P53-Y220C selective small-molecule reactivator compound, pharmaceutical composition and application of p53-Y220C selective small-molecule reactivator compound

Abstract

The invention provides a compound as shown in a formula (I), which has a good p53-Y220C mutant activation effect as a drug, can effectively recover the activity of p53-Y220C mutant protein, is used for treating tumor diseases containing the p53-Y220C mutant protein, and is used for preparing drugs for the diseases or the diseases.

Description

p53-Y220C selective small molecule reactivator compound, pharmaceutical composition and use thereof

[0001] This application claims priority to prior applications filed with the State Intellectual Property Office of China on September 11, 2023, with patent application number 202311166450.3, entitled “p53-Y220C Selective Small Molecule Reactivator Compounds, Pharmaceutical Compositions, and Uses Thereof”; and prior application number 202311566620.7, entitled “p53-Y220C Selective Small Molecule Reactivator Compounds, Pharmaceutical Compositions, and Uses Thereof”, filed with the State Intellectual Property Office of China on November 22, 2023. The entire contents of the aforementioned prior applications are incorporated herein by reference. Technical Field

[0002] The present invention belongs to the field of pharmaceutical compounds, and in particular relates to p53-Y220C selective small molecule reactivator compounds, pharmaceutical compositions and uses thereof. Background Art

[0003] Cancer is a collection of related diseases characterized by the uncontrolled proliferation of tumor cells, which may metastasize throughout the body. Tumor cell proliferation relies on oncogenes and tumor suppressor genes. Genetic mutations can lead to abnormal activation or inhibition of these genes, further promoting uncontrolled cell division. TP53, a well-known tumor suppressor gene, is known as the "guardian of the genome." TP53 belongs to the p53-like transcription factor family and consists of three members: the TP53, TP63, and TP73 genes, which encode three proteins, p53, p63, and p73, respectively. They share structural similarities: their DNA-binding domains are nearly identical, binding to similar DNA-specific sequences and regulating the transcription of the same and some different genes. Their C-terminal domains differ in size, sequence, and function, regulating DNA binding and transcription and mediating protein-protein interactions. Their N-terminal sequences encode at least two distinct transcriptional activation domains. Similar to p53, p63 is also a key transcription factor that responds to DNA damage by inducing apoptosis. It acts in the skull, face, limbs, and central nervous system, participating in the generation and regeneration of squamous cell epithelial cells throughout the body. p73 is required for the production of ciliated epithelial cells, and acts on male germ cells, the immune system, the hearing system, the trachea, the lungs, the central nervous system, etc.

[0004] The p53 tumor suppressor is mainly distributed in the nucleoplasm of cells. As a receptor for cell stress, it can respond to a variety of cell stresses, including ultraviolet radiation, hypoxia, oncogene activation and DNA damage. After activation, p53 binds to a specific DNA sequence in a tetrameric form, mediates downstream gene transcription, and inhibits cancer progression through multiple mechanisms, such as including cell cycle regulation, apoptosis, aging, stem cell differentiation, metabolism (reducing sugar synthesis), ROS and mitochondria, DNA damage repair, etc. p53 can activate proteins involved in the above-mentioned pathways, including, for example, Fas / Apol, KILLER / DR5, Bax, Puma, Noxa, Bid, caspase-3, caspase6, caspase 7, caspase-8, caspase-9 and p21. In addition, p53 can also inhibit the transcription of a variety of genes, including c-MYC, Cyclin B, VEGF, RAD51, and hTERT.

[0005] Due to its critical role in tumor suppression, the TP53 gene is the most frequently mutated gene in human cancers, with mutations occurring in nearly 50% of malignant tumors. In contrast to mutations in other tumor suppressor genes such as RB1, APC, and PTEN, the majority of TP53 mutations are missense mutations, accounting for up to 75%. Missense mutations are primarily located within the DNA-binding domain of p53, leading to abnormal folding of the protein sequence required for DNA recognition and binding. Numerous amino acid positions within p53 are potentially susceptible to mutation, including Val143, His168, Arg175, Tyr220, Gly245, Arg248, Arg299, Phe270, Arg273, and Arg282. Accordingly, p53 mutations that can abrogate wild-type p53 activity include, for example, R175H, Y220C, G245S, R248Q, R248W, R273H, and R282H. These p53 mutations can both distort the structure of the DNA binding site and disrupt its thermodynamic stability. TP53 mutations can result in the loss of wild-type p53's tumor suppressor transcriptional activity through homozygous deletions (LOHs); dominant negative effects through heterozygous deletions; and gain-of-function effects, such as increased binding to p63 and p73, which promote tumorigenesis and progression. Therefore, mutant p53 proteins can be considered a heterogeneous group of proteins, exhibiting varying degrees of loss of normal tumor suppressor function and gain of oncogenic properties (GOF).

[0006] Although different p53 mutants exhibit varying activities, missense TP53 mutants can be considered proto-oncogenes, promoting tumor migration and drug resistance, leading to poor prognosis, and thus becoming therapeutic targets for drug development. Among them, the oncogenic p53 Y220C mutant exhibits a particularly suitable structure for the development of small-molecule stabilizers. It is the ninth most common missense p53 mutation found in cancer, accounting for approximately 100,000 new cancer cases worldwide each year. The mutation of Tyr220 to Cys creates a narrow hydrophobic cleft on the p53 DBD surface, reducing its thermal stability by approximately 4 kcal / mol. While wild-type p53 is moderately stable, melting and denaturing at 44°C, the Y220C mutant protein rapidly unfolds from its folded state and denatures at physiological body temperature, effectively abolishing the tumor suppressor signaling of wild-type p53 and driving tumorigenesis. Importantly, the hydrophobic cleft created by the Y220C mutation is located away from p53 protein surfaces involved in DNA recognition or protein-protein interactions, allowing for the development of small-molecule drugs without interfering with its binding to its natural DNA substrate. The Y220C mutant is a temperature-sensitive mutant that binds to DNA at lower temperatures and denatures at body temperature. When a small molecule compound selectively binds to the p53-Y220C mutant, it stabilizes the Y220C mutant, reducing the likelihood of p53 protein denaturation at body temperature. This compound converts p53 from its unfolded state to a folded state, restoring its wild-type conformation and transcriptional activity.

[0007] Studies have shown that several small molecules, such as PK083, PK7088, PK5196, and PC14586, can bind to the hydrophobic cleft of the Y220C mutant. However, these molecules, such as PK083, PK7088, PK5196, and PC14586, are effective at relatively high concentrations, and therefore, off-target effects cannot be ruled out. Therefore, there is a need to develop better small molecules that can more specifically bind to the Y220C mutant, better restore the structure and transcriptional activity of wild-type p53, and more effectively suppress tumors.

[0008] Summary of the Invention

[0009] To solve the problems existing in the prior art, the present invention provides a compound represented by formula (I) and its racemate, stereoisomer, tautomer, isotope-labeled substance, nitrogen oxide, solvate, polymorph, metabolite, ester, prodrug or pharmaceutically acceptable salt:

[0010] in, It means that the ring in which it exists is aromatic;

[0011] X1, X2, X3 are the same or different and are independently selected from CH or N, and at least two of X1, X2, X3 are CH;

[0012] X4 is selected from CR4, N, NR4;

[0013] X5 is selected from CR5, N, NR5, O, S;

[0014] R1, R2 are the same or different and are independently selected from H, halogen, C 1-12 Alkyl, C 1-12 alkoxy;

[0015] R3 is selected from H, C 1-12 alkyl;

[0016] R4, R5 are the same or different and are independently selected from H, halogen, cyano, C 1-12 Alkyl, C 1-12 Alkoxy, C 3-12 Cycloalkyl, halogenated C 1-12 Alkyl, halogenated C 1-12 Alkoxy, halogenated C 3-12 Cycloalkyl, cyano C 1-12 Alkyl, cyano C 1-12 alkoxy;

[0017] W is selected from C 2-6 Alkenylene, C 2-6 Alkynylidene, C 3-14 Cycloalkylene, C 6-10 Arylene, 5-10 membered heteroarylene;

[0018] Ring A is selected from C 3-14 Cycloalkyl, 3-14 membered heterocyclic group, C 6-10 Aryl, 5-10 membered heteroaryl;

[0019] R a selected from H, CN, oxo (=O), halogen, OH, unsubstituted or optionally substituted with one, two or more R a1 Substituted with the following groups: C 1-12 Alkyl, C 1-12 Alkoxy, C 3-12 Cycloalkyl, 3-14 membered heterocyclic group, C 6-10 Aryl, 5-10 membered heteroaryl, -C(O)N(R a11 )(R a12 )、-N(R a13 )(R a14 )、-S(O)2-R a15 、-S(O)(=NR a16 )(R a17 )、-P(O)(R a18 )(R a19 );

[0020] Or, two Ra The atom to which it is attached is unsubstituted or optionally substituted with one, two or more R a1 Substituted with the following groups: C 3-12 Cycloalkyl, 3-14 membered heterocyclic group;

[0021] Each R a1 The same or different, independently selected from H, OH, CN, halogen, unsubstituted or optionally substituted by one, two or more R a2 Substituted with the following groups: C 1-12 Alkyl, C 1-12 Alkoxy, C 3-12 Cycloalkyl, 3-14 membered heterocyclic group, C 6-10 Aryl, 5-10 membered heteroaryl, -NH2, -S(O)2-C 1-12 Alkyl; each R a2 The same or different, independently selected from H, OH, NH2, CN, halogen, C 1-6 Alkyl, C 1-6 Alkoxy; R a11 、R a12 、R a13 、R a14 、R a15 、R a16 、R a17 、R a18 、R a19 The same or different, independently selected from H, C 1-12 Alkyl, C 3-7 Cycloalkyl, 3-8 membered heterocyclic group; m is selected from 0, 1, 2, 3, 4, 5;

[0022] Z is absent or selected from NH, S, O, C 1-6 Alkylene, C 1-6 Alkylene-NH;

[0023] Ring E is selected from 3-14 membered heterocyclic group, C 3-12 Cycloalkyl;

[0024] R e Selected from H, CN, halogen, unsubstituted or optionally substituted by one, two or more R e1 Substituted with the following groups: C 1-12 Alkyl, C 1-12 Alkoxy, C 3-12 Cycloalkyl, 3-14 membered heterocyclic group, C 1-12 Alkyl-NH-, (C 1-12 Alkyl) 2-N-; or, two R e and their respective atoms to form C 3-12Cycloalkyl, 3-14 membered heterocyclic group; or, two R e The atoms to which they are attached form a 3-14 membered heterocyclic group; p is selected from 0, 1, 2, 3, 4, 5;

[0025] Each R e1 The same or different, independently selected from H, OH, CN, halogen, C 1-12 Alkyl, C 1-12 alkoxy;

[0026] R x Selected from H, CN, halogen, C 1-12 Alkyl, C 1-12 Alkoxy; n is selected from 0, 1, 2 or 3;

[0027] Y is selected from C(O), C(O)NH, C(S), SO2.

[0028] According to some embodiments, the Selected from

[0029] According to some embodiments, R1, R2, and R3 are the same or different and are independently selected from H, C 1-6 alkyl.

[0030] According to some embodiments, R1, R2, and R3 are all H.

[0031] According to some embodiments, R4, R5 are the same or different and are independently selected from C 1-6 Alkyl, halogenated C 1-6 Alkyl, cyano C 1-6 alkyl;

[0032] According to some embodiments, R4 is selected from methyl, ethyl, propyl, isopropyl, cyanomethyl, 2-fluoroethyl, 2,2-difluoroethyl, 2,2,2-trifluoroethyl.

