WO2022214009A1

WO2022214009A1 - High-activity hpk1 kinase inhibitor

Info

Publication number: WO2022214009A1
Application number: PCT/CN2022/085446
Authority: WO
Inventors: 陈宇锋; 时永强; 武朋; 温俏冬; 杨寒; 吕萌; 刘灿丰; 陈凯旋; 王友平; 何南海
Original assignee: 杭州阿诺生物医药科技有限公司
Priority date: 2021-04-08
Filing date: 2022-04-07
Publication date: 2022-10-13
Also published as: TW202304434A; CN116888100A

Abstract

Provided are a compound, which has a structure of formula (I) and is capable of inhibiting HPK1 kinase activity, and a pharmaceutical composition comprising the compound. Further provided is the use of the compound in the prevention and/or treatment of cancers, tumors, inflammatory diseases, autoimmune diseases or immune-mediated diseases.

Description

A highly active HPK1 kinase inhibitor

This application claims the priority of Chinese Patent Application No. 202110378424.1 filed on April 8, 2021 with the State Intellectual Property Office of the People's Republic of China, entitled "A Highly Active HPK1 Kinase Inhibitor", the contents of which are hereby incorporated by reference in their entirety. .

technical field

The present invention relates to a heterocyclic compound, in particular to a highly active HPK1 kinase inhibitor and use thereof.

Background technique

HPK1, a member of the MAP4K family, is mainly expressed in cells of the hematopoietic system and acts as an intracellular negative regulator of T cell proliferation and signaling. After antigen stimulation of T cells, the adaptor protein SLP-76 in the cytoplasm is recruited to the lipid membrane TCR complex, providing binding sites for signal transduction-related kinases to achieve TCR-mediated signal transmission and induce T cell activation. In this process, HPK1 is activated by phosphorylation by the tyrosine kinases Lck and Zap70, and is involved in the regulation of T cell receptor protein interactions. HPK1 phosphorylates the Ser376 site of the adaptor protein SLP-76, so that SLP-76 binds to the scaffold protein 14-3-3ε and is degraded by the proteasome, and this effect reduces the binding of SLP-76 to signal transduction-related kinases It blocked TCR signal transduction, which in turn inhibited T cell activation and proliferation. On the other hand, HPK1 is also involved in regulating the maturation and activation of dendritic cells (DCs), especially inhibiting the expression of T-cell activation-related proteins such as CD80, CD86 and MHC complexes in DCs, thereby affecting DC regulation The role of T cell activation; the presentation of tumor antigens by activated DCs and the mutual cooperation between DCs and T cells is one of the most important links in the anti-tumor immune system. In addition, there are a large number of immunosuppressive molecules such as PGE2 and TGF-β in the tumor microenvironment, and the immunosuppressive effects mediated by these factors are also related to HPK1. Overall, small-molecule compounds that specifically target and inhibit HPK1 can improve T cell function, enhance DC cell function, and at the same time reverse the tumor immunosuppressive microenvironment. effect.

Therefore, there is still an urgent need for effective inhibitors of HPK1 kinase activity in the prior art, in order to provide more effective options for anti-tumor.

SUMMARY OF THE INVENTION

The present invention has unexpectedly discovered a compound of formula (I) and a pharmaceutically acceptable salt thereof or a stereoisomer thereof having HPK1 kinase inhibitory activity. Accordingly, in a first aspect, the present invention provides a compound having the structure of formula (I), or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof:

in

X represents N or CR ₁₀ ;

R ₁ represents hydrogen, C ₁ -C ₆ alkyl, halogenated C ₁ -C ₆ alkyl, C ₃ -C ₆ cycloalkyl, halogenated C ₃ -C ₆ cycloalkyl, halogen, cyano or -C (O)NH ₂ ;

R ₂ represents hydrogen, halogen, hydroxy, (C ₁ -C ₆ )alkyl, (C ₂ -C ₆ )alkenyl, -(C ₀ -C ₆ alkylene)(C ₃ -C ₈ )cycloalkyl , -(C ₀ -C ₆ alkylene) (4-8 membered) heterocycloalkyl, (C ₁ -C ₆ ) alkoxy, (C ₃ -C ₈ ) cycloalkyl (C ₀ -C ₆ ) alkylene)oxy, (4-8 membered)heterocycloalkyl(C ₀ -C ₆ alkylene)oxy;

R ₃ represents hydrogen or C ₁ -C ₆ alkyl, C ₃ -C ₆ alkenyl, C ₃ -C ₆ cycloalkyl, hydroxy (C ₁ -C ₆ alkyl), halogenated C ₁ -C ₆ alkyl , 4-8 membered heterocycloalkyl;

R ₄ represents hydrogen or C ₁ -C ₆ alkyl; and the C ₁ -C ₆ alkyl may be optionally substituted with a substituent selected from halogen, -OR ^a , -SR ^a , -P(O )R ^a R ^b , -S(O) ₂ R ^a , -S(O)(NH)R ^a , -S(O)R ^a , -S(O) ₂ NR ^a R ^b , -C(O) NR ^a R ^b , -C(O)OH, -OC(O)NR ^a R ^b , -NR ^a C(O) R ^b , -NR ^a S(O) ₂ R ^b ;

R ₅ and R _5' each independently represent hydrogen, C ₁ -C ₆ alkyl, (C ₂ -C ₆ )alkenyl, halogen, halogenated C ₁ -C ₆ alkyl or hydroxy (C ₁ -C ₆ alkane) base);

Or R ₅ and R _5' together form a 3-6 membered ring with the carbon atom connected to it, and the ring can also optionally contain 0, 1 or 2 heteroatoms selected from N, O, S;

R ₆ and R _6' each independently represent hydrogen, C ₁ -C ₆ alkyl, (C ₂ -C ₆ )alkenyl, halogen, halogenated C ₁ -C ₆ alkyl, hydroxy (C ₁ -C ₆ alkane) base);

Or R ₆ and R _6' together form a 3-6 membered ring with the carbon atom connected to it, and the ring can also optionally contain 0, 1 or 2 heteroatoms selected from N, O, and S;

p represents 1 or 2;

R ₇ and R _7' each independently represent hydrogen, C ₁ -C ₆ alkyl, (C ₂ -C ₆ )alkenyl, halogen, halogenated C ₁ -C ₆ alkyl, hydroxy (C ₁ -C ₆ alkane) base);

Or R ₇ and R _7' together form a 3-6 membered ring with the carbon atom connected to it, and the ring can also optionally contain 0, 1 or 2 heteroatoms selected from N, O, and S;

W means

Among them, A means

Indicates that the ring contains a single bond or a double bond;

When the chemical bond between A and A' is a double bond, A' represents CR ^a or N, and B represents CR ^b ;

When the chemical bond between A and A' is a single bond, A' represents CHR ^a , B represents CHR ^b or does not exist;

R ₈ and R ₉ each independently represent hydrogen or C ₁ -C ₆ alkyl; or R ₈ and R ₉ may together with W ₁ and W ₂ form a 3-6 membered ring;

W ₁ means C, W ₂ means CH or N;

R ^M and R ^N each independently represent hydrogen or C ₁ -C ₆ alkyl;

R ₁₀ represents hydrogen, halogen, (C ₁ -C ₆ )alkyl, (C ₂ -C ₆ )alkenyl, -(C ₀ -C ₆ alkylene)(C ₃ -C ₈ )cycloalkyl, - (C ₀ -C ₆ alkylene) 4-10 membered heterocycloalkyl, -(C ₀ -C ₆ alkylene) (C ₆ -C ₁₀ ) aryl, -(C ₀ -C ₆ alkylene ) 5-10 membered heteroaryl,

Alternatively, when X represents CR ₁₀ , R ₁₀ may together with the adjacent R ₂ form (5-10 membered) cycloalkyl or 5-10 membered heterocycloalkyl;

For cycloalkyl, heterocycloalkyl, aryl, heteroaryl as defined above, it may be optionally substituted with 0, 1, 2 or 3 substituents selected from the group consisting of: (C ₁ -C ₆ ) alkyl, (C ₂ -C ₆ )alkenyl, halo(C ₁ -C ₆ )alkyl, halo(C ₁ -C ₆ )alkoxy, -(C ₁ -C ₆ alkylene) -O-(C ₁ -C ₆ )alkyl, (C ₃ -C ₈ )cycloalkyl, halo(C ₃ -C ₈ )cycloalkyl, halogen, -CN, oxo, -NR ^a R ^b , -OR ^a , -SR ^a , -(C ₁ -C ₆ alkylene) hydroxyl, -C(O)R ^a , -N(R _a )C(O)R ^a , -NR _a C(O) OR _a , -NR ^a SO ₂ R ^a , -C(O)OR ^a , -C(O)N R ^a R ^b , -S(O) ₂ N R ^a R ^b , -S(O)R ^a , -S (O) ₂ R ^a , -P(O)R ^a R ^b ;

R ^a and R ^b each independently represent hydrogen, C ₁ -C ₆ alkyl, hydroxyl (C ₁ -C ₆ alkyl), halogenated C ₁ -C ₆ alkyl;

m and n represent 0, 1 or 2, respectively.

In a technical solution of the present invention, W represents

Among them, A means

Indicates that the ring contains a single bond or a double bond;

o each independently represents 0 or 1;

R ₈ represents hydrogen or C ₁ -C ₆ alkyl;

R ₉ each independently represents hydrogen or C ₁ -C ₆ alkyl;

Or R ₈ together with adjacent R ₉ and the carbon atoms to which they are attached form a 3-6 membered ring;

W ₂ , ^RM and ^RN are as described above.

In a technical solution of the present invention, W represents

or

Indicates that the ring contains a single bond or a double bond;

o each independently represents 0 or 1;

R ₈ represents hydrogen or C ₁ -C ₆ alkyl;

R ₉ each independently represents hydrogen or C ₁ -C ₆ alkyl;

W ₂ , ^RM and ^RN are as described in claim 1 .

In one embodiment of the present invention, W is an aromatic heterocycle, preferably an aromatic lactam.

In one embodiment of the present invention, W is a saturated heterocycle, preferably a saturated lactam.

In the preferred technical solution of the present invention, wherein the ring W represents

or

In a further preferred technical solution of the present invention, the ring W represents

In a further preferred technical solution of the present invention, the ring W represents W represents

In a preferred technical solution of the present invention, wherein, R ₁ represents hydrogen, halogen, C ₁ -C ₆ alkyl, C ₁ -C ₆ haloalkyl, cyano or -C(O)NH ₂ ; more preferably, R ₁ represents hydrogen, halogen, C ₁ -C ₆ alkyl or C ₁ -C ₆ haloalkyl.

In the preferred technical solution of the present invention, wherein, R ₂ represents hydroxyl, (C ₁ -C ₆ )alkyl, -(C ₀ -C ₆ alkylene) (C ₃ -C ₈ )cycloalkyl, -( C ₀ -C ₆ alkylene) (4-8 membered) heterocycloalkyl, (C ₁ -C ₆ )alkoxy; more preferably, R ₂ represents C ₁ -C ₆ alkyl, C ₁ -C _6alkoxy or C3 _- _C6cycloalkyl .

In a preferred technical solution of the present invention, wherein, R ₃ represents hydrogen or C ₁ -C ₆ alkyl, C ₃ -C ₆ cycloalkyl, halogenated C ₁ -C ₆ alkyl; more preferably, R ₃ represents hydrogen or C ₁ -C ₆ alkyl.

In the preferred technical solution of the present invention, wherein, R ₄ represents hydrogen or C ₁ -C ₆ alkyl; and the C ₁ -C ₆ alkyl can be optionally substituted by a substituent selected from the following: halogen, - OR ^a , -SR ^a , -P(O)R ^a R ^b , -S(O) ₂ R ^a , -S(O)(NH)R ^a , -S(O)R ^a , -S(O) _2NR ^a R ^b , -C(O)NR ^a R ^b , -C(O)OH; more preferably, R ₄ represents hydrogen or C ₁ -C ₆ alkyl.

In a preferred technical solution of the present invention, wherein, R ₅ and R _5' each independently represent hydrogen, halogen, C ₁ -C ₆ alkyl or halogenated C ₁ -C ₆ alkyl.

In a preferred technical solution of the present invention, wherein, R ₆ and R _6′ each independently represent hydrogen, halogen, C ₁ -C ₆ alkyl or halogenated C ₁ -C ₆ alkyl.

In a preferred technical solution of the present invention, wherein, R ₇ and R _7' each independently represent hydrogen, halogen, C ₁ -C ₆ alkyl, halogenated C ₁ -C ₆ alkyl or hydroxyl (C ₁ -C ₆ alkyl).

In a preferred technical solution of the present invention, wherein, R ₈ represents hydrogen or C ₁ -C ₆ alkyl.

In a preferred technical solution of the present invention, wherein, R ₉ represents hydrogen or C ₁ -C ₆ alkyl.

In a preferred technical solution of the present invention, wherein, R ₁₀ represents hydrogen, halogen, (C ₁ -C ₆ )alkyl, -(C ₀ -C ₆ alkylene) (C ₃ -C ₈ )cycloalkyl, -(C ₀ -C ₆ alkylene)4-10 membered heterocycloalkyl; more preferably, R ₁₀ represents hydrogen, halogen or (C ₁ -C ₆ )alkyl.

In the preferred technical solution of the present invention, wherein, R ^M represents hydrogen or C ₁ -C ₆ alkyl.

In a preferred technical solution of the present invention, R ^a and R ^b each independently represent hydrogen, C ₁ -C ₆ alkyl or halogenated C ₁ -C ₆ alkyl.

Preferably, the compounds of the present invention have the following structure:

In addition, the present invention also provides a pharmaceutical composition comprising the compound of the present invention and a pharmaceutically acceptable carrier.

In addition, the present invention also provides the use of a compound or pharmaceutical composition of the present invention in the preparation of a medicament for preventing and/or treating cancer, tumor, inflammatory disease, autoimmune disease or immune-mediated disease.

In addition, the present invention also provides the use of a compound of the present invention for preventing and/or treating cancer, tumor, inflammatory disease, autoimmune disease or immune-mediated disease.

It is particularly noted that in this text, when referring to a "compound" having a specific structural formula, its stereoisomers, diastereomers, enantiomers, racemic mixtures and isotopes are generally also encompassed derivative.

It is well known to those skilled in the art that salts, solvates and hydrates of a compound are alternative existing forms of the compound, and they can all be converted into the compound under certain conditions. When referring to a compound, it generally includes its pharmaceutically acceptable salts, and further includes its solvates and hydrates.

Similarly, when referring to a compound herein, prodrugs, metabolites and nitrogen oxides thereof are generally also included.

The pharmaceutically acceptable salts of the present invention can be formed using, for example, the following inorganic or organic acids: "Pharmaceutically acceptable salts" refers to salts that, within the scope of sound medical judgment, are suitable for use in contact with humans and lower levels of and other animal tissues, without undue toxicity, irritation, allergic reactions, etc., can be called a reasonable benefit/risk ratio. The salts can be prepared in situ during the final isolation and purification of the compounds of the invention, or separately by reacting the free base or free acid with a suitable reagent, as outlined below. For example, the free base functionality can be reacted with a suitable acid. In addition, when the compounds of the present invention bear an acidic moiety, suitable pharmaceutically acceptable salts thereof may include metal salts, such as alkali metal salts (eg, sodium or potassium salts); and alkaline earth metal salts (eg, calcium or magnesium salts) . Examples of pharmaceutically acceptable non-toxic acid addition salts are amino groups with inorganic acids (eg, hydrochloric, hydrobromic, phosphoric, sulfuric, and perchloric) or organic acids (eg, acetic, oxalic, maleic, tartaric, citric acid, succinic acid or malonic acid), or by using other methods in the art such as ion exchange. Other pharmaceutically acceptable salts include adipate, sodium alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, Camphorsulfonate, citrate, cyclopentane propionate, digluconate, lauryl sulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerin Phosphate, gluconate, hernisulfate, heptanoate, caproate, hydroiodate, 2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malic acid salt, maleate, malonate, mesylate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectic acid Salt, persulfate, 3-phenylpropionate, phosphate, bitter salt, pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, p-toluene Sulfonate, undecanoate, valerate, etc. Representative alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, and the like. Other pharmaceutically acceptable salts include, where appropriate, nontoxic ammonium salts, quaternary ammonium salts, and ammonium cations formed with counter ions, for example, halides, hydroxides, carboxylates, sulfates, phosphates, nitrates, lower Alkyl sulfonates and aryl sulfonates.

The pharmaceutically acceptable salts of the present invention can be prepared by conventional methods, for example, by dissolving a compound of the present invention in a water-miscible organic solvent (eg, acetone, methanol, ethanol, and acetonitrile), adding thereto an excess of an organic acid or inorganic An aqueous acid solution to precipitate the salt from the resulting mixture, the solvent and remaining free acid removed therefrom, and the precipitated salt isolated.

The precursors or metabolites described in the present invention may be those known in the art, as long as the precursors or metabolites can be metabolized in vivo to form the target compound. For example, "prodrugs" refer to those prodrugs of the compounds of the present invention which, within the scope of sound medical judgment, are suitable for use in contact with human and lower animal tissues without undue toxicity, irritation, allergic response, etc., Have a reasonable benefit/risk ratio and be valid for its intended use. The term "prodrug" refers to a compound that is rapidly transformed in vivo to yield the parent compound of the above formula, eg, by in vivo metabolism, or N-demethylation of a compound of the present invention.

