C-3 pyrazole substituted indazole derivative and application thereof

Abstract

The invention belongs to the field of medicinal chemistry, and particularly relates to a C-3 pyrazole substituted indazole derivative (a compound of a general formula, a stereoisomer, a pharmaceutically acceptable salt, a hydrate, a solvate or a prodrug thereof) and application (application in preparation of a therapeutic agent, especially a TRK inhibitor). The derivative is a C-3 pyrazole substituted indazole compound with a structure as shown in a general formula (I), or a stereoisomer, a pharmaceutically acceptable salt, a hydrate, a solvate or a prodrug of the C-3 pyrazole substituted indazole compound, the C-3 pyrazole substituted indazole derivative with the structure as shown in the general formula (I), and the stereoisomer, the pharmaceutically acceptable salt, the hydrate, the solvate or the prodrug of the derivative have the activity as a protein kinase inhibitor, in particular to TRK kinase. # imgabs0 #

Description

Technical Field

[0001] The present invention belongs to the field of pharmaceutical chemistry, and specifically relates to a C-3 pyrazole-substituted indazole derivative (a general compound, a stereoisomer thereof, a pharmaceutically acceptable salt, a hydrate, a solvate or a prodrug) and its use (use in the preparation of a therapeutic agent, especially a TRK inhibitor). Background Art

[0002] TRKs, the full name of which is tropomyosin receptor kinase, is a member of the transmembrane receptor tyrosine kinase family. TRK is divided into three subtypes, TRKA, TRKB and TRKC, according to the genes encoding them, which are encoded by NTRK1, NTRK2 and NTRK3 respectively. TRK receptor proteins play a vital role in the development and differentiation of neurons, and the types and numbers of neurotrophic factors and neurotrophic factor receptors vary according to the different sites. The main endogenous ligand of TRKA is nerve growth factor (NGF), the ligand of TRKB mainly includes brain-derived neurotrophic factor (BDNF) and neurotrophin 4 (NT-4), and the only endogenous ligand of TRKC is neurotrophin 3 (NT-3). In normal physiological processes, TRK is mainly activated by its endogenous ligands, thereby activating its downstream signaling pathways such as RAS / ERK, PI3K / AKT and PLC-γ1, regulating cell proliferation, differentiation, migration and apoptosis, and the signal transduction process is regulated by its transmembrane extracellular receptor binding region. Under abnormal circumstances, unregulated overactivation of TRK will continuously induce cell differentiation and proliferation, leading to the generation and progression of cancer.

[0003] To date, people have found overactivation of TRK in many types of cancer, with specific characteristics such as NTRK mutation, TRKs overexpression and NTRK gene fusion, which have been found in common and rare cancers, including colorectal cancer, lung cancer, large cell neuroendocrine carcinoma and non-small cell lung cancer (NSCLC), melanoma, acute myeloid leukemia, breast cancer, skin cancer (such as basal cell carcinoma) and lung cancer, neuroblastoma, cylindroma and other tumors. NTRK gene fusion is the most likely to cause cancer. Similar to the oncogenic fusion of ALK and ROS1 genes, the gene feature generally refers to the in-frame fusion of the 3' end of the NTRK gene containing the catalytic tyrosine kinase domain with the 5' end of the partner gene. Since the extracellular domain of the TRK fusion protein has certain structural or functional deficiencies, antibodies targeting TRKs or their ligands will not be effective anticancer drugs. Blocking the abnormal signal transduction pathway of TRKs through small molecule inhibitors is an effective means to treat NTRK gene fusion cancers. The successful launch of larotrectinib proves the feasibility of this strategy. Its clinical data on 17 NTRK fusion cancers, including salivary gland mammary secretory carcinoma (MASC), infantile fibrosarcoma, melanoma, gastrointestinal stromal tumor (GIST), thyroid cancer, colorectal adenocarcinoma or lung adenocarcinoma, show that larotrectinib has a significant therapeutic effect on patients regardless of gene fusion type (NTRK1, NTRK2 or NTRK3 and partner gene) and histology, and is unrelated to age (4 months to 76 years old). However, after long-term use, patients will develop drug resistance, which reduces the efficacy of larotrectinib. Summary of the invention

[0004] The object of the present invention is to provide a novel C-3 pyrazole-substituted indazole derivative and its use in the preparation of therapeutic agents, especially TRK inhibitors.

[0005] In order to achieve the above object, the present invention adopts the following technical solution:

[0006] A C-3 pyrazole-substituted indazole derivative, wherein the derivative is a C-3 pyrazole-substituted indazole compound having a structure represented by general formula (I), or a stereoisomer, a pharmaceutically acceptable salt, a hydrate, a solvate or a prodrug thereof;

[0007]

[0008] Wherein, L is selected from amide Urea Fragment;

[0009] Ring A is selected from aryl or heteroaryl which is unsubstituted or substituted with 1 to 3 R a which may be the same or different;

[0010] Ra is selected from halogen, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, C1-C6 haloalkoxy, C1-C6 alkylamino, C3-C6 cycloalkyl, 0-2 aryl groups or heteroaryl groups which may be the same or different from Rb; Rb is selected from H, halogen, C1-C6 alkyl;

[0011] Preferably, the derivative is a C-3 pyrazole-substituted indazole compound having a structure represented by general formula (I), or a stereoisomer, a pharmaceutically acceptable salt, a hydrate, a solvate or a prodrug thereof;

[0012] wherein L is selected from amide and urea fragments;

[0013] Ring A is selected from aryl or heteroaryl which is unsubstituted or substituted by 1-3 identical or different Ra; Ra is selected from halogen, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 alkoxy, C1-C4 haloalkoxy, amine, C3-C6 cycloalkyl, phenyl, piperazinyl, morpholinyl which are substituted by 0-2 identical or different Rb; Rb is selected from H, halogen, C1-C6 alkyl;

[0014] Further preferably, the derivative is a C-3 pyrazole-substituted indazole compound having a structure represented by general formula (I), or a stereoisomer, a pharmaceutically acceptable salt, a hydrate, a solvate or a prodrug thereof;

[0015] wherein L is selected from amide and urea fragments;

[0016] Ring A is selected from aryl or heteroaryl which is unsubstituted or substituted by 1-3 identical or different Ra; Ra is selected from halogen, methyl, ethyl, isopropyl, tert-butyl, cyclohexyl, trifluoromethyl, dimethylamino, methylpiperazinyl, morpholinyl, C6 cycloalkyl, phenyl which is substituted by 0-2 identical or different Rb; Rb is selected from H, fluorine, chlorine, bromine;

[0017] Still more preferably, the derivative is a C-3 pyrazole-substituted indazole compound having a structure represented by general formula (I), or a stereoisomer, a pharmaceutically acceptable salt, a hydrate, a solvate or a prodrug thereof;

[0018] wherein L is selected from amide and urea fragments;

[0019] Ring A is selected from phenyl, isoxazolyl, pyrazolyl which are unsubstituted or substituted with 1-3 identical or different Ra; Ra is selected from halogen, methyl, ethyl, isopropyl, tert-butyl, cyclohexyl, trifluoromethyl, dimethylamino, methylpiperazinyl, morpholinyl, C6 cycloalkyl, phenyl which is substituted with 0-2 identical or different Rb; Rb is selected from H, fluorine, chlorine, bromine;

[0020] More preferably, a C-3 pyrazole-substituted indazole derivative having a structure represented by general formula (I), or a stereoisomer, a pharmaceutically acceptable salt, a hydrate, a solvate or a prodrug thereof;

[0021] N-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)benzamide;

[0022] N-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-fluorobenzamide;

[0023] N-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-chlorobenzamide;

[0024] N-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-bromobenzamide;

[0025] N-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-methylbenzamide;

[0026] N-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-(trifluoromethyl)benzamide;

[0027] 1-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-phenylurea;

[0028] 1-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-(3-fluorophenyl)urea;

[0029] 1-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-(3-chlorophenyl)urea;

[0030] 1-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-(3-bromophenyl)urea;

[0031] 1-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-(m-tolyl)urea;