[0033] According to some embodiments, R5 is selected from H.

[0034] According to some embodiments, W is selected from C 2-4 Alkenylene, C 2-4 Alkynylidene, C 3-6 cycloalkylene;

[0035] According to some embodiments, W is selected from:

[0036] According to some embodiments, ring A is selected from phenyl, 5-6 membered heteroaryl, 8-9 membered heteroaryl, 6-9 membered heterocyclyl, C 3-6 cycloalkyl;

[0037] According to some embodiments, Ring A is selected from:

[0038] According to some embodiments, R a selected from H, CN, oxo (=O), halogen, OH, unsubstituted or optionally substituted with one, two or more R a1 Substituted with the following groups: C 1-6 Alkyl, C 1-6 Alkoxy, C 3-6 Cycloalkyl, 4-7 membered heterocyclic group, phenyl, 5-6 membered heteroaryl, -C(O)(NHC 1-6 Alkyl), -S(O)2-C 1-6 Alkyl, -S(O)(=NH)(C 1-6 alkyl), -S(O)(=NC 1-6 Alkyl)(C 1-6 alkyl), -P(O)(C 1-6 Alkyl)(C 1-6 Alkyl), -NH2,

[0039] Or, two R a The atom to which it is attached is unsubstituted or optionally substituted with one, two or more R a1 Substituted with the following groups: C 3-6 Cycloalkyl; e.g.

[0040] According to some embodiments, each R a1 The same or different, independently selected from H, OH, CN, halogen, C 1-6 Alkyl, halogenated C 1-6 Alkyl, C 1-6 Alkoxy, C 3-6 Cycloalkyl, 5-6 membered heterocyclic group, phenyl, 5-6 membered heteroaryl, cyano 1-6 Alkyl, cyano C 1-6 Alkoxy, C 1-6 Alkyl-NH-, (C 1-6 alkyl)2-N-, -S(O)2-C 1-6 Alkyl, C 1-6 Alkyl-OC 1-6 Alkyl, hydroxyl C 1-6 alkyl;

[0041] According to some embodiments, R a Selected from H, F, Cl, Br, oxo (=O), CN, NH2, Methylamino, dimethylamino, methyl, ethyl, isopropyl, tert-butyl, cyclopropyl, cyclobutyl, cyclopentyl, methoxy, difluoromethyl, trifluoromethyl, 2,2-difluoroethyl, 2,2,2-trifluoroethyl, phenyl, pyridyl, benzyl, morpholinyl,

[0042] According to some embodiments, Selected from

[0043] According to some embodiments, Z is absent or is selected from NH, S, O, CH2.

[0044] According to some embodiments, ring E is selected from 6-9 membered heterocyclyl or C 5-6 Cycloalkyl;

[0045] According to some embodiments, ring E is selected from

[0046] According to some embodiments, R e Selected from H, halogen, C 1-6 Alkyl, halogenated C 1-6 Alkyl, cyano C 1-6 Alkyl, hydroxyl C 1-6 Alkyl, -C 1-6 Alkyl-OC 1-6 Alkyl, C 1-6 Alkoxy, C 3-6 Cycloalkyl, 5-6 membered heterocyclic group, (C 1-6 Alkyl) 2-N-; or, two R e and their respective atoms to form C 3-6 Cycloalkyl;

[0047] According to some embodiments, R e Selected from H, F, Cl, CN, methyl, methoxy, ethyl, isopropyl, cyclopropyl, dimethylamino,

[0048] Alternatively, two R attached to the same carbon atom e The atoms to which they are attached form a cyclopropane ring.

[0049] According to some embodiments, Selected from

[0050] According to some embodiments, X1, X2, and X3 are all CH, or one of X1, X2, and X3 is N.

[0051] According to some embodiments, when X4 is selected from CR4, X5 is selected from O, S, NR5; when X4 is selected from NR4, X5 is selected from CR5, N; when X4 is selected from N, X5 is selected from CR5, NR5.

[0052] According to some embodiments, R x Selected from H, CN, halogen, C 1-6 Alkyl, C 1-6 alkoxy;

[0053] According to some embodiments, R x Selected from H, F, Cl, CN, methoxy, methyl.

[0054] According to some embodiments, the compound represented by formula (I) has the structure shown below:

[0055] Among them, ring A, ring E, X1, X2, X3, X4, X5, Y, R1, R2, R3, R a 、R e 、R x 、m、n、p、 Has the definition described in the present invention.

[0056] According to some embodiments, the compound represented by formula (I) has the structure shown below:

[0057] Among them, ring A, ring E, X1, X2, X3, Y, R1, R2, R3, R4, R a 、R e 、R x 、m、n、p、 Has the definition described in the present invention.

[0058] According to some embodiments, the compound represented by formula (I) has the structure shown below:

[0059] Among them, R a 、R x , m, and n have the definitions described in the present invention.

[0060] According to some embodiments, among the compounds represented by formula (I) and their racemates, stereoisomers, tautomers, isotope-labeled substances, nitrogen oxides, solvates, polymorphs, metabolites, esters, prodrugs or pharmaceutically acceptable salts, illustrative, non-limiting specific examples of the compounds represented by formula (I) are as follows:

[0061] The present invention also provides a method for preparing the compound represented by formula (I), comprising the following steps: reacting compound 1 with compound 2 to obtain the compound represented by formula (I);

[0062] Among them, ring A, ring E, X1, X2, X3, X4, X5, W, Y, Z, R1, R2, R3, R a 、R e 、R x 、m、n、p、 Has the definition described in the present invention.

[0063] The present invention also provides a pharmaceutical composition comprising a therapeutically effective amount of at least one of the compound represented by formula (I), its racemate, stereoisomer, tautomer, isotope-labeled substance, nitrogen oxide, solvate, polymorph, metabolite, ester, prodrug or pharmaceutically acceptable salt thereof.

[0064] According to an embodiment of the present invention, the pharmaceutical composition further comprises one or more pharmaceutically acceptable excipients.

[0065] According to an embodiment of the present invention, the pharmaceutical composition may further contain one or more other therapeutic agents.

[0066] The present invention also provides a method for treating tumor diseases, comprising administering to a patient a preventive or therapeutically effective amount of a compound represented by formula (I), its racemate, stereoisomer, tautomer, isotope-labeled substance, nitrogen oxide, solvate, polymorph, metabolite, ester, prodrug or pharmaceutically acceptable salt thereof.

[0067] According to an embodiment of the present invention, the tumor is a tumor containing a p53-Y220C mutant.

[0068] The present invention also provides a method for treating tumor diseases, comprising administering to a patient a preventive or therapeutically effective amount of the above-mentioned pharmaceutical composition.

[0069] In some embodiments, the patient comprises a mammal, preferably a human.

[0070] The present invention also provides a compound represented by formula (I), at least one of its racemates, stereoisomers, tautomers, isotope-labeled substances, nitrogen oxides, solvates, polymorphs, metabolites, esters, prodrugs or pharmaceutically acceptable salts for treating tumor diseases, or a pharmaceutical composition thereof.

[0071] The present invention also provides the use of at least one of the compound represented by formula (I), its racemate, stereoisomer, tautomer, isotope-labeled substance, nitrogen oxide, solvate, polymorph, metabolite, ester, prodrug or pharmaceutically acceptable salt thereof in the preparation of a drug.

[0072] According to an embodiment of the present invention, the use can be in the preparation of an anti-tumor drug containing a p53-Y220C mutant, such as in the preparation of a p53-Y220C reactivator drug.

[0073] According to an embodiment of the present invention, the tumor containing the p53-Y220C mutant includes acute lymphoblastic leukemia, acute myeloid leukemia, adrenocortical carcinoma, AIDS-related cancer, AIDS-related lymphoma, anal cancer, appendix cancer, astrocytoma, basal cell carcinoma, bile duct cancer, bladder cancer, bone cancer, brain tumors such as cerebellar astrocytoma, cerebral astrocytoma / malignant glioma, ependymoma, medulloblastoma, Supratentorial blastoma, neuroectodermal tumor, optic pathway and hypothalamic glioma, breast cancer, bronchial adenoma, Burkitt lymphoma, cancer of unknown primary, central nervous system lymphoma, cerebellar astrocytoma, cervical cancer, childhood cancer, chronic lymphocytic leukemia, chronic myeloproliferative disease, colon cancer, cutaneous T-cell lymphoma, desmoplastic small round cell tumor, endometrial cancer, ependymoma, esophageal cancer, Ewing's sarcoma, germ cell tumors, gallbladder cancer, gastric cancer, gastrointestinal tract cancer Carcinoid tumors, gastrointestinal stromal tumors, gliomas, hairy cell leukemia, head and neck cancer, heart cancer, hepatocellular carcinoma, Hodgkin lymphoma, hypopharyngeal cancer, intraocular melanoma, islet cell carcinoma, Kaposi's sarcoma, kidney cancer, laryngeal cancer, lip and oral cancer, liposarcoma, liver cancer, lung cancer (non-small cell lung cancer and small cell lung cancer), lymphoma, leukemia, macroglobulinemia, malignant fibrous histiocytoma / osteosarcoma of bone, medulloblastoma, melanoma, mesothelioma, metastatic squamous cell carcinoma with occult primary, oral cancer, multiple endocrine tumors Cancer syndrome, myelodysplastic syndrome, myeloid leukemia, nasal cavity and paranasal sinus cancer, nasopharyngeal cancer, neuroblastoma, non-Hodgkin lymphoma, non-small cell lung cancer, oral cancer, oropharyngeal cancer, osteosarcoma / malignant fibrous histiocytoma of bone, ovarian cancer, ovarian epithelial cancer, ovarian germ cell tumor, pancreatic cancer, pancreatic islet cell cancer, paranasal sinus and nasal cavity cancer, parathyroid cancer, penile cancer, pharyngeal cancer, pheochromocytoma, pineal astrocytoma, pineal germ cell tumor, pituitary adenoma, pleuropulmonary blastoma, plasma cell tumor, prostate cancer Primary central nervous system lymphoma, prostate cancer, rectal cancer, renal cell carcinoma, transitional cell carcinoma of the renal pelvis and ureter, retinoblastoma, rhabdomyosarcoma, salivary gland cancer, sarcoma, skin cancer, Merkel cell carcinoma of the skin, small intestine cancer, soft tissue sarcoma, squamous cell carcinoma, stomach cancer, T-cell lymphoma, laryngeal cancer, thymoma, thymic carcinoma, thyroid cancer, trophoblastic tumor (pregnancy), cancer of unknown primary site, urethral cancer, uterine sarcoma, vaginal cancer, vulvar cancer, Waldenstrom's macroglobulinemia, and Wilms' tumor. Beneficial effects

[0074] The compound of the present invention has good p53-Y220C mutant activation effect and can effectively restore the activity of the p53-Y220C mutant protein. It is used to treat tumor diseases containing the p53-Y220C mutant protein and to prepare drugs for such diseases or disorders.

[0075] Definitions and Explanations of Terms

[0076] Unless otherwise indicated, the definitions of groups and terms in this specification and claims, including definitions used as examples, exemplary definitions, preferred definitions, definitions in tables, and definitions of specific compounds in the Examples, may be arbitrarily combined and coupled with one another. The group definitions and compound structures resulting from such combinations and couplings should be understood to be within the scope of this specification and / or claims.

[0077] Unless otherwise indicated, numerical ranges recited in this specification and claims are equivalent to reciting at least each specific integer value therein. For example, the numerical range "1-14" is equivalent to reciting each integer value in the numerical range "1-14", namely, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, and 14.