"Solvate" as used herein means a physical association of a compound of the present invention with one or more solvent molecules (whether organic or inorganic). This physical association includes hydrogen bonding. In certain instances, such as when one or more solvent molecules are incorporated into the crystal lattice of the crystalline solid, the solvate will be capable of isolation. Solvent molecules in a solvate may exist in regular and/or disordered arrangements. Solvates may contain stoichiometric or non-stoichiometric amounts of solvent molecules. "Solvate" encompasses both solution phase and isolatable solvates. Exemplary solvates include, but are not limited to, hydrates, ethanolates, methanolates, and isopropanolates. Solvation methods are well known in the art.

The "stereoisomerism" mentioned in the present invention is divided into conformational isomerism and configurational isomerism. Configurational isomerism can also be divided into cis-trans isomerism and optical isomerism (ie optical isomerism). A certain configuration of organic molecules due to the rotation or distortion of carbon and carbon single bonds, a stereoisomerism phenomenon in which each atom or atomic group of the molecule has different arrangements in space. The common structures of alkane and cycloalkane compounds Such as the chair conformation and boat conformation appearing in the cyclohexane structure. "Stereoisomer" means when the compounds of the present invention contain one or more asymmetric centers and are thus available as racemates and racemic mixtures, single enantiomers, diastereomeric mixtures and single diastereomers. The compounds of the present invention have asymmetric centers, each of which produces two optical isomers, and the scope of the present invention includes all possible optical isomers and diastereoisomeric mixtures and pure or partially pure compounds . The compounds described herein may exist in tautomeric forms having different points of attachment of the hydrogen through displacement of one or more double bonds. For example, a ketone and its enol form are keto-enol tautomers. Each tautomer and mixtures thereof are included in the compounds of the present invention. Enantiomers, diastereomers, racemates, mesomers, cis-trans isomers, tautomers, geometric isomers, epimers of all compounds of formula (I) Constructs and mixtures thereof, etc., are all included in the scope of the present invention.

"Isotopic derivatives" of the present invention refer to molecules in which compounds are isotopically labeled in this patent. Isotopes commonly used as isotopic labels are: hydrogen isotopes, ² H and ³ H; carbon isotopes: ¹¹ C, ¹³ C and ¹⁴ C; chlorine isotopes: ³⁵ Cl and ³⁷ Cl; fluorine isotopes: ¹⁸ F; iodine isotopes: ¹²³ I and ¹²⁵ I; nitrogen isotopes: ¹³ N and ¹⁵ N; oxygen isotopes: ¹⁵ O, ¹⁷ O and ¹⁸ O and sulfur isotopes ³⁵ S. These isotopically labeled compounds can be used to study the distribution of medicinal molecules in tissues. In particular, deuterium ³ H and carbon ¹³ C are more widely used because of their ease of labeling and detection. Substitution of certain heavy isotopes, such as deuterium ( ² H), can enhance metabolic stability, prolong half-life and thus provide therapeutic advantages for the purpose of reducing dosage. Isotopically-labeled compounds are generally synthesized from labeled starting materials, and their synthesis is accomplished using known synthetic techniques as for non-isotopically-labeled compounds.

The present invention also provides the use of the compound of the present invention in the preparation of a medicament for preventing and/or treating cancer, tumor, inflammatory disease, autoimmune disease or immune-mediated disease.

Furthermore, the present invention provides a pharmaceutical composition for preventing and/or treating cancer, tumor, inflammatory disease, autoimmune disease, neurodegenerative disease, attention-related disease or immune-mediated disease, comprising the present invention compound as active ingredient.

In addition, the present invention provides a method for preventing and/or treating cancer, tumor, inflammatory disease, autoimmune disease, neurodegenerative disease, attention-related disease or immune-mediated disease, comprising: A mammal in need thereof is administered a compound of the present invention.

Representative examples of inflammatory, autoimmune, and immune-mediated diseases may include, but are not limited to, arthritis, rheumatoid arthritis, spondyloarthritis, gouty arthritis, osteoarthritis, juvenile arthritis , other arthritic conditions, lupus, systemic lupus erythematosus (SLE), skin-related disorders, psoriasis, eczema, dermatitis, atopic dermatitis, pain, lung disease, lung inflammation, adult respiratory distress syndrome (ARDS) , pulmonary sarcoidosis, chronic pulmonary inflammatory disease, chronic obstructive pulmonary disease (COPD), cardiovascular disease, atherosclerosis, myocardial infarction, congestive heart failure, myocardial ischemia-reperfusion injury, inflammatory bowel disease, Crohn's disease, ulcerative colitis, irritable bowel syndrome, asthma, Sjögren's syndrome, autoimmune thyroid disease, urticaria (rubella), multiple sclerosis, scleroderma, organ transplant rejection, xenotransplantation, idiopathic Thrombocytopenic purpura (ITP), Parkinson's disease, Alzheimer's disease, diabetes-related diseases, inflammation, pelvic inflammatory disease, allergic rhinitis, allergic bronchitis, allergic sinusitis, leukemia, lymphoma, B cell lymphoma, T-cell lymphoma, myeloma, acute lymphoblastic leukemia (ALL), chronic lymphocytic leukemia (CLL), acute myeloid leukemia (AML), chronic myeloid leukemia (CML), hairy cell leukemia, Ho Jie King's disease, non-Hodgkin's lymphoma, multiple myeloma, myelodysplastic syndromes (MDS), myeloproliferative neoplasms (MPN), diffuse large B-cell lymphoma, and follicular lymphoma.

Representative examples of cancers or tumors may include, but are not limited to, skin cancer, bladder cancer, ovarian cancer, breast cancer, stomach cancer, pancreatic cancer, prostate cancer, colon cancer, lung cancer, bone cancer, brain cancer, neurocytoma, rectal cancer , colon cancer, familial adenomatous polyposis cancer, hereditary non-polyposis colorectal cancer, esophageal cancer, lip cancer, laryngeal cancer, hypopharyngeal cancer, tongue cancer, salivary gland cancer, gastric cancer, adenocarcinoma, medullary thyroid cancer, papillary thyroid cancer, kidney cancer, renal parenchymal cancer, ovarian cancer, cervical cancer, endometrial cancer, endometrial cancer, choriocarcinoma, pancreatic cancer, prostate cancer, testicular cancer, urinary cancer, melanoma, brain tumors such as Glioblastoma, astrocytoma, meningioma, medulloblastoma and peripheral neuroectodermal tumors, Hodgkin lymphoma, non-Hodgkin lymphoma, Burkitt lymphoma, acute lymphoblastic leukemia ( ALL), chronic lymphocytic leukemia (CLL), acute myeloid leukemia (AML), chronic myeloid leukemia (CML), adult T-cell leukemia lymphoma, diffuse large B-cell lymphoma (DLBCL), hepatocellular carcinoma, gallbladder carcinoma, bronchial carcinoma, small cell lung cancer, non-small cell lung cancer, multiple myeloma, basal cell tumor, teratoma, retinoblastoma, choroidal melanoma, seminoma, rhabdomyosarcoma, craniopharyngioma, Osteosarcoma, chondrosarcoma, sarcoma, liposarcoma, fibrosarcoma, Ewing's sarcoma, or plasmacytoma.

Representative examples of therapeutic agents for the treatment of inflammatory diseases, autoimmune diseases, and immune-mediated diseases may include, but are not limited to, steroidal drugs (eg, prednisone, prednisone, methylhydroponil) sone, cortisone, hydroxycortisone, betamethasone, dexamethasone, etc.), methotrexate, leflunomide, anti-TNFα agents (eg, etanercept, infliximab, adalix monoclonal antibodies, etc.), calcineurin inhibitors (eg, tacrolimus, pimecrolimus, etc.), and antihistamines (eg, diphenhydramine, hydroxyzine, loratadine, ebaz) Cetirizine, ketotifen, cetirizine, levocetirizine, fexofenadine, etc.), and at least one therapeutic agent selected from them may be included in the pharmaceutical composition of the present invention.

The compound of the present invention or a pharmaceutically acceptable salt thereof can be administered orally or parenterally as an active ingredient in an effective amount ranging from 0.1 to 2,000 mg/kg body weight/day, in mammals including humans (about 70 kg body weight), Preferably from 1 to 1,000 mg/kg body weight/day, and administered in single or 4 divided doses per day, or on/off schedule. The dosage of the active ingredient can be adjusted according to a number of relevant factors, such as the condition of the subject to be treated, the type and severity of the disease, the rate of administration, and the opinion of the physician. In some cases, amounts less than the above doses may be appropriate. An amount greater than the above dose may be used if it does not cause adverse side effects and the amount may be administered in divided doses per day.

In addition, the present invention also provides a method for preventing and/or treating tumors, cancer, viral infections, organ transplant rejection, neurodegenerative diseases, attention-related diseases or autoimmune diseases, comprising A compound of the present invention or a pharmaceutical composition of the present invention is administered to a mammal in need thereof.

The pharmaceutical compositions of the present invention may be formulated according to any of conventional methods into dosage forms such as tablets, granules, powders for oral administration or parenteral administration (including intramuscular, intravenous and subcutaneous routes, intratumoral injection) , capsules, syrups, emulsions, microemulsions, solutions or suspensions.

Other features of this invention will become apparent in the course of describing exemplary embodiments of this invention, which are given to illustrate the invention and not to limit it, and the following examples were prepared using the methods disclosed in this invention. , isolation and characterization.

The compounds of the present invention can be prepared in a variety of ways known to those skilled in the art of organic synthesis, using the methods described below as well as synthetic methods known in the art of synthetic organic chemistry or by variations thereof known to those skilled in the art. Compounds of the present invention were synthesized. Preferred methods include, but are not limited to, those described below. Reactions are carried out in a solvent or solvent mixture suitable for the kit materials used and for the transformation being effected. Those skilled in the art of organic synthesis will understand that the functionality present on the molecule is consistent with the proposed transformation. This sometimes requires the discretion to alter the order of synthetic steps or starting materials to obtain the desired compounds of the invention.

Detailed ways

the term

Unless otherwise stated, terms used in the present application, including the specification and claims, are defined as follows. It must be noted that, in the specification and the appended claims, the singular form "a" includes the plural unless the context clearly dictates otherwise. Conventional methods of mass spectrometry, nuclear magnetic resonance, HPLC, protein chemistry, biochemistry, recombinant DNA techniques and pharmacology were used unless otherwise stated. In this application, the use of "or" or "and" means "and/or" unless stated otherwise.

In the specification and claims, a given chemical formula or name shall encompass all stereo and optical isomers and racemates in which such isomers exist. Unless otherwise indicated, all chiral (enantiomeric and diastereomeric) and racemic forms are within the scope of the present invention. Numerous geometric isomers of C=C double bonds, C=N double bonds, ring systems, etc. may also exist in the compounds, and all such stable isomers are encompassed by the present invention. The present invention describes cis- and trans- (or E- and Z-) geometric isomers of the compounds of the present invention, and which may be isolated as a mixture of isomers or as separate isomeric forms. The compounds of the present invention can be isolated in optically active or racemic forms. All methods used to prepare the compounds of the present invention and intermediates prepared therein are considered part of this invention. In the preparation of enantiomeric or diastereomeric products, they can be separated by conventional methods, eg by chromatography or fractional crystallization. Depending on the process conditions, the final product of the invention is obtained in free (neutral) or salt form. Both free forms and salts of these final products are within the scope of the present invention. If desired, one form of a compound can be converted into another. Free bases or acids can be converted into salts; salts can be converted into free compounds or another salt; mixtures of isomeric compounds of the present invention can be separated into individual isomers. The compounds of the present invention, their free forms and salts, may exist in various tautomeric forms in which hydrogen atoms are transposed to other parts of the molecule and the chemical bonds between the atoms of the molecule are thereby rearranged. It is to be understood that all tautomeric forms that may exist are encompassed by the present invention.

Unless otherwise defined, the definitions of the substituents of the present invention are each independent rather than interrelated, for example, for R ^a (or R ^a ') in a substituent, it is independent in the definitions of different substituents . Specifically, selecting a definition for ^Ra (or ^Ra ') in one substituent does not mean that ^Ra (or ^Ra ') has the same definition in other substituents. More specifically, for example (to list only non-exhaustive) for NR ^a R ^a ', when the definition of R ^a (or R ^a ') is selected from hydrogen, it does not mean that in -C(O)-NR In ^a R ^a ', R ^a (or R ^a ') must be hydrogen.

Unless otherwise defined, when a substituent is annotated as "optionally substituted", the substituent is selected from, for example, substituents such as alkyl, cycloalkyl, aryl, heterocyclyl, halo, hydroxy, alkane oxy, oxo, alkanoyl, aryloxy, alkanoyloxy, amino, alkylamino, arylamino, arylalkylamino, disubstituted amine groups (wherein the 2 amino substituents are selected from alkyl, aryl or arylalkyl), alkanoylamino, aroylamino, aralkanoylamino, substituted alkanoylamino, substituted arylamino, substituted aralkanoylamino, thio, alkylthio group, arylthio, arylalkylthio, arylthiocarbonyl, arylalkylthiocarbonyl, alkylsulfonyl, arylsulfonyl, arylalkylsulfonyl, sulfonamido such as -SO ₂ _NH2 , substituted sulfonamido, nitro, cyano, carboxyl, carbamoyl such as _-CONH2 , substituted carbamoyl such as -CONH alkyl, -CONH aryl, -CONH arylalkyl or at nitrogen In the case of having two substituents selected from alkyl, aryl or arylalkyl, alkoxycarbonyl, aryl, substituted aryl, guanidino, heterocyclic groups such as indolyl, imidazolyl, Furyl, thienyl, thiazolyl, pyrrolidinyl, pyridyl, pyrimidinyl, pyrrolidinyl, piperidinyl, morpholinyl, piperazinyl, homopiperazinyl, etc. and substituted heterocyclyl.

The terms "alkyl" or "alkylene" as used herein are intended to include branched and straight chain saturated aliphatic hydrocarbon groups having the indicated number of carbon atoms. For example, " _C1 - _C6 alkyl" means an alkyl group having 1 to 6 carbon atoms. Examples of alkyl groups include, but are not limited to, methyl (Me), ethyl (Et), propyl (eg n-propyl and isopropyl), butyl (eg n-butyl, isobutyl, tert-butyl) and Pentyl (eg n-pentyl, isopentyl, neopentyl).

The term "alkenyl" refers to a straight or branched chain hydrocarbon group containing one or more double bonds and usually 2 to 20 carbon atoms in length. For example, "C2-C6 alkenyl" contains two to six carbon atoms. Alkenyl groups include, but are not limited to, for example, vinyl, propenyl, butenyl, 1-methyl-2-buten-1-yl, and the like.

The term "alkynyl" refers to a straight or branched chain hydrocarbon group containing one or more triple bonds and usually 2 to 20 carbon atoms in length. For example, "C2 _- _C6alkynyl " contains two to six carbon atoms. Representative alkynyl groups include, but are not limited to, for example, ethynyl, 1-propynyl, 1-butynyl, and the like.

The term "alkoxy" or "alkyloxy" refers to -O-alkyl. "C ₁ -C ₆ alkoxy" (or alkyloxy) is intended to include C ₁ , C ₂ , C ₃ , C ₄ , C ₅ , C ₆ alkoxy. Examples of alkoxy groups include, but are not limited to, methoxy, ethoxy, propoxy (eg, n-propoxy and isopropoxy), and t-butoxy. Similarly, "alkylthio" or "thioalkoxy" represents an alkyl group, as defined above, having the indicated number of carbon atoms attached through a sulfur bridge; eg, methyl-S- and ethyl-S-.

The term "carbonyl" refers to an organic functional group (C=O) composed of two atoms, carbon and oxygen, linked by a double bond.

The term "aryl", alone or as part of a larger moiety such as "aralkyl", "aralkoxy" or "aryloxyalkyl", refers to a single ring having a total of 5 to 12 ring members , bicyclic or tricyclic ring systems, wherein at least one ring in the system is aromatic and wherein each ring in the system contains from 3 to 7 ring members. In certain embodiments of the present invention, "aryl" refers to an aromatic ring system including, but not limited to, phenyl, biphenyl, indanyl, 1-naphthyl, 2-naphthyl, and tetrahydronaphthalene base. The term "aralkyl" or "arylalkyl" refers to an alkyl residue attached to an aryl ring. Non-limiting examples include benzyl, phenethyl, and the like. A fused aryl group can be attached to another group at a suitable position on the cycloalkyl or aromatic ring. For example, dashed lines drawn from a ring system indicate that the bond may be attached to any suitable ring atom.

The term "cycloalkyl" refers to a monocyclic or bicyclic cyclic alkyl group, preferably having 3 to 8 ring members. A monocyclic cyclic alkyl group refers to a C3 _- _C8 cyclic alkyl group, including but not limited to cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and norbornyl. Branched cycloalkyl groups such as 1-methylcyclopropyl and 2-methylcyclopropyl are included in the definition of "cycloalkyl". Bicyclic cyclic alkyl groups include bridged, spiro or fused ring cycloalkyl groups.

The term "cycloalkenyl" refers to a monocyclic or bicyclic cyclic alkenyl group, preferably having 3 to 8 ring members. _Monocyclic cyclic alkenyl refers to C3 _- C8 cyclic alkenyl, including but not limited to cyclopropenyl, cyclobutenyl, cyclopentenyl, cyclohexenyl and norbornyl. Branched cycloalkenyl groups such as 1-methylcyclopropenyl and 2-methylcyclopropenyl are included in the definition of "cycloalkenyl". Bicyclic cyclic alkenyl groups include bridged, spiro or fused ring cyclic alkenyl groups.