[0032] 1-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-(3-(trifluoromethyl)phenyl)urea;

[0033] 1-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-(3-isopropylphenyl)urea;

[0034] 1-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-(3-(tert-butyl)phenyl)urea;

[0035] 1-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-(3-(dimethylamino)phenyl)urea;

[0036] 1-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-(3-morpholinophenyl)urea;

[0037] 1-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-(3-(tert-butyl)isoxazol-5-yl)urea;

[0038] 1-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-(5-(tert-butyl)isoxazol-3-yl)urea;

[0039] 1-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-(3-(tert-butyl)-1-methyl-1H-pyrazol-5-yl)urea;

[0040] 1-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-(3-(tert-butyl)-1-isopropyl-1H-pyrazol-5-yl)urea;

[0041] 1-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-(1,3-di-tert-butyl-1H-pyrazol-5-yl)urea;

[0042] 1-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-(3-(tert-butyl)-1-cyclohexyl-1H-pyrazol-5-yl)urea;

[0043] 1-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-(3-(tert-butyl)-1-phenyl-1H-pyrazol-5-yl)urea;

[0044] 1-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-(3-(tert-butyl)-1-(4-fluorophenyl)-1H-pyrazol-5-yl)urea;

[0045] 1-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-(3-(tert-butyl)-1-(4-chlorophenyl)-1H-pyrazol-5-yl)urea;

[0046] 1-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-(1-(4-bromophenyl)-3-(tert-butyl)-1H-pyrazol-5-yl)urea;

[0047] 1-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-(3-(tert-butyl)-1-(2-fluorophenyl)-1H-pyrazol-5-yl)urea;

[0048] 1-((3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-(3-(tert-butyl)-1-(3-fluorophenyl)-1H-pyrazol-5-yl)urea;

[0049] 1-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-(3-(tert-butyl)-1-(3,4-difluorophenyl)-1H-pyrazol-5-yl)urea;

[0050] 1-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-(3-(tert-butyl)-1-(3,5-difluorophenyl)-1H-pyrazol-5-yl)urea;

[0051] 1-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-(3-(tert-butyl)-1-(2,5-difluorophenyl)-1H-pyrazol-5-yl)urea;

[0052] 1-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-(3-(tert-butyl)-1-(2,4-difluorophenyl)-1H-pyrazol-5-yl)urea;

[0053] The compound represented by the general formula I, and its geometric isomers or pharmaceutically acceptable salts, hydrates, solvates or prodrugs thereof are used in the preparation of drugs for preventing or treating diseases related to the expression or activity of TRK kinase.

[0054] The compound represented by the general formula I, and its geometric isomers or pharmaceutically acceptable salts, hydrates, solvates or prodrugs thereof are used in the preparation of preventive or anti-tumor drugs.

[0055] A pharmaceutical composition comprising a therapeutically effective amount of a C-3 pyrazole-substituted indazole derivative or a stereoisomer thereof, a pharmaceutically acceptable salt, hydrate, solvate or prodrug according to any one of claims 1 to 5 and a pharmaceutically acceptable carrier or excipient.

[0056] The composition is used in preparing medicine for preventing or treating tumors, cancers or severe pain caused by different reasons.

[0057] The composition is used in preparing preventive or anti-tumor drugs.

[0058] The following is an explanation of some of the terms involved in the present invention:

[0059] Halogen: refers to fluorine, chlorine, bromine or iodine. Alkyl: straight chain or branched chain alkyl, such as methyl, ethyl, propyl, isopropyl, n-butyl or tert-butyl. Cycloalkyl: substituted or unsubstituted cyclic alkyl, such as cyclopropyl, cyclopentyl or cyclohexyl. Substituents such as methyl, halogen, etc. Haloalkyl: straight chain or branched chain alkyl, the hydrogen atoms on these alkyl groups can be partially or completely replaced by halogen atoms, for example, chloromethyl, dichloromethyl, trichloromethyl, fluoromethyl, difluoromethyl, trifluoromethyl, etc. Alkoxy: straight chain or branched chain alkyl, the hydrogen atoms of the hydroxyl group can be replaced by these straight chain or branched chain alkyl groups, for example, methyloxy, ethyloxy, propyloxy, isopropyloxy, etc.

[0060] In a second aspect, the present invention provides a method for preparing a C-3 pyrazole-substituted indazole derivative or a stereoisomer, a pharmaceutically acceptable salt, a hydrate, a solvate or a prodrug thereof represented by the general formula (I) described in the first aspect above:

[0061]

[0062] 6-Bromo-1H-indazole is used as a starting material, and undergoes an iodination reaction with iodosuccinimide to obtain intermediate 2. Intermediate 2 reacts with 3,4-dihydro-2H-pyran to obtain 3. Intermediate 3 undergoes a Suzuki coupling reaction with 1-THP-4-pyrazoleboronic acid pinacol ester to obtain 4. Intermediate 4 undergoes a Buchwald-Hartwig reaction with m-nitroaniline to obtain 5. Intermediate 5 is reduced with hydrazine hydrate and Pd / C to obtain intermediate 6. Intermediate 6 undergoes a condensation reaction with the corresponding aromatic amine to obtain intermediate 7 or 9. Intermediate 7 or 9 is deprotected with p-toluenesulfonic acid to obtain the final product 8 or 10.

[0063] The raw material 11 undergoes an acylation reaction with phenyl chloroformate to obtain an intermediate 12, and the intermediate 12 undergoes a condensation reaction with the intermediate 6 to obtain an intermediate 13. The intermediate 13 is deprotected with toluenesulfonic acid to obtain the final product 14.

[0064] In the above preparation process, R 1 is H, methyl, trifluoromethyl, fluorine, chlorine, bromine, R 2 is phenyl or isoxazolyl which is unsubstituted or substituted with one Ra; Ra is H, fluorine, chlorine, bromine, methyl, isopropyl, tert-butyl, trifluoromethyl, dimethylamino, methylpiperazinyl or morpholinyl; R 3 It is methyl, isopropyl, tert-butyl, cyclohexyl, or phenyl which is unsubstituted or substituted by one Rb; Rb is H, fluorine, chlorine, or bromine.

[0065] Further, 6-bromo-1H-indazole is used as a starting material, and an iodination reaction is carried out with iodosuccinimide under high temperature conditions to obtain intermediate 2, the high temperature reaction temperature is 50-80°C, preferably 65°C, and the reaction solvent can be dichloromethane, acetonitrile, acetone, N,N-dimethylformamide, preferably N,N-dimethylformamide; intermediate 2 is reacted with 3,4-dihydro-2H-pyran under low temperature conditions to obtain intermediate 3, the reaction solvent can be acetonitrile, tetrahydrofuran, dichloromethane, preferably dichloromethane, and the reaction temperature is 0-30°C, preferably 30°C; intermediate 31-THP-4-pyrazoleboronic acid pinacol ester undergoes Suzuki coupling reaction to obtain intermediate 4, the reaction temperature is 60-120°C, preferably 80°C, and the reaction solvent can be N,N-dimethylformamide, toluene, 1,4-dioxane, preferably N,N-dimethylformamide; raw material 5 undergoes reduction reaction to obtain intermediate 6, and the reduction conditions can be Fe / NH 4 Cl,H 2 and Pd / C, hydrazine hydrate and Pd / C, etc., preferably hydrazine hydrate and Pd / C, the reaction solvent can be methanol, ethanol, acetonitrile, etc., preferably ethanol, the reaction temperature is 45-80°C, preferably 80°C; intermediate 6 undergoes acylation reaction with corresponding aromatic amine under low temperature conditions to obtain intermediate 7, the low temperature reaction temperature is 0-30°C, preferably 30°C, the reaction solvent can be N,N-dimethylformamide, tetrahydrofuran, preferably N,N-dimethylformamide; intermediate 6 undergoes condensation reaction with corresponding aromatic amine to obtain final product 9, the reaction temperature is 0-30°C, preferably 30°C, the reaction solvent can be tetrahydrofuran, dichloromethane, etc., preferably tetrahydrofuran; intermediate 7 or 9 reacts with p-toluenesulfonic acid to obtain final product 8 or 10, the reaction solvent can be ethanol:ethyl acetate:water=3:3:1, the reaction temperature is 40-80°C, preferably 50°C.