[0078] The term "optional" (or "optionally", "optionally") in the general formula definitions of this application means the situation of being substituted by zero, one or more substituents, for example, "optionally substituted by one, two or more R" means that it may not be substituted by R (unsubstituted) or may be optionally substituted by one, two or more R.

[0079] "More" means three or more.

[0080] The term "C 1-12 "Alkyl" is understood to mean straight-chain and branched alkyl groups having 1 to 12 carbon atoms, "C 1-8 "Alkyl" means straight chain and branched chain alkyl groups having 1, 2, 3, 4, 5, 6, 7, or 8 carbon atoms, "C 1-6 The term "alkyl" refers to straight-chain and branched alkyl groups having 1, 2, 3, 4, 5 or 6 carbon atoms. The alkyl group is, for example, methyl, ethyl, propyl, butyl, pentyl, hexyl, isopropyl, isobutyl, sec-butyl, tert-butyl, isopentyl, 2-methylbutyl, 1-methylbutyl, 1-ethylpropyl, 1,2-dimethylpropyl, neopentyl, 1,1-dimethylpropyl, 4-methylpentyl, 3-methylpentyl, 2-methylpentyl, 1-methylpentyl, 2-ethylbutyl, 1-ethylbutyl, 3,3-dimethylbutyl, 2,2-dimethylbutyl, 1,1-dimethylbutyl, 2,3-dimethylbutyl, 1,3-dimethylbutyl or 1,2-dimethylbutyl, or the like or isomers thereof.

[0081] The term "C 1-12 "Alkoxy" should be understood as "C 1-12 Alkyl-O-", C 1-12 Alkyl is as defined above.

[0082] The term "C 3-12"Cycloalkyl" is understood to mean a saturated monovalent monocyclic, bicyclic (such as condensed, bridged, spiro) hydrocarbon ring or tricyclic alkane having 3 to 12 carbon atoms, preferably "C 3-10 Cycloalkyl", more preferably "C 3-8 Cycloalkyl". The term "C 3-12 "Cycloalkyl" is understood to mean a saturated monovalent monocyclic, bicyclic (eg bridged, spiro) hydrocarbon ring or tricyclic alkane having 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 carbon atoms. 3-12 The cycloalkyl group may be a monocyclic hydrocarbon group such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl or cyclodecyl, or a bicyclic hydrocarbon group such as borneol, indolyl, hexahydroindolyl, tetrahydronaphthyl, decahydronaphthyl, bicyclo[2.1.1]hexyl, bicyclo[2.2.1]heptyl, bicyclo[2.2.1]heptenyl, 6,6-dimethylbicyclo[3.1.1]heptyl, 2,6,6-trimethylbicyclo[3.1.1]heptyl, bicyclo[2.2.2]octyl, 2,7-diazaspiro[3,5]nonyl, 2,6-diazaspiro[3,4]octyl, or a tricyclic hydrocarbon group such as adamantyl.

[0083] The term "C 6-14 "Aryl" is understood to mean preferably a monovalent aromatic or partially aromatic monocyclic, bicyclic or tricyclic hydrocarbon ring having 6 to 14 carbon atoms, which may be a single aromatic ring or polyaromatic rings fused together, preferably "C 6-10 Aryl". The term "C 6-14 "Aryl" is understood to mean preferably a monovalent aromatic or partially aromatic monocyclic, bicyclic or tricyclic hydrocarbon ring ("C 6-14 or a ring having 9 carbon atoms ("C9 aryl"), for example indanyl or indenyl, or a ring having 10 carbon atoms ("C 10 aryl) such as tetrahydronaphthyl, dihydronaphthyl or naphthyl, or a ring having 13 carbon atoms ("C 13 aryl), such as fluorenyl, or a ring having 14 carbon atoms ("C 14 aryl”), such as anthracenyl. When the C 6-20 When the aryl group is substituted, it may be monosubstituted or polysubstituted. Furthermore, there is no limitation on the position of substitution, and for example, substitution may be at the ortho, para or meta position.

[0084] The term "5-14 membered heteroaryl" is understood to include monovalent monocyclic, bicyclic (e.g. fused, bridged, spiro) or tricyclic aromatic ring systems having 5 to 14 ring atoms and containing 1 to 5 heteroatoms independently selected from N, O and S, for example "5-10 membered heteroaryl". The term "5-14 membered heteroaryl" is understood to include monovalent monocyclic, bicyclic or tricyclic aromatic ring systems having 5, 6, 7, 8, 9, 10, 11, 12, 13 or 14 ring atoms, in particular 5 or 6 or 9 or 10 carbon atoms, and containing 1 to 5, preferably 1 to 3, heteroatoms each independently selected from N, O and S and, in each case, may additionally be benzofused. "Heteroaryl" also refers to a radical in which a heteroaromatic ring is fused to one or more aryl, alicyclic or heterocyclyl rings, wherein the radical or point of attachment is on the heteroaromatic ring. Non-limiting examples include 1-, 2-, 3-, 5-, 6-, 7-, or 8-indolizinyl, 1-, 3-, 4-, 5-, 6-, or 7-isoindolyl, 2-, 3-, 4-, 5-, 6-, or 7-indolyl, 2-, 3-, 4-, 5-, 6-, or 7-indazolyl, 2-, 4-, 5-, 6-, 7-, or 8-purinyl, 1-, 2-, 3-, 4-, 6-, 7-, 8-, or 9-quinolizinyl, 2-, 3-, 4-, 5-, 6-, 7-, or 8-quinolinyl, 1-, 3-, 4- , 5-, 6-, 7- or 8-isoquinolyl, 1-, 4-, 5-, 6-, 7- or 8-phthalazinyl, 2-, 3-, 4-, 5- or 6-naphthyridinyl, 2-, 3-, 5-, 6-, 7- or 8-quinazolinyl, 3-, 4-, 5-, 6-, 7- or 8-cinnolinyl, 2-, 4-, 6- or 7-pteridinyl, 1-, 2-, 3-, 4-, 5-, 6-, 7- or 8-4aHcarbazolyl, 1-, 2-, 3-, 4-, 5-, 6-, 7- or 8-4aHcarbazolyl, 1-, 2-, 3-, 4-, 5-, 6-, 7-, 8-, or 9-oxo-1-carbazolyl, 1-, 2-, 3-, 4-, 5-, 6-, 7-, 8-, or 9-oxo-1-carbazolyl, 1-, 2-, 3-, 4-, 5-, 6-, 7-, 8-, or 9-oxo-1-carbazolyl, 1-, 2-, 3-, 4-, 5-, 6-, 7-, 8-, or 9-oxo-1-carbazolyl, 1-, 2-, 3-, 4-, 5-, 6-, 7-, 8-, or 9-oxo-1-carbazolyl, -phenazinyl, 1-, 2-, 3-, 4-, 6-, 7-, 8-, 9-, or 10-phenothiazinyl, 1-, 2-, 3-, 4-, 6-, 7-, 8-, 9-, or 10-phenazinyl, 2-, 3-, 4-, 5-, 6-, or 1-, 3-, 4-, 5-, 6-, 7-, 8-, 9-, or 10-benzoisoquinolinyl, 2-, 3-, 4-, or thieno[2,3-b]furanyl, 2-, 3-, 5-, 6-, 7-, 8-, 9-, 10-, or 11-7H-pyrazino[2,3-c]carbazolyl, 2-, 3-, 5-, 6- or 7-2H-furo[3,2-b]-pyranyl, 2-, 3-, 4-, 5-, 7- or 8-5H-pyrido[2,3-d]-o-oxazinyl, 1-, 3- or 5-1H-pyrazolo[4,3-d]oxazolyl, 2-, 4- or 54H-imidazo[4,5-d]thiazolyl, 3-, 5- or 8-pyrazino[2,3-d]pyridazinyl, 2-, 3-, 5- or 6-imidazo[2,1-b]thiazolyl, 1-, 3-, 6-, 7-, 8- or 9-furo[3,4-c]cinnolinyl, 1-, 2-, 3-, 4-, 5-, 6-, 8-, 9-, 10 or 11-4H-pyrido[2,3

[0015] In some embodiments, the present invention further comprises carbazolyl, 2-, 3-, 6-, or 7-imidazo[1,2-b][1,2,4]triazinyl, 7-benzo[b]thienyl, 2-, 4-, 5-, 6-, or 7-benzoxazolyl, 2-, 4-, 5-, 6-, or 7-benzimidazolyl, 2-, 4-, 4-, 5-, 6-, or 7-benzothiazolyl, 1-, 2-, 4-, 5-, 6-, 7-, 8-, or 9-benzoxapinyl, 2-, 4-, 5-, 6-, 7-, or 8-benzoxazinyl, 1-, 2-, 3-, 5-, 6-, 7-, 8-, 9-, 10-, or 11-4H-pyrrolo[1,2-b][2]benzazepinyl. Typical fused heteroaryl groups include, but are not limited to, 2-, 3-, 4-, 5-, 6-, 7-, or 8-quinolyl, 1-, 3-, 4-, 5-, 6-, 7-, or 8-isoquinolyl, 2-, 3-, 4-, 5-, 6-, or 7-indolyl, 2-, 3-, 4-, 5-, 6-, or 7-benzo[b]thienyl, 2-, 4-, 5-, 6-, or 7-benzoxazolyl, 2-, 4-, 5-, 6-, or 7-benzimidazolyl, and 2-, 4-, 5-, 6-, or 7-benzothiazolyl. When the 5- to 14-membered heteroaryl group is linked to other groups to form a compound of the present invention, the carbon atoms on the 5- to 14-membered heteroaryl ring may be linked to the other groups, or heteroatoms on the 5- to 14-membered heteroaryl ring may be linked to the other groups. When the 5- to 14-membered heteroaryl group is substituted, it may be monosubstituted or polysubstituted. Furthermore, there is no limitation on the substitution site, for example, a hydrogen atom connected to a carbon atom on a heteroaryl ring may be substituted, or a hydrogen atom connected to a heteroatom on a heteroaryl ring may be substituted.

[0085] Unless otherwise defined, the term "3-14 membered heterocyclyl" refers to a saturated or unsaturated non-aromatic ring or ring system, for example, a 4-, 5-, 6- or 7-membered monocyclic ring, a 7-, 8-, 9-, 10-, 11- or 12-membered bicyclic ring (such as a fused ring, a bridged ring, a spirocyclic ring) or a 10-, 11-, 12-, 13- or 14-membered tricyclic ring system, and contains at least one, for example 1, 2, 3, 4, 5 or more heteroatoms selected from O, S and N, wherein N and S may also be optionally oxidized to various oxidation states to form nitrogen oxides, -S(O)- or -S(O)2- states. Preferably, the heterocyclyl may be selected from "3-10 membered heterocyclyl". The term "3-10 membered heterocyclyl" means a saturated or unsaturated non-aromatic ring or ring system, and contains at least one heteroatom selected from O, S and N. The heterocyclic group can be connected to the rest of the molecule by any one of the carbon atoms or nitrogen atom (if present). The heterocyclic group can include fused or bridged rings and spirocyclic rings. In particular, the heterocyclic group can include but is not limited to: 4-membered rings, such as azetidinyl, oxetane; 5-membered rings, such as tetrahydrofuranyl, dioxolyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl, pyrrolinyl; or 6-membered rings, such as tetrahydropyranyl, piperidinyl, morpholinyl, dithianyl, thiomorpholinyl, piperazinyl or trithianyl; or 7-membered rings, such as diazepanyl. Optionally, the heterocyclic group can be benzo-fused. The heterocyclic group may be bicyclic, such as, but not limited to, a 5,5-membered ring, such as a hexahydrocyclopenta[c]pyrrol-2(1H)-yl ring, or a 5,6-membered bicyclic ring, such as a hexahydropyrrolo[1,2-a]pyrazin-2(1H)-yl ring. The heterocyclic group may be partially unsaturated, i.e., it may contain one or more double bonds, such as, but not limited to, dihydrofuranyl, dihydropyranyl, 2,5-dihydro-1H-pyrrolyl, 4H-[1,3,4]thiadiazinyl, 1,2,3,5-tetrahydrooxazolyl, or 4H-[1,4]thiazinyl, or it may be benzo-fused, such as, but not limited to, dihydroisoquinolinyl. When the 3-14-membered heterocyclic group is linked to other groups to form the compounds of the present invention, the linking may be to a carbon atom of the 3-14-membered heterocyclic group or to a heterocyclic atom on the 3-14-membered heterocyclic group ring. For example, when the 3-14 membered heterocyclic group is selected from piperazinyl, the nitrogen atom on the piperazinyl group may be connected to the other group. Or when the 3-14 membered heterocyclic group is selected from piperidinyl, the nitrogen atom on the piperidinyl ring and the carbon atom at the para position thereof may be connected to the other group.

