 0(Cw烷基)， - C(0)(Cw烷基)或 3- 7元环 烷基； R 5 represents a hydrogen atom, C 2-4 alkenyl, C 2-4 alkynyl, C 1-4 alkyl, amino , - NH (C 1-3 alkyl), - N (C alkyl) 2 , - OH, - 0(Cw alkyl), - C(0)(Cw alkyl) or 3-7 membered cycloalkyl;
m表示 0, 1或 2。 m represents 0, 1 or 2.
3、 如权利要求 2所述的化合物、 其药学上可接受的盐或其立体异 构体： 3. The compound of claim 2, its pharmaceutically acceptable salt or its stereoisomer:
其中， 环 A和环 B分别独立的表示苯基， 5- 6元环烷基， 含有 N、 0、 S杂原子的 5- 6元杂环烷基， 5- 6元杂芳基或 8- 10元二环结构； Among them, Ring A and Ring B independently represent phenyl, 5-6-membered cycloalkyl, 5-6-membered heterocycloalkyl containing N, 0, S heteroatoms, 5-6-membered heteroaryl or 8- 10-membered bicyclic structure;
和 L2分别独立的表示共价键， -NH-, -N(CH3)-, -Ο-, -S(0)m -, and L 2 independently represent covalent bonds, -NH-, -N(CH 3 )-, -Ο-, -S(0) m -,
-N(CH3)C(0)-, -C(0)N(CH3) -, - N(CH3)S(0)2-或- S(0)2N(CH3) -； -N(CH 3 )C(0)-, -C(0)N(CH 3 ) -, - N(CH 3 )S(0) 2 -or- S(0) 2 N(CH 3 ) -;
a表示共价键， 未被取代或被 CH3取代的亚氨基； b表示- CO-或- S02-； a represents a covalent bond, unsubstituted or imino substituted by CH 3 ; b means - CO- or - S0 2 -;
c表示未被取代或被一或两个甲基取代的 1,2-亚乙烯基或亚乙炔基； d表示共价键或亚甲基； c represents 1,2-vinylidene or ethynylene group that is unsubstituted or substituted by one or two methyl groups; d represents a covalent bond or methylene group;
e表示氢原子， 甲氧基， 氨基， 哌啶基， 吗啉基， 吡咯烷基， 哌嗪 基， 甲基哌嗪基， 6- 9元螺环结构， 6- 8元并环结构， 6- 8元桥环结构， 甲基氨基或二- (甲基）氨基； e represents a hydrogen atom, methoxy group, amino group, piperidinyl group, morpholinyl group, pyrrolidinyl group, piperazinyl group, methylpiperazinyl group, 6-9 membered spiro ring structure, 6-8 membered parallel ring structure, 6 - 8-membered bridged ring structure, methylamino or di-(methyl)amino;
和 分别独立的表示氢原子， 卤素原子， 硝基或 - L3- R4 , and independently represent hydrogen atom, halogen atom, nitro group or - L 3 - R4,
L3表示共价键， -NH-, - N(C1-3烷基) -， -0-, - 0- C1-3亚烷基-， -S-Ci-3 亚烷基， - S(0)m- , - C(O)- , -NHC(O)-, - C(0)NH- , - NHS(0)2- ,― S(0)2NH- , - oc(o)-或- c(o)o- , L 3 represents a covalent bond, -NH-, - N(C 1-3 alkyl) -, -0-, - 0- C 1-3 alkylene-, -S-Ci -3 alkylene, - S(0) m - , - C(O)- , -NHC(O)-, - C(0)NH- , - NHS(0) 2 - ,― S(0) 2 NH- , - oc(o )-or- c(o)o-,
R4表示氢原子， 烷基， - N(Cw烷基 )2 , - NHC(0)0- (Cw烷基)， -OH, - 0(C1-4烷基)， - S(0)2(C1-3烷基）， 5- 6元环烷基， 苯基或 5- 6元杂 芳基； R4 represents a hydrogen atom, alkyl group, - N(Cw alkyl)2, - NHC(0)0- (Cw alkyl), -OH, - 0(C 1-4 alkyl), - S(0) 2 (C 1-3 alkyl), 5-6 membered cycloalkyl, phenyl or 5-6 membered heteroaryl;