"Halo" or "halogen" includes fluorine, chlorine, bromine and iodine. "Haloalkyl" is intended to include branched and straight chain saturated aliphatic hydrocarbon groups having the indicated number of carbon atoms and substituted with 1 or more halogens, preferably 1, 2 or 3 halogens. Examples of haloalkyl include, but are not limited to, fluoromethyl, difluoromethyl, trifluoromethyl, trichloromethyl, pentafluoroethyl, pentachloroethyl, 2,2,2-trifluoroethyl, heptafluoroethyl propyl and heptachloropropyl. Examples of haloalkyl groups also include "fluoroalkyl groups" intended to include branched and straight chain saturated aliphatic hydrocarbon groups having the specified number of carbon atoms (preferably 1 to 6 carbon atoms) substituted with 1 or more fluorine atoms.

"Haloalkoxy" or "haloalkyloxy" means a haloalkyl group, as defined above, having the indicated number of carbon atoms (preferably 1 to 6 carbon atoms) attached through an oxygen bridge. For example, " _haloC1 - _C6alkoxy " is intended to include _C1 , _C2 , C3, _C4 , _C5 , _C6 _haloalkoxy . Examples of haloalkoxy include, but are not limited to, trifluoromethoxy, 2,2,2-trifluoroethoxy, and pentafluoroethoxy. Similarly, "haloalkylthio" or "thiohaloalkoxy" represents a haloalkyl group as defined above having the indicated number of carbon atoms (preferably 1 to 6 carbon atoms) attached through a sulfur bridge; eg trifluoromethyl base-S- and pentafluoroethyl-S-.

In the present disclosure, the expression C _x1 -C _x2 is used when referring to some substituent groups, which means that the number of carbon atoms in the substituent groups may be x1 to x2. For example, C ₀ -C ₈ indicates that the group contains 0, 1, 2, 3, 4, 5, 6, 7 or 8 carbon atoms, and C ₁ -C ₈ indicates that the group contains 1, 2, 3 , 4, 5, 6, 7 or 8 carbon atoms, C ₂ -C ₈ means the group contains 2, 3, 4, 5, 6, 7 or 8 carbon atoms, C ₃ -C ₈ means the group The group contains 3, 4, 5, 6, 7 or 8 carbon atoms, C ₄ -C ₈ means the group contains 4, 5, 6, 7 or 8 carbon atoms, and C ₀ -C ₆ means the A group contains 0, 1, 2, 3, 4, 5 or 6 carbon atoms, C ₁ -C ₆ means the group contains 1, 2, 3, 4, 5 or 6 carbon atoms, C ₂ -C ₆ indicates that the group contains 2, 3, 4, 5 or 6 carbon atoms and C3 _- _C6 indicates that the group contains 3, 4, 5 or 6 carbon atoms.

In this disclosure, the expression "x1-x2 membered ring" is used when referring to cyclic groups (eg, aryl, heteroaryl, cycloalkyl, and heterocycloalkyl), which refers to the ring atoms of the group The number may be x1 to x2. For example, the 3-12 membered cyclic group can be a 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 membered ring, and the number of ring atoms can be 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12; 3-6 membered ring means that the cyclic group can be a 3, 4, 5 or 6 membered ring, and the number of ring atoms can be 3, 4, 5 or 6 ; 3-8 membered ring means that the cyclic group can be a 3, 4, 5, 6, 7 or 8 membered ring, and the number of ring atoms can be 3, 4, 5, 6, 7 or 8; 3-9 Ring member means that the cyclic group can be a 3, 4, 5, 6, 7, 8 or 9 membered ring, and the number of ring atoms can be 3, 4, 5, 6, 7, 8 or 9; 4-7 Ring member means that the cyclic group can be 4, 5, 6 or 7 membered ring, and the number of ring atoms can be 4, 5, 6 or 7; 5-8 membered ring means that the cyclic group can be 5, 6, 7 or 8 membered ring, the number of ring atoms can be 5, 6, 7 or 8; 5-12 membered ring means that the cyclic group can be 5, 6, 7, 8, 9, 10, 11 or 12-membered ring, the number of ring atoms can be 5, 6, 7, 8, 9, 10, 11 or 12; 6-12-membered ring means that the cyclic group can be 6, 7, 8, 9, 10, 11- or 12-membered ring, the number of ring atoms may be 6, 7, 8, 9, 10, 11 or 12. The ring atoms may be carbon atoms or heteroatoms, eg heteroatoms selected from N, O and S. When the ring is a heterocycle, the heterocycle may contain 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more ring heteroatoms, eg selected from N, O and S of heteroatoms.

In the present disclosure, the one or more halogens may each be independently selected from fluorine, chlorine, bromine, and iodine.

The term "heteroaryl" means a stable 3-, 4-, 5-, 6-, or 7-membered aromatic monocyclic or aromatic bicyclic or 7-, 8-, 9-, 10-, 11-, 12-membered Aromatic polycyclic heterocycles, which are fully unsaturated, partially unsaturated, and which contain carbon atoms and 1, 2, 3, or 4 heteroatoms independently selected from N, O, and S; and include Any of the following polycyclic groups wherein any heterocycle as defined above is fused to a benzene ring. Nitrogen and sulfur heteroatoms can optionally be oxidized. Nitrogen atoms are substituted or unsubstituted (ie, N or NR, where R is H or, if defined, another substituent). Heterocycles can be attached to their pendant groups at any heteroatom or carbon atom that results in a stable structure. The heterocyclyl groups described herein may be substituted on a carbon or nitrogen atom if the resulting compound is stable. The nitrogens in the heterocycle may be optionally quaternized. Preferably, when the total number of S and O atoms in the heterocycle exceeds 1, these heteroatoms are not adjacent to each other. Preferably, the total number of S and O atoms in the heterocycle is not greater than one. When the term "heterocycle" is used, it is intended to include heteroaryl groups. Examples of heteroaryl groups include, but are not limited to, acridinyl, azetidinyl, acridine, benzimidazolyl, benzofuranyl, benzothiofuranyl, benzothienyl, benzoxanyl azolyl, benzoxazolinyl, benzothiazolyl, benzotriazolyl, benzotetrazolyl, benzisoxazolyl, benzisothiazolyl, benzimidazolinyl, carbazolyl, 4aH-carbazolyl, carboline, chromanyl, chromenyl, cinnoline, decahydroquinolinyl, 2H,6H-1,5,2-dithiazinyl, dihydrofuro[2, 3-b] tetrahydrofuranyl, furanyl, furanyl, imidazolidinyl, imidazolinyl, imidazolyl, 1H-indazolyl, imidazopyridyl, indolenyl, indolyl, Indolyl, indolyl, 3H-indolyl, isatinoyl, isobenzofuranyl, isochromanyl, isoindazolyl, isoindolyl, isoindolyl, iso Quinolinyl, isothiazolyl, isothiazolopyridyl, isoxazolyl, isoxazolopyridyl, methylenedioxyphenyl, morpholinyl, naphthyl, octahydroisoquinoline base, oxadiazolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl, 1,3,4-oxadiazolyl , oxazolidinyl, oxazolyl, oxazolopyridyl, oxazolidinyl, rylene, oxindole, pyrimidinyl, phenanthridine, phenanthroline, phenazinyl , phenothiazinyl, phenoxthiyl, phenoxazinyl, phthalazinyl, piperazinyl, piperidinyl, piperidinone, 4-piperidinone, piperonyl, pteridyl, purinyl, Pyranyl, pyrazinyl, pyrazolidine, pyrazolinyl, pyrazolopyridyl, pyrazolyl, pyridazinyl, pyridooxazolyl, pyridoimidazolyl, pyridothiazolyl, pyridyl , pyrimidinyl, pyrrolidinyl, pyrrolinyl, 2-pyrrolidone, 2H-pyrrolyl, pyrrolyl, quinazolinyl, quinolinyl, 4H-quinuclidine, quinoxalinyl, quinuclidinyl, Tetrazolyl, tetrahydrofuranyl, tetrahydroisoquinolyl, tetrahydroquinolyl, 6H-1,2,5-thiadiazinyl, 1,2,3-thiadiazolyl, 1,2,4- Thiadiazolyl, 1,2,5-thiadiazolyl, 1,3,4-thiadiazolyl, thianthyl, thiazolyl, thienyl, thiazolopyridyl, thienothiazolyl, thienooxa azolyl, thienoimidazolyl, thienyl, triazinyl, 1,2,3-triazolyl, 1,2,4-triazolyl, 1,2,5-triazolyl, 1,3,4 - Triazolyl and xanthyl, quinolinyl, isoquinolinyl, phthalazinyl, quinazolinyl, indolyl, isoindolyl, indoline, 1H-indazolyl, benzo Imidazolyl, 1,2,3,4-tetrahydroquinolinyl, 1,2,3,4-tetrahydroisoquinolinyl, 5,6,7,8-tetrahydro-quinolinyl, 2,3 - Dihydro-benzofuranyl, chromanyl, 1,2,3,4-tetrahydro-quinoxalinyl and 1,2,3,4-tetrahydro-quinazolinyl. The term "heteroaryl" may also include biaryl structures formed by the above-defined "aryl" and a monocyclic "heteroaryl", such as, but not limited to, "-phenylbipyridyl-", "- Phenylbipyrimidinyl", "-pyridylbiphenyl", "-pyridylbipyrimidinyl-", "-pyrimidinylbiphenyl-"; wherein the present invention also includes fused rings containing, for example, the above heterocycles and Spiro compounds.

The term "heterocycloalkyl" as used herein refers to a monocyclic heterocycloalkyl system, or a bicyclic heterocycloalkyl system, and also includes spiroheterocycles or bridged heterocycloalkyls. Monocyclic heterocycloalkyl refers to a 3-8 membered cyclic alkyl system containing at least one saturated or unsaturated but non-aromatic alkyl group selected from O, N, S, P. A bicyclic heterocycloalkyl system refers to a heterocycloalkyl group fused to a phenyl group, or a cycloalkyl group, or a cycloalkenyl group, or a heterocycloalkyl group, or a heteroaryl group.

The term "bridged cycloalkyl" as used herein refers to polycyclic compounds that share two or more carbon atoms. It can be divided into bicyclic bridged cyclic hydrocarbons and polycyclic bridged cyclic hydrocarbons. The former consists of two alicyclic rings sharing more than two carbon atoms; the latter is a bridged cyclic hydrocarbon consisting of three or more rings.

The term "spirocycloalkyl" as used herein refers to polycyclic hydrocarbons in which a single carbon atom (called a spiro atom) is shared between the monocyclic rings.

The term "bridged heterocyclic group" as used herein refers to a polycyclic compound sharing two or more carbon atoms, and the ring contains at least one atom selected from O, N and S. It can be divided into bicyclic bridged heterocycles and polycyclic bridged heterocycles.

The term "heterospirocyclyl" as used herein refers to polycyclic hydrocarbons in which a single carbon atom (called a spiro atom) is shared between single rings, and the ring contains at least one atom selected from O, N and S.

The term "substituted" as used herein means that at least one hydrogen atom is replaced by a non-hydrogen group, provided that normal valences are maintained and the substitution results in a stable compound. As used herein, a ring double bond is a double bond formed between two adjacent ring atoms (eg, C=C, C=N, or N=N).

Where nitrogen atoms (eg, amines) are present on the compounds of the present invention, these nitrogen atoms can be converted to N-oxides by treatment with oxidizing agents (eg, mCPBA and/or hydrogen peroxide) to obtain other compounds of the present invention . Accordingly, both shown and claimed nitrogen atoms are considered to encompass both the shown nitrogen and its N-oxides to obtain the derivatives of the present invention.

When any variable occurs more than once in any composition or formula of a compound, its definition at each occurrence is independent of its definition at each other occurrence. Thus, for example, if a group is shown to be substituted with 0-3 R groups, the group may be optionally substituted with up to three R groups, and at each occurrence R is independently selected from the definition of R. Furthermore, combinations of substituents and/or variables are only permissible if such combinations result in stable compounds.

The term "patient" as used herein refers to an organism treated by the methods of the present invention. Such organisms preferably include, but are not limited to, mammals (eg, murine, simian/monkey, equine, bovine, porcine, canine, feline, etc.) and most preferably refer to humans.

As used herein, the term "effective amount" means the amount of a drug or agent (ie, a compound of the invention) that will elicit the biological or medical response of a tissue, system, animal or human, eg, sought by a researcher or clinician. Furthermore, the term "therapeutically effective amount" means an amount that results in improved treatment, cure, prevention or alleviation of a disease, disorder or side effect, or a reduction in the incidence of a disease, as compared to a corresponding subject not receiving such amounts or the rate of progression of the disease. An effective amount can be administered in one or more administrations, administrations or doses and is not intended to be limited by a particular formulation or route of administration. The term also includes within its scope an amount effective to enhance normal physiology.

As used herein, the term "treating" includes any effect that results in amelioration of a condition, disease, disorder, etc., eg, alleviation, reduction, modulation, amelioration or elimination, or amelioration of symptoms thereof.

The term "pharmaceutically acceptable" is used herein to refer to those compounds, substances, compositions and/or dosage forms which, within the scope of sound medical judgment, are suitable for use in contact with human and animal tissues without unduly toxic, irritating sexual, allergic reactions and/or other problems or complications and are commensurate with a reasonable benefit/risk ratio.

The phrase "pharmaceutically acceptable carrier" as used herein means a pharmaceutical substance, composition or vehicle such as a liquid or solid filler, diluent, excipient, manufacturing aid (eg lubricants, talc, magnesium stearate, calcium stearate or zinc stearate or stearic acid) or a solvent encapsulating material which is involved in carrying or transporting a subject compound from one organ or part of the body to another organ or part of the body. Each carrier must be "acceptable" in the sense of being compatible with the other ingredients of the formulation and not injurious to the patient.

The term "pharmaceutical composition" means a composition comprising a compound of the present invention and at least one other pharmaceutically acceptable carrier. "Pharmaceutically acceptable carrier" refers to a medium generally accepted in the art for delivering a biologically active agent to an animal, particularly a mammal, including (ie) adjuvants, excipients or vehicles such as diluents, preservatives , fillers, flow regulators, disintegrants, wetting agents, emulsifiers, suspending agents, sweeteners, flavoring agents, perfuming agents, antibacterial agents, antifungal agents, lubricating and dispersing agents, depending on The mode of administration and the nature of the dosage form.

Certain Pharmacy and Medical Terms

The term "acceptable", as used herein, refers to a formulation component or active ingredient that does not have undue deleterious effects on the health of the general target of treatment.

The term "cancer", as used herein, refers to an abnormal growth of cells that is uncontrollable and, under certain conditions, is capable of metastasizing (spreading). Cancers of this type include, but are not limited to, solid tumors (eg, bladder, bowel, brain, chest, uterus, heart, kidney, lung, lymphoid tissue (lymphoma), ovary, pancreas or other endocrine organs (eg, thyroid), prostate , skin (melanoma), or blood tumor (eg, nonleukemic leukemia).

The term "co-administration" or similar terms, as used herein, refers to the administration of several selected therapeutic agents to a patient, administered in the same or different administrations at the same or different times.

The term "enhancing" or "enhancing", as used herein, refers to the ability of a desired result to be increased or prolonged, either in potency or duration. Thus, in enhancing the therapeutic effect of a drug, the term "enhancer" refers to the drug's ability to increase or prolong the potency or duration of the drug in the system. As used herein, "potentiation value" refers to the ability to maximize the ability of another therapeutic agent in an ideal system.

The term "immune disease" refers to a disease or condition of an adverse or deleterious response to an endogenous or exogenous antigen. The result is usually the dysfunction of cells, or the destruction and dysfunction of the cells, or the destruction of organs or tissues that may produce immune symptoms.

The terms "kit" and "product packaging" are synonymous.

The term "subject" or "patient" includes mammals and non-mammals. Mammals include, but are not limited to, mammals: humans, non-human primates such as orangutans, apes and monkeys; agricultural animals such as cattle, horses, goats, sheep, pigs; livestock such as rabbits, dogs; laboratory animals including rodents, Such as rats, mice and guinea pigs. Non-mammalian animals include, but are not limited to, birds, fish, and the like. In a preferred embodiment, the selected mammal is a human.

The terms "treatment", "course of treatment" or "therapy" as used herein include alleviating, inhibiting or ameliorating the symptoms or conditions of a disease; inhibiting the development of complications; ameliorating or preventing the underlying metabolic syndrome; inhibiting the development of a disease or symptom, Such as controlling the development of a disease or condition; alleviating a disease or symptom; reducing a disease or symptom; alleviating complications caused by a disease or symptom, or preventing and/or treating symptoms caused by a disease or symptom.

As used herein, a compound or pharmaceutical composition, when administered, results in amelioration, especially improvement in severity, delay in onset, slow progression, or reduction in duration of a disease, symptom or condition. Whether fixed or temporary, continuous or intermittent, conditions may be attributable to or associated with the administration.

Route of administration

Suitable routes of administration include, but are not limited to, oral, intravenous, rectal, aerosol, parenteral, ocular, pulmonary, transdermal, vaginal, ear canal , nasal administration and topical administration. In addition, by way of example only, parenteral administration includes intramuscular, subcutaneous, intravenous, intramedullary, ventricular, intraperitoneal, intralymphatic, and intranasal.

In one aspect, the compounds described herein are administered locally rather than systemically. In certain specific embodiments, the depot formulation is administered by implantation (eg, subcutaneously or intramuscularly) or by intramuscular injection. Furthermore, in another embodiment, the drug is administered by a targeted drug delivery system. For example, liposomes encapsulated by organ-specific antibodies. In this particular embodiment, the liposomes are selectively targeted to specific organs and absorbed.