[0086] The term "spirocyclic" refers to a ring system in which two rings share one ring atom.

[0087] The term "fused ring" refers to a ring system in which two rings share two ring atoms.

[0088] The term "bridged ring" refers to a ring system in which two rings share three or more ring atoms.

[0089] The term "halogen" refers to fluorine, chlorine, bromine and iodine.

[0090] "Halo" means substituted with one or more halogens.

[0091] Unless otherwise stated, the definitions of terms herein also apply to groups containing the term, e.g. 1-12 The definition of alkyl also applies to C 1-12 Alkyloxy (ie C 1-12 alkoxy).

[0092] The term "alkylene" refers to a divalent group, which is as defined above. For example, the term "alkylene" refers to a divalent alkyl group, wherein alkyl is as defined above, and alkylene is preferably an alkylene group having 1 to 12 carbon atoms (i.e., C 1-12 alkylene), more preferably an alkylene containing 1 to 6 carbon atoms (i.e., C 1-6 alkylene).

[0093] It will be appreciated by those skilled in the art that the compounds of formula (I) may exist in the form of various pharmaceutically acceptable salts. If these compounds have a basic center, they may form acid addition salts; if these compounds have an acidic center, they may form base addition salts; if these compounds contain both an acidic center (e.g., a carboxyl group) and a basic center (e.g., an amino group), they may also form internal salts.

[0094] The compounds of the present invention may exist in the form of solvates (e.g., hydrates), wherein the compounds of the present invention contain a polar solvent as a structural element of the crystal lattice of the compound, in particular water, methanol or ethanol. The amount of polar solvent, in particular water, may be present in a stoichiometric or non-stoichiometric ratio.

[0095] Depending on their molecular structure, the compounds of the present invention may be chiral and therefore may exist in various enantiomeric forms. Thus, these compounds may exist in racemic or optically active forms. The compounds of the present invention encompass isomers or mixtures thereof, racemates, in which each chiral carbon is in the R or S configuration. The compounds of the present invention or their intermediates can be separated into enantiomeric compounds by chemical or physical methods well known to those skilled in the art, or used in this form for synthesis. In the case of racemic amines, diastereomers are prepared from the mixture by reaction with an optically active resolving agent. Examples of suitable resolving agents are optically active acids, such as R and S forms of tartaric acid, diacetyltartaric acid, dibenzoyltartaric acid, mandelic acid, malic acid, lactic acid, appropriate N-protected amino acids (e.g., N-benzoylproline or N-phenylsulfonylproline) or various optically active camphorsulfonic acids. Chromatographic enantiomer resolution can also be advantageously performed with the aid of optically active resolving agents (e.g., dinitrobenzoylphenylglycine, cellulose triacetate or other carbohydrate derivatives or chirally derivatized methacrylate polymers immobilized on silica gel). Suitable eluents for this purpose are aqueous or alcoholic solvent mixtures, e.g., hexane / isopropanol / acetonitrile.

[0096] The corresponding stable isomers can be separated according to known methods, for example by extraction, filtration or column chromatography.

[0097] The term "patient" refers to any animal including mammals, preferably mice, rats, other rodents, rabbits, dogs, cats, pigs, cows, sheep, horses or primates, and most preferably humans.

[0098] The term "therapeutically effective amount" refers to that amount of an active compound or drug that will elicit the biological or medical response that a researcher, veterinarian, physician, or other clinician is seeking in a tissue, system, animal, individual, or human, and includes one or more of the following: (1) prevents disease, e.g., prevents a disease, disorder, or condition in an individual who is susceptible to the disease, disorder, or condition but who is not yet experiencing or developing the pathology or symptoms of the disease. (2) inhibits disease, e.g., inhibits the disease, disorder, or condition (i.e., prevents further development of the pathology and / or symptoms) in an individual who is experiencing or developing the pathology or symptoms of the disease, disorder, or condition. (3) alleviates disease, e.g., alleviates the disease, disorder, or condition (i.e., reverses the pathology and / or symptoms) in an individual who is experiencing or developing the pathology or symptoms of the disease, disorder, or condition. DETAILED DESCRIPTION

[0099] The technical solutions of the present invention will be described in further detail below with reference to specific embodiments. It should be understood that the following embodiments are merely illustrative and explanations of the present invention and should not be construed as limiting the scope of protection of the present invention. All technologies implemented based on the above content of the present invention are encompassed within the scope of protection that the present invention is intended to protect.

[0100] Unless otherwise specified, the raw materials and reagents used in the following examples are commercially available or can be prepared by known methods.

[0101] The structures of the compounds of the present invention are determined by nuclear magnetic resonance (NMR) and / or liquid chromatography-mass spectrometry (LC-MS). NMR chemical shifts (δ) are given in parts per million (ppm). NMR measurements are performed using a Bruker AVANCE-400 NMR spectrometer, using deuterated dimethyl sulfoxide (DMSO-d6), deuterated methanol (CD3OD), or deuterated chloroform (CDCl3) as the solvent, and tetramethylsilane (TMS) as the internal standard.

[0102] Liquid chromatography-mass spectrometry (LC-MS) was performed using an Agilent 1200 Infinity Series mass spectrometer. HPLC was performed using an Agilent 1200DAD high-pressure liquid chromatograph (Sunfire C18 150×4.6 mm column) and a Waters 2695-2996 high-pressure liquid chromatograph (Gimini C18 150×4.6 mm column).

[0103] Thin-layer chromatography silica gel plates use Yantai Huanghai HSGF254 or Qingdao Ocean Chemical GF254 silica gel plates. The specifications used for TLC are 0.15mm-0.20mm, and the specifications used for thin-layer chromatography separation and purification products are 0.4mm-0.5mm. Column chromatography generally uses Yantai Huanghai 200-300 mesh silica gel as the carrier.

[0104] Unless otherwise stated, the raw materials and reagents used in the following examples are commercially available or can be prepared by known methods. Unless otherwise stated, all reactions of the present invention were carried out under a dry nitrogen or argon atmosphere with continuous magnetic stirring, with dry solvents, and reaction temperatures are reported in degrees Celsius.

[0105] Example 1

[0106] Step 1: Synthesis of ethyl 7-bromobenzofuran-3-carboxylate (Compound 001-2)

[0107] Under nitrogen protection, 3-bromo-2-hydroxybenzaldehyde 001-1 (10 g, 0.063 mol, 1 eq) was dissolved in dichloromethane (20 ml), and fluoroboric acid (0.57 g, 0.007 mol, 0.01 eq) was added, followed by ethyl diazoacetate (17.12 g, 0.15 mol, 2.4 eq). After no more gas was generated, sulfuric acid (10 ml) was added. After completion of the reaction, sodium carbonate solution was added dropwise to quench the reaction. The aqueous phase was extracted with ethyl acetate (3 × 100 ml), and the organic phases were combined, backwashed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give compound 001-2 (5 g, 75.56%).

[0108] LCMS: (ESI, m / z): 268.95 [M+H] + .

[0109] 1 H NMR (400MHz, DMSO-d6) δ8.88(s,1H),7.98(dd,J=7.9,1.1Hz,1H),7.68(dd,J=7.9 ,1.1Hz,1H),7.37(t,J=7.8Hz,1H),4.36(q,J=7.1Hz,2H),1.35(t,J=7.1Hz,3H).

[0110] Step 2: Synthesis of 7-bromobenzofuran-3-methanol (Compound 001-3)

[0111] Under nitrogen protection, compound 7-bromobenzofuran-3-carboxylic acid ethyl ester 001-2 (5 g, 0.018 mol, 1 eq) was dissolved in dichloromethane (20 ml), and a toluene solution of diisobutylaluminum hydride (40 ml, 0.037 mol, 2.0 eq) was added dropwise at -78°C. The mixture was warmed to room temperature and stirred for 1 hour. After the reaction was completed, the reaction mixture was quenched with an aqueous solution of sodium sulfate decahydrate at room temperature, filtered, and extracted with ethyl acetate (3×100 ml). The organic phases were combined, backwashed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 1:1) to obtain compound 001-3 (3.6 g, 99.25%).

[0112] 1 H NMR(400MHz,DMSO-d6)δ8.00(s,1H),7.71(dd,J=7.8,1.1Hz,1H),7.55(dd,J=7.8,1 .0Hz,1H),7.22(t,J=7.8Hz,1H),5.24(t,J=5.5Hz,1H),4.63(dd,J=5.5,1.1Hz,2H).

[0113] Step 3: Synthesis of 7-bromo-3-bromomethylbenzofuran (Compound 001-4)

[0114] Under nitrogen, compound 7-bromobenzofuran-3-methanol 001-3 (3.5 g, 0.015 mol, 1 eq) was dissolved in anhydrous ether (10 mL). Phosphorus tribromide (12.19 g, 0.045 mol, 3 eq) was added dropwise at 0°C, and the mixture was stirred at 0°C for 30 minutes. The reaction mixture was quenched with ice water and extracted with ethyl acetate (3 × 50 mL). The organic phases were combined, backwashed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by silica gel column chromatography (petroleum ether as eluent) to obtain compound 001-4 (1.2 g, 99.18%).

[0115] 1 H NMR (400MHz, DMSO-d6) δ8.29(s,1H),7.76(dd,J=7.8,1.0Hz,1H),7.62(dd,J=7.8,1.1Hz,1H),7.30(t,J=7.8Hz,1H),4.90(d,J=0.8Hz,2H).

[0116] Step 4: Synthesis of 7-bromo-3-(2,2,2-trifluoroethyl)benzofuran (Compound 001-5)

[0117] Under nitrogen, compound 7-bromo-3-bromomethylbenzofuran 001-4 (1.1 g, 0.004 mol, 1 eq), cuprous iodide (88 mg, 0.458 mmol, 0.12 eq), and methyl fluorosulfonyldifluoroacetate (1.54 g, 0.008 mol, 2 eq) were dissolved in N,N-dimethylformamide (5 mL) and stirred at 80°C for 1 hour. After completion of the reaction, the reaction solution was quenched with water and extracted with ethyl acetate (3 × 50 mL). The organic phases were combined, backwashed with saturated sodium chloride solution (3 × 50 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (petroleum ether as eluent) to afford compound 001-5 (530 mg, 91.81%).

[0118] 1 H NMR (400MHz, DMSO-d6) δ8.18(s,1H),7.75(d,J=7.7Hz,1H),7.61(dd,J=7.8,1.1Hz,1H),7.28(t,J=7.8Hz,1H),3.87(qd,J=11.4,0.9Hz,2H).