R2表示- L4- R5 , R 2 means - L 4 - R 5 ,
L4表示- S- C1-3亚烷基 -或- S(0)m- , L 4 represents - S- C 1-3 alkylene - or - S(0) m -,
R5表示氢原子， C1-4烷基， 氨基， - NH(C1-3烷基)， - N(C1-3烷基 )2, -OH, - 0(Cw烷基)， - C(0)(Cw烷基)或 4-7元环烷基， R 5 represents a hydrogen atom, C 1-4 alkyl group, amino group, - NH (C 1-3 alkyl group), - N (C 1-3 alkyl group) 2 , -OH, - 0 (Cw alkyl group), - C(0)(Cw alkyl) or 4-7 membered cycloalkyl,
所述 烷基部分、 环烷基、 苯基、 杂芳基、 螺环结构、 并环结构、 桥环结构可以进一步被 1- 4个（^取代， The alkyl part, cycloalkyl, phenyl, heteroaryl, spiro ring structure, paracyclic structure, and bridged ring structure can be further substituted by 1-4 (^,
Qi表示 素原子、 C 烷基、 氨基、 烷基氨基、 二 -（C 烷基） 氨基、 羟基、 烷氧基、 烷氧羰基、 氨基甲酰基、 烷基氨基甲 酰基、 二 -（C 烷基）氨基甲酰基或 5- 6元环烷基， 其中（^可以相同 或不同； Qi represents a primary atom, C alkyl, amino, alkylamino, di-(C alkyl) amino, hydroxyl, alkoxy, alkoxycarbonyl, carbamoyl, alkylcarbamoyl, di-(C alkyl) ) carbamoyl or 5-6 membered cycloalkyl, where (^ can be the same or different;
所述环烷基、二环结构上碳原子可以被 1 -4个相同或不同的 N、 NH、 N(C1-3烷基)、 0、 S(0)m、 C(O)替换； The carbon atoms on the cycloalkyl and bicyclic structures can be replaced by 1-4 identical or different N, NH, N(C 1-3 alkyl), 0, S(0) m , C(O);
所述杂芳基、 螺环结构、 并环结构、 桥环结构含有 1-4个杂原子， 分别独立的选自 N、 0或 S; The heteroaryl group, spiro ring structure, parallel ring structure, and bridged ring structure contain 1-4 heteroatoms, each independently selected from N, 0 or S;
m表示 0, 1或 2; m represents 0, 1 or 2;
p和 q分别独立的表示 0, 1 , 2 , 3或 4。 p and q independently represent 0, 1, 2, 3 or 4.
4、 如权利要求 3 所述的化合物、 其药学上可接受的盐或其立体异 构体： 4. The compound of claim 3, its pharmaceutically acceptable salt or its stereoisomer:
其中， 环 A和环 B分别独立的表示苯基， 含有 N杂原子的 5- 6元 杂环烷基或 5-6元杂芳基； Among them, Ring A and Ring B independently represent phenyl, 5-6 membered containing N heteroatoms Heterocycloalkyl or 5-6 membered heteroaryl;
Li和 L2分别独立的表示- NH- , — 0-或- S(0)m-； Li and L 2 independently represent - NH- , — 0- or - S(0) m -;
a表示共价键或亚氨基； a represents covalent bond or imino group;
b表示- CO-； b means - CO-;
c表示 1,2-亚乙烯基； c represents 1,2-vinylidene;
d表示共价键或亚甲基； d represents covalent bond or methylene;
e表示氢原子， 哌啶基， 吗啉基， 吡咯烷基， 哌嗪基或二-（甲基） 氨基； e represents a hydrogen atom, piperidinyl, morpholinyl, pyrrolidinyl, piperazinyl or di-(methyl)amino;