Pharmaceutical composition and dosage

The present invention also provides pharmaceutical compositions comprising a therapeutically effective amount of one or more compounds of the present invention formulated together with one or more pharmaceutically acceptable carriers (additives) and/or diluents, and optionally a one or more of the other therapeutic agents described above. The compounds of the present invention may be administered for any of the above uses by any suitable means, eg orally, such as tablets, pills, powders, granules, elixirs, tinctures, suspensions (including nanosuspensions, microsuspensions, spray-dried dispersions), syrups and emulsions; sublingual; buccal; parenterally, such as by subcutaneous, intravenous, intramuscular or intrasternal injection or infusion techniques (eg, in sterile injectable aqueous or non-aqueous solutions or suspensions liquid); nasally, including administration to nasal membranes, such as by inhalation spray; topically, such as in cream or ointment; or rectally, such as in suppository; or by intratumoral injection. They can be administered alone, but are usually administered using a pharmaceutical carrier selected on the basis of the chosen route of administration and standard pharmaceutical practice.

Pharmaceutically acceptable carriers are formulated according to a number of factors within the purview of those skilled in the art. These factors include, but are not limited to: the type and nature of the active agent being formulated; the subject to which the composition containing the active agent is to be administered; the intended route of administration of the composition; and the therapeutic indication being targeted. Pharmaceutically acceptable carriers include aqueous and non-aqueous liquid media and various solid and semisolid dosage forms.

Such carriers may include many different ingredients and additives in addition to the active agent, which are included in the formulation for various reasons known to those skilled in the art, such as stabilizing the active agent, binders, and the like. A description of suitable pharmaceutical carriers and factors involved in carrier selection can be found in a number of readily available sources such as Allen L.V.Jr. et al. Remington: The Science and Practice of Pharmacy (2 Volumes), 22nd Edition (2012 ), Pharmaceutical Press.

Of course, dosage regimens for the compounds of the present invention will vary depending on known factors, such as the pharmacodynamic properties of the particular agent and its mode and route of administration; the species, age, sex, health, medical condition and weight of the recipient. nature and extent of symptoms; type of concomitant treatment; frequency of treatment; route of administration, renal and hepatic function of the patient, and desired effect. According to general guidance, the daily oral dose of each active ingredient should be from about 0.001 mg/day to about 10-5000 mg/day, preferably from about 0.01 mg/day to about 1000 mg/day, and most preferably, when used for the indicated effect Typically from about 0.1 mg/day to about 250 mg/day. During constant rate infusion, the most preferred intravenous dose should be from about 0.01 mg/kg/minute to about 10 mg/kg/minute. The compounds of the present invention may be administered in a single daily dose, or the total daily dose may be administered in divided doses of two, three or four times daily.

The compounds are usually in the form of suitable pharmaceutical diluents, excipients or carriers (herein) appropriately selected according to the intended form of administration (eg, oral tablets, capsules, elixirs and syrups) and consistent with conventional pharmaceutical practice. are administered in the form of a mixture of drug carriers).

Dosage forms (pharmaceutical compositions) suitable for administration may contain from about 1 mg to about 2000 mg of active ingredient per dosage unit. In these pharmaceutical compositions, the active ingredient will generally be present in an amount of about 0.1-95% by weight, based on the total weight of the composition.

A typical capsule for oral administration contains at least one compound of the invention (250 mg), lactose (75 mg) and magnesium stearate (15 mg). The mixture was passed through a 60 mesh screen and packaged into size 1 gelatin capsules.

A typical injectable formulation can be prepared by aseptically placing at least one compound of the invention (250 mg) in a vial, aseptically lyophilizing and sealing. For use, the contents of the vial are mixed with 2 mL of physiological saline to produce an injectable formulation.

Within the scope of the present invention are pharmaceutical compositions comprising, either alone or in combination with a pharmaceutical carrier, a therapeutically effective amount of at least one compound of the present invention as an active ingredient. Optionally, the compounds of the present invention may be used alone, in combination with other compounds of the present invention, or in combination with one or more other therapeutic agents (eg, anticancer agents or other pharmaceutically active substances).

Regardless of the chosen route of administration, the compounds of the present invention (which may be used in a suitable hydrated form) and/or the pharmaceutical compositions of the present invention are formulated into pharmaceutical dosage forms by conventional methods known to those skilled in the art.

The actual dosage level of the active ingredient in the pharmaceutical compositions of the present invention can be varied to obtain an amount of active ingredient that is effective in achieving the desired therapeutic response, composition, and mode of administration for a particular patient, without being toxic to the patient.

The dose level selected will depend on a variety of factors, including the activity of the particular compound of the invention or its ester, salt or amide employed; the route of administration; the time of administration; the rate of excretion of the particular compound employed; the rate and extent of absorption duration of treatment; other drugs, compounds and/or substances used in combination with the particular compound used; factors well known in the medical arts such as age, sex, weight, condition, general health and previous medical history of the patient being treated.

A physician or veterinarian having ordinary skill in the art can readily determine and prescribe an effective amount of the desired pharmaceutical composition. For example, in order to achieve the desired therapeutic effect, a physician or veterinarian may initiate a dose of a compound of the present invention used in a pharmaceutical composition at a level lower than that required and gradually increase the dosage until the desired effect is achieved. In general, a suitable daily dose of a compound of the present invention will be that amount of the compound that is the lowest dose effective to produce a therapeutic effect. Such an effective dose will generally depend on the factors discussed above. Typically, oral, intravenous, intracerebroventricular and subcutaneous doses of the compounds of the invention for patients will range from about 0.01 to about 50 mg/kg body weight/day. If desired, an effective daily dose of the active compound may be administered separately in two, three, four, five, six or more sub-doses at appropriate intervals throughout the day, optionally in unit dosage form. In certain aspects of the invention, the dosing is once a day.

Although the compounds of the present invention may be administered alone, it is preferred to administer the compounds in the form of a pharmaceutical formulation (composition). Kit/Product Packaging

Kits/product packaging are also described herein for use in the treatment of the above-mentioned indications. These kits may consist of a transporter, a pack, or a case of containers, which may be divided into compartments to accommodate one or more containers, such as vials, test tubes, and the like, each container containing the a single component of the method described above. Suitable containers include bottles, vials, syringes and test tubes, among others. Containers are made of acceptable materials such as glass or plastic.

For example, the container may contain one or more of the compounds described herein, which may be present as pharmaceutical components or in admixture with other ingredients described herein. The container may have a sterile outlet (eg, the container may be an IV pack or bottle, the stopper being pierced by a hypodermic needle). Such kits may carry a compound, along with instructions for use, labeling, or operating instructions as described herein.

A typical kit may include one or more containers, each containing one or more materials (such as reagents, or concentrated stock solutions, and/ or equipment). These materials include, but are not limited to, buffers, diluents, filters, needles, syringes, dispensers, bags, containers, vials and/or tubes, with a list of contents and/or instructions for use, and instructions for the inner packaging. The entire set of instructions is to be included.

Labels can be displayed on or closely associated with the container. The presence of a label on a container means that the letters, numbers or other features of the label are affixed, molded, or engraved on the container; the label may also appear in a container box or shipping box containing a variety of containers, such as in product inserts. A label may be used to indicate a specific therapeutic use of the contents. The label may also indicate instructions for use of the contents, such as described in the above method.

All features described in this specification (including any stated claims, abstract and figures), and/or all steps involved in any method or process, may be present in any combination, unless certain features Or steps are mutually exclusive in the same combination.

The above features mentioned in the present invention or the features mentioned in the embodiments can be combined arbitrarily. All the features disclosed in this specification can be used in combination with any composition, and each feature disclosed in the specification can be replaced by any alternative features that serve the same, equivalent or similar purpose. Therefore, unless otherwise stated, the disclosed features are only general examples of equivalent or similar features.

The present invention will be further described below in conjunction with specific embodiments. It should be understood that these examples are only used to illustrate the present invention and not to limit the scope of the present invention. In the following examples, the experimental methods without specific conditions are usually in accordance with conventional conditions or in accordance with the conditions suggested by the manufacturer. All percentages, ratios, ratios, or parts are by weight unless otherwise indicated.

The unit in the weight volume percentage in the present invention is well known to those skilled in the art, for example, it refers to the weight of the solute in 100 ml of the solution. Unless otherwise defined, all professional and scientific terms used herein have the same meanings as those familiar to those skilled in the art. In addition, any methods and materials similar or equivalent to those described can be used in the methods of the present invention. Methods and materials for preferred embodiments described herein are provided for illustrative purposes only.

Example

Generic process

When the preparation route is not included, the raw materials and reagents used in the present invention are all known products, which can be synthesized according to methods known in the art, or can be obtained by purchasing commercially available products. None of the commercially available reagents were used without further purification.

Room temperature refers to 20-30°C.

Unless otherwise specified in the reaction examples, the reactions were all carried out under nitrogen atmosphere. Nitrogen atmosphere means that the reaction flask is connected to a nitrogen balloon of about 1 L.

The hydrogenation reaction is usually evacuated and filled with hydrogen, and the operation is repeated 3 times. Hydrogen atmosphere means that the reaction flask is connected to a hydrogen balloon of about 1 L.

microwave reaction use

Initiator+ microwave reactor.

The structures of the compounds of the present invention were determined by nuclear magnetic resonance (NMR) and mass spectrometry (MS). NMR shifts ([delta]) are given in units of 10<" ⁶ > (ppm). NMR was measured using a (Bruker Ascend ^TM 500 type) nuclear magnetic instrument, and the solvent was deuterated dimethyl sulfoxide (DMSO-d6), deuterated chloroform (CDCl ₃ ), deuterated methanol (CD ₃ OD), and internal standard. For tetramethylsilane (TMS). The following abbreviations are used for the multiplicity of NMR signals: s=singlet, brs=broad, d=doublet, t=triplet, m=multiplet. Coupling constants are listed in J values, measured in Hz.

The measurement of LC-MS was performed using a Thermo liquid mass spectrometer (UltiMate 3000+MSQ PLUS). For the HPLC measurement, a Thermo high pressure liquid chromatograph (UltiMate 3000) was used. Preparative Reversed Phase Chromatography A Thermo (UltiMate 3000) Preparative Reversed Phase Chromatograph was used. Fast column chromatography uses AIJER (FS-9200T) automatic column passing machine, and silica gel prepacked column uses Santai

Prepacked columns. The thin layer chromatography silica gel plate is made of Yantai Huanghai HSGF254 or Qingdao GF254 silica gel plate, and the specifications of the thin layer chromatography separation and purification products are 0.4mm ~ 0.5mm.

The synthetic methods of some intermediates in the present invention are as follows:

Intermediate 1

Intermediate 1 was prepared by the following steps:

The first step: 1-methyl-3,5-dinitropyridin-2-one Int-1a (1.0 g, 5.02 mmol) was dissolved in methanol (50 mL), followed by adding ammonia methanol solution (7 mol/L, 8.61 mL, 60.27 mmol) and 1-methylpiperidin-4-one Int-1b (625 mg, 5.52 mmol). The reaction mixture was heated to 50°C and stirred for 5 hours. After cooling to room temperature, it was left to stand for 48 hours, the reaction solution was concentrated under reduced pressure, and the residue was added with ethyl acetate (50 mL), followed by filtration. The filtrate was concentrated under reduced pressure to obtain Int-1c (1.0 g) as a red solid, which was directly used in the next reaction. ESI-MS (m/z): 194.4 [M+H] ⁺ ; ¹ H NMR (500 MHz, DMSO-d ₆ ) δ 9.14 (d, J=2.5 Hz, 1 H), 8.36 (d, J=2.5 Hz) , 1H), 3.64(s, 2H), 3.02(t, J=6.0Hz, 2H), 2.74(t, J=6.0Hz, 2H), 2.39(s, 3H).

The second step: The compound Int-1c (1.0 g) obtained in the previous step was dissolved in methanol (30 mL), 10% Pd-C (400 mg) was added, and the reaction was carried out at room temperature for 6 hours under a hydrogen atmosphere. The palladium carbon was removed by filtration, and the filtrate was concentrated to obtain Int-1d as a yellow solid (800 mg, yield 94.70%). ESI-MS (m/z): 164.2 [M+H] ⁺ .

The third step: Compound Int-1d (100 mg, 0.61 mmol) was dissolved in acetic acid (3 mL), N-bromosuccinimide (109 mg, 0.61 mmol) was added, and the reaction mixture was stirred at room temperature for 1 hour. Saturated aqueous sodium bicarbonate solution was added to quench the reaction until no bubbles were generated, the aqueous phase was extracted with methanol/dichloromethane (1/20, 50 mL×2), the organic phases were combined, dried over anhydrous sodium sulfate, filtered and concentrated to obtain compound Int-1e (38 mg, 25% yield). ESI-MS (m/z): 242.3 [M+H] ⁺ ; ¹ H NMR (500 MHz, DMSO-d ₆ ) δ 6.77 (s, 1H), 5.25 (s, 2H), 3.37 (s, 2H) , 2.69(t, J=6.0Hz, 2H), 2.60(t, J=6.0Hz, 2H), 2.32(s, 3H).

The fourth step: Compound Int-1e (37mg, 0.15mmol) was dissolved in methanol (1mL), cuprous iodide (3mg, 0.015mmol), 1,10-phenanthroline (3mg, 0.03mmol) and carbonic acid were added. Cesium (99 mg, 0.30 mmol). The reaction mixture was replaced with nitrogen and heated to 100°C with microwave and stirred for 2 hours. The reaction was cooled to room temperature, the reaction solution was concentrated, and the residue was purified by preparative thin layer chromatography (methanol/dichloromethane/triethylamine=1/10/0.1) to obtain a yellow solid Int-1 (20 mg, yield 67%) . ESI-MS (m/z): 194.5 [M+H] ⁺ ; ¹ H NMR (500 MHz, DMSO-d ₆ ) δ 6.54 (s, 1H), 4.68 (s, 2H), 3.80 (s, 3H) , 3.30(s, 2H), 2.64(t, J=5.6Hz, 2H), 2.59(t, J=5.7Hz, 2H), 2.31(s, 3H).

Intermediate 2

Intermediate 2 was prepared by the following steps:

The first step: Compound Int-1e (230mg, 0.94mmol) was dissolved in ethanol (2mL), cuprous iodide (18mg, 0.095mmol), 1,10-phenanthroline (34mg, 0.18mmol) and carbonic acid were added Cesium (619 mg, 1.90 mmol). The reaction mixture was heated to 100°C by microwave and stirred for 5 hours after displacing nitrogen. The reaction was cooled to room temperature, filtered, the filtrate was concentrated, and the residue was purified by silica gel column chromatography (methanol/dichloromethane/triethylamine=1/50/0.1) to give yellow solid Int-2 (113 mg, yield 57%). ESI-MS (m/z): 208.5 [M+H] ⁺ .

Intermediate 3

Intermediate 3 was prepared by the following steps:

The first step: Compound Int-1e (100 mg, 0.41 mmol) and trimethylcyclotriboroxane (148 mg, 1.19 mmol) were dissolved in dioxane (1.5 mL) and water (0.15 mL), and carbonic acid was added Potassium (171 mg, 1.24 mmol), Pd(dppf)Cl2 (30 _mg , 0.041 mmol). After the reaction system was replaced with nitrogen, it was heated to 140° C. with a microwave and stirred for 1 hour. The reaction was cooled to room temperature, the reaction mixture was filtered through celite, and the filtrate was concentrated. The residue was purified by silica gel column chromatography (methanol/dichloromethane=1/20) to obtain Int-3 as a yellow solid (50 mg, yield 68%). ESI-MS (m/z): 178.6 [M+H] ⁺ .

Intermediate 4

Intermediate 4 was prepared by the following steps:

The first step: Compound Int-1e (350 mg, 1.45 mmol) and potassium vinyl trifluoroborate (387 mg, 2.89 mmol) were dissolved in 1,4-dioxane (1.5 mL) and water (0.15 mL), Potassium carbonate (399 mg, 2.89 mmol) and Pd(dppf)Cl2 (105 _mg , 0.14 mmol) were added. After the reaction system was replaced with nitrogen, it was heated to 120° C. with a microwave reactor and stirred for 1 hour. After the reaction was cooled to room temperature, it was filtered through celite, the filtrate was concentrated, and the residue was separated by column chromatography (methanol/dichloromethane=1/20) to obtain yellow solid Int-4a (136 mg, yield 49%). ESI-MS (m/z): 190.7 [M+H] ⁺ ; ¹ H NMR (500 MHz, CDCl ₃ ) δ 6.85 (dd, J=17.2, 11.0 Hz, 1H), 6.66 (s, 1H), 6.16 (dd,J=17.3,1.9Hz,1H),5.49(dd,J=11.0,1.9Hz,1H),3.67-3.63(m,2H),3.55(s,2H),3.00(t,J=6.1 Hz, 2H), 2.80 (t, J=6.1 Hz, 2H), 2.49 (s, 3H).

The second step: Compound Int-4a (60 mg, 0.31 mmol) was dissolved in methanol (5 mL), 10% palladium on carbon (20 mg) was added, and the mixture was stirred at room temperature for 1 hour under a hydrogen atmosphere. The reaction solution was filtered through celite, and the filtrate was concentrated to obtain Intermediate 4 (37 mg, yield 61%). ESI-MS (m/z): 192.7 [M+H] ⁺ .

Intermediate 5

Intermediate 5 was prepared by the following steps:

The first step: Compound Int-1e (100mg, 0.41mmol) was dissolved in a mixed solvent of toluene (3mL) and water (0.3mL), cyclopropylboronic acid (42mg, 0.49mmol), potassium phosphate (306mg, 1.45 mmol), tricyclohexylphosphine (23 mg, 0.082 mmol) and palladium acetate (9 mg, 0.041 mmol). After the reaction system was replaced with nitrogen, it was heated to 100°C and stirred for 18 hours. After the reaction was cooled to room temperature, the reaction solution was filtered through celite, the filtrate was concentrated, and the residue was separated by column chromatography (methanol/dichloromethane=1/20) to obtain yellow solid Int-5 (61 mg, yield 72%). ESI-MS (m/z): 204.2 [M+H] ⁺ .