[0119] Step 5: Synthesis of 7-bromo-2-iodo-3-(2,2,2-trifluoroethyl)-1-benzofuran (Compound 001-6)

[0120] Under nitrogen, compound 7-bromo-3-(2,2,2-trifluoroethyl)benzofuran 001-5 (250 mg, 0.899 mmol, 1 eq) and N-iodosuccinimide (243 mg, 1.079 mmol, 1.2 eq) were dissolved in trifluoroacetic acid (4 mL) and stirred at room temperature for 2 hours. After completion of the reaction, the reaction solution was concentrated under reduced pressure, 10 mL of water was added, and the mixture was extracted with ethyl acetate (3 x 20 mL). The organic phases were combined, backwashed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by reverse-phase column chromatography (C18 column, mobile phase: water and acetonitrile, gradient from 10% to 60% over 10 minutes, UV 254 nm detector). The product was extracted with ethyl acetate, and the organic phase was concentrated under reduced pressure to yield compound 001-6 (103 mg, 72.53%).

[0121] GCMS:(ESI,m / z):403.9[M] +

[0122] 1 H NMR (400MHz, DMSO-d6) δ7.71(d,J=7.9Hz,1H),7.55(d,J=7.8Hz,1H),7.26(t,J=7.8Hz,1H),3.75(q,J=11.1Hz,2H).

[0123] Step 6: Synthesis of N-{3-[7-bromo-3-(2,2,2-trifluoroethyl)-1-benzofuran-2-yl]prop-2-yn-1-yl}-1-tert-butyl-4-carboxamide (Compound 001-7)

[0124] Under nitrogen protection, compound 7-bromo-2-iodo-3-(2,2,2-trifluoroethyl)-1-benzofuran 001-6 (100 mg, 0.247 mmol, 1 eq), 1-tert-butyl-N-(propyl-2-yn-1-yl)pyrazole-4-carboxamide (60.83 mg, 0.296 mmol, 1.2 eq), cuprous iodide (4.7 mg, 0.025 mmol, 0.1 eq), diisopropylamine (249.89 mg, 2.470 mmol, 10 eq), tetrakis(triphenylphosphine)palladium (28.54 mg, 0.025 mmol, 0.1 eq) were dissolved in tetrahydrofuran (0.2 mL), the temperature was raised to 50° C., and the mixture was stirred for 1 hour. After the reaction was completed, the mixture was concentrated by rotary evaporation, and the residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 3:1) to give compound 001-7 (80 mg, 64.55%).

[0125] LCMS: (ESI, m / z): 481.85 [M+H]+.

[0126] Step 7: Synthesis of 1-(tert-butyl)-N-(3-(7-(((3S,4R)-3-fluoro-1-methylpiperidin-4-yl)amino)-3-(2,2,2-trifluoroethyl)benzofuran-2-yl)prop-2-yn-1-yl)-1H-pyrazole-4-carboxamide (Compound 001)

[0127] Under nitrogen protection, N-{3-[7-bromo-3-(2,2,2-trifluoroethyl)-1-benzofuran-2-yl]prop-2-yn-1-yl}-1-tert-butyl-4-carboxamide 001-7 (70 mg, 0.145 mmol, 1 eq), (3S,4R)-3-fluoro-1-methylpiperidin-4-amine dihydrochloride (35.74 mg, 0.174 mmol, 1.2 eq), (SP-4-1)-[1,3-bis[2,6-bis(1-ethylpropyl)phenyl]-4,5-dichloro-1,3-dimethoxy-1-yl]-1-[[[2,6-bis(1-ethylpropyl)phenyl]-1-[[[2,6-bis(1-ethylpropyl)phenyl]-1-[[[2,6-bis(1-ethylpropyl)phenyl]-1-[[[2,6-bis(1-ethylpropyl)phenyl]-1-[[[2,6-bis(1-ethylpropyl)phenyl]-1-[[ [H-2H-imidazol-2-ylidene] dichloro(2-methylpyridine)palladium (12.21 mg, 0.014 mmol, 0.1 eq) and cesium carbonate (189.15 mg, 0.580 mmol, 4 eq) were dissolved in 1,4-dioxane (2.5 mL), heated to 100 ° C, and stirred for 2 hours. After the reaction was completed, the reaction mixture was diluted with ethyl acetate, washed with saturated sodium chloride solution (20 mL × 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by high performance liquid chromatography under the following conditions (chromatographic column specifications: Kinetex 5μm EVOC18, 30mm*150mm; mobile phase A: water (10 mmol / L ammonium bicarbonate), mobile phase B: acetonitrile; flow rate: 60 ml / min; elution gradient: 32% B to 56% B in 10 minutes; detection wavelength: UV 254nm / 220nm; retention time (minutes): 8.15), to obtain compound 001 (6.66 mg, 8.57%).

[0128] LCMS: (ESI, m / z): 533.90 [M+H]+.

[0129] 1 H NMR (400MHz, DMSO-d6, ppm) δ8.66(t,J=5.6Hz,1H),8.32(s,1H),7.90(s,1H),7.10(t,J=7.8Hz,1H),6. 92(d,J=7.7Hz,1H),6.75(d,J=7.7Hz,1H),5.34(d,J=8.9Hz,1H),4.78(d,J=49.4Hz,1H),4.39(d,J=5.6 Hz,2H),3.76(q,11.1Hz,2H),3.71–3.57(m,1H),3.03(t,J=11.9Hz,1H),2.78(d,J=11.2Hz,1H),2.26(d ,J=13.1Hz,1H),2.18(s,3H),2.08(t,J=11.5Hz,1H),1.95–1.83(m,1H),1.75–1.68(m,1H),1.53(s,9H)

[0130] Example 2

[0131] Step 1: Synthesis of 2-bromo-4-chloro-1-(2,2,2-trifluoroethyl)-1H-benzo[d]imidazole (Compound 002-2)

[0132] A solution of 2-bromo-4-chloro-1H-benzo[d]imidazole 002-1 (350 mg, 1.512 mmol, 1 eq), trifluoroethyl trifluoromethanesulfonate (421.12 mg, 1.814 mmol, 1.2 eq), and potassium carbonate (626.9 mg, 4.536 mmol, 3 eq) in N,N-dimethylformamide (4 mL) was stirred at room temperature for 2 hours. After completion of the reaction, the mixture was quenched with water and extracted with ethyl acetate (3 × 20 mL). The organic phases were combined, backwashed with saturated sodium chloride solution (3 × 20 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 5:1) to afford compound 002-2 (200 mg, 42.19%).

[0133] LCMS: (ESI, m / z): 312.65 [M+H] + .

[0134] Step 2: Synthesis of 1-tert-butyl-N-{3-[4-chloro-1-(2,2,2-trifluoroethyl)-1H-benzo[d]imidazol-2-yl]prop-2-yn-1-yl}-1H-pyrazole-4-carboxamide (Compound 002-3)

[0135] Under nitrogen protection, at 80 ° C, compound 2-bromo-4-chloro-1-(2,2,2-trifluoroethyl)-1H-benzo[d]imidazole 002-2 (200 mg, 0.638 mmol, 1 eq), 1-tert-butyl-N-(prop-2-yn-1-yl)-1H-pyrazole-4-carboxamide (196.42 mg, 0.957 mmol, 1.5 eq), cuprous iodide (12.15 mg, 0.064 mmol, 0.1 eq), diisopropylamine (645.57 mg, 6.380 mmol, 1 0eq) and a dimethyl sulfoxide solution (3 mL) of tetrakistriphenylphosphine palladium (73.72 mg, 0.064 mmol, 0.1 eq) were stirred for 4 hours. After the reaction, the reaction mixture was quenched with water at room temperature and extracted with ethyl acetate (3×30 mL). The organic phases were combined, backwashed with saturated sodium chloride solution (3×30 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 1:3) to obtain compound 002-3 (150 mg, 53.70%).

[0136] LCMS: (ESI, m / z): 438.00 [M+H] + .

[0137] Step 3: Synthesis of 1-(tert-butyl)-N-(3-(4-(((3S,4R)-3-fluoro-1-methylpiperidin-4-yl)amino)-1-(2,2,2-trifluoroethyl)-1H-benzo[d]imidazol-2-yl)prop-2-yn-1-yl)-1H-pyrazole-4-carboxamide (Compound 002)

[0138] Under nitrogen protection, at 120 ° C, compound 1-tert-butyl-N-{3-[4-chloro-1-(2,2,2-trifluoroethyl)-1H-benzo[d]imidazol-2-yl]prop-2-yn-1-yl}-1H-pyrazole-4-carboxamide 002-3 (80 mg, 0.183 mmol, 1 eq), (3S,4R)-3-fluoro-1-methylpiperidin-4-amine dihydrochloride (44.97 mg, 0.220 mmol, 1.2 eq), (SP-4-1)-[1,3-bis[2,6-bis(1-ethylpropyl)phenyl]-4,5-dichloro-1,3-dihydro- A solution of 2H-imidazole-2-ylidene]dichloro(2-methylpyridine)palladium (11.53 mg, 0.014 mmol, 0.1 eq) and cesium carbonate (178.59 mg, 0.549 mmol, 3 eq) in 1,4-dioxane (2 mL) was stirred for 4 hours. After the reaction, the reaction mixture was quenched with water at room temperature and extracted with ethyl acetate (3 × 10 mL). The organic phases were combined, backwashed with saturated sodium chloride solution (3 × 10 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by high performance liquid chromatography under the following conditions (chromatographic column specifications: Sunfire C18 5μm, 30mm x 150mm; mobile phase A: water (0.1% ammonium bicarbonate), mobile phase B: acetonitrile; flow rate: 60 ml / min; elution gradient: 60% to 82% in 10 minutes; detection wavelength: UV 254 nm / 220 nm; retention time (min): 7.88) to obtain compound 002 (40.9 mg, 41.95%).

[0139] LCMS: (ESI, m / z): 534.15 [M+H] + .

[0140] 1H NMR (400MHz, DMSO-d6, ppm) δ8.31(s,1H),7.89(s,1H),7.11(s,1H),7.03(t,J=7.9Hz,1H), 6.85(d,J=8.1Hz,1H),6.49(d,J=7.8Hz,1H),5.31(q,J=9.1Hz,2H),5.13(d,J=9.6Hz,1H),4 .84–4.65(m,1H),4.45(s,2H),3.93(dq,J=27.7,9.4,8.7Hz,1H),3.03–2.86(m,1H),2.80–2 .64(m,1H),2.09(s,4H),2.00–1.86(m,1H),1.72(td,J=10.6,9.4,3.5Hz,2H),1.54(s,9H).

[0141] Example 3

[0142] Step 1: Synthesis of tert-butyl (3-(7-bromo-3-(2,2,2-trifluoroethyl)benzofuran-2-yl)prop-2-yn-1-yl)carbamate (Compound 006-1)

[0143] Under nitrogen protection, compound 7-bromo-2-iodo-3-(2,2,2-trifluoroethyl)benzofuran 001-6 (300 mg, 0.741 mmol, 1 eq), tert-butyl prop-2-yn-1-ylcarbamate (137.97 mg, 0.889 mmol, 1.2 eq), cuprous iodide (14.11 mg, 0.074 mmol, 0.1 eq), diisopropylamine (749.66 mg, 7.410 mmol, 10 eq), and tetrakis(triphenylphosphine)palladium (85.61 mg, 0.074 mmol, 0.1 eq) were dissolved in tetrahydrofuran (5 mL), heated to 50 ° C and stirred for 1 hour. After the reaction was completed, the reaction mixture was concentrated under reduced pressure and the crude product was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 12:1) to obtain compound 006-1 (250 mg, 65.58%). LCMS: (ESI, m / z): 376.90 [M+H-56] + .