Ri表示氢原子， 素原子， 未被取代或被一至三个 素原子取代的 甲基、 甲氧基， 甲基氨基或二 -（甲基）氨基； Ri represents a hydrogen atom, a prime atom, a methyl group, a methoxy group, a methylamino group or a di-(methyl)amino group that is unsubstituted or substituted by one to three prime atoms;
R3表示氢原子， 卤素原子或- L3- R4, R 3 represents a hydrogen atom, a halogen atom or - L 3 - R 4 ,
L3表示共价键， -NH-, - N(C1-3烷基) -， -0-, - 0- C1-3亚烷基-， -S-Ci-3 亚烷基-， - S(0)m- , - C(O)- , -NHC(O)-, - C(0)NH- , - OC(O)-或- C(0)0- , R4表示氢原子，甲基，乙基， - N(C1-3烷基 )2, - NHC(0)0- CH3，- 0(CH3), -0(CH2CH3), - 0(C(CH3)3), -S(0)2-CH3 , 环戊烷基， 环己烷基， 吡咯烷 基， 四氢呋喃基， 哌啶基， 吗啉基， 哌嗪基， 苯基， 吡咯基， 咪唑基， 噻唑基， 噁唑基， 噻二唑基， 吡啶基或嘧啶基； L 3 represents a covalent bond, -NH-, - N(C 1-3 alkyl) -, -0-, - 0- C 1-3 alkylene-, -S-Ci -3 alkylene-, - S(0) m - , - C(O)- , -NHC(O)-, - C(0)NH- , - OC(O)- or - C(0)0- , R4 represents hydrogen atom, Methyl, ethyl, - N(C 1-3 alkyl) 2 , - NHC(0)0- CH 3 , - 0(CH 3 ), -0(CH 2 CH 3 ), - 0(C(CH 3 ) 3 ), -S(0) 2 -CH 3 , cyclopentyl, cyclohexyl, pyrrolidinyl, tetrahydrofuranyl, piperidyl, morpholinyl, piperazinyl, phenyl, pyrrolyl, imidazolyl, thiazolyl, oxazolyl, thiadiazolyl, pyridyl or pyrimidinyl;
表示甲基硫基， 乙基硫基， 甲基磺酰基， 氨基磺酰基， 甲基亚磺 酰基或氨基亚磺酰基， Represents methylthio, ethylthio, methylsulfonyl, aminosulfonyl, methylsulfinyl or aminosulfinyl,
所述甲基硫基、 乙基硫基、 甲基磺酰基、 氨基磺酰基、 甲基亚磺酰 基、 氨基亚磺酰基可以进一步被一至两个相同或不同的甲氧基、 吡咯烷 基、 四氢咪唑基、 哌啶基、 吗啉基、 哌嗪基取代， The methylthio group, ethylthio group, methylsulfonyl group, aminosulfonyl group, methylsulfinyl group, and aminosulfinyl group can be further replaced by one to two identical or different methoxy groups, pyrrolidinyl groups, or tetrahydrofuranyl groups. Hydrogen imidazolyl, piperidinyl, morpholinyl, piperazinyl substituted,
所述吡咯烷基、 四氢咪唑基、 哌啶基、 吡啶基、 吗啉基、 哌嗪基、 苯基、 吡咯基、 咪唑基、 噻唑基、 噁唑基、 噻二唑基、 吡啶基或嘧啶基 可以进一步被一至两个相同或不同的 取代， The pyrrolidinyl, tetrahydroimidazolyl, piperidinyl, pyridyl, morpholinyl, piperazinyl, phenyl, pyrrolyl, imidazolyl, thiazolyl, oxazolyl, thiadiazolyl, pyridyl or The pyrimidinyl group may be further substituted by one to two identical or different substitutes,
表示 素原子、 甲基、 氨基、 甲基氨基、 二- (甲基）氨基、 羟 基、 甲氧基、 甲氧羰基、 氨基甲酰基、 甲基氨基甲酰基或二— （甲基） 氨基甲酰基； Represents the prime atom, methyl, amino, methylamino, di-(methyl)amino, hydroxyl, methoxy, methoxycarbonyl, carbamoyl, methylcarbamoyl or di-(methyl)carbamoyl ;
m表示 0, 1或 2; m represents 0, 1 or 2;
p和 q分别独立的表示 0, 1或 2。 p and q independently represent 0, 1 or 2.