Intermediate 6

Intermediate 6 was prepared by the following steps:

The first step: N-tert-butoxycarbonyl-4-piperidinone Int-6a (4.4 g, 22.1 mmol) and 1-methyl-3,5-dinitro-2-pyridone Int-1a (4.0 g, 20.1 mmol) was dissolved in methanol (150 mL) and ammonia in methanol (7N, 34.4 mL, 240.8 mmol) was added. Stir at 60°C for 6 hours under nitrogen protection. The reaction solution was cooled to room temperature, and stirring was continued for 2 days. The reaction was monitored by LCMS, the reaction solution was concentrated, ethyl acetate (150 mL) was added, stirred for half an hour, filtered, and the filtrate was concentrated to obtain yellow solid Int-6b (5.1 g, yield 91%). ESI-MS (m/z): 280.1 [M+H] ⁺ .

The second step: Compound Int-6b (5.0 g, 17.9 mmol) was dissolved in methanol (50 mL), and 10% palladium/carbon (500 mg) was added. The mixture was stirred at room temperature under a hydrogen atmosphere (hydrogen balloon) for 16 hours. After the reaction, the reaction solution was filtered, and the filtrate was concentrated to obtain Int-6c (3.7 g, yield 84%) as a light yellow solid. ESI-MS (m/z): 250.2 [M+H] ⁺ .

The third step: Compound Int-6c (3.7 g, 14.8 mmol) was dissolved in DMF (20 mL), and N-bromosuccinimide (2.78 g, 15.6 mmol) and acetic acid (370 mg) were added. The reaction mixture was stirred at room temperature for 2 hours and completed by LCMS. Water (100 mL) was added, the aqueous phase (150 mL*3) was extracted with ethyl acetate, the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated, and the residue was separated by silica gel column chromatography to obtain a yellow solid Int- 6d (3.6 g, 74% yield). ESI-MS (m/z): 328.2 [M+H] ⁺ .

The fourth step: Compound Int-6d (500 mg, 1.53 mmol) was dissolved in methanol (5 mL), and sodium methoxide methanol solution (5N, 0.33 mL, 1.65 mmol) was added. The reaction mixture was heated to 100°C with microwave and stirred for 3 hours. The reaction was cooled to room temperature, the reaction solution was concentrated, and the residue was separated by silica gel column chromatography to obtain Int-6 as a yellow solid (330 mg, yield 77%). ESI-MS (m/z): 280.2 [M+H] ⁺ .

Example 1

3-(((5-Chloro-2-((2-methoxy-6-methyl-5,6,7,8-tetrahydro-1,6-naphthalen-3-yl)amino) pyrimidin-4-yl)amino)methyl)-1-methylpyridin-2(1H)-one

Compound 1 was prepared by the following steps:

The first step: 2,4,5-trichloropyrimidine 1a (57 mg, 0.31 mmol) and 3-(aminomethyl)-1-methyl-1,2-dihydropyridin-2-one hydrochloride 1b (50 mg, 0.28 mmol) and dissolved in isopropanol (2 mL) and N,N-diisopropylethylamine (85 mg, 0.65 mmol, 0.11 mL) was added. The reaction mixture was stirred at room temperature for 18 hours. The reaction solution was filtered, the solid was washed with isopropanol, and dried to obtain a white solid 1c (62 mg, yield 75%). ESI-MS (m/z): 285.2 [M+H] ⁺ .

The second step: Compound 1c (62 mg, 0.21 mmol) and Int-1 (42 mg, 0.21 mmol) were dissolved in 1,4-dioxane (5 mL), and BrettPhos Pd G3 (19 mg, 0.021 mmol) was added, BrettPhos (23 mg, 0.043 mmol) and cesium carbonate (141 mg, 0.43 mmol), the reaction system was heated to 100° C. and stirred for 18 hours after replacing nitrogen. After the reaction solution was cooled to room temperature, the reaction solution was filtered through celite, and the filtrate was concentrated. The residue was purified by silica gel column chromatography (methanol/dichloromethane=1/30) to obtain a crude product, which was then separated by reverse-phase preparative HPLC to obtain white solid 1 (25 mg, yield 26%). ESI-MS (m/z): 442.3 [M+H] ⁺ ; ¹ H NMR (500 MHz, DMSO-d6) δ 8.00 (s, 1H), 7.86 (s, 1H), 7.69-7.61 (m, 2H) ),7.56(s,1H),7.08(d,J＝6.8Hz,1H),6.19(t,J＝6.8Hz,1H),4.37(d,J＝5.9Hz,2H),3.86(s,3H ), 3.50(s, 3H), 3.12(s, 2H), 2.70(t, J=6.0Hz, 2H), 2.60(t, J=5.9Hz, 2H), 2.33(s, 3H).

Example 2

3-(((5-Chloro-2-((2-ethoxy-6-methyl-5,6,7,8-tetrahydro-1,6-naphthalen-3-yl)amino) pyrimidin-4-yl)amino)methyl)-1-methylpyridin-2(1H)-one

Substituting Int-2 for Int-1 in the second step of Example 1, and using similar methods and reaction steps, compound 2 can be obtained. ESI-MS (m/z): 456.1[M+H] ⁺ ; ¹ HNMR (500MHz, DMSO-d6) δ 8.00(s, 1H), 7.87(s, 1H), 7.69(t, J=5.9Hz ,1H),7.64(dd,J＝6.7,2.0Hz,1H),7.48(s,1H),7.12-7.06(m,1H),6.18(t,J＝6.8Hz,1H),4.38(d, J=5.9Hz, 2H), 4.30(q, J=7.0Hz, 2H), 3.50(s, 3H), 3.09(s, 2H), 2.67(d, J=5.9Hz, 2H), 2.59(d, J=5.7Hz, 2H), 2.32 (s, 3H), 1.32 (t, J=7.0Hz, 3H).

Example 3

3-(((2-((2-Ethoxy-6-methyl-5,6,7,8-tetrahydro-1,6-naphthalen-3-yl)amino)-5-( Trifluoromethyl)pyrimidin-4-yl)amino)methyl)-1-methylpyridin-2(1H)-one

Substitute 2,4-dichloro-5-trifluoromethylpyrimidine for 2,4,5-trichloropyrimidine 1a in the first step of Example 1, and then substitute Int-2 for Int in the second step of Example 1 -1, Compound 3 can be obtained by a similar method and reaction procedure. ESI-MS (m/z): 490.2[M+H] ⁺ ; ¹ H NMR (500MHz, DMSO-d6) δ 8.23(s, 1H), 7.95(s, 1H), 7.76(s, 1H), 7.64(dd,J=6.7,2.0Hz,1H),7.59(br s,1H),7.02(d,J=7.0Hz,1H),6.19(t,J=6.8Hz,1H),4.40(d, J=5.6Hz, 2H), 4.30(q, J=7.0Hz, 2H), 3.50(s, 3H), 3.10(br s, 2H), 2.69(t, J=5.9Hz, 2H), 2.59(t , J=5.9Hz, 2H), 2.33 (s, 3H), 1.31 (t, J=7.0Hz, 3H).

Example 4

5-(((5-Chloro-2-((2-methoxy-6-methyl-5,6,7,8-tetrahydro-1,6-naphthalen-3-yl)amino) pyrimidin-4-yl)amino)methyl)pyridin-2(1H)-one

Substitute 5-(aminomethyl)-1,2-dihydropyridin-2-one for 3-(aminomethyl)-1-methyl-1,2-dihydropyridine- 2-keto hydrochloride 1b, compound 4 can be obtained by a similar method and reaction procedure. ESI-MS (m/z): 428.2[M+H] ⁺ ; ¹ H NMR (500MHz, DMSO-d6) δ 7.99(s, 1H), 7.96(s, 1H), 7.74(s, 1H), 7.68(t, J=5.9Hz, 1H), 7.40(dd, J=9.4, 2.6Hz, 1H), 7.20(br s, 1H), 6.27(d, J=9.4Hz, 1H), 4.29(d, J=5.6Hz, 2H), 3.88(s, 3H), 3.34-3.30(m, 2H), 2.74(t, J=5.8Hz, 2H), 2.63(t, J=5.9Hz, 2H), 2.33( s, 3H).

Example 5

3-(((5-Chloro-2-((2,6-dimethyl-5,6,7,8-tetrahydro-1,6-naphthalen-3-yl)amino)pyrimidine-4 -yl)amino)methyl)-1-methylpyridin-2(1H)-one

Substituting Int-3 for Int-1 in the second step of Example 1, and using similar methods and reaction steps, compound 5 can be obtained. ESI-MS (m/z): 426.2 [M+H] ⁺ ; ¹ H NMR (500 MHz, DMSO-d6) δ 8.36 (s, 1H), 7.91 (s, 1H), 7.63 (dd, J=6.8 ,2.0Hz,1H),7.50-7.40(m,2H),7.08-6.96(m,1H),6.18(t,J=6.8Hz,1H),4.29(d,J=5.9Hz,2H),3.47 (s,3H),3.19(s,2H),2.76(t,J=6.0Hz,2H),2.62(t,J=6.0Hz,2H),2.31(s,3H),2.30(s,3H) .

Example 6

3-(((5-Chloro-2-((2-methoxy-6-methyl-5,6,7,8-tetrahydro-1,6-naphthalen-3-yl)amino) pyrimidin-4-yl)amino)methyl)-1,4,6-trimethylpyridin-2(1H)-one

Compound 6 was prepared by the following steps:

Step 1: To a suspension of sodium hydride (202 mg, 5.06 mmol, content 60%) in DMF (5 mL) at 0°C in an ice bath and nitrogen atmosphere, compound 6a (500 mg, 3.37 mmol) in DMF (5 mL) was added solution. After stirring for 30 minutes, iodomethane (718 mg, 5.06 mmol, 0.31 mL) was added, and the mixture was stirred at room temperature for 16 hours. The reaction solution was diluted with ethyl acetate, and washed with water and saturated brine in this order. The organic phase was dried over anhydrous sodium sulfate, filtered and concentrated, and the residue was separated by silica gel column chromatography (petroleum ether/ethyl acetate=1/1) to obtain yellow solid 6b (230 mg, yield 42%). ESI-MS (m/z): 163.1 [M+H] ⁺ .

Second step: Compound 6b (230 mg, 1.42 mmol) was dissolved in methanol (5 mL), Raney Ni (0.5 mL, aqueous suspension) and ammonia (0.5 mL) were added. The mixture was stirred at room temperature under an atmosphere of hydrogen (hydrogen balloon) for 16 hours. The reaction solution was filtered with celite, the filter cake was washed with methanol, and the filtrate was concentrated to obtain compound 6c (200 mg), which was directly used in the next reaction. ESI-MS (m/z): 167.1 [M+H] ⁺ .

The third step: Compound 6c (135 mg) was dissolved in isopropanol (2 mL), and N,N-diisopropylethylamine (159 mg, 1.23 mmol, 0.21 mL) and 2,4,5-trichloropyrimidine were added 1a (150 mg, 0.81 mmol) and the mixture was stirred at room temperature for 16 hours. The reaction solution was concentrated, and the residue was separated by silica gel column chromatography (dichloromethane/methanol=10/1) to obtain 6d (130 mg) as a white solid. ESI-MS (m/z): 313.2 [M+H] ⁺ .

The fourth step: Compound 6d (71 mg, 0.22 mmol) and Int-1 (40 mg, 0.20 mmol) were dissolved in 1,4-dioxane (4 mL), cesium carbonate (202 mg, 0.62 mmol) was added, BrettPhos ( 22 mg, 0.041 mmol) and BrettPhos Pd G3 (18 mg, 0.020 mmol). After the reaction system was replaced with nitrogen, it was heated to 100°C and stirred for 16 hours. After the reaction solution was cooled to room temperature, the reaction solution was concentrated, and the residue was purified by preparative thin layer chromatography (dichloromethane/methanol=10/1) to obtain a crude product, which was then separated by reverse preparative HPLC to obtain compound 6 (10 mg, yield 10%). ESI-MS (m/z): 470.2[M+H] ⁺ ; ¹ H NMR (500MHz, DMSO-d6) δ 8.07(s, 1H), 7.94(s, 1H), 7.69(s, 1H), 7.07(br s,1H),6.05(s,1H),4.44(d,J=5.5Hz,2H),3.88(s,3H),3.46-3.40(m,5H),2.75(br s,2H) , 2.65(t, J=5.5Hz, 2H), 2.34(s, 3H), 2.30(s, 3H), 2.12(s, 3H).

Example 7

3-(((5-Chloro-2-((2-methoxy-6-methyl-5,6,7,8-tetrahydro-1,6-naphthalen-3-yl)amino) Pyrimidin-4-yl)amino)methyl)-4,6-lutidine-2(1H)-one

Substitute 3-(aminomethyl)-4,6-dimethyl-1,2-dihydropyridin-2-one for 3-(aminomethyl)-1-methyl- 1,2-Dihydropyridin-2-one hydrochloride 1b, compound 7 can be obtained by a similar method and reaction procedure. ESI-MS(m/z): 456.0[M+H] ⁺ ; ¹ HNMR (500MHz, DMSO-d6)δ11.59(br s,1H),8.09(s,1H),7.94(s,1H), 7.69(s, 1H), 7.20(br s, 1H), 5.89(s, 1H), 4.42(br s, 2H), 3.89(s, 3H), 3.42(s, 2H), 2.75(br s, 2H ), 2.65(br s, 2H), 2.34(s, 3H), 2.12(s, 6H).

Example 8

4-(((5-Chloro-2-((2-methoxy-6-methyl-5,6,7,8-tetrahydro-1,6-naphthalen-3-yl)amino) pyrimidin-4-yl)amino)methyl)pyridin-2(1H)-one

Substitute 4-(aminomethyl)-1,2-dihydropyridin-2-one for 3-(aminomethyl)-1-methyl-1,2-dihydropyridine- 2-keto hydrochloride 1b, compound 8 can be obtained by a similar method and reaction procedure. ESI-MS (m/z): 428.1 [M+H] ⁺ ; ¹ H NMR (500 MHz, DMSO-d6) δ 8.01 (s, 1H), 7.90 (s, 1H), 7.83 (t, J=6.1 Hz, 1H), 7.62(s, 1H), 7.31(d, J=6.7Hz, 1H), 6.11(dd, J=6.8, 1.7Hz, 1H), 6.08(s, 1H), 4.41(d, J ＝5.8Hz, 2H), 3.86(s, 3H), 3.22(s, 2H), 2.71(t, J＝5.9Hz, 2H), 2.61(t, J＝5.9Hz, 2H), 2.33(s, 3H) ).

Example 9

3-(((5-Chloro-2-((2-methoxy-6-methyl-5,6,7,8-tetrahydro-1,6-naphthalen-3-yl)amino) Pyrimidine-4-yl)amino)methyl)-1-ethylpyridin-2(1H)-one

Compound 9 was prepared by the following steps:

The first step: Compound 9a (700 mg, 5.83 mmol) was dissolved in DMF (10 mL), cesium carbonate (2.28 g, 6.99 mmol) was added, and then iodoethane (1.36 g, 8.74 mmol, 0.70 mL) was added dropwise, and the reaction was carried out. The mixture was stirred at room temperature overnight. The reaction solution was diluted with water, and the aqueous phase was extracted with ethyl acetate. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated. The residue was separated by silica gel column chromatography (petroleum ether/ethyl acetate=1/1) to obtain 9b (730 mg, yield 84%) as a white solid. ESI-MS (m/z): 149.4 [M+H] ⁺ ; ¹ H NMR (500 MHz, Chloroform-d) δ 7.80 (dd, J=7.0, 2.0 Hz, 1 H), 7.57 (dd, J=7.0, 2.0Hz, 1H), 6.28 (t, J=7.0Hz, 1H), 4.05 (q, J=7.0Hz, 2H), 1.40 (t, J=7.0Hz, 3H).

Second step: Compound 9b (100 mg, 0.67 mmol) was dissolved in methanol (5 mL), Raney Ni (0.3 mL, aqueous suspension) and ammonia (1 mL) were added. The mixture was stirred at room temperature under a hydrogen atmosphere (hydrogen balloon) overnight. The reaction solution was filtered with celite, the filter cake was washed with methanol, and the filtrate was concentrated to obtain compound 9c (100 mg), which was directly used in the next reaction.

The third step: Compound 9c (100 mg) was dissolved in isopropanol (5 mL), and N,N-diisopropylethylamine (169 mg, 1.31 mmol, 0.23 mL) and 2,4,5-trichloropyrimidine were added 1a (180 mg, 0.98 mmol), the reaction mixture was stirred at room temperature overnight. The reaction solution was filtered, and the filter cake was washed with isopropanol and dried to obtain 9d (100 mg) as a white solid. ESI-MS (m/z): 299.2 [M+H] ⁺ .

The fourth step: Compound 9d (50 mg, 0.16 mmol) and Int-1 (32 mg, 0.16 mmol) were dissolved in 1,4-dioxane (5 mL), cesium carbonate (108 mg, 0.33 mmol) was added, BrettPhos ( 9 mg, 0.016 mmol) and BrettPhos Pd G3 (15 mg, 0.016 mmol). The reaction system was replaced with nitrogen and then heated to 100°C and stirred overnight. After the reaction solution was cooled to room temperature, the reaction solution was concentrated, and the residue was purified by preparative thin layer chromatography (dichloromethane/methanol=10/1) to obtain a crude product, which was then separated by reverse preparative HPLC to obtain compound 9 (8 mg, yield 11%). ESI-MS (m/z): 456.1 [M+H] ⁺ ; ¹ H NMR (500 MHz, DMSO-d6) δ 8.00 (s, 1H), 7.91 (s, 1H), 7.66-7.62 (m, 2H ), 7.56(s, 1H), 7.08(dd, J=7.0, 2.0Hz, 1H), 6.20(t, J=7.0Hz, 1H), 4.39(d, J=6.0Hz, 2H), 3.98(q , J＝7.0Hz, 2H), 3.85(s, 3H), 3.18(s, 2H), 2.70(t, J＝6.0Hz, 2H), 2.60(t, J＝6.0Hz, 2H), 2.32(s , 3H), 1.25 (t, J=7.0Hz, 3H).