[0066] The raw material 11 undergoes an acylation reaction with phenyl chloroformate to obtain an intermediate 12, the reaction temperature is 0-30°C, preferably 30°C, and the reaction solvent can be N,N-dimethylformamide, dichloromethane, tetrahydrofuran, preferably tetrahydrofuran; the intermediate 12 undergoes a condensation reaction with the intermediate 6 to obtain the intermediate 13, the reaction temperature is 30-60°C, preferably 50°C, and the reaction solvent can be N,N-dimethylformamide, dichloromethane, tetrahydrofuran, preferably tetrahydrofuran; the intermediate 13 is deprotected with toluenesulfonic acid to obtain the final product 14, the reaction solvent can be ethanol:ethyl acetate:water=3:3:1, and the reaction temperature is 40-80°C, preferably 50°C.

[0067] In the third aspect, the present invention provides the use of C-3 pyrazole-substituted indazole derivatives, the compounds represented by the general formula I, and their geometric isomers or pharmaceutically acceptable salts, hydrates, solvates or prodrugs in the preparation of drugs for preventing or treating diseases related to the expression or activity of TRK kinase.

[0068] In a fourth aspect, the compound represented by the general formula I, and its geometric isomers or pharmaceutically acceptable salts, hydrates, solvates or prodrugs thereof are used in the preparation of preventive or anti-tumor drugs.

[0069] In a fifth aspect, the present invention provides a pharmaceutical composition comprising a therapeutically effective amount of the C-3 pyrazole-substituted indazole derivative or its stereoisomer, pharmaceutically acceptable salt, hydrate, solvate or prodrug described in the first aspect above and a pharmaceutically acceptable carrier or excipient.

[0070] In the sixth aspect, the present invention provides the use of the composition described in the first aspect above in the preparation of a drug for preventing or treating a disease related to the expression or activity of TRK kinase, characterized in that: preferably, the disease is a tumor, cancer or severe pain caused by different reasons.

[0071] In a seventh aspect, the composition is used in the preparation of preventive or anti-tumor drugs.

[0072] Compared with the prior art, the present invention has the following beneficial effects:

[0073] The present invention designs a series of novel C-3 pyrazole-substituted indazole derivatives for NTRK gene fusion cancers, and finds that compounds with such structures exhibit good inhibitory activity against wild-type enzymes and various mutants of TRK kinases, and can be used to treat tumors caused by NTRK gene fusions, overcome clinical drug resistance caused by TRK mutations, or other diseases related to abnormal TRK expression. DETAILED DESCRIPTION

[0074] The present invention is further described below with reference to specific examples. It should be understood that the specific examples described herein are only used to explain the present invention and are not used to limit the scope of the present invention. The H NMR spectrum of the compound was measured using a Bruker ARX-400; all reagents used were analytically pure or chemically pure.

[0075] If no specific techniques or conditions are specified in the examples, the techniques or conditions described in the literature in the field or the product instructions are used. If no manufacturer is specified for the reagents or instruments used, they are all conventional products that can be purchased through regular channels.

[0076] The experimental methods in the following examples are conventional methods unless otherwise specified. The experimental materials used in the following examples are commercially available products unless otherwise specified.

[0077] The preparation route of Example 1 is as follows:

[0078]

[0079] The specific synthesis steps are as follows:

[0080] Synthesis of 6-bromo-3-iodo-1H-indazole (2)

[0081] Dissolve 6-bromo-1H-indazole (4.67 g, 23.63 mmol) in N,N-dimethylformamide, add iodosuccinimide (5.32 g, 23.63 mmol) at room temperature. Heat the reaction solution to 65°C and stir for 5 hours. Cool to room temperature, monitor the reaction of the raw materials by TLC, slowly add 300 mL of water, a large amount of solid precipitates, stir at room temperature for 30 minutes, filter, and filter to obtain 7.59 g of white solid, with a yield of 99%.

[0082] Synthesis of 6-bromo-3-iodo-1-(tetrahydro-2H-pyran-2-yl)-1H-indazole (3)

[0083] Intermediate 2 (7.59 g, 23.5 mmol) and p-toluenesulfonic acid (0.2 g, 1.17 mmol) were dissolved in DCM. DHP (5.93 g, 70.51 mmol) in DCM was slowly dripped into the reaction solution at 0°C, and the mixture was moved to room temperature for 5 h. After the reaction of the raw materials was completed, the reaction solution was diluted with DCM and then washed with H 2 O (50 mL x 3) and saturated sodium chloride for extraction. The organic layer was collected and purified by silica gel column chromatography (1%-5% ethyl acetate / petroleum ether) to obtain 5.39 g of a white solid with a yield of 72%.

[0084] Synthesis of 6-bromo-1-(tetrahydro-2H-pyran-2-yl)-3-(1-(tetrahydro-1H-pyran-2-yl)-1H-pyrazol-4-yl)-1-indazole (4)

[0085] Intermediate 3 (5.39 g, 13.24 mmol), 1-THP-4-pyrazoleboronic acid pinacol ester (4.42 g, 15.89 mmol) and Cs 2 CO 3 (8.6 g, 26.48 mmol) was dissolved in DMF solution and purged with argon for 10 min before adding Pd(PPh 3 ) 4 (0.76 g, 0.66 mmol). The reaction solution was heated to 80°C and reacted overnight. After TLC monitoring, the reaction of the raw material was complete, the solvent was removed, the mixture was dissolved in ethyl acetate, and then H 2 O (100 mL×3) and saturated sodium chloride (100 mL×2) were used for extraction. The organic layer was collected and purified by silica gel column chromatography (5%-15% ethyl acetate / petroleum ether) to obtain 3.6 g of a white solid with a yield of 63.1%.

[0086] Synthesis of N-(3-nitrophenyl)-1-(tetrahydro-2H-pyran-2-yl)-3-(1-(tetrahydro-1H-pyran-2-yl)-1H-pyrazol-4-yl)-1H-indazol-6-amine (5)

[0087] Intermediate 4 (1.88 g, 4.37 mmol), m-nitroaniline (0.6 g, 4.37 mmol), Xantphos (0.5 g, 0.87 mmol) and Cs 2 CO 3 (2.99 g, 9.19 mmol) was dissolved in 1,4-dioxane and purged with argon for 10 min before adding Pd(OAc) 2 (0.05 g, 0.22 mmol). The reaction solution was heated to 80°C for 4 hours. After TLC monitoring, the reaction of the raw material was complete, the solvent was removed, and the solution was dissolved with EA, and then H 2 The mixture was extracted with 100 mL x 3 and saturated sodium chloride (100 mL x 2). The organic phase was collected and purified by silica gel column chromatography (10%-30% ethyl acetate / petroleum ether) to obtain 1.4 g of a medium white solid with a yield of 65.7%.

[0088] N 1 Synthesis of -(1-(tetrahydro-2H-pyran-2-yl)-3-(1-(trihydro-2H-pyridin-2-yl)-1H-pyrazol-4-yl)-1-indazol-6-yl)benzene-1,3-diamine (6)

[0089] Intermediate 5 (1.4 g, 2.95 mmol) was dissolved in ethanol, and hydrazine hydrate (0.75 g, 14.7 mmol) and Pd / C (0.18 g) were added under stirring at room temperature. The reaction solution was heated to 60°C and stirred for 2 hours. After the reaction of the raw material was completed by TLC monitoring, Pd / C was filtered out, the filtrate was collected, the solvent was removed by rotation, and it was directly used in the next step without further purification.