[0144] Step 2: Synthesis of tert-butyl (3-(7-((3S,4R)-3-fluoro-1-methylpiperidin-4-yl)amino)-3-(2,2,2-trifluoroethyl)benzofuran-2-yl)prop-2-yn-1-yl)carbamate (Compound 006-2)

[0145] Under nitrogen protection, compound (tert-butyl 3-(7-bromo-3-(2,2,2-trifluoroethyl)benzofuran-2-yl)prop-2-yn-1-yl)carbamate 006-1 (240 mg, 0.555 mmol, 1 eq), (3S,4R)-3-fluoro-1-methylpiperidin-4-amine (88.07 mg, 0.666 mmol, 1.2 eq), cesium carbonate (723.65 mg, 2.220 mmol, 4 eq), (SP-4-1)-[1, 3-Bis[2,6-bis(1-ethylpropyl)phenyl]-4,5-dichloro-1,3-dihydro-2H-imidazol-2-ylidene]dichloro(2-methylpyridine)palladium (46.71 mg, 0.056 mmol, 0.1 eq) was dissolved in 1,4-dioxane (8 mL), heated to 100°C and stirred for 2 hours. After the reaction was completed, the reaction mixture was concentrated under reduced pressure and the residue was purified by silica gel column chromatography (ethyl acetate as eluent) to give compound 006-2 (200 mg, 71.74%).

[0146] LCMS: (ESI, m / z): 484.10 [M+H] + .

[0147] Step 3: Synthesis of dihydrochloride salt of (3S,4R)-N-[2-(3-aminopropyl-1-ynyl)-3-(2,2,2-trifluoroethyl)benzofuran-7-yl]-3-fluoro-1-methylpiperidin-4-amine (Compound 006-3)

[0148] Under nitrogen protection, compound (3-(7-((3S,4R)-3-fluoro-1-methylpiperidin-4-yl)amino)-3-(2,2,2-trifluoroethyl)benzofuran-2-yl)prop-2-yn-1-yl)carbamic acid tert-butyl ester 006-2 (200 mg, 0.414 mmol, 1 eq) was added to the reaction flask and dissolved in dichloromethane (3 mL). A 4 mol / L solution of hydrogen chloride in 1,4-dioxane (3 mL) was added and the mixture was stirred at room temperature for 1 hour. After the reaction was completed, the reaction mixture was concentrated under reduced pressure to obtain compound 006-3 (110 mg, 55.19%).

[0149] LCMS: (ESI, m / z): 383.95 [M+H] + .

[0150] Step 4: Synthesis of 5-amino-1-(tert-butyl)N-(3-(7-((3S,4R)-3-fluoro-1-methylpiperidin-4-yl)amino)-3-(2,2,2-trifluoroethyl)benzofuran-2-yl)propyl-2-yn-1-yl)-1H-pyrazole-4-carboxamide (Compound 006)

[0151] Under nitrogen protection, 5-amino-1-tert-butyl-1H-pyrazole-4-carboxylic acid (48.18 mg, 0.263 mmol, 1.2 eq), 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (83.33 mg, 0.219 mmol, 1 eq), N,N-diisopropylethylamine (141.62 mg, 1.095 mmol, 5 eq) were added to the reaction flask, dissolved in N,N-dimethylformamide (5 mL), stirred at room temperature for 10 minutes, and then compound (3S,4R)-N-[ 2-(3-Aminopropyl-1-ynyl)-3-(2,2,2-trifluoroethyl)benzofuran-7-yl]-3-fluoro-1-methylpiperidin-4-amine dihydrochloride 006-3 (100 mg, 0.219 mmol, 1 eq) was stirred at room temperature for 1 hour. After the reaction, water (10 mL) was added and extracted with ethyl acetate (20 mL × 2). The organic phases were combined and backwashed with saturated sodium chloride solution (30 mL × 1), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by high performance liquid chromatography under the following conditions (chromatographic column specifications: Sunfire C18 5μm, 30mm*150mm; mobile phase A: water (0.1% formic acid), mobile phase B: acetonitrile; flow rate: 60 ml / min; elution gradient: 15% B to 33% B in 7 min; detection wavelength: UV 254nm / 220nm; retention time (min): 6.68) to obtain the formate salt of compound 006 (50.47 mg, 41.85%).

[0152] LCMS: (ESI, m / z): 549.10 [M+H] + .

[0153] 1H NMR (400MHz, DMSO-d6, ppm) δ8.34(t,J=5.7Hz,1H),7.64(s,1H),7.10(t,J=7.8Hz,1H),6.92(d,J=7.8Hz,1 H),6.74(d,J=7.8Hz,1H),6.19(s,2H),5.36(d,J=9.0Hz,1H),4.78(d,J=49.2Hz,1H),4.33(d,J=5.6Hz,2H) ,3.76(q,J=11.1Hz,2H),3.70–3.56(m,1H),3.03(t,J=10.9Hz,1H),2.79(d,J=11.4Hz,1H),2.27(d,J=13.0 Hz,1H),2.19(s,3H),2.08(t,J=11.0Hz,1H),1.90(qd,J=12.3,3.7Hz,1H),1.75–1.68(m,1H),1.52(s,9H).

[0154] Example 4

[0155] Step 1: Synthesis of tert-butyl (3-(4-bromo-1-(2,2,2-trifluoroethyl)-indol-2-yl)prop-2-yn-1-yl)carbamate (Compound 173-2)

[0156] Under nitrogen protection, the reaction flask was added with compound 4-bromo-2-iodo-1-(2,2,2-trifluoroethyl)indole 173-1 (1 g, 2.475 mmol, 1 eq), N-(tert-butoxycarbonyl)propargylamine (422.6 mg, 2.723 mmol, 1.1 eq), tetrakistriphenylphosphine palladium (286.06 mg, 0.248 mmol, 0.1 eq), cuprous iodide (47.14 mg, 0.248 mmol, 0.1 eq), diisopropylamine (2.50 g, 24.750 mmol, 10 eq), and dissolved in tetrahydrofuran (10.0 mL). The reaction was carried out at 50 ° C. for 2 hours. After the reaction was completed, the solvent was directly dried and the residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 1:1) to give compound 173-2 (900 mg, 75.88%).

[0157] LCMS: (ESI, m / z): 430.70 [M+H] + .

[0158] Step 2: Synthesis of tert-butyl [3-(4-([(3S,4R)-3-fluoro-1-methylpiperidin-4-yl]amino-1-(2,2,2-trifluoroethyl)indol-2-yl)prop-2-yn-1-yl]carbamate (Compound 173-3)

[0159] Under nitrogen protection, the reaction flask was added with compound (tert-butyl 3-(4-bromo-1-(2,2,2-trifluoroethyl)-indol-2-yl)prop-2-yn-1-yl)carbamate 173-2 (215 mg, 0.499 mmol, 1 eq), (3S,4R)-3-fluoro-1-methylpiperidin-4-amine dihydrochloride (131.8 mg, 0.997 mmol, 2.00 eq), (SP-4-1)-[1,3-bis[2,6-bis(1-ethylpropyl)phenyl]-4,5- -dichloro-1,3-dihydro-2H-imidazol-2-ylidene]dichloro(2-methylpyridine)palladium (41.94 mg, 0.050 mmol, 0.1 eq) and cesium carbonate (649.74 mg, 1.996 mmol, 4 eq) were dissolved in dioxane (6.0 mL) and reacted at 120 ° C for 4 hours. After the reaction was completed, the mixture was filtered and the filtrate was dried. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 1:1) to obtain compound 173-3 (130 mg, 70.87%).

[0160] LCMS: (ESI, m / z): 482.90 [M+H] + .

[0161] Step 3: Synthesis of 2-(3-aminoprop-1-yn-1-yl)-N-[(3S,4R)-3-fluoro-1-methylpiperidin-4-yl]-1-(2,2,2-trifluoroethyl)indole-4-amine dihydrochloride (Compound 173-4)

[0162] To the reaction flask was added tert-butyl [3-(4-([(3S,4R)-3-fluoro-1-methylpiperidin-4-yl]amino-1-(2,2,2-trifluoroethyl)indol-2-yl)prop-2-yn-1-yl]carbamate 173-3 (119 mg, 0.247 mmol, 1 eq), dissolved in dichloromethane (3 mL), and then added with a 4 mol / L hydrogen chloride 1,4-dioxane solution (1.5 mL). The reaction was allowed to react at room temperature overnight. After the reaction was complete, the solvent was directly dried to give compound 173-4 (100 mg, 95.43%).

[0163] LCMS: (ESI, m / z): 383.00 [M+H] + .

[0164] Step 4: Synthesis of 5-amino-1-tert-butyl-N-[3-(4-{[(3S,4R)-3-fluoro-1-methylpiperidin-4-yl]amino}-1-(2,2,2-trifluoroethyl)indol-2-yl)prop-2-yn-1-yl]-1H-pyrazole-4-carboxamide (Compound 173)

[0165] Compound 2-(3-aminoprop-1-yn-1-yl)-N-[(3S,4R)-3-fluoro-1-methylpiperidin-4-yl]-1-(2,2,2-trifluoroethyl)indole-4-amine dihydrochloride 173-4 (70 mg, 0.183 mmol, 1 eq), 5-amino-1-tert-butylpyrazole-4-carboxylic acid (50.31 mg, 0.274 mmol, 1.5 eq), tetramethyl chlorouronium hexafluorophosphate (205.44 mg) were added to the reaction flask in sequence. , 0.732 mmol, 4 eq), N-methylimidazole (150.3 mg, 1.830 mmol, 10 eq), acetonitrile (5 mL) was added to dissolve, and the reaction was allowed to react at room temperature overnight. After the reaction was completed, water was added to quench the reaction mixture, and the reaction mixture was extracted with ethyl acetate (3×10 mL). The organic phases were combined, backwashed with saturated sodium chloride solution (1×10 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by high performance liquid chromatography under the following conditions (chromatographic column specifications: Xselect CSH™ Prep C18 5μm 30*150mm OBD; mobile phase A: water (0.1% formic acid), mobile phase B: acetonitrile; flow rate: 60 ml / min; elution gradient: 11% B to 31% B in 10 minutes; detection wavelength: UV 254 nm / 220 nm; retention time (minutes): 9.1) to give the formate salt of compound 173 (7.47 mg, 7.34%).

[0166] LCMS: (ESI, m / z): 548.00 [M+H] + .

[0167] 1 H NMR (400MHz, DMSO-d6, ppm): δ8.28(t,J=5.6Hz,1H),7.64(s,1H),7.21(s,1H),7.03(t,J=8.0H z,1H),6.77(d,J=8.2Hz,1H),6.26(d,J=7.8Hz,1H),6.17(s,2H),5.52(d,J=8.6Hz,1H),5.03(q ,J=9.1Hz,2H),4.87(d,J=49.0Hz,1H),4.28(d,J=5.5Hz,2H),3.72–3.55(m,1H),3.27–3.10(m ,1H),2.99–2.84(m,1H),2.38–2.12(m,5H),2.04–1.90(m,1H),1.82–1.69(m,1H),1.52(s,9H).