Example 10

3-(((5-Chloro-2-((2-methoxy-6-methyl-5,6,7,8-tetrahydro-1,6-naphthalen-3-yl)amino) pyrimidin-4-yl)amino)methyl)-1,5,6-trimethylpyridin-2(1H)-one

Substituting 2-hydroxy-3-cyano-5,6-lutidine for 3-cyano-2-hydroxy-4,6-lutidine in the first step of Example 9, using a similar method and In the reaction step, compound 10 can be obtained. ESI-MS (m/z): 470.1 [M+H] ⁺ ; ¹ HNMR (500 MHz, DMSO-d6) δ 7.98 (s, 1H), 7.93 (s, 1H), 7.58 (t, J=6.0 Hz ,1H),7.56(s,1H),6.94(s,1H),4.36(d,J=6.0Hz,2H),3.86(s,3H),3.52(s,3H),3.19(s,2H) , 2.71(t, J=6.0Hz, 2H), 2.60(t, J=6.0Hz, 2H), 2.33(s, 3H), 2.27(s, 3H), 1.99(s, 3H).

Example 11

3-(((5-Chloro-2-((2-methoxy-6-methyl-5,6,7,8-tetrahydro-1,6-naphthalen-3-yl)amino) pyrimidin-4-yl)amino)methyl)pyridin-2(1H)-one

Substitute 3-(aminomethyl)pyridin-2(1H)-one for the 3-(aminomethyl)-1-methyl-1,2-dihydropyridin-2-one salt in the first step of Example 1 Compound 11 can be obtained using a similar method and reaction procedure for the acid salt 1b. ESI-MS(m/z): 428.3[M+H] ⁺ ; ¹ H NMR (500MHz, DMSO-d6) δ 11.68(br s, 1H), 8.00(s, 1H), 7.90(s, 1H) ,7.65-7.62(m,1H),7.56(s,1H),7.33-7.28(m,1H),7.13-7.09(m,1H),6.15(t,J＝6.6Hz,1H),4.35(d , J＝5.9Hz, 2H), 3.86(s, 3H), 3.17(s, 2H), 2.69(d, J＝5.9Hz, 2H), 2.60(d, J＝5.7Hz, 2H), 2.33(s , 3H).

Example 12

3-(((5-Chloro-2-((2-cyclopropyl-6-methyl-5,6,7,8-tetrahydro-1,6-naphthalen-3-yl)amino) pyrimidin-4-yl)amino)methyl)-1-methylpyridin-2(1H)-one

Substituting Int-5 for Int-1 in the second step of Example 1, and using similar methods and reaction steps, compound 12 can be obtained. ESI-MS (m/z): 452.2 [M+H] ⁺ ; ¹ H NMR (500 MHz, DMSO-d6) δ 8.55 (s, 1H), 7.90 (s, 1H), 7.62 (dd, J=6.7 ,2.0Hz,1H),7.36(t,J＝6.0Hz,1H),7.34(s,1H),6.97(d,J＝6.9Hz,1H),6.15(t,J＝6.8Hz,1H), 4.29(d,J＝6.0Hz,2H),3.46(s,3H),3.20(s,2H),2.71(t,J＝6.0Hz,2H),2.61(t,J＝6.0Hz,2H), 2.31(s, 3H), 2.23-2.12(m, 1H), 0.85-.075(m, 4H).

Example 13

3-(((5-Chloro-2-((2-ethyl-6-methyl-5,6,7,8-tetrahydro-1,6-naphthalen-3-yl)amino)pyrimidine -4-yl)amino)methyl)-1-methylpyridin-2(1H)-one

Substituting Int-4 for Int-1 in the second step of Example 1, and using similar methods and reaction steps, compound 13 can be obtained. ESI-MS (m/z): 440.2 [M+H] ⁺ ; ¹ H NMR (500 MHz, DMSO-d6) δ 8.36 (s, 1H), 7.89 (s, 1H), 7.62 (dd, J=6.8, 2.0Hz, 1H), 7.37(d, J＝6.0Hz, 1H), 7.36(s, 1H), 6.98(d, J＝6.9Hz, 1H), 6.16(t, J＝6.8Hz, 1H), 4.27 (d, J＝6.0Hz, 2H), 3.46(s, 3H), 3.22(s, 2H), 2.78(t, J＝6.0Hz, 2H), 2.69-2.61(m, 4H), 2.32(s, 3H), 1.07 (t, J=7.5Hz, 3H).

Example 14

3-(((5-Chloro-2-((2-methoxy-6-methyl-5,6,7,8-tetrahydro-1,6-naphthalen-3-yl)amino) pyrimidin-4-yl)amino)methyl)-1-methylpyrrolidin-2-one

Substitute 3-(aminomethyl)-1-methylpyrrolidin-2-one for 3-(aminomethyl)-1-methyl-1,2-dihydropyridine-2 in the first step of Example 1 - Ketone hydrochloride 1b, compound 14 can be obtained by a similar method and reaction procedure. ESI-MS (m/z): 432.3[M+H] ⁺ ; ¹ H NMR (500MHz, DMSO-d6) δ 8.13(s, 1H), 7.96(s, 1H), 7.61(s, 1H), 7.38-7.33(m, 1H), 3.88(s, 3H), 3.65-3.58(m, 1H), 3.50-3.40(m, 3H), 3.27-3.24(m, 2H), 2.79-2.73(m, 6H) ), 2.66-2.62(m, 2H), 2.34(s, 3H), 2.08-2.02(m, 1H), 1.78-1.72(m, 1H).

Example 15

5-(((5-Chloro-2-((2-methoxy-6-methyl-5,6,7,8-tetrahydro-1,6-naphthalen-3-yl)amino) pyrimidin-4-yl)amino)methyl)pyrrolidin-2-one

Substitute 5-aminomethyl-2-pyrrolidone for 3-(aminomethyl)-1-methyl-1,2-dihydropyridin-2-one hydrochloride 1b in the first step of Example 1, and use similar According to the method and reaction steps, compound 15 can be obtained. ESI-MS(m/z): 418.4[M+H] ⁺ ; ¹ HNMR (500MHz, DMSO-d6)δ8.10(s,1H),7.95(s,1H),7.75(br s,1H), 7.67(s, 1H), 7.27(t, J=5.9Hz, 1H), 3.89(s, 3H), 3.82-3.77(m, 1H), 3.53-3.47(m, 1H), 3.41(s, 2H) ,3.36-3.30(m,1H),2.75(t,J＝5.9Hz,2H),2.64(t,J＝6.3Hz,2H),2.34(s,3H),2.13-2.03(m,3H), 1.78-1.71 (m, 1H).

Example 16

3-(((2-((2-Methoxy-6-methyl-5,6,7,8-tetrahydro-1,6-naphthalen-3-yl)amino)-5-methyl pyrimidin-4-yl)amino)methyl)-1-methylpyridin-2(1H)-one

Substituting 2,4-dichloro-5-methylpyrimidine for 2,4,5-trichloropyrimidine 1a in the first step of Example 1, and using a similar method and reaction procedure, compound 16 can be obtained. ESI-MS (m/z): 422.3 [M+H] ⁺ ; ¹ H NMR (500 MHz, DMSO-d6) δ 8.19 (s, 1H), 7.74 (s, 1H), 7.63 (dd, J=6.8, 1.9Hz, 1H), 7.35(br s, 1H), 7.31(br s, 1H), 7.15(d, J=6.8Hz, 1H), 6.19(t, J=6.8Hz, 1H), 4.41(d, J=5.9Hz, 2H), 3.96-3.85(m, 5H), 3.52(s, 3H), 3.30-3.22(m, 2H), 2.91(br s, 2H), 2.74(br s, 2H), 2.49 (s, 3H), 2.02 (s, 3H).

Example 17

3-(((5-Fluoro-2-((2-methoxy-6-methyl-5,6,7,8-tetrahydro-1,6-naphthalen-3-yl)amino) pyrimidin-4-yl)amino)methyl)-1-methylpyridin-2(1H)-one

Substituting 2,4-dichloro-5-fluoropyrimidine for 2,4,5-trichloropyrimidine 1a in the first step of Example 1, and using a similar method and reaction procedure, compound 17 can be obtained. ESI-MS (m/z): 426.3 [M+H] ⁺ ; ¹ H NMR (500 MHz, DMSO-d6) δ 8.03 (s, 1H), 8.00-7.91 (m, 2H), 7.64 (dd, J =6.7,2.0Hz,1H),7.43(s,1H),7.19(dd,J=7.0,1.8Hz,1H),6.19(t,J=6.8Hz,1H),4.37(d,J=5.9Hz ,2H),3.87(s,3H),3.50(s,3H),3.47(br s,2H),2.88(br s,2H),2.79(t,J＝5.8Hz,2H),2.49(s, 3H).

Example 18

4-(((5-Chloro-2-((2-methoxy-6-methyl-5,6,7,8-tetrahydro-1,6-naphthalen-3-yl)amino) pyrimidin-4-yl)amino)methyl)-1-methylpyridin-2(1H)-one

Substitute 4-(aminomethyl)-1-methyl-1,2-dihydropyridin-2-one for 3-(aminomethyl)-1-methyl-1,2 in the first step of Example 1 - Dihydropyridin-2-one hydrochloride 1b, compound 33 can be obtained by a similar method and reaction procedure. ESI-MS (m/z): 442.3 [M+H] ⁺ ; ¹ H NMR (500 MHz, DMSO-d6) δ 8.01 (s, 1H), 7.89 (s, 1H), 7.84 (t, J=5.5 Hz,1H),7.66-7.60(m,2H),6.17-6.13(m,2H),4.41(d,J=5.9Hz,2H),3.85(s,3H),3.37(s,3H),3.21 (br s, 2H), 2.71 (t, J=5.3 Hz, 2H), 2.61 (t, J=5.7 Hz, 2H), 2.33 (s, 3H).

Example 19

3-(((5-Chloro-2-((2-methoxy-5,6,7,8-tetrahydro-1,6-naphthalen-3-yl)amino)pyrimidin-4-yl )amino)methyl)-1-methylpyridin-2(1H)-one

Compound 19 was prepared by the following steps:

The first step: Compound 1b (244 mg, 0.85 mmol) and Int-7 (200 mg, 0.71 mmol) were dissolved in 1,4-dioxane (5 mL), cesium carbonate (700 mg, 2.15 mmol) was added, BrettPhos ( 76 mg, 0.14 mmol) and BrettPhos Pd G3 (64 mg, 0.071 mmol). After the reaction system was replaced with nitrogen, it was heated to 120°C and stirred for 16 hours. After the reaction solution was cooled to room temperature, the reaction solution was concentrated, and the residue was purified by silica gel column chromatography (dichloromethane/methanol=20/1) to obtain the crude compound 19a (300 mg), which was directly used in the next reaction. ESI-MS (m/z): 528.1 [M+H] ⁺ .

The second step: The crude 19a obtained in the previous step (300 mg) was dissolved in acetonitrile (5 mL), and p-toluenesulfonic acid monohydrate (323 mg, 1.70 mmol) was added. The reaction mixture was warmed to 70°C and stirred for 2 hours. The reaction solution was concentrated to obtain a crude product of compound 19 (240 mg), 20 mg of which was purified by reverse-phase preparative HPLC to obtain compound 19 (1 mg). ESI-MS (m/z): 428.3 [M+H] ⁺ ; ¹ H NMR (500 MHz, DMSO-d6) δ 8.00 (s, 1H), 7.84 (s, 1H), 7.69-7.60 (m, 2H ), 7.53(s, 1H), 7.09(d, J＝6.9Hz, 1H), 6.18(t, J＝6.8Hz, 1H), 4.37(d, J＝5.9Hz, 2H), 3.85(s, 3H ), 3.52(s, 2H), 3.49(s, 3H), 2.96(t, J=5.8Hz, 2H), 2.59(t, J=5.8Hz, 2H).

Example 20

2-((2-Methoxy-6-methyl-5,6,7,8-tetrahydro-1,6-naphthalen-3-yl)amino)-4-(((1-methyl yl-2-carbonyl-1,2-dihydropyridin-3-yl)methyl)amino)pyrimidine-5-carbonitrile

Substituting 2,4-dichloro-5-cyanopyrimidine for 2,4,5-trichloropyrimidine 1a in the first step of Example 1, and using a similar method and reaction procedure, compound 20 can be obtained. ESI-MS (m/z): 433.2 [M+H] ⁺ ; ¹ H NMR (500 MHz, DMSO-d6) δ 8.38 (s, 1H), 8.24-8.19 (m, 1H), 8.16 (s, 1H) ), 7.71(s, 1H), 7.63-7.60(m, 1H), 6.98-6.94(m, 1H), 6.15-6.12(m, 1H), 4.16(d, J=6.1Hz, 2H), 3.85( s, 3H), 3.47 (s, 3H), 3.14 (br s, 2H), 2.73 (d, J=6.1 Hz, 2H), 2.63-2.60 (m, 2H), 2.33 (s, 3H).

Example 21

3-(((5-Chloro-2-((2-methoxy-6-methyl-5,6,7,8-tetrahydro-1,6-naphthalen-3-yl)amino) Pyrimidine-4-yl)amino)methyl)-1,5-lutidine-2(1H)-one

Compound 21 was prepared by the following steps:

Step 1: Dissolve 3-bromo-2-hydroxy-5-methylpyridine 21a (200mg, 1.06mmol) in tetrahydrofuran (5mL), add NaH (50mg, 1.28mmol, content 60%) at 0°C, The reaction solution was stirred for 30 minutes. Iodomethane (227 mg, 1.60 mmol) was added dropwise to the reaction solution, and the reaction solution was further stirred at 0°C for 2 hours. The reaction solution was quenched with saturated aqueous ammonium chloride solution and extracted with ethyl acetate. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate=5/3) to obtain compound 21b (200 mg, yield 93%). ESI-MS (m/z): 202.5 [M+H] ⁺ .

The second step: Compound 21b (180 mg, 0.89 mmol) was dissolved in DMF (5 mL), Pd(PPh ₃ ) ₄ (103 mg, 0.09 mmol) and Zn(CN) ₂ (105 mg, 0.89 mmol) were added, and the reaction system was replaced After nitrogen, the reaction was heated to 130°C by microwave for 4 hours. After the reaction solution was cooled to room temperature, the reaction solution was filtered through celite, and the filtrate was concentrated. The residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate=5/4) to obtain 21c (122 mg, yield 92%) as a white solid. ESI-MS (m/z): 149.4 [M+H] ⁺ .

The third step: Compound 21c (120 mg, 0.81 mmol) was dissolved in methanol (5 mL) and ammonia water (0.5 mL), Raney Nickel (0.3 mL, aqueous suspension) and BOC anhydride (212 mg, 0.97 mmol) were added, and the mixture was Stir overnight at room temperature under a hydrogen atmosphere (hydrogen balloon). The reaction solution was filtered through Celite, and the filtrate was concentrated. The residue was dissolved in 4N hydrochloric acid dioxane solution (5 mL), and the reaction solution was stirred at room temperature for 1 hour. The reaction solution was concentrated to obtain the crude product of compound 21d, which was directly used in the next reaction. ESI-MS (m/z): 153.8 [M+H] ⁺ .

The fourth step: The crude compound 21d obtained in the previous step and 2,4,5-trichloropyrimidine 1a (169 mg, 0.92 mmol) were dissolved in i-PrOH (5 mL), and N,N-diisopropylethyl was added. Amine (357 mg, 2.76 mmol), the reaction was stirred at 90°C overnight. After the reaction solution was cooled to room temperature, the reaction solution was concentrated. The residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate=5/4) to obtain 21e (98 mg, yield 36%) as a white solid. ESI-MS (m/z): 300.2 [M+H] ⁺ .

The fifth step: Compound 21e (50mg, 0.17mmol) and Int-1 (36mg, 0.18mmol) were dissolved in 1,4-dioxane (5mL), BrettPhos G3Pd (15mg, 17umol), BrettPhos (9mg , 17umol) and cesium carbonate (109mg, 0.34mmol). The reaction system was replaced with nitrogen and then heated to 100°C and stirred overnight. After the reaction solution was cooled to room temperature, the reaction solution was filtered through celite, and the filtrate was concentrated. The residue was purified by preparative thin layer chromatography (petroleum ether/ethyl acetate = 1/1) to obtain the crude product of compound 21, which was purified by reverse phase preparative HPLC to obtain compound 21 (6 mg, yield 8%). ESI-MS (m/z): 456.2 [M+H] ⁺ . ¹ H NMR (500MHz, DMSO-d ₆ )δ7.99(s,1H),7.89(s,1H),7.62(t,J=6.0Hz,1H),7.57(s,1H),7.43(s,1H) 1H), 6.97(s, 1H), 4.37(d, J=5.8Hz, 2H), 3.86(s, 3H), 3.45(s, 3H), 3.16(s, 2H), 2.70(t, J=5.7 Hz, 2H), 2.60 (t, J=5.9 Hz, 2H), 2.33 (s, 3H), 1.96 (s, 3H).