5、 如权利要求 4所述的化合物、 其药学上可接受的盐或其立体异 构体： 其中， 环 A和环 B分别独立的表示苯基， 吡啶基， 哌啶基， 咪唑 基， 吡唑基， 噻唑基， 噻哈基， 1,2,3-三唑基或 1,2,4-三唑基； 5. The compound of claim 4, its pharmaceutically acceptable salt or its stereoisomer: Wherein, Ring A and Ring B independently represent phenyl, pyridyl, piperidinyl, imidazolyl, pyrazolyl, thiazolyl, thiahyl, 1,2,3-triazolyl or 1,2,4 -triazolyl;
Li和 L2分别独立的表示- NH-或- 0-； Li and L 2 independently represent - NH- or - 0-;
a表示共价键或亚氨基； a represents covalent bond or imino group;
b表示- CO-； b means - CO-;
c表示 1,2-亚乙烯基； c represents 1,2-vinylidene;
d表示共价键或亚甲基； d represents covalent bond or methylene;
e表示氢原子， 哌啶基， 吡咯烷基， 吗啉基， 哌嗪基或二-（甲基） 氨基； e represents a hydrogen atom, piperidinyl, pyrrolidinyl, morpholinyl, piperazinyl or di-(methyl)amino;
表示氢原子， 氟原子， 氯原子，三氟甲基， 甲氧基或三氟甲氧基； Represents a hydrogen atom, a fluorine atom, a chlorine atom, a trifluoromethyl group, a methoxy group or a trifluoromethoxy group;
R3表示氢原子， 氟原子， 氯原子或- L3- R4, R 3 represents a hydrogen atom, a fluorine atom, a chlorine atom or - L 3 - R 4 ,
L3表示共价键， -NH-, - N (Ci-3烷基） -， - 0- , - 0- CH2CH2- , -S-CH2CH2- 或- s(o)m -, L 3 represents a covalent bond, -NH-, - N (Ci -3 alkyl) -, - 0- , - 0- CH 2 CH 2 - , -S-CH 2 CH 2 - or - s(o) m -,
R4表示氢原子，甲基，乙基， - N(C1-3烷基 )2, - NHC(0)0- CH3，- 0(CH3), - 0(C(CH3)3), -S(0)2-CH3 , 吡咯烷基， 四氢呋喃基， 哌啶基， 吗啉基， 苯基， 吡咯基， 咪唑基， 噻唑基， 噁唑基， 噻二唑基或吡啶基， R4 represents a hydrogen atom, methyl, ethyl, - N(C 1-3 alkyl) 2 , - NHC(0)0- CH 3 , - 0(CH 3 ), - 0(C(CH 3 ) 3 ) , -S(0) 2 -CH 3 , pyrrolidinyl, tetrahydrofuranyl, piperidinyl, morpholinyl, phenyl, pyrrolyl, imidazolyl, thiazolyl, oxazolyl, thiadiazolyl or pyridyl,
所述苯基、 吡咯基、 咪唑基、 噻唑基、 噁唑基、 噻二唑基、 吡啶基 可以进一步被一至两个相同或不同的 取代， The phenyl group, pyrrolyl group, imidazolyl group, thiazolyl group, oxazolyl group, thiadiazolyl group, and pyridyl group can be further substituted by one to two identical or different ones,
表示氨基甲酰基、 甲基氨基甲酰基或二- (甲基）氨基甲酰基； R2表示未被取代或被甲氧基、 哌啶基、 吗啉基取代的乙基硫基， 甲 基硫基， 甲基磺酰基， 氨基磺酰基， 甲基亚磺酰基， 氨基亚磺酰基； m表示 0, 1或 2; represents carbamoyl, methylcarbamoyl or di-(methyl)carbamoyl; R 2 represents unsubstituted or ethylthio substituted by methoxy, piperidinyl or morpholinyl, methylthio base, methylsulfonyl group, aminosulfonyl group, methylsulfinyl group, aminosulfinyl group; m represents 0, 1 or 2;
p和 q分别独立的表示 0或 1。 p and q independently represent 0 or 1.