[0090] Synthesis of N-(3-((1-(tetrahydro-2H-pyran-2-yl)-3-(1-(trihydro-2H-pyran-2-yl)-1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)benzamide (7)

[0091] Intermediate 6 (0.10 g, 0.22 mmol) and benzoic acid (0.1 g, 0.26 mmol) were dissolved in DMF, and DIPEA (0.09 g, 0.66 mmol) and HATU (0.03 g, 0.26 mmol) were added. The reaction solution was stirred at room temperature for 12 hours. After TLC monitoring, the reaction of the raw materials was completed, and the reaction solution was slowly poured into water, and a white solid precipitated. Filter, collect the filter cake, and dry to obtain 0.1 g of a white solid with a yield of 81.3%.

[0092] Preparation of N-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)benzamide (Example 1)

[0093]

[0094] Intermediate 7 (0.1 g, 0.18 mmol) was dissolved in a mixed solvent of ethanol: ethyl acetate: water = 3:3:1, and p-toluenesulfonic acid (0.09 g, 0.53 mmol) was added. The reaction solution was heated to 50 °C and stirred for 2 hours. After TLC monitoring, the reaction of the raw material was completed, diluted with EA, and then washed with H 2 The organic phase was collected and purified by silica gel column chromatography (1%-5% methanol / dichloromethane) to obtain 0.03 g of compound 8 (ie, the compound of Example 1). The yield was 42.8%. 1 HNMR (600 MHz, DMSO-d 6)δ13.04(s,1H),12.45(s,1H),10.19(s,1H),8.39(s,1H),8.31(s,1H),8. 01(s,1H),7.96-7.93(m,2H),7.88(d,J=8.7Hz,1H),7.79(t,J=2.2Hz,1H), 7.61-7.57(m,1H),7.55-7.51(m,2H),7.30-7.27(m,1H),7.24(t,J=8.0Hz, 1H),7.14(d,J=2.0Hz,1H),6.94(dd,J=8.7,1.9Hz,1H),6.91-6.87(m,1H). 13 C NMR (151 MHz, DMSO-d 6 )δ166.03,144.14,143.01,142.79,140.56,138.15,137.09,135.56,131.99,129.71,128.84 (2C),128.13(2C),126.16,121.65,115.33,115.01,114.55,113.53,112.76,109.55,94.54.

[0095] Preparation of N-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-fluorobenzamide (Example 2)

[0096]

[0097] Referring to the method for preparing Example 1, the benzoic acid in step f is replaced with m-fluorobenzoic acid in equal proportion to obtain Example 2. 1 H NMR (600 MHz, DMSO-d 6 )δ13.04(s,1H),12.46(s,1H),10.25(s,1H),8.40(s,1H),8.31(s,1H),8.01(s,1H) ,7.88(d,J=8.7Hz,1H),7.81(d,J=7.7Hz,1H),7.76(q,J=4.5,2.9Hz,2H),7.59(td, J=8.0,5.7Hz,1H),7.45(td,J=8.6,2.7Hz,1H),7.28(d,J=8.2Hz,1H),7.25(t,J=7. 9Hz, 1H), 7.14 (d, J = 1.9Hz, 1H), 6.94 (dd, J = 8.7, 1.9Hz, 1H), 6.90 (d, J = 7.8Hz, 1H). 13 C NMR (151 MHz, DMSO-d6 )δ164.58,162.39(d,J=244.5Hz),144.22,142.99,142.71,140.28,138.16,137.83(d,J=6.5Hz),131.05(d,J=7.8Hz),129. 77,126.17,124.37,121.66,118.97,118.83,115.38,115.01(d,J=3.8Hz),114.88,114.57,113.70,112.75,109.49,94.68.

[0098] Preparation of N-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-chlorobenzamide (Example 3)

[0099]

[0100] Referring to the method for preparing Example 1, the benzoic acid in step f is replaced with m-chlorobenzoic acid in equal proportion to obtain Example 3. 1 H NMR (600 MHz, DMSO-d 6 )δ13.07(s,1H),12.48(s,1H),10.31(s,1H),8.43(s,1H),8.31(s,1H),8.01(t,J=2.1Hz,2 H),7.92(dt,J=7.9,1.2Hz,1H),7.88(d,J=8.7Hz,1H),7.76(t,J=2.1Hz,1H),7.66(ddd,J= 8.0,2.2,1.0Hz,1H),7.57(t,J=7.9Hz,1H),7.29(dt,J=8.4,1.4Hz,1H),7.24(t,J=8.0Hz, 1H), 7.14 (d, J=1.9Hz, 1H), 6.94 (dd, J=8.7, 1.9Hz, 1H), 6.90 (ddd, J=7.9, 2.2, 1.1Hz, 1H). 13 C NMR (151 MHz, DMSO-d 6 )δ164.50,144.22,142.99,142.71,140.28,138.14,137.48,137.07,133.65,131.83,130.87,12 9.75,127.89,127.00,126.14,121.65,115.36,115.00,114.56,113.71,112.75,109.50,94.69.

[0101] Preparation of N-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-bromobenzamide (Example 4)

[0102]

[0103] Referring to the method for preparing Example 1, the benzoic acid in step f is replaced with m-bromobenzoic acid in equal proportion to obtain Example 4. 1 H NMR (600 MHz, DMSO-d 6 )δ13.04(s,1H),12.47(s,1H),10.29(s,1H),8.41(s,1H),8.14(t,J=1.9Hz,3H ),7.95(d,J=7.8Hz,1H),7.88(d,J=8.7Hz,1H),7.79(dd,J=8.0,2.0Hz,1H),7. 76(t,J=2.1Hz,1H),7.50(t,J=7.9Hz,1H),7.28(d,J=8.1Hz,1H),7.25(t,J=7. 9Hz,1H),7.14(d,J=1.8Hz,1H),6.94(dd,J=8.8,1.9Hz,1H),6.92-6.87(m,1H). 13 CNMR (151MHz, DMSO-d 6 )δ164.42,144.22,143.00,142.70,140.27,138.14,137.67,134.73,131.13(2C),130.70(2C ),129.77,127.36,122.14,121.67,115.37,115.00,114.59,113.72,112.73,109.47,94.70.

[0104] Preparation of N-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-methylbenzamide (Example 5)

[0105]

[0106] Referring to the method for preparing Example 1, the benzoic acid in step f is replaced with m-toluic acid in equal proportion to obtain Example 5. 1 H NMR (600 MHz, DMSO-d 6)δ13.03(s,1H),12.45(s,1H),10.14(s,1H),8.38(s,1H),8.30(s,1H),8.02(s ,1H),7.87(d,J=8.8Hz,1H),7.80-7.75(m,2H),7.74(dt,J=6.9,2.1Hz,1H),7.4 4-7.37(m,2H),7.28(dt,J=8.1,1.5Hz,1H),7.23(t,J=8.0Hz,1H),7.13(d,J=1. 9Hz, 1H), 6.94 (dd, J=8.7, 1.9Hz, 1H), 6.88 (dd, J=7.3, 2.1Hz, 1H), 2.40 (s, 3H). 13 C NMR (151 MHz, DMSO-d 6 )δ166.12,144.13,143.01,142.79,140.60,138.14,137.05,135.56,132.56,130.13,129.69,128.7 4,128.60,126.24,125.29,121.65,115.32,115.01,114.55,113.48,112.74,109.53,94.53,21.44.

[0107] Preparation of N-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-(trifluoromethyl)benzamide (Example 6)

[0108]

[0109] Referring to the method for preparing Example 1, the benzoic acid in step f is replaced with m-trifluoromethylbenzoic acid in equal proportion to obtain Example 6. 1 H NMR (600 MHz, DMSO-d 6 )δ13.05(s,1H),12.47(s,1H),10.42(s,1H),8.42(s,1H),8.31(s,1H),8.28(d,J=1 .8Hz,1H),8.28-8.25(m,1H),8.01(s,1H),7.98-7.95(m,1H),7.88(d,J=8.7Hz,1H) ,7.79(t,J=7.9Hz,1H),7.76(t,J=2.0Hz,1H),7.29(dt,J=8.0,1.6Hz,1H),7.26(t, J=7.8Hz,1H),7.14(d,J=2.0Hz,1H),6.95(dd,J=8.7,1.9Hz,1H),6.93-6.90(m,1H).13 C NMR (151 MHz, DMSO-d 6 )δ164.55,144.23,142.98,142.72,140.14,138.15,137.10,136.34,132.30,130.22,129.83,129.74,129.53,128.61, 128.58,126.20,125.36,124.70(q,J=3.9Hz),123.55,121.68,115.37,114.96,114.65,113.86,112.87,109.60,94.75.