[0168] Example 5

[0169] Step 1: Synthesis of tert-butyl 3-[7-bromo-3-(2,2,2-trifluoroethyl)-1-benzothiophen-2-yl]prop-2-yn-1-yl}carbamate (Compound 005-2)

[0170] Under nitrogen protection, a solution of compound 7-bromo-2-iodo-3-(2,2,2-trifluoroethyl)-1-benzothiophene 005-1 (500 mg, 1.188 mmol, 1 eq), N-(tert-butoxycarbonyl)propargylamine (202.75 mg, 1.307 mmol, 1.1 eq), cuprous iodide (22.62 mg, 0.119 mmol, 0.1 eq), diisopropylamine (1.201 g, 11.880 mmol, 10 eq) and tetrakistriphenylphosphine palladium (137.24 mg, 0.119 mmol, 0.1 eq) in tetrahydrofuran (5 mL) was stirred at 50°C for 1 hour. After the reaction was completed, the reaction solution was concentrated in vacuo and the residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 5:1) to give compound 005-2 (500 mg, 93.91%).

[0171] LCMS: (ESI, m / z): 393.75 [M+H-56] + .

[0172] Step 2: Synthesis of tert-butyl 3-(7-{[(3S,4R)-3-fluoro-1-methylpiperidin-4-amino}-3-(2,2,2-trifluoroethyl)-1-benzothiophen-2-yl]prop-2-yn-1-yl]carbamate (Compound 005-3)

[0173] Under nitrogen protection, at 100 ° C, compound 3-[7-bromo-3-(2,2,2-trifluoroethyl)-1-benzothiophen-2-yl]prop-2-yn-1-yl}carbamic acid tert-butyl ester 005-2 (150 mg, 0.335 mmol, 1 eq), (3S,4R)-3-fluoro-1-methylpiperidin-4-amine dihydrochloride (53.07 mg, 0.402 mmol, 1.2 eq), (SP-4-1)-[1,3-bis[2,6-bis(1-ethylpropyl)phenyl]-4,5 A solution of [-dichloro-1,3-dihydro-2H-imidazol-2-ylidene]dichloro(2-methylpyridine)palladium (9.38 mg, 0.011 mmol, 0.1 eq) and cesium carbonate (327.05 mg, 1.005 mmol, 3 eq) in 1,4-dioxane (3.0 mL) was stirred for 3 hours. After the reaction was completed, the reaction solution was concentrated under reduced pressure and the resulting residue was purified by silica gel column chromatography (dichloromethane / methanol = 10:1) to give compound 005-3 (150 mg, 89.74%).

[0174] LCMS: (ESI, m / z): 500.05 [M+H] + .

[0175] Step 3: Synthesis of (3S,4R)-N-[2-(3-aminoprop-1-yn-1-yl)-3-(2,2,2-trifluoroethyl)-1-benzothiophen-7-yl]-3-fluoro-1-methylpiperidin-4-amine dihydrochloride (Compound 005-4)

[0176] At room temperature, a solution of compound 3-(7-{[(3S,4R)-3-fluoro-1-methylpiperidin-4-amino}-3-(2,2,2-trifluoroethyl)-1-benzothiophen-2-yl]prop-2-yn-1-yl]carbamic acid tert-butyl ester 005-3 (150 mg, 0.300 mmol, 1 eq) and 4 mol / L hydrogen chloride in 1,4-dioxane (3 mL) in methanol (3 mL) was stirred for 2 hours. After the reaction was completed, the reaction solution was concentrated under reduced pressure to obtain compound 005-4 (110 mg, 91.71%).

[0177] LCMS: (ESI, m / z): 399.95 [M+H] + .

[0178] Step 3: Synthesis of 5-amino-1-tert-butyl-N-[3-(7-{[(3S,4R)-3-fluoro-1-methylpiperidin-4-yl]amino}-3-(2,2,2-trifluoroethyl)-1-benzothiophen-2-yl)prop-2-yn-1-yl]-1H-pyrazole-4-carboxamide (Compound 005)

[0179] At room temperature, a solution of 5-amino-1-tert-butyl-1H-pyrazole-4-carboxylic acid (38.79 mg, 0.212 mmol, 1 eq), O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (80.5 mg, 0.212 mmol, 1 eq) and N,N-diisopropylethylamine (136.81 mg, 1.060 mmol, 5 eq) in N,N-dimethylformamide (5 mL) was stirred for 15 minutes, and then the compound (3S,4R)-N-[2-(3-aminoprop-1-yn- 1-yl)-3-(2,2,2-trifluoroethyl)-1-benzothiophen-7-yl]-3-fluoro-1-methylpiperidin-4-amine dihydrochloride 005-4 (100 mg, 0.212 mmol, 1 eq) was stirred at room temperature for 1 hour. After the reaction, the reaction mixture was quenched with water at room temperature, and the reaction mixture was extracted with ethyl acetate (3×10 mL). The organic phases were combined, backwashed with saturated sodium chloride solution (3×10 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by high performance liquid chromatography under the following conditions (chromatographic column specifications: Sunfire C18 5μm, 30mm×150mm; mobile phase A: water (0.1% formic acid), mobile phase B: acetonitrile; flow rate: 60 ml / min; elution gradient: 15% to 30% in 10 minutes; detection wavelength: UV 254nm / 220nm; retention time (minutes): 8.18) to obtain compound 005 (15 mg, 11.60%).

[0180] LCMS: (ESI, m / z): 565.15 [M+H] + .

[0181] 1 H NMR (400MHz, DMSO-d6, ppm) δ8.31(t,J=5.6Hz,1H),7.64(s,1H),7.33–7.19(m,2H),6.80(d,J=7.4 Hz,1H),6.18(s,2H),5.16(d,J=8.6Hz,1H),4.81(d,J=49.3Hz,1H),4.32(d,J=5.6Hz,2H),3.92(q ,J=11.1Hz,2H),3.74-3.58(m,1H),3.04(t,J=11.6Hz,1H),2.80(d,J=11.2Hz,1H),2.29(d,J=13. 0Hz,1H),2.19(s,3H),2.10(t,J=11.3Hz,1H),2.01–1.89(m,1H),1.81–1.68(m,1H),1.52(s,9H).

[0182] Biological evaluation

[0183] Test Example 1: In vitro DNA binding activity assay

[0184] Time-Resolved Fluorescence Resonance Energy Transfer (TR-FREt) was used to detect the ability of compounds to modulate Y220C DNA binding. First, recombinant Y220C p53 DBD protein with a his-tag and a biotin-labeled DNA sequence were prepared. The donor, Eu(Europium)-SA (Streptavidin), binds to the DNA sequence via biotin. The acceptor, a his-antibody linked to allophycocyanin (APC), binds to the his-tagged Y220C p53 DBD protein. When excited with 340nm wavelength light, the donor, Eu(Europium)-SA (Streptavidin), emits fluorescence at 620nm. When the donor and acceptor are in close proximity, the donor transfers some of their energy to the acceptor, generating fluorescence at 665nm. That is, if the p53 mutant protein is reactivated by the compound and binds to DNA, the donor energy is transferred to the acceptor, generating fluorescence at 620 and 665 nm. If there is no interaction, only fluorescence at 615 nm is generated.

[0185] a. Add 4 μL of diluted compound solution to each well of a 384-well microplate.

[0186] b. Add 4 μL of 4X P53 working solution to each well of a 384-well microplate.

[0187] c. Seal the 384-well microplate and equilibrate at room temperature for 60 minutes.

[0188] d. Add 4 μl of biotinylated DNA solution to a 384-well microplate.

[0189] e. Add 4 μl of 4X detection solution (MAb Anti-6His-Eu and Streptavidin-d2) to each well of a 384-well microplate.

[0190] f. Incubate overnight, protect from light.

[0191] g. Read the data at 665nm and 615nm wavelengths on the BMG.

[0192] The representative compounds in this article were tested for their reactivation activity on p53-Y220C mutant protein by the above experiments. 150 See Table 1 for values.

[0193] Table 1 SC activity of representative compounds in this article against p53-Y220C mutant protein 150 value

[0194] Note: SC 150 Indicates the concentration of compound required to increase protein-DNA binding activity by 50%

[0195] Experimental conclusion: The above representative compounds can effectively restore the activity of p53-Y220C mutant protein.

[0196] The above is an exemplary description of the implementation methods of the technical solution of the present invention. It should be understood that the scope of protection of the present invention is not limited to the above implementation methods. Any modifications, equivalent substitutions, improvements, etc. made by those skilled in the art within the spirit and principles of the present invention shall be included in the scope of protection of the claims of this application.

Claims

1. A compound represented by formula (I) and its racemate, stereoisomer, tautomer, isotope-labeled substance, nitrogen oxide, solvate, polymorph, metabolite, ester, prodrug or pharmaceutically acceptable salt: in, It means that the ring in which it exists is aromatic; X1, X2, X3 are the same or different, independently selected from CH or N, and at least two of X1, X2, X3 are CH; X4 is selected from CR4, N, NR4; X5 is selected from CR5, N, NR5, O, S; R1 and R2 are the same or different and are independently selected from H, halogen, C 1-12 Alkyl, C 1-12 Alkoxy; R3 is selected from H, C 1-12 alkyl; R4 and R5 are the same or different and are independently selected from H, halogen, cyano, C 1-12 Alkyl, C 1-12 Alkoxy, C 3-12 Cycloalkyl, halogenated C 1-12 Alkyl, halogenated C 1-12 Alkoxy, halogenated C 3-12 Cycloalkyl, cyano C 1-12 Alkyl, cyano C 1-12 Alkoxy; W is selected from C 2-6 Alkenylene, C 2-6 Alkynylidene, C 3-14 Cycloalkylene, C 6-10 Arylene, 5-10 membered heteroarylene; Ring A is selected from C 3-14 Cycloalkyl, 3-14 membered heterocyclic group, C 6-10 Aryl, 5-10 membered heteroaryl; R a is selected from H, CN, oxo (=O), halogen, OH, unsubstituted or optionally substituted with one, two or more R a1 Substituted with the following groups: C 1-12 Alkyl, C 1-12 Alkoxy, C 3-12 Cycloalkyl, 3-14 membered heterocyclic group, C 6-10 Aryl, 5-10 membered heteroaryl, -C(O)N(R a11 )(R a12 )、-N(R a13 )(R a14 )、-S(O)2-R a15 、-S(O)(=NR a16 )(R a17 )、-P(O)(R a18 )(R a19 ); Or, two R a The atom to which it is attached is unsubstituted or optionally substituted with one, two or more R a1 Substituted with the following groups: C 3-12 Cycloalkyl, 3-14 membered heterocyclic group; Each R a1 are the same or different and are independently selected from H, OH, CN, halogen, unsubstituted or optionally substituted by one, two or more R a2 Substituted with the following groups: C 1-12 Alkyl, C 1-12 Alkoxy, C 3-12 Cycloalkyl, 3-14 membered heterocyclic group, C 6-10 Aryl, 5-10 membered heteroaryl, -NH2, -S(O)2-C 1-12 Alkyl; each R a2 are the same or different and are independently selected from H, OH, NH2, CN, halogen, C 1-6 Alkyl, C 1-6 Alkoxy; R a11 , R a12 , R a13 , R a14 , R a15 , R a16 , R a17 , R a18 , R a19 The same or different, independently selected from H, C 1-12 Alkyl, C 3-7 Cycloalkyl, 3-8 membered heterocyclic group; m is selected from 0, 1, 2, 3, 4, 5; Z is absent or selected from NH, S, O, C 1-6 Alkylene, C 1-6 Alkylene-NH; Ring E is selected from 3-14 membered heterocyclic group, C 3-12 Cycloalkyl; R e is selected from H, CN, halogen, unsubstituted or optionally substituted with one, two or more R e1 Substituted with the following groups: C 1-12 Alkyl, C 1-12 Alkoxy, C 3-12 Cycloalkyl, 3-14 membered heterocyclic group, C 1-12 Alkyl-NH-, (C 1-12 Alkyl)2-N-; or, two R e With their respective attached atoms form C 3-12 Cycloalkyl, 3-14 membered heterocyclic group; or, two R connected to different carbon atoms e The atoms to which they are attached form a 3-14 membered heterocyclic group; p is selected from 0, 1, 2, 3, 4, 5; Each R e1 are the same or different and are independently selected from H, OH, CN, halogen, C 1-12 Alkyl, C 1-12 Alkoxy; R x Selected from H, CN, halogen, C 1-12 Alkyl, C 1-12 Alkoxy; n is selected from 0, 1, 2 or 3; Y is selected from C(O), C(O)NH, C(S), SO2.