Example 22

3-(((2-((2-Methoxy-6-methyl-5,6,7,8-tetrahydro-1,6-naphthalen-3-yl)amino)pyrimidine-4- yl)amino)methyl)-1-methylpyridin-2(1H)-one

Substituting 2,4-dichloropyrimidine for 2,4,5-trichloropyrimidine 1a in the first step of Example 1, and using similar methods and reaction steps, compound 22 can be obtained. ESI-MS (m/z): 408.3[M+H] ⁺ ; ¹ HNMR (500MHz, DMSO-d6) δ 8.08 (br s, 1H), 7.83 (d, J=5.8Hz, 1H), 7.68 ( br s,1H),7.63(dd,J=6.7,1.9Hz,1H),7.28(s,1H),7.19(br s,1H),6.19(t,J=6.8Hz,1H),6.10(br s, 1H), 4.30(br s, 2H), 3.88(s, 3H), 3.49(s, 3H), 3.19(br s, 2H), 2.70(t, J＝6.6Hz, 2H), 2.62(t , J=5.7Hz, 2H), 2.33 (s, 3H).

Example 23

3-(((5-Chloro-2-((2-methoxy-6-methyl-5,6,7,8-tetrahydro-1,6-naphthalen-3-yl)amino) Pyrimidin-4-yl)amino)methyl)-1,4-lutidine-2(1H)-one

Compound 23 was prepared by the following steps:

Step 1: Dissolve 3-bromo-2-hydroxy-4-methylpyridine 23a (200mg, 1.06mmol) in tetrahydrofuran (5mL), add NaH (50mg, 1.28mmol, content 60%) at 0°C, The reaction solution was stirred for 30 minutes. Iodomethane (227 mg, 1.60 mmol) was added dropwise to the reaction solution, and the reaction solution was further stirred at 0°C for 2 hours. The reaction solution was quenched with saturated aqueous ammonium chloride solution and extracted with ethyl acetate. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate=5/3) to obtain 23b as a yellow oil (210 mg, yield 98%). ESI-MS (m/z): 202.5 [M+H] ⁺ .

The second step: Compound 23b (210 mg, 1.04 mmol) was dissolved in DMF (5 mL), Pd(PPh ₃ ) ₄ (120 mg, 0.11 mmol) and Zn(CN) ₂ (122 mg, 1.04 mmol) were added, and the reaction system was replaced After nitrogen, the reaction was heated to 130°C by microwave for 4 hours. After the reaction solution was cooled to room temperature, the reaction solution was filtered through celite, and the filtrate was concentrated. The residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate=5/4) to obtain 23c (143 mg, yield 92%) as a white solid. ESI-MS (m/z): 149.4 [M+H] ⁺ .

The third step: Compound 23c (143 mg, 0.97 mmol) was dissolved in methanol (5 mL) and ammonia water (0.5 mL), Raney Nickel (0.3 mL, aqueous suspension) and BOC anhydride (253 mg, 1.16 mmol) were added, and the mixture was Stir overnight at room temperature under a hydrogen atmosphere (hydrogen balloon). The reaction solution was filtered through Celite, and the filtrate was concentrated. The residue was dissolved in 4N hydrochloric acid dioxane solution (5 mL), and the reaction solution was stirred at room temperature for 1 hour. The reaction solution was concentrated to obtain the crude product of compound 23d, which was directly used in the next reaction. ESI-MS (m/z): 153.5 [M+H] ⁺ .

The fourth step: The crude compound 23d obtained in the previous step and 2,4,5-trichloropyrimidine (176 mg, 0.96 mmol) were dissolved in i-PrOH (5 mL), DIEA (372 mg, 2.88 mmol) was added, and the reaction solution was Stir overnight at 90°C. After the reaction solution was cooled to room temperature, the reaction solution was concentrated. The residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate=5/4) to obtain 23e (161 mg, yield 56%) as a white solid. ESI-MS (m/z): 300.2 [M+H] ⁺ .

The fifth step: Compound 23e (50mg, 0.17mmol) and Int-1 (32mg, 0.17mmol) were dissolved in 1,4-dioxane (5mL), BrettPhos G3Pd (15mg, 17umol), BrettPhos (9mg , 17umol) and cesium carbonate (109mg, 0.34mmol). The reaction system was replaced with nitrogen and then heated to 100°C and stirred overnight. After the reaction solution was cooled to room temperature, the reaction solution was filtered through celite, and the filtrate was concentrated. The residue was purified by preparative thin layer chromatography (petroleum ether/ethyl acetate=1/1) to obtain the crude product of compound 23, which was purified by reverse phase preparative HPLC to obtain compound 23 (7 mg, yield 9%). ESI-MS (m/z): 456.1 [M+H] ⁺ ; ¹ H NMR (500 MHz, DMSO-d ₆ ) δ 8.07 (s, 1H), 7.95 (s, 1H), 7.71 (s, 1H) ,7.56(d,J＝6.9Hz,1H),7.14(t,J＝5.4Hz,1H),6.12(d,J＝7.0Hz,1H),4.47(d,J＝5.4Hz,2H),3.88 (s, 3H), 3.43(s, 5H), 2.76(t, J=5.8Hz, 2H), 2.65(t, J=5.9Hz, 2H), 2.34(s, 3H), 2.16(s, 3H) .

Example 24

3-(((5-Chloro-2-((2-methoxy-6-methyl-5,6,7,8-tetrahydro-1,6-naphthalen-3-yl)amino) pyrimidin-4-yl)amino)methyl)pyrrolidin-2-one

Compound 24 was prepared by the following steps:

The first step: compound 3-(hydroxymethyl)pyrrolidin-2-one 24a (258 mg, 2.24 mmol), phthalimide (362 mg, 2.4, 7 mmol) and triphenylphosphine (646 mg, 2.47 mmol) mmol) was dissolved in THF (5 mL), and diisopropyl azodicarboxylate (498 mg, 2.47 mmol, 0.48 mL) was slowly added dropwise at 0°C. The reaction mixture was warmed to room temperature and stirred for 18 hours. The reaction was complete by LCMS. The reaction solution was concentrated, and the residue was purified by silica gel column chromatography to obtain white solid 24b (335 mg, yield 61%). ESI-MS (m/z): 245.5 [M+H] ⁺ .

The second step: compound 24b (335 mg, 1.37 mmol) was dissolved in ethanol (5 mL), and hydrazine hydrate ((171 mg, 2.74 mmol, 0.17 mL, content 80%) was added. The reaction mixture was raised to 85° C. and stirred for 2 hours and then cooled. The reaction solution was filtered, and the filtrate was concentrated to obtain a crude product of compound 24c (156 mg), which was directly used in the next reaction.

The third step: The crude compound 24c obtained in the previous step (156 mg) and 2,4,5-trichloropyrimidine 1a (501 mg, 2.73 mmol) were dissolved in i-PrOH (5 mL), and DIPEA (529 mg, 4.10 mmol) was added. . The reaction mixture was stirred at room temperature for 18 hours. The reaction solution was concentrated. The residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate=1/1) to obtain 24d as a white solid (300 mg, yield 84%). ESI-MS (m/z): 261.2 [M+H] ⁺ .

The fourth step: Compound 24d (54 mg, 0.20 mmol) and Int-1 (40 mg, 0.20 mmol) were dissolved in 1,4-dioxane (4 mL), cesium carbonate (134 mg, 0.41 mmol) was added, BrettPhos ( 22 mg, 0.041 mmol) and BrettPhos Pd G3 (18 mg, 0.020 mmol). After the reaction system was replaced with nitrogen, it was heated to 110°C and stirred for 16 hours. After the reaction solution was cooled to room temperature, the reaction solution was concentrated, and the residue was purified by preparative thin layer chromatography to obtain the crude product, which was then separated by reverse preparative HPLC to obtain compound 24 (17 mg, yield 20%). ESI-MS(m/z): 418.2[M+H] ⁺ ; ¹ HNMR (500MHz, DMSO-d6)δ8.15(s,1H),7.96(s,1H),7.74(br s,1H), 7.61(s, 1H), 7.35-7.31(m, 1H), 3.88(s, 3H), 3.65-3.59(m, 1H), 3.48-3.37(m, 3H), 3.21-3.11(m, 2H), 2.74 (t, J=5.9 Hz, 2H), 2.69-2.61 (m, 3H), 2.34 (s, 3H), 2.14-2.06 (m, 1H), 1.85-1.76 (m, 1H).

Example 25

4-(((5-Chloro-2-((2-methoxy-6-methyl-5,6,7,8-tetrahydro-1,6-naphthalen-3-yl)amino) pyrimidin-4-yl)amino)methyl)piperidin-2-one

Replacing 3-(aminomethyl)-1-methyl-1,2-dihydropyridin-2-one hydrochloride in the first step of Example 1 with 4-(aminomethyl)piperidin-2-one 1b, using a similar method and reaction procedure, compound 25 can be obtained. ESI-MS (m/z): 432.2[M+H] ⁺ ; ¹ H NMR (500MHz, DMSO-d6) δ 8.07(s, 1H), 7.94(s, 1H), 7.64(s, 1H), 7.43(s, 1H), 7.39(t, J=5.9Hz, 1H), 3.88(s, 3H), 3.41(s, 2H), 3.31-3.27(m, 2H), 3.21-3.14(m, 1H) ,3.10-3.03(m,1H),2.75(t,J＝5.9Hz,2H),2.64(t,J＝5.9Hz,2H),2.34(s,3H),2.23-2.15(m,2H), 1.94-1.83 (m, 1H), 1.81-1.74 (m, 1H), 1.36-1.27 (m, 1H).

Example 26

3-(((2-((2-Methoxy-6-methyl-5,6,7,8-tetrahydro-1,6-naphthalen-3-yl)amino)-5-( Trifluoromethyl)pyrimidin-4-yl)amino)methyl)-1-methylpyridin-2(1H)-one

Substituting 2,4-dichloro-5-trifluoromethylpyrimidine for 2,4,5-trichloropyrimidine 1a in the first step of Example 1, and using a similar method and reaction procedure, compound 26 can be obtained. ESI-MS (m/z): 476.2[M+H] ⁺ ; ¹ H NMR (500MHz, DMSO-d6) δ 8.23(s, 1H), 8.06(s, 1H), 7.74(s, 1H), 7.64(d,J＝6.5Hz,1H),7.57(br s,1H),7.01(d,J＝6.8Hz,1H),6.19(t,J＝6.8Hz,1H),4.39(d,J＝ 5.2Hz, 2H), 3.84(s, 3H), 3.50(s, 3H), 3.12(s, 2H), 2.71(d, J=6.9Hz, 2H), 2.60(t, J=5.9Hz, 2H) , 2.33(s, 3H).

Example 27

(R)-3-(1-((5-Chloro-2-((2-methoxy-6-methyl-5,6,7,8-tetrahydro-1,6-naphthalene- 3-yl)amino)pyrimidin-4-yl)amino)ethyl)-1-methylpyridin-2(1H)-one

Compound 27 was prepared by the following steps:

The first step: Compound 27a (500 mg, 4.06 mmol) was dissolved in DMF (5 mL), cesium carbonate (1.59 g, 4.87 mmol) and methyl iodide (692 mg, 4.87 mmol) were added under ice bath at 0 °C, and the reaction rose to Stir at room temperature for 16 hours. After the reaction was completed, the reaction mixture was diluted with water (50 mL) and extracted with ethyl acetate (40 mL*2). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated. The residue was separated by silica gel column chromatography (petroleum ether/ethyl acetate=2/1) to obtain compound 27b (290 mg, yield 52%) as a white solid. ¹ H NMR (500MHz, CDCl ₃ ) δ 10.37 (s, 1H), 8.04 (dd, J=7.0.4.5Hz, 1H), 7.63 (dd, J=6.5, 2, 0Hz, 1H), 6.32 (t , J=7.0Hz, 1H), 3.64 (s, 3H).

The second step: Compound 27b (200 mg, 1.46 mmol) was dissolved in DCE (5 mL), cesium carbonate (950 mg, 2.92 mmol) and S-tert-butylsulfinamide (212 mg, 1.75 mmol) were added, and the mixture was heated at 65 °C under stirring for 16 hours. After the reaction was completed, the reaction mixture was diluted with water (30 mL) and extracted with ethyl acetate (30 mL*2). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to give compound 27c (330 mg, yield 94%) as a pale yellow oil. ESI-MS (m/z): 241.4 [M+H] ⁺ ; ¹ H NMR (500 MHz, DMSO-d6) δ 8.71 (s, 1H), 8.15 (dd, J=6.0.2.0 Hz, 1H), 8.05 (dd, J=6.5, 2, 5Hz, 1H), 6.41 (t, J=7.0Hz, 1H), 3.52 (s, 3H), 1.15 (s, 9H).

The third step: Compound 27c (35 mg, 0.14 mmol) was dissolved in DCM (3 mL), methylmagnesium bromide (1 mol/L in THF, 0.29 mL) was added under nitrogen protection at -78 °C, and the mixture was heated at -78 °C under stirring for 2 hours. After the reaction was completed, saturated ammonium chloride solution (20 mL) was added to quench the reaction, and the mixture was extracted with ethyl acetate (20 mL*2). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain compound 27d (30 mg, yield 80%) as a pale yellow oil. ESI-MS (m/z): 257.5 [M+H] ⁺ ; ¹ H NMR (500 MHz, DMSO-d6) δ 7.61 (dd, J=7.0.2.0 Hz, 1 H), 7.46 (dd, J=6.5 ,2,0Hz,1H),6.25-6.20(m,1H),5.68(d,J＝10.5Hz,1H),4.42-4.37(m,1H), 3.44(s,3H),1.29(d,J = 6.5Hz, 3H), 1.10 (s, 9H). The stereoconfiguration of the newly formed chiral center of compound 27d is assumed to be R-form.

The fourth step: Compound 27d (30 mg, 0.11 mmol) was dissolved in MeOH (2 mL), dioxane hydrochloride solution (4 mol/L, 0.12 mL) was added, and the reaction mixture was stirred at room temperature for 2 hours. After the reaction was completed, the reaction solution was concentrated to obtain a white solid compound 27e (22 mg, yield 99%). ESI-MS (m/z): 153.1 [M+H] ⁺ .

The fifth step: Compound 27e (22 mg, 0.11 mmol) was dissolved in isopropanol (2 mL), 2,4,5-trichloropyrimidine 1a (32 mg, 0.17 mmol) and N,N-diisopropylethyl acetate were added. Amine (30 mg, 0.23 mmol). The reaction mixture was stirred at room temperature for 16 hours. After the reaction was completed, the reaction solution was concentrated, and the residue was purified by preparative thin layer chromatography (petroleum ether/ethyl acetate=1/1) to obtain white solid compound 27f (31 mg, yield 88%). ESI-MS (m/z): 299.1 [M+H] ⁺ .

The sixth step: Compound Int-1 (20 mg, 0.10 mmol) and compound 27f (31 mg, 0.10 mmol) were dissolved in 1,4-dioxane (3 mL), cesium carbonate (67 mg, 0.20 mmol) was added, BrettPhos (11 mg, 0.020 mmol) and BrettPhos Pd G3 (9 mg, 0.010 mmol). The reaction mixture was replaced with nitrogen and then heated to 100°C with stirring for 18 hours. The reaction solution was filtered with celite, the filtrate was concentrated, and the residue was purified by preparative thin layer chromatography (dichloromethane/methanol=10/1) to obtain a crude product, which was then separated by reverse preparative HPLC to obtain a white solid compound 27 (3 mg, yield 6%). ESI-MS (m/z): 299.1 [M+H] ⁺ ; ¹ H NMR (400 MHz, DMSO-d6) δ 8.00 (s, 1H), 7.99 (s, 1H), 7.67-7.63 (m, 1H) ), 7.57(s, 1H), 7.53(d, J=8.4Hz, 1H), 7.37-7.33(m, 1H), 6.26-6.19(m, 1H), 5.35-5.26(m, 1H), 3.87( s, 3H), 3.49(s, 3H), 3.45-3.43(m, 2H), 2.77-2.71(m, 2H), 2.68-2.60(m, 2H), 2.37(s, 3H), 1.44(d, J=6.8Hz, 3H).

According to the synthetic route and the synthetic method of the intermediate described in the above examples, the following examples were prepared.

Biological Screening and Results of HPK1 Inhibitors

Test Example 1: Detection of the ability of compounds to inhibit HPK1 kinase activity (Method 1)

The required reagents are as follows

Experimental procedure

The specific operations are as follows: configure the enzymatic reaction system buffer (10mM MOPS, pH 7.2, 5mM β-glycerol-phosphate, 10mM MgCl2, 0.8mM EDTA, 2mM EGTA, 0.1mM DTT); test compounds (1mM in DMSO) Compound stock solution) was diluted with buffer to a maximum concentration of 60uM (containing 6% DMSO), and prepared a 60 μM concentration starting with a 5-fold dilution with a buffer containing 6% DMSO for a total of 8 point gradient concentrations of compounds; then The HPK1 kinase was diluted to 30 nM with buffer. Add 2 μl of HPK1 kinase dilution to each well of a Greiner 384-well microplate (Cat. No. 784075), and add 2 μl of buffer to control wells; add 1 μl of diluted compound to reaction wells after brief centrifugation, and add 1 μl of control wells containing 6% DMSO buffer; briefly centrifuged and then placed in a 25°C constant temperature incubator (Shanghai Yiheng Scientific Instrument Co., Ltd., product number: LRH-150) for 20 min. Add 3 μl of reaction substrate (10 μM MBP and 20 μM ATP dissolved in distilled water) to each well, centrifuge briefly and then incubate in a constant temperature incubator at 25°C for 60 min. The enzymatic reaction activity was detected by ADP-Glo Kinase Assay Kit, ADP - Glo Kinase Assay Kit assays are performed according to the kit's operating instructions. Data are described using the compound's median inhibitory concentration IC50.