[0110] The preparation route of Example 7 is as follows:

[0111]

[0112] The specific synthesis steps are as follows:

[0113] Synthesis of 1-phenyl-3-(3-(((1-(tetrahydro-2H-pyran-2-yl)-3-(1-(trihydro-2H-pyran-2-yl]-1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)urea (9)

[0114] Aniline (0.04 g, 0.44 mmol) was dissolved in dry dichloromethane, and DIPEA (0.09 g, 0.66 mmol) and triphosgene (0.04 g, 0.13 mmol) were added in sequence at 0°C, with argon protection. After stirring at 0°C for 3 hours, intermediate 6 (0.1 g, 0.22 mmol) was added to the reaction solution, which was moved to room temperature and stirred at room temperature for 12 hours. TLC detection showed that there was a slight residue of raw materials, and the reaction solution was diluted with DCM, and then H 2 O (100 mL×3) and saturated sodium chloride (100 mL×2) were used for extraction. The organic phase was collected and purified by silica gel column chromatography (1%-4% methanol / dichloromethane) to obtain 0.08 g of intermediate 9 with a yield of 63.4%.

[0115] Preparation of 1-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-phenylurea (Example 7)

[0116]

[0117] Compound 10 (Example 7) was obtained by referring to step g of Preparation Example 1. 1 H NMR (600 MHz, DMSO-d 6)δ13.03(s,1H),12.44(s,1H),8.62(d,J=11.3Hz,2H),8.33(s,1H),8.30 (s,1H),8.00(s,1H),7.86(d,J=8.7Hz,1H),7.48-7.39(m,3H),7.27(t,J =7.8Hz,2H),7.16(t,J=8.0Hz,1H),7.11(d,J=2.0Hz,1H),6.96(t,J=7.3Hz,1H),6.90(ddd,J=14.3,8.4,2.0Hz,2H),6.77(dd,J=8.0,2.2Hz,1H). 13 C NMR (151 MHz, DMSO-d 6 )δ152.94,144.31,143.00,142.80,141.08,140.18,138.14,130.13,129.91,129.27(2C ),122.27,121.64,118.61(2C),115.29,115.00,114.57,111.58,110.63,107.44,94.48.

[0118] Preparation of 1-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-(3-fluorophenyl)urea (Example 8)

[0119]

[0120] Referring to the method for preparing Example 7, the aniline in step h was replaced with m-fluoroaniline in equal proportion to obtain Example 8. 1 H NMR (600 MHz, DMSO-d 6 )δ13.03(s,1H),12.44(s,1H),8.85(s,1H),8.70(s,1H),8.34(s,1H),8.30( s,1H),8.00(s,1H),7.86(d,J=8.7Hz,1H),7.48(dt,J=12.0,2.3Hz,1H),7.4 1(t,J=2.1Hz,1H),7.29(td,J=8.2,6.9Hz,1H),7.17(t,J=8.0Hz,1H),7.11( dd,J=8.5,2.0Hz,2H),6.91(ddd,J=9.8,8.1,1.9Hz,2H),6.82-6.74(m,2H).

[0121] Preparation of 1-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-(3-chlorophenyl)urea (Example 9)

[0122]

[0123] Referring to the method for preparing Example 7, the aniline in step h was replaced with m-chloroaniline in equal proportion to obtain Example 9. 1 H NMR (600 MHz, DMSO-d 6 )δ13.04(s,1H),12.44(s,1H),8.95(s,1H),8.82(s,1H),8.34(s,1H),8.31(s,1 H),8.01(s,1H),7.87(d,J=8.7Hz,1H),7.71(t,J=2.1Hz,1H),7.43(t,J=2.2Hz, 1H),7.31-7.25(m,2H),7.17(t,J=8.0Hz,1H),7.12(d,J=1.9Hz,1H),7.01(dt,J =7.5,1.8Hz,1H),6.91(ddd,J=7.3,5.4,2.0Hz,2H),6.79(dd,J=8.1,2.2Hz,1H). 13 C NMR (151 MHz, DMSO-d 6 )δ152.83,144.32,143.00,142.79,141.79,140.80,138.15,137.10,133.67,130.88,129.94,12 6.18,121.86,121.65,117.95,117.06,115.30,114.98,114.59,111.82,110.83,107.64,94.51.

[0124] Preparation of 1-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-(3-bromophenyl)urea (Example 10)

[0125]

[0126] Referring to the method for preparing Example 7, the aniline in step h was replaced with m-bromoaniline in equal proportion to obtain Example 10. 1 H NMR (600 MHz, DMSO-d 6)δ12.99(s,1H),12.45(s,1H),9.06(s,1H),8.92(s,1H),8.34(s,1H),8.16(s,2H),7.90-7.81(m,2H),7.43(s,1H),7.32( d,J=8.1Hz,1H),7.23(t,J=8.0Hz,1H),7.17(t,J=8.0Hz,1H),7.15-7.09(m,2H),6.94-6.90(m,2H),6.79(d,J=8.0Hz,1H). 13 C NMR (151 MHz, DMSO-d 6 )δ152.84,144.31,143.00,142.79,142.00,140.86,138.15,131.18,130.13,129.91,124.7 1,122.20,121.65,120.76,117.41,115.30,115.00,114.56,111.77,110.80,107.64,94.48.

[0127] Preparation of 1-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-(3-bromophenyl)urea (Example 11)

[0128]

[0129] Referring to the method for preparing Example 7, the aniline in step h was replaced with m-methylaniline in equal proportions to obtain Example 11. 1 H NMR (600 MHz, DMSO-d 6 )δ13.03(s,1H),12.44(s,1H),8.63(s,1H),8.55(s,1H),8.33(s,1H),8.31(s,1H),8.0 1(s,1H),7.86(d,J=8.8Hz,1H),7.43(t,J=2.1Hz,1H),7.28(d,J=2.0Hz,1H),7.23-7.20 (m,1H),7.15(dt,J=9.0,7.9Hz,2H),7.11(d,J=1.8Hz,1H),6.91(dd,J=8.7,1.9Hz,1H), 6.89(ddd,J=8.0,2.1,0.9Hz,1H), 6.77(dddd,J=8.9,6.2,2.0,0.9Hz,2H), 2.27(s,3H). 13 C NMR (151 MHz, DMSO-d 6)δ152.93,144.30,143.00,142.83,141.11,140.10,138.44,138.15,137.10,129.91,129.12,126.1 7,123.02,121.65,119.12,115.79,115.29,115.00,114.57,111.57,110.61,107.43,94.46,21.69.

[0130] Preparation of 1-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-(3-(trifluoromethyl)phenyl)urea (Example 12)

[0131]

[0132] Referring to the method for preparing Example 7, the aniline in step h was replaced with m-trifluoromethylaniline in equal proportions to obtain Example 12. 1 H NMR (600 MHz, DMSO-d 6 )δ13.03(s,1H),12.44(s,1H),9.14(s,1H),8.88(s,1H),8.35(s,1H),8.30(s,Z 1H),8.01(s,2H),7.87(d,J=8.7Hz,1H),7.57(d,J=8.3Hz,1H),7.50(t,J=7.9Hz,1H),7.43(d,J=2.3Hz,1H),7.3 0(d,J=7.7Hz,1H),7.18(t,J=8.0Hz,1H),7.12(d,J=1.8Hz,1H),6.98-6.89(m,2H),6.81(dd,J=8.0,2.2Hz,1H). 13 C NMR (151 MHz, DMSO-d 6 )δ152.94,144.33,143.01,142.80,141.06,140.73,138.14,130.42,130.10,129.95,129.89,125.59,123. 78,122.24,121.66,118.51,115.30,114.96,114.61,114.50(d,J=4.4Hz),111.82,110.91,107.80,94.51.