2. The compound according to claim 1 and its racemate, stereoisomer, tautomer, isotope-labeled substance, nitrogen oxide, solvate, polymorph, metabolite, ester, prodrug or pharmaceutically acceptable salt thereof, characterized in that: Preferably, R1, R2, and R3 are the same or different and are independently selected from H, C 1-6 alkyl; Preferably, R1, R2, and R3 are all H; Preferably, the Selected from Preferably, R4 and R5 are the same or different and are independently selected from C 1-6 Alkyl, halogenated C 1-6 Alkyl, cyano C 1-6 alkyl; Preferably, R4 is selected from methyl, ethyl, propyl, isopropyl, cyanomethyl, 2-fluoroethyl, 2,2-difluoroethyl, 2,2,2-trifluoroethyl; Preferably, R5 is selected from H.

3. The compound according to claim 1 or 2, and its racemate, stereoisomer, tautomer, isotope-labeled substance, nitrogen oxide, solvate, polymorph, metabolite, ester, prodrug or pharmaceutically acceptable salt thereof, characterized in that: W is selected from C 2-4 Alkenylene, C 2-4 Alkynylidene, C 3-6 Cycloalkylene; Preferably, W is selected from:

4. The compound according to any one of claims 1 to 3 and its racemate, stereoisomer, tautomer, isotope-labeled substance, nitrogen oxide, solvate, polymorph, metabolite, ester, prodrug or pharmaceutically acceptable salt thereof, characterized in that: Ring A is selected from phenyl, 5-6 membered heteroaryl, 8-9 membered heteroaryl, 6-9 membered heterocyclyl, C 3-6 cycloalkyl; Preferably, ring A is selected from: Preferably, R a is selected from H, CN, oxo (=O), halogen, OH, unsubstituted or optionally substituted with one, two or more R a1 Substituted with the following groups: C 1-6 Alkyl, C 1-6 Alkoxy, C 3-6 Cycloalkyl, 4-7 membered heterocyclyl, phenyl, 5-6 membered heteroaryl, -C(O)(NHC 1-6 Alkyl), -S(O)2-C 1-6 Alkyl, -S(O)(=NH)(C 1-6 alkyl), -S(O)(=NC 1-6 Alkyl)(C 1-6 alkyl), -P(O)(C 1-6 Alkyl)(C 1-6 Alkyl), -NH2, Or, two R a The atom to which it is attached is unsubstituted or optionally substituted with one, two or more R a1 Substituted with the following groups: C 3-6 Cycloalkyl; e.g. Preferably, each R a1 are the same or different and are independently selected from H, OH, CN, halogen, C 1-6 Alkyl, halogenated C 1-6 Alkyl, C 1-6 Alkoxy, C 3-6 Cycloalkyl, 5-6 membered heterocyclic group, phenyl, 5-6 membered heteroaryl, cyano 1-6 Alkyl, cyano C 1-6 Alkoxy, C 1-6 Alkyl-NH-, (C 1-6 Alkyl)2-N-, -S(O)2-C 1-6 Alkyl, C 1-6 Alkyl-OC 1-6 Alkyl, hydroxyl C 1-6 alkyl; Preferably, R a Selected from H, F, Cl, Br, oxo (=O), CN, NH2, Methylamino, dimethylamino, methyl, ethyl, isopropyl, tert-butyl, cyclopropyl, cyclobutyl, cyclopentyl, methoxy, difluoromethyl, trifluoromethyl, 2,2-difluoroethyl, 2,2,2-trifluoroethyl, phenyl, pyridyl, benzyl, morpholinyl, 5. The compound according to any one of claims 1 to 4 and its racemate, stereoisomer, tautomer, isotope-labeled substance, nitrogen oxide, solvate, polymorph, metabolite, ester, prodrug or pharmaceutically acceptable salt thereof, characterized in that: Selected from Preferably, Z is absent or selected from NH, S, O, CH2; Preferably, ring E is selected from a 6-9 membered heterocyclic group or a C 5-6 Cycloalkyl; Preferably, ring E is selected from Preferably, R e Selected from H, halogen, C 1-6 Alkyl, halogenated C 1-6 Alkyl, cyano C 1-6 Alkyl, hydroxyl C 1-6 Alkyl, -C 1-6 Alkyl-OC 1-6 Alkyl, C 1-6 Alkoxy, C 3-6 Cycloalkyl, 5-6 membered heterocyclic group, (C 1-6 Alkyl)2-N-; or, two R e With their respective attached atoms form C 3-6 Cycloalkyl; Preferably, R e Selected from H, F, Cl, CN, methyl, methoxy, ethyl, isopropyl, cyclopropyl, dimethylamino, Alternatively, two R attached to the same carbon atom e to the atoms to which they are attached respectively, forming a cyclopropane ring; Preferably, Selected from Preferably, X1, X2, and X3 are all CH, or one of X1, X2, and X3 is N; Preferably, when X4 is selected from CR4, X5 is selected from O, S, NR5; when X4 is selected from NR4, X5 is selected from CR5, N; when X4 is selected from N, X5 is selected from CR5, NR5; Preferably, R x Selected from H, CN, halogen, C 1-6 Alkyl, C 1-6 Alkoxy; Preferably, R x Selected from H, F, Cl, CN, methoxy, methyl.

6. The compound according to any one of claims 1 to 5 and its racemate, stereoisomer, tautomer, isotope-labeled substance, nitrogen oxide, solvate, polymorph, metabolite, ester, prodrug or pharmaceutically acceptable salt thereof, characterized in that: The compound represented by formula (I) has the structure shown below: Among them, ring A, ring E, X1, X2, X3, X4, X5, Y, R1, R2, R3, R a , R e , R x ,m,n,p, Having the definition of any one of claims 1 to 5; Preferably, the compound represented by formula (I) has the structure shown below: Among them, ring A, ring E, X1, X2, X3, Y, R1, R2, R3, R4, R a , R e , R x ,m,n,p, Having the definition of any one of claims 1 to 5; Preferably, the compound represented by formula (I) has the structure shown below: Among them, R a , R x , m, n have the definitions as described in any one of claims 1-5.

7. The compound according to any one of claims 1 to 6 and its racemate, stereoisomer, tautomer, isotope-labeled substance, nitrogen oxide, solvate, polymorph, metabolite, ester, prodrug or pharmaceutically acceptable salt thereof, characterized in that: Among the compounds represented by formula (I) and their racemates, stereoisomers, tautomers, isotope-labeled substances, nitrogen oxides, solvates, polymorphs, metabolites, esters, prodrugs or pharmaceutically acceptable salts, illustrative, non-limiting specific examples of the compounds represented by formula (I) are as follows:

8. A method for preparing the compound according to any one of claims 1 to 7, comprising the following steps: Compound 1 reacts with compound 2 to obtain a compound represented by formula (I); Among them, ring A, ring E, X1, X2, X3, X4, X5, W, Y, Z, R1, R2, R3, R a , R e , R x ,m,n,p, It has the definition as described in any one of claims 1 to 7.

9. A pharmaceutical composition comprising a therapeutically effective amount of at least one of the compound of any one of claims 1 to 7, its racemate, stereoisomer, tautomer, isotope-labeled substance, nitrogen oxide, solvate, polymorph, metabolite, ester, prodrug or a pharmaceutically acceptable salt thereof.

10. Use of at least one of the compound according to any one of claims 1 to 7, its racemate, stereoisomer, tautomer, isotope-labeled substance, nitrogen oxide, solvate, polymorph, metabolite, ester, prodrug or pharmaceutically acceptable salt thereof in the preparation of a medicament; Preferably, the use may be for preparing a drug for treating a tumor containing a p53-Y220C mutant, such as for preparing a drug for treating a p53-Y220C reactivator; Preferably, the tumor containing the p53-Y220C mutant includes acute lymphocytic leukemia, acute myeloid leukemia, adrenocortical carcinoma, AIDS-related cancer, AIDS-related lymphoma, anal cancer, appendix cancer, astrocytoma, basal cell carcinoma, bile duct cancer, bladder cancer, bone cancer, brain tumors such as cerebellar astrocytoma, cerebral astrocytoma / glioblastoma, ependymoma, medulloblastoma, supratentorial primitive cell tumor, neuroectodermal tumor, visual pathway and hypothalamic glioma, breast cancer, bronchial adenoma, Burkitt's lymphoma, unknown primary cancer, central nervous system lymphoma, cerebellar astrocytoma, cervical cancer, childhood cancer, chronic lymphocytic leukemia, chronic myeloproliferative disease, colon cancer, cutaneous T-cell lymphoma, desmoplastic small round cell tumor, endometrial cancer, ependymoma, food Tubular cancer, Ewing's sarcoma, germ cell tumors, gallbladder cancer, gastric cancer, gastrointestinal carcinoid tumors, gastrointestinal stromal tumors, gliomas, hairy cell leukemia, head and neck cancer, heart cancer, hepatocellular carcinoma, Hodgkin's lymphoma, hypopharyngeal cancer, intraocular melanoma, islet cell carcinoma, Kaposi's sarcoma, kidney cancer, laryngeal cancer, lip cancer and oral cancer, liposarcoma, liver cancer, lung cancer such as non-small cell lung cancer and small cell lung cancer, lymphoma, leukemia, macroglobulinemia, malignant bone fibrous histiocytoma / osteosarcoma, medulloblastoma, melanoma, mesothelioma, metastatic squamous cell carcinoma with occult primary, oral cancer, multiple endocrine neoplasms syndrome, myelodysplastic syndrome, myeloid leukemia, nasal cancer and paranasal sinus cancer, nasopharyngeal cancer, neuroblastoma, non-Hodgkin's lymphoma, non-small cell lung cancer, oral cancer, oropharyngeal cancer, osteosarcoma / malignant fibrous histiocytoma of bone, ovarian cancer, Ovarian epithelial cancer, ovarian germ cell tumor, pancreatic cancer, pancreatic islet cell cancer, paranasal sinus and nasal cavity cancer, parathyroid cancer, penile cancer, pharyngeal cancer, pheochromocytoma, pineal astrocytoma, pineal germ cell tumor, pituitary adenoma, pleuropulmonary blastoma, plasma cell tumor, primary central nervous system lymphoma, prostate cancer, rectal cancer, renal cell carcinoma, transitional cell carcinoma of the renal pelvis and ureter, retinoblastoma, rhabdomyosarcoma, salivary gland cancer, sarcoma, skin cancer, cutaneous Merkel cell carcinoma, small intestine cancer, soft tissue sarcoma, squamous cell carcinoma, gastric cancer, T-cell lymphoma, laryngeal cancer, thymoma, thymic carcinoma, thyroid cancer, trophoblastic tumor (pregnancy), cancer of unknown primary site, urethral cancer, uterine sarcoma, vaginal cancer, vulvar cancer, Waldenstrom macroglobulinemia, and Wilms tumor.

Images