6、 如权利要求 5 所述的化合物、 其药学上可接受的盐或其立体异 构体： 6. The compound of claim 5, its pharmaceutically acceptable salt or its stereoisomer:
其中， 环 A和环 B分别独立的表示苯基， 吡啶基， 吡唑基或哌啶 基； Among them, Ring A and Ring B independently represent phenyl, pyridyl, pyrazolyl or piperidinyl;
Li和 L2分别独立的表示- NH-； Li and L 2 independently represent - NH-;
a表示亚氨基； a represents imino;
b表示- CO-； b means - CO-;
c表示 1,2-亚乙烯基； c represents 1,2-vinylidene;
d表示共价键或亚甲基； e表示氢原子， 吡咯烷基， 吗啉基， 哌嗪基或二-（甲基）氨基； Ri表示氢原子； d represents covalent bond or methylene; e represents a hydrogen atom, pyrrolidinyl, morpholinyl, piperazinyl or di-(methyl)amino group; Ri represents a hydrogen atom;
R3表示- L3- L3表示共价键， - 0-或- 0- CH2CH2- , 表示- N(CH3)2, -0(CH3), 四氢呋喃基， 吗啉基， 苯基或吡啶基， R 3 represents - L 3 - L 3 represents covalent bond, - 0- or - 0- CH 2 CH 2 - , represents - N(CH 3 ) 2 , -0(CH 3 ), tetrahydrofuranyl, morpholinyl, phenyl or pyridyl,
所述苯基、 吡啶基可以进一步被一至两个相同或不同的 取代， 表示氨基甲酰基、 甲基氨基甲酰基或二- (甲基）氨基甲酰基； The phenyl and pyridyl groups can be further substituted by one to two identical or different ones, representing carbamoyl, methylcarbamoyl or di-(methyl)carbamoyl;
R2表示未被取代或被甲氧基、 吗啉基取代的乙基硫基， 甲基硫基， 甲基磺酰基， 氨基磺酰基， 甲基亚磺酰基， 氨基亚磺酰基； R 2 represents an ethylthio group, a methylthio group, a methylsulfonyl group, an aminosulfonyl group, a methylsulfinyl group, or an aminosulfinyl group that is unsubstituted or substituted by a methoxy group or a morpholinyl group;
p表示 1 ; p means 1;
q表示 1。 q means 1.
7、 如权利要求 1 所述的化合物、 其药学上可接受的盐或其立体异 构体： 7. The compound of claim 1, its pharmaceutically acceptable salt or its stereoisomer:
其中环 A表示苯基； Ring A represents phenyl;
环 B表示苯基， 吡唑基或吡啶基； Ring B represents phenyl, pyrazolyl or pyridyl;
Li和 L2分别独立的表示- NH-； Li and L 2 independently represent - NH-;
a表示亚氨基； a represents imino;
b表示- CO-； b means - CO-;
c表示 1,2-亚乙烯基； c represents 1,2-vinylidene;
d表示共价键； d represents covalent bond;
e表示氢原子； e represents hydrogen atom;
Ri表示氢原子； Ri represents hydrogen atom;
L3表示共价键， -NH-, - N(C1-3烷基) -， -0-, - 0- C1-3亚烷基-， -S-Ci-3 亚烷基-， 或- C(0)NH- , L 3 represents a covalent bond, -NH-, - N(C 1-3 alkyl) -, -0-, - 0- C 1-3 alkylene-, -S-Ci -3 alkylene-, or - C(0)NH- ,
R4表示氢原子， 甲基， 乙基， 甲氧基乙基， -0(CH3), -0(C(CH3)3) 或吗啉基； R4 represents a hydrogen atom, methyl, ethyl, methoxyethyl, -0(CH 3 ), -0(C(CH 3 ) 3 ) or morpholinyl;
R2表示- L4- R5 , R 2 means - L 4 - R 5 ,
L4表示- S- C1-3亚烷基 -或- S(0)m- , L 4 represents - S- C 1-3 alkylene - or - S(0) m -,
R5表示氢原子或 d_4烷基； R 5 represents a hydrogen atom or d_4 alkyl group;
p表示 1； p means 1;
q表示 1。 q means 1.