[0133] Preparation of 1-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-(3-isopropylphenyl)urea (Example 13)

[0134]

[0135] Referring to the method for preparing Example 7, the aniline in step h was replaced with 3-isopropylaniline in equal proportion to obtain Example 13. 1 H NMR (600 MHz, DMSO-d 6 )δ13.03(s,1H),12.44(s,1H),8.92(d,J=21.5Hz,2H),8.34(s,1H),8.15(s,2H),7.86(d, J=8.6Hz,1H),7.46(t,J=2.2Hz,1H),7.36(t,J=2.0Hz,1H),7.26(dt,J=8.3,1.3Hz,1H),7. 16(q,J=8.1Hz,2H),7.11(d,J=1.9Hz,1H),6.91(ddd,J=8.5,4.9,2.1Hz,2H),6.84(dt,J= 7.7,1.3Hz,1H),6.78(dd,J=8.1,2.2Hz,1H),2.83(p,J=6.9Hz,1H),1.19(d,J=6.9Hz,6H). 13 C NMR (151 MHz, DMSO-d 6 )δ153.10,149.46,144.23,143.02,142.91,141.32,140.33,138.14,130.13,129.82,129.11,121.62 ,120.21,116.58,116.22,115.24,115.02,114.52,111.40,110.70,107.63,94.31,33.97,24.36(2C).

[0136] Preparation of 1-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-(3-(tert-butyl)phenyl)urea (Example 14)

[0137]

[0138] Referring to the method for preparing Example 7, the aniline in step h was replaced with 3-tert-butylaniline in equal proportion to obtain Example 14. 1 H NMR (600 MHz, DMSO-d 6)δ13.04(s,1H),12.44(s,1H),8.58(d,J=3.7Hz,2H),8.35(s,1H),8.30(s,1H),8.01( s,1H),7.87(d,J=8.6Hz,1H),7.44(dt,J=14.7,2.1Hz,2H),7.28(ddd,J=8.0,2.1,1.0H z,1H),7.18(dt,J=17.8,7.9Hz,2H),7.12(d,J=2.0Hz,1H),7.00(ddd,J=7.8,1.8,1.0H z,1H),6.92(dd,J=8.7,1.9Hz,1H),6.90-6.86(m,1H),6.83-6.77(m,1H),1.27(s,9H). 13 C NMR (151 MHz, DMSO-d 6 )δ153.01,151.76,144.29,143.01,142.86,141.16,139.90,138.15,137.11,129.88,128.91,126.19,121 .64,119.31,115.93,115.63,115.28,115.01,114.55,111.42,110.67,107.61,94.40,34.85,31.60(3C).

[0139] Preparation of 1-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-(3-(dimethylamino)phenyl)urea (Example 15)

[0140]

[0141] Referring to the method for preparing Example 7, the aniline in step h was replaced with 3-(dimethylamino)aniline in equal proportion to obtain Example 15. 1 H NMR (600 MHz, DMSO-d 6)δ13.02(s,1H),12.43(s,1H),8.81(s,1H),8.71(s,1H),8.34(s,1H),8.27(s,1H),8.13-7 .93(m,1H),7.86(d,J=8.7Hz,1H),7.43(d,J=2.2Hz,1H),7.15(t,J=8.0Hz,1H),7.11(d,J= 1.9Hz,1H),7.05(t,J=8.1Hz,1H),6.93-6.90(m,2H),6.88(dd,J=8.1,2.0Hz,1H),6.78(dd ,J=7.9,2.2Hz,1H),6.70(dd,J=7.8,2.0Hz,1H),6.35(dd,J=8.3,2.5Hz,1H),2.87(s,6H).

[0142] Preparation of 1-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-(3-morpholinophenyl)urea (Example 16)

[0143]

[0144] Referring to the method for preparing Example 7, the aniline in step h was replaced with 3-morpholineaniline in equal proportion to obtain Example 16. 1 H NMR (600 MHz, DMSO-d 6 )δ13.03(s,1H),12.43(s,1H),8.83(s,1H),8.76(s,1H),8.34(s,1H),8.30-7.95(m,2H),7.86 (d,J=8.7Hz,1H),7.44(t,J=2.2Hz,1H),7.16(t,J=2.1Hz,1H),7.15(d,J=8.0Hz,1H),7.13-7.0 8(m,2H),6.91(dd,J=8.8,1.9Hz,1H),6.88(dd,J=7.9,2.0Hz,1H),6.82(dd,J=7.8,1.9Hz,1H), 6.78(dd,J=8.0,2.1Hz,1H),6.56(dd,J=8.3,2.4Hz,1H),3.75-3.71(m,4H),3.08-3.04(m,4H). 13 C NMR (151 MHz, DMSO-d 6)δ153.02,152.09,144.23,143.04,142.91,141.16,140.99,138.09,129.87,129.66,121.64,115.2 2,114.96,114.57,111.49,110.68,109.88,109.59,107.63,105.38,94.32,66.57(2C),49.02(2C).

[0145] Preparation of 1-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-(3-(tert-butyl)isoxazol-5-yl)urea (Example 17)

[0146]

[0147] Referring to the method for preparing Example 7, the aniline in step h was replaced with 3-(tert-butyl)isoxazole-5-amine in equal proportions to obtain Example 17. 1 H NMR (600 MHz, DMSO-d 6 )δ13.04(s,1H),12.46(s,1H),10.01(s,1H),8.82(s,1H),8.37(s,1H),8.3 1(s,1H),8.01(s,1H),7.87(d,J=8.7Hz,1H),7.44(t,J=2.1Hz,1H),7.19(t ,J=8.1Hz,1H),7.12(d,J=2.0Hz,1H),6.91(dd,J=8.8,1.9Hz,1H),6.89(dd ,J=7.9,2.0Hz,1H),6.81(dd,J=8.1,2.2Hz,1H),6.04(s,1H),1.25(s,9H). 13 CNMR (151MHz, DMSO-d 6 )δ173.03,162.15,150.10,144.46,142.97,142.62,140.10,138.17,137.08,130.06,126.17 ,121.69,115.40,114.98,114.61,112.35,110.82,107.54,94.72,83.97,32.40,29.52(3C).

[0148] Preparation of 1-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-(5-(tert-butyl)isoxazol-3-yl)urea (Example 18)

[0149]

[0150] Referring to the method for preparing Example 7, the aniline in step h was replaced with 5-(tert-butyl)isoxazole-3-amine in equal proportions to obtain Example 18. 1 H NMR (600 MHz, DMSO-d 6 )δ13.04(s,1H),12.45(s,1H),9.71(s,1H),9.12(s,1H),8.36(s,1H),8.29(s,1H),8.03(s,1H),7.87(d,J=8.7Hz,1H),7.45(t,J=2.2Hz,1 H),7.18(t,J=8.0Hz,1H),7.12(d,J=1.9Hz,1H),6.90(ddd,J=10.1,8.3,2.0Hz,2H),6.81(dd,J=8.0,2.1Hz,1H),6.49(s,1H),1.29(s,9H). 13 C NMR (151 MHz, DMSO-d 6 )δ180.59,158.91,151.88,144.36,142.99,142.73,140.45,138.14,137.05,129.99,126.21 ,121.67,115.32,114.97,114.60,112.12,110.84,107.68,94.55,92.95,32.92,28.82(3C).

[0151] The preparation route of Example 19 is as follows:

[0152]

[0153] The specific synthesis steps are as follows:

[0154] Synthesis of phenyl (3-(tert-butyl)-1-methyl-1H-pyrazol-5-yl) carbamate (12)

[0155] Dissolve 3-(tert-butyl)-1-methyl-1H-pyrazole-5-amine (0.2 g, 1.3 mmol) in dry tetrahydrofuran, add pyridine (0.3 g, 3.9 mmol) and phenyl chloroformate (0.3 g, 1.95 mmol), and stir the reaction solution at room temperature for 3 hours. TLC monitoring shows that the reaction is complete. Remove the solvent, add ethyl acetate to dissolve, and extract with 1N HCl (aq) (50 mL × 3) and saturated sodium chloride. Collect the organic layer and spin dry it, and use it directly in the next step without treatment.