化合物编号Compound number	IC50(nM)IC50(nM)	化合物编号Compound number	IC50(nM)IC50(nM)
11	<0.1<0.1	22	8888
33	45.7345.73	55	18.5218.52
66	2.62.6	77	<0.1<0.1
88	5.435.43	99	16.1216.12
1010	98.5498.54	1111	4.984.98
1313	82.5982.59	1414	18.9918.99
1616	34.4234.42	1717	82.8282.82
1919	8.528.52	21twenty one	94.7594.75
23twenty three	0.160.16	24twenty four	0.710.71
2525	33.9933.99	2626	1.351.35
2727	1.921.92	2828	18.3518.35
2929	19.4319.43	3030	29.7829.78
3131	11.8011.80	3232	0.850.85
3333	13.5613.56	3434	43.5143.51
3535	42.2842.28	3636	3.443.44
3737	8.038.03	3838	12.8212.82
3939	19.0119.01	4040	2.062.06
4242	81.0681.06	4343	4.264.26
4444	0.480.48	4545	29.4029.40
4646	44.9744.97	4747	20.9720.97
4848	17.9417.94	4949	5.785.78
5050	9.679.67	5151	14.6114.61

52

9.00

53

2.69

Test Example 2: Detection of agonistic ability of compounds to secrete cytokine interleukin-2 (IL-2) by Jurkat cells (Method 2)

The required reagents and cells are as follows

Experimental reagents:

试剂reagent	品牌brand	货号article number	保存条件Storage Conditions
RPMI1640RPMI1640	BIBI	01-100-1ACS01-100-1ACS	4℃4℃
FBSFBS	BIBI	04-002-1A04-002-1A	-80℃-80℃
BSABSA	AmrescoAmresco	0332-1KG0332-1KG	4℃4℃
磷酸盐缓冲溶液Phosphate buffer solution	BIBI	02-024-1ACS02-024-1ACS	4℃4℃
Tween-20Tween-20	SolarbioSolarbio	T8220T8220	RTRT
Anti-human CD3 AntibodyAnti-human CD3 Antibody	invitrogeninvitrogen	16-0037-8516-0037-85	4℃4℃
Anti-human CD28 AntibodyAnti-human CD28 Antibody	invitrogeninvitrogen	16-0289-8516-0289-85	4℃4℃
Human IL-2 DuoSet ELISAHuman IL-2 DuoSet ELISA	R&DR&D	DY202DY202	4℃4℃

Experimental cells:

细胞cell	细胞类型cell type	品牌brand
Jurkat E6-1Jurkat E6-1	人T淋巴细胞白血病细胞human T-lymphocytic leukemia cells	中科院细胞库Chinese Academy of Sciences Cell Bank

Experimental procedure

The specific operations are as follows: dissolve the compound powder to 10 mM with DMSO, add 2 μl of the compound to 998 μl of RPMI 1640 medium (both in this test containing 10% FBS), and vortex to mix the highest concentration point. The compound solution was gradually diluted 3-fold with 0.2% DMSO medium for a total of 8 concentration points. The control was treated with RPMI 1640 medium solution containing 0.1% DMSO. 1 × 105 Jurkat E6-1 cells were added to each well of a Corning 96-well cell culture plate (Cat. No. 3599), followed by an equal volume of compound dilutions. The control group was added RPMI 1640 medium containing 0.2% DMSO, and placed in 37 Incubate for 1 h in a cell incubator (Thermo Fisher Scientific, model: 3111). Then, the final concentrations of 1 μg/ml Anti-human CD3 Antibody and 1 μg/ml Anti-human CD28 Antibody were added and incubated in a 37°C cell incubator for 24 hours. Human IL-2 DuoSet ELISA KIT was used to detect the IL-2 content in the cell supernatant, and the Human IL-2 DuoSet ELISA detection was carried out according to the operating instructions of the kit. Data are described as the highest fold ratio of the stimulation signal of the compound to the signal of 0.1% DMSO.

Test Example 3: Detection of the agonistic ability of compounds to secrete cytokine interleukin-2 (IL-2) by human PBMC cells (Method 3)

The required reagents are as follows

试剂reagent	品牌brand	货号article number	保存条件Storage Conditions
RPMI1640RPMI1640	BIBI	01-100-1ACS01-100-1ACS	4℃4℃
FBSFBS	BIBI	04-002-1A04-002-1A	-80℃-80℃

BSABSA	AmrescoAmresco	0332-1KG0332-1KG	4℃4℃
磷酸盐缓冲溶液Phosphate buffer solution	BIBI	02-024-1ACS02-024-1ACS	4℃4℃
Tween-20Tween-20	SolarbioSolarbio	T8220T8220	RTRT
Anti-human CD3 AntibodyAnti-human CD3 Antibody	invitrogeninvitrogen	16-0037-8516-0037-85	4℃4℃
Anti-human CD28 AntibodyAnti-human CD28 Antibody	invitrogeninvitrogen	16-0289-8516-0289-85	4℃4℃
Human IL-2 DuoSet ELISAHuman IL-2 DuoSet ELISA	R&DR&D	DY202DY202	4℃4℃

Experimental cell source information:

Experimental procedure

The specific operations are as follows: human PBMCs were taken out from liquid nitrogen according to standard operations, thawed and thawed in a water bath at 37°C, resuspended in RPMI 1640 medium (both containing 10% FBS in this test), and washed twice by centrifugation. ; Human PBMC cells were subsequently resuspended in RPMI 1640 medium for later use. The compound powder was dissolved to 10 mM with DMSO, and 2 μl of the compound was added to 998 μl of RPMI 1640 medium, and the highest concentration point was obtained after vortexing and mixing. The compound solution was gradually diluted 3-fold with 0.2% DMSO medium for a total of 8 concentration points. The control was treated with RPMI 1640 medium solution containing 0.1% DMSO. 1×105 human PBMC cells were added to each well of a Corning 96-well cell culture plate (Cat. No. 3599), followed by an equal volume of compound diluents. For the control group, RPMI 1640 medium containing 0.2% DMSO was added, and the cells were placed at 37°C. The cells were incubated in a cell incubator (Thermo Fisher Scientific, model: 3111) for 1 h. Then, the final concentrations of 0.01μg/ml Anti-human CD3 Antibody and 1μg/ml Anti-human CD28 Antibody were added, and incubated at 37°C for 24h. Human IL-2 DuoSet ELISA KIT was used to detect the IL-2 content in the cell supernatant, and the Human IL-2 DuoSet ELISA detection was carried out according to the operating instructions of the kit. Data are described as the highest fold ratio of the stimulation signal of the compound to the signal of 0.1% DMSO.

The above results show that the compounds of the present invention can significantly increase IL-2.

Claims

A compound having the structure of formula (I) or a pharmaceutically acceptable salt, isotopic derivative or stereoisomer thereof:

in

X represents N or CR 10 ;

R 1 represents hydrogen, C 1 -C 6 alkyl, halogenated C 1 -C 6 alkyl, C 3 -C 6 cycloalkyl, halogenated C 3 -C 6 cycloalkyl, halogen, cyano or -C (O)NH 2 ;

R 2 represents hydrogen, halogen, hydroxy, (C 1 -C 6 )alkyl, (C 2 -C 6 )alkenyl, -(C 0 -C 6 alkylene)(C 3 -C 8 )cycloalkyl , -(C 0 -C 6 alkylene) (4-8 membered) heterocycloalkyl, (C 1 -C 6 ) alkoxy, (C 3 -C 8 ) cycloalkyl (C 0 -C 6 ) alkylene)oxy or (4-8 membered heterocycloalkyl)(C 0 -C 6 alkylene)oxy;

R 3 represents hydrogen or C 1 -C 6 alkyl, C 3 -C 6 alkenyl, C 3 -C 6 cycloalkyl, hydroxy (C 1 -C 6 alkyl), halogenated C 1 -C 6 alkyl , 4-8 membered heterocycloalkyl;

R 4 represents hydrogen or C 1 -C 6 alkyl; and the C 1 -C 6 alkyl may be optionally substituted with a substituent selected from halogen, -OR a , -SR a , -P(O )R a R b , -S(O) 2 R a , -S(O)(NH)R a , -S(O)R a , -S(O) 2 NR a R b , -C(O) NR a R b , -C(O)OH, -OC(O)NR a R b , -NR a C(O) R b , -NR a S(O) 2 R b ;

R 5 and R 5' each independently represent hydrogen, C 1 -C 6 alkyl, C 2 -C 6 alkenyl, halogen, halogenated C 1 -C 6 alkyl or hydroxy (C 1 -C 6 alkyl) ;

Or R 5 and R 5' together form a 3-6 membered ring with the carbon atom connected to it, and the ring can also optionally contain 0, 1 or 2 heteroatoms selected from N, O, S;

R 6 and R 6' each independently represent hydrogen, C 1 -C 6 alkyl, C 2 -C 6 alkenyl, halogen, halogenated C 1 -C 6 alkyl or hydroxy (C 1 -C 6 alkyl) ;

Or R 6 and R 6' together form a 3-6 membered ring with the carbon atom connected to it, and the ring can also optionally contain 0, 1 or 2 heteroatoms selected from N, O, and S;

p represents 1 or 2;

R 7 and R 7' each independently represent hydrogen, C 1 -C 6 alkyl, C 2 -C 6 alkenyl, halogen, halogenated C 1 -C 6 alkyl or hydroxy (C 1 -C 6 alkyl) ;

Or R 7 and R 7' together form a 3-6 membered ring with the carbon atom connected to it, and the ring can also optionally contain 0, 1 or 2 heteroatoms selected from N, O, and S;

W means

where A means

Indicates that the ring contains a single bond or a double bond;

When the chemical bond between A and A' is a double bond, A' represents CR a or N, and B represents CR b ;

When the chemical bond between A and A' is a single bond, A' represents CHR a , B represents CHR b or does not exist;

R 8 and R 9 each independently represent hydrogen or C 1 -C 6 alkyl; or R 8 and R 9 may together with W 1 and W 2 form a 3-6 membered ring;

W 1 means C, W 2 means CH or N;

R M and R N each independently represent hydrogen or C 1 -C 6 alkyl;

R 10 represents hydrogen, halogen, (C 1 -C 6 )alkyl, (C 2 -C 6 )alkenyl, -(C 0 -C 6 alkylene)(C 3 -C 8 )cycloalkyl, - (C 0 -C 6 alkylene) 4-10 membered heterocycloalkyl, -(C 0 -C 6 alkylene) (C 6 -C 10 ) aryl or -(C 0 -C 6 alkylene ) 5-10 membered heteroaryl,

Alternatively, when X represents CR 10 , R 10 may together with the adjacent R 2 form a 5-10 membered cycloalkyl or 5-10 membered heterocycloalkyl;

wherein, R a and R b each independently represent hydrogen, C 1 -C 6 alkyl, hydroxyl (C 1 -C 6 alkyl) or halogenated C 1 -C 6 alkyl;

m and n represent 0, 1 or 2, respectively.
The compound of claim 1 or a pharmaceutically acceptable salt, isotopic derivative or stereoisomer thereof, wherein,

W means

Among them, A means

Indicates that the ring contains a single bond or a double bond;

o each independently represents 0 or 1;

R 8 represents hydrogen or C 1 -C 6 alkyl;

R 9 each independently represents hydrogen or C 1 -C 6 alkyl;

Or R 8 together with adjacent R 9 and the carbon atoms to which they are attached form a 3-6 membered ring;

W 2 , RM and RN are as described in claim 1 .
The compound of claim 2 or a pharmaceutically acceptable salt, isotopic derivative or stereoisomer thereof, wherein,

W means
or

Indicates that the ring contains a single bond or a double bond;

o each independently represents 0 or 1;

R 8 represents hydrogen or C 1 -C 6 alkyl;

R 9 each independently represents hydrogen or C 1 -C 6 alkyl;

Or R 8 together with adjacent R 9 and the carbon atoms to which they are attached form a 3-6 membered ring;

W 2 , RM and RN are as described in claim 1 .
The compound of claim 3 or a pharmaceutically acceptable salt, isotopic derivative or stereoisomer thereof, wherein W is an aromatic heterocycle.
The compound of claim 3 or a pharmaceutically acceptable salt, isotopic derivative or stereoisomer thereof, wherein W is a saturated heterocycle.
The compound of claim 1 or a pharmaceutically acceptable salt, isotopic derivative or stereoisomer thereof, wherein ring W represents

or
The compound of claim 6 or a pharmaceutically acceptable salt, isotopic derivative or stereoisomer thereof, wherein ring W represents
The compound of claim 7 or a pharmaceutically acceptable salt, isotopic derivative or stereoisomer thereof, wherein ring W represents
The compound of any preceding claim, or a pharmaceutically acceptable salt, isotopic derivative or stereoisomer thereof, wherein R 1 represents hydrogen, halogen, C 1 -C 6 alkyl, C 1 -C 6 haloalkyl, cyano or -C(O) NH2 .
The compound of claim 9 or a pharmaceutically acceptable salt isotopic derivative or stereoisomer thereof, wherein R 1 represents hydrogen, halogen, C 1 -C 6 alkyl or C 1 -C 6 haloalkyl .
The compound of any preceding claim, or a pharmaceutically acceptable salt isotopic derivative or stereoisomer thereof, wherein R 2 represents hydroxy, (C 1 -C 6 )alkyl, -(C 0 - C 6 alkylene) (C 3 -C 8 )cycloalkyl, -(C 0 -C 6 alkylene)(4-8 membered)heterocycloalkyl, (C 1 -C 6 )alkoxy.
The compound of claim 11 or a pharmaceutically acceptable salt, isotopic derivative or stereoisomer thereof, wherein R 2 represents C 1 -C 6 alkyl, C 1 -C 6 alkoxy or C 3 -C 6 cycloalkyl.
The compound of any preceding claim, or a pharmaceutically acceptable salt isotopic derivative or stereoisomer thereof, wherein R 3 represents hydrogen or C 1 -C 6 alkyl, C 3 -C 6 cycloalkane base, halogenated C 1 -C 6 alkyl;
The compound of claim 13 or a pharmaceutically acceptable salt, isotopic derivative or stereoisomer thereof, wherein R 3 represents hydrogen or C 1 -C 6 alkyl.
The compound of any preceding claim, or a pharmaceutically acceptable salt isotopic derivative or stereoisomer thereof, wherein R 4 represents hydrogen or C 1 -C 6 alkyl; and the C 1 -C 6Alkyl can be optionally substituted by a substituent selected from the group consisting of halogen, -OR a , -SR a , -P(O)R a R b , -S(O) 2 R a , -S(O) (NH)R a , -S(O)R a , -S(O) 2 NR a R b , -C(O)NR a R b , -C(O)OH.
The compound of claim 15 or a pharmaceutically acceptable salt, isotopic derivative or stereoisomer thereof, wherein R 4 represents hydrogen or C 1 -C 6 alkyl. 17. The compound of any preceding claim, or a pharmaceutically acceptable salt isotopic derivative or stereoisomer thereof, wherein R 5 , R 5' each independently represent hydrogen, halogen, C 1 -C 6 alkyl or halogenated C 1 -C 6 alkyl.
The compound of any preceding claim, or a pharmaceutically acceptable salt isotopic derivative or stereoisomer thereof, wherein R 6 , R 6′ each independently represent hydrogen, halogen, C 1 -C 6 alkane or halogenated C 1 -C 6 alkyl.
The compound of any preceding claim, or a pharmaceutically acceptable salt isotopic derivative or stereoisomer thereof, wherein R 7 and R 7' each independently represent hydrogen, halogen, C 1 -C 6 Alkyl, halogenated C1 - C6 alkyl or hydroxy ( C1 - C6 alkyl). 20. The compound of any preceding claim, or a pharmaceutically acceptable salt isotopic derivative or stereoisomer thereof, wherein R8 represents hydrogen or C1 - C6 alkyl.
A compound as claimed in any preceding claim, or a pharmaceutically acceptable salt isotopic derivative or stereoisomer thereof, wherein R 9 represents hydrogen or C 1 -C 6 alkyl.
The compound of any preceding claim, or a pharmaceutically acceptable salt isotopic derivative or stereoisomer thereof, wherein R 10 represents hydrogen, halogen, (C 1 -C 6 )alkyl, -(C 0 - C6alkylene )(C3 - C8 )cycloalkyl, -( C0 - C6alkylene )4-10 membered heterocycloalkyl.
The compound of claim 22, or a pharmaceutically acceptable salt isotopic derivative or stereoisomer thereof, wherein R 10 represents hydrogen, halogen or (C 1 -C 6 )alkyl.
A compound as claimed in any preceding claim, or a pharmaceutically acceptable salt isotopic derivative or stereoisomer thereof, wherein R M represents hydrogen or C 1 -C 6 alkyl.
The compound of any preceding claim, or a pharmaceutically acceptable salt isotopic derivative or stereoisomer thereof, wherein R a , R b each independently represent hydrogen, C 1 -C 6 alkyl or halo Substituted C 1 -C 6 alkyl.
A compound with the following structure:
A pharmaceutical composition comprising a compound as claimed in any preceding claim, or a pharmaceutically acceptable salt, isotopic derivative or stereoisomer thereof, and a pharmaceutically acceptable carrier.
The compound of any one of claims 1-26 or a pharmaceutically acceptable salt, isotopic derivative, stereoisomer or the pharmaceutical composition of claim 27 is prepared for the prevention and/or treatment of cancer, Use in the medicament of tumors, inflammatory diseases, autoimmune diseases or immune-mediated diseases.
The compounds of claims 1-26 or their pharmaceutically acceptable salts, isotopic derivatives or stereoisomers thereof for preventing and/or treating cancer, tumors, inflammatory diseases, autoimmune diseases or immune mediators Use in leading diseases.