8、 如权利要求 1 所述的化合物、 其药学上可接受的盐或其立体异 8. The compound of claim 1, its pharmaceutically acceptable salt or its stereoisomer
Figure imgf000078_0001
Figure imgf000078_0001
-77-
Figure imgf000079_0001
Figure imgf000080_0001
-77-
Figure imgf000079_0001
Figure imgf000080_0001
其药学上可接受的盐或其立体异构体。 Its pharmaceutically acceptable salt or its stereoisomer.
9、 权利要求 1-8 任一项所述的化合物、 其药学上可接受的盐或其 立体异构体的制备方法， 所述方法包括以下步骤： 9. A method for preparing the compound according to any one of claims 1 to 8, its pharmaceutically acceptable salt or its stereoisomer, the method comprising the following steps:
Figure imgf000080_0002
Figure imgf000080_0002
式 (i)化合物 Compounds of formula (i)
其中环 A、 环 B、 Li、 L2、 Ri 、 R2、 R3、 a、 b、 c、 d、 e、 p和 q 如权利要求 1所定义。 Wherein Ring A, Ring B, Li, L 2 , Ri , R 2 , R 3 , a, b, c, d, e, p and q are as defined in claim 1.
10、 药物组合物， 所述组合物含有权利要求 1-8任一项所述的化合 物、 其药学上可接受的盐或其立体异构体和可药用载体。 10. Pharmaceutical composition, said composition containing the compound described in any one of claims 1 to 8, its pharmaceutically acceptable salt or its stereoisomer and a pharmaceutically acceptable carrier.
11、 药物组合物， 含有权利要求 1-7任一项所述的化合物、 其药学 上可接受的盐或其立体异构体， 以及选自抗肿瘤剂、免疫抑制剂和 /或抗 炎药的第二治疗剂。 11. Pharmaceutical composition, containing the compound according to any one of claims 1 to 7, its pharmaceutically acceptable salt or its stereoisomer, and an anti-tumor agent, an immunosuppressive agent and/or an anti-inflammatory agent. of second therapeutic agents.
12、 权利要求 10- 11任一项所述的药物组合物， 其中所述药物组合 物为药学上可接受的任一剂型。 12. The pharmaceutical composition according to any one of claims 10 to 11, wherein the pharmaceutical composition is in any pharmaceutically acceptable dosage form.
13、 如权利要求 1-8任一项所述的化合物、 其药学上可接受的盐或 其立体异构体或者如权利要求 10—11任一项所述的药物组合物在制备用 于预防和 /或治疗 B细胞相关的血癌、 炎性和 /或自身免疫性疾病的药物 中的应用。 13. The compound according to any one of claims 1 to 8, its pharmaceutically acceptable salt or its stereoisomer or the pharmaceutical composition according to any one of claims 10 to 11 when prepared for the prevention of and/or application in drugs to treat B cell-related blood cancers, inflammatory and/or autoimmune diseases.
14、 预防和 /或治疗个体中 B细胞相关的血癌、 炎性和 /或自身免疫 性疾病的方法， 所述方法包括给所述个体施用如权利要求 1-8任一项所 述的化合物、 其药学上可接受的盐或其立体异构体或者如权利要求 14. A method for preventing and/or treating B cell-related blood cancer, inflammatory and/or autoimmune diseases in an individual, the method comprising administering to the individual a compound according to any one of claims 1-8, Its pharmaceutically acceptable salts or its stereoisomers or as claimed in the claims
10-11任一项所述的药物组合物。 The pharmaceutical composition according to any one of 10-11.
PCT/CN2013/083429 2012-09-12 2013-09-12 Nitrogen-containing heteroaromatic ring derivative as tyrosine kinase inhibitor WO2014040555A1 (en)

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