[0156] Synthesis of 1-(3-(tert-butyl)-1-methyl-1H-pyrazol-5-yl)-3-(3-((1-(tetrahydro-2H-pyran-2-yl)-3-(1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)urea (13)

[0157] Intermediate 6 (0.2 g, 0.43 mmol) was dissolved in dry THF, and DIPEA (0.17 g, 1.29 mmol) and intermediate 12 (0.36 g, 1.3 mmol) were added in sequence. The reaction solution was heated to 50°C and stirred for 12 hours. TLC monitoring showed that the reaction was almost complete. The solvent was removed and purified by silica gel column chromatography (1%-4% methanol / dichloromethane) to obtain 0.12 g of intermediate 13 with a yield of 44%.

[0158] Preparation of 1-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-(3-(tert-butyl)-1-methyl-1H-pyrazol-5-yl)urea (Example 19)

[0159]

[0160] Compound 14 (ie, Example 19) was obtained by referring to step g of Preparation Example 1. 1 H NMR (600 MHz, DMSO-d 6 )δ13.02(s,1H),12.44(s,1H),8.87(s,1H),8.47(s,1H),8.33(s,1H), 8.27(s,1H),8.03(s,1H),7.86(d,J=8.7Hz,1H),7.42(t,J=2.2Hz,1H), 7.16(t,J=8.0Hz,1H),7.11(d,J=1.9Hz,1H),6.91(dt,J=8.6,2.3Hz,2H),6.77(dd,J=8.0,2.2Hz,1H),6.05(s,1H),3.60(s,3H),1.20(s,9H). 13 C NMR (151 MHz, DMSO-d 6 )δ159.06,152.21,144.34,142.99,142.75,140.85,138.13,137.59,137.02,129.98,126.22,12 1.66,115.30,114.97,114.61,111.84,110.58,107.32,94.56,94.00,35.36,32.26,30.83(3C).

[0161] Preparation of 1-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-(3-(tert-butyl)-1-isopropyl-1H-pyrazol-5-yl)urea (Example 20)

[0162]

[0163] Referring to the method for preparing Example 19, the 3-(tert-butyl)-1-methyl-1H-pyrazole-5-amine in step i was replaced with 3-(tert-butyl)-1-isopropyl-1H-pyrazole-5-amine in equal proportions to obtain Example 20. 1 H NMR (600 MHz, DMSO-d 6 )δ13.03(s,1H),12.44(s,1H),8.90(s,1H),8.39(s,1H),8.33(s,1H),8.30(s,1H) ,8.01(d,J=6.2Hz,1H),7.86(d,J=8.7Hz,1H),7.40(t,J=2.1Hz,1H),7.16(t,J=8. 0Hz,1H),7.11(d,J=1.8Hz,1H),6.91(dd,J=8.7,1.9Hz,2H),6.77(dd,J=7.9,2.2H z,1H),6.01(s,1H),4.37(hept,J=6.5Hz,1H),1.34(d,J=6.6Hz,6H),1.21(s,9H). 13 C NMR (151 MHz, DMSO-d 6 )δ157.83,151.43,143.24,141.89,141.67,139.89,137.04,135.87,134.87,128.84,125.08,120.55 ,114.20,113.90,113.44,110.60,109.41,106.22,93.43(2C),47.04,45.03,29.82(3C),21.57(2C).

[0164] Preparation of 1-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-(1,3-di-tert-butyl-1H-pyrazol-5-yl)urea (Example 21)

[0165]

[0166] Referring to the method for preparing Example 19, the 3-(tert-butyl)-1-methyl-1H-pyrazole-5-amine in step i was replaced with 1,3-di-tert-butyl-1H-pyrazole-5-amine in equal proportions to obtain Example 21.1 H NMR (600 MHz, DMSO-d 6 )δ13.03(s,1H),12.43(s,1H),8.90(s,1H),8.30(s,1H),8.00(s,1H),7.86(d,J=8.7Hz,1H),7.78(s,1H),7.38(t,J=2.1Hz,1 H),7.15(t,J=8.1Hz,1H),7.10(d,J=1.9Hz,1H),6.93-6.89(m,2H),6.77-6.74(m,1H),6.02(s,1H),1.54(s,9H),1.21(s,9H).

[0167] Preparation of 1-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-(3-(tert-butyl)-1-cyclohexyl-1H-pyrazol-5-yl)urea (Example 22)

[0168]

[0169] Referring to the method for preparing Example 19, the 3-(tert-butyl)-1-methyl-1H-pyrazole-5-amine in step i was replaced with 3-(tert-butyl)-1-cyclohexyl-1H-pyrazole-5-amine in equal proportions to obtain Example 22. 1 H NMR (600 MHz, DMSO-d 6 )δ13.03(s,1H),12.43(s,1H),8.81(s,1H),8.32(d,J=10.5Hz,3H),8.00(s,1H),7.86(d, J=8.7Hz,1H),7.40(t,J=2.2Hz,1H),7.16(t,J=8.1Hz,1H),7.11(d,J=1.8Hz,1H),6.91(d d,J=8.7,1.8Hz,2H),6.77(dd,J=8.1,2.1Hz,1H),6.02(s,1H),3.93(tt,J=11.1,3.9Hz,1 H),1.86-1.73(m,7H),1.64(d,J=12.8Hz,1H),1.33(td,J=12.9,6.6Hz,2H),1.20(s,9H). 13 C NMR (151 MHz, DMSO-d 6)δ158.81(2C),152.43,144.35,143.02,142.77,140.94,138.10,136.05,130.13,129.97,121.66,115.30,11 4.94,114.60,111.76,110.55,107.33,94.59,94.32,55.58,32.70(2C),32.34,30.90(3C),25.63(2C),25.39.

[0170] Preparation of 1-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-(3-(tert-butyl)-1-phenyl-1H-pyrazol-5-yl)urea (Example 23)

[0171]

[0172] Referring to the method for preparing Example 19, the 3-(tert-butyl)-1-methyl-1H-pyrazole-5-amine in step i was replaced with 3-(tert-butyl)-1-phenyl-1H-pyrazole-5-amine in equal proportions to obtain Example 23. 1 H NMR (600 MHz, DMSO-d 6 )δ13.06(s,1H),12.46(s,1H),9.30(s,1H),8.64(s,1H),8.35(s,1H),8.29(s ,1H),8.00(s,0H),7.85(d,J=8.7Hz,1H),7.57-7.47(m,4H),7.42-7.35(m,2H ),7.14(t,J=8.0Hz,1H),7.10(d,J=1.8Hz,1H),6.90(dd,J=8.7,1.9Hz,1H),6 .87(ddd,J=8.2,2.1,0.9Hz,1H),6.78-6.75(m,1H),6.35(s,1H),1.28(s,9H). 13 C NMR (151 MHz, DMSO-d 6 )δ161.26,152.20,144.34,142.98,142.75,140.84,139.05,138.11,137.64,129.93,129.75(2C),129.45,127.69 ,124.68(2C),122.95,121.63,115.28,114.97,114.60,111.79,110.47,107.25,96.26,94.54,32.49,30.66(3C).

[0173] Preparation of 1-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-(3-(tert-butyl)-1-(4-fluorophenyl)-1H-pyrazol-5-yl)urea (Example 24)

[0174]

[0175] Referring to the method for preparing Example 19, the 3-(tert-butyl)-1-methyl-1H-pyrazole-5-amine in step i was replaced with 3-(tert-butyl)-1-(4-fluorophenyl)-1H-pyrazole-5-amine in equal proportions to obtain Example 24. 1 H NMR (600 MHz, DMSO-d 6 )δ13.08(s,1H),12.48(s,1H),9.49(s,1H),8.84(s,1H),8.38(s,1H),8.29(s, 1H),8.00(s,1H),7.85(d,J=8.7Hz,1H),7.61-7.55(m,2H),7.37(t,J=2.1Hz,1H ),7.35-7.31(m,2H),7.13(t,J=8.1Hz,1H),7.10(d,J=1.8Hz,1H),6.91(dd,J=8 .8,1.9Hz,1H),6.89-6.86(m,1H),6.78-6.75(m,1H),6.32(s,1H),1.27(s,9H).

[0176] Preparation of 1-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-(3-(tert-butyl)-1-(4-chlorophenyl)-1H-pyrazol-5-yl)urea (Example 25)

[0177]

[0178] Referring to the method for preparing Example 19, the 3-(tert-butyl)-1-methyl-1H-pyrazole-5-amine in step i was replaced with 3-(tert-butyl)-1-(4-chlorophenyl)-1H-pyrazole-5-amine in equal proportions to obtain Example 25. 1 H NMR (600 MHz, DMSO-d 6)δ13.03(s,1H),12.44(s,1H),9.00(s,1H),8.42(s,1H),8.32(s,2H),8.01( s,1H),7.86(d,J=8.7Hz,1H),7.61-7.55(m,4H),7.36(t,J=2.2Hz,1H),7.15 (t,J=8.0Hz,1H),7.09(d,J=1.9Hz,1H),6.90(dd,J=8.7,1.9Hz,1H),6.86(d d,J=8.1,2.0Hz,1H),6.77(dd,J=8.0,2.2Hz,1H),6.37(s,1H),1.27(s,9H). 13 C NMR (151 MHz, DMSO-d 6 )δ161.67,152.05,147.83,144.37,142.98,142.70,140.73,138.13,137.90,132.00,129.97,129.72(2C),129.35 ,126.31(2C),124.36,121.66,115.33,114.96,114.60,111.86,110.50,107.24,96.46,94.65,32.53,30.60(3C).

[0179] Preparation of 1-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-(1-(4-bromophenyl)-3-(tert-butyl)-1H-pyrazol-5-yl)urea (Example 26)

[0180]

[0181] Referring to the method for preparing Example 19, the 3-(tert-butyl)-1-methyl-1H-pyrazole-5-amine in step i was replaced with 1-(4-bromophenyl)-3-(tert-butyl)-1H-pyrazole-5-amine in equal proportions to obtain Example 26. 1 H NMR (600 MHz, DMSO-d 6)δ13.03(s,1H),12.44(s,1H),9.11(s,1H),8.53(s,1H),8.33(s,2H),8.01( s,1H),7.85(d,J=8.7Hz,1H),7.75-7.68(m,2H),7.55-7.49(m,2H),7.36(t, J=2.2Hz,1H),7.14(t,J=8.0Hz,1H),7.09(d,J=1.8Hz,1H),6.90(dd,J=8.7, 1.9Hz,1H),6.88-6.85(m,1H),6.78-6.74(m,1H),6.36(s,1H),1.27(s,9H). 13 C NMR (151 MHz, DMSO-d 6 )δ161.69,152.12,144.37,142.97,142.71,140.79,138.35,138.13,137.88,132.63(2C),132.25,129.94,126.51 (2C),124.60,121.65,120.31,115.32,114.98,114.57,111.80,110.48,107.25,96.63,94.61,32.54,30.59(3C).

[0182] Preparation of 1-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-(3-(tert-butyl)-1-(2-fluorophenyl)-1H-pyrazol-5-yl)urea (Example 27)

[0183]

[0184] Referring to the method for preparing Example 19, the 3-(tert-butyl)-1-methyl-1H-pyrazole-5-amine in step i was replaced with 3-(tert-butyl)-1-(2-fluorophenyl)-1H-pyrazole-5-amine in equal proportions to obtain Example 27. 1 H NMR (600 MHz, DMSO-d 6)δ13.03(s,1H),12.43(s,1H),8.86(s,1H),8.32(s,3H),8.00(s,1H),7.86(d,J=8.7Hz, 1H),7.61-7.53(m,2H),7.49(ddd,J=10.0,8.3,1.4Hz,1H),7.39(td,J=7.6,1.4Hz,1H),7 .36(t,J=2.1Hz,1H),7.14(t,J=8.0Hz,1H),7.10(d,J=1.8Hz,1H),6.90(dd,J=8.7,1.9Hz ,1H),6.84(dd,J=8.2,2.0Hz,1H),6.76(dd,J=8.0,2.1Hz,1H),6.38(s,1H),1.26(s,9H). 13 C NMR (151 MHz, DMSO-d 6 )δ162.11,157.08(d,J=249.8Hz),151.33,144.41,142.98,142.66,140.60,139.41,138.13,131.36(d,J=7.8Hz),130.21,130.02,126.30 ,126.22,125.74(d,J=3.9Hz),121.68,117.41,117.28,115.34,114. 96,114.63,111.88,110.34,107.04,94.64,93.19,32.57,30.63(3C).

[0185] Preparation of 1-((3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-(3-(tert-butyl)-1-(3-fluorophenyl)-1H-pyrazol-5-yl)urea (Example 28)

[0186]

[0187] Referring to the method for preparing Example 19, the 3-(tert-butyl)-1-methyl-1H-pyrazole-5-amine in step i was replaced with 3-(tert-butyl)-1-(3-fluorophenyl)-1H-pyrazole-5-amine in equal proportions to obtain Example 28. 1 H NMR (600 MHz, DMSO-d 6)δ13.04(s,1H),12.44(s,1H),9.09(s,1H),8.50(s,1H),8.33(s,1H),8.30(s,1H),8.05-7.98(m ,1H),7.86(d,J=8.7Hz,1H),7.56(q,J=7.6Hz,1H),7.42(dd,J=8.9,1.7Hz,2H),7.37(t,J=2.2Hz, 1H),7.24(td,J=8.0,7.1,1.8Hz,1H),7.15(t,J=8.1Hz,1H),7.10(d,J=1.9Hz,1H),6.90(dd,J=8. 7,1.9Hz,1H),6.88(dd,J=7.9,2.0Hz,1H),6.77(dd,J=8.1,2.1Hz,1H),6.38(s,1H),1.28(s,9H). 13 C NMR (151 MHz, DMSO-d 6 )δ163.37,161.75,152.14,147.96,144.37,142.97,142.71,140.75,1 40.54(d,J=10.3Hz),138.14,137.94,131.46(d,J=9.1Hz),129.96,121 .65,120.42(d,J=3.3Hz),115.33,114.98,114.59,114.41,114.27,111 .86,111.64,111.48,110.55,107.30,96.92,94.63,32.54,30.56(3C).

[0188] Preparation of 1-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-(3-(tert-butyl)-1-(3,4-difluorophenyl)-1H-pyrazol-5-yl)urea (Example 29)

[0189]

[0190] Referring to the method for preparing Example 19, the 3-(tert-butyl)-1-methyl-1H-pyrazole-5-amine in step i was replaced with 3-(tert-butyl)-1-(3,4-difluorophenyl)-1H-pyrazole-5-amine in equal proportions to obtain Example 29. 1 H NMR (600 MHz, DMSO-d 6)δ13.04(s,1H),12.46(s,1H),9.31(s,1H),8.74(s,1H),8.35(s,1H),8.27(s,1H),7.99(s,1H) ,7.85(d,J=8.7Hz,1H),7.68(ddd,J=11.6,7.2,2.6Hz,1H),7.59(dt,J=10.6,8.8Hz,1H),7.45-7 .40(m,1H),7.37(t,J=2.1Hz,1H),7.14(t,J=8.0Hz,1H),7.09(d,J=1.9Hz,1H),6.90(dd,J=8.7 ,1.9Hz,1H),6.87(dd,J=8.2,2.0Hz,1H),6.77(dd,J=8.3,2.1Hz,1H),6.36(s,1H),1.27(s,9H). 13 C NMR (151 MHz, DMSO-d 6 )δ160.63,151.19,148.55(dd,J=247.0,14.1Hz),147.76(dd,J=245.8,12. 8Hz),143.28,141.90,141.65,139.74,137.05,134.74(d,J=7.9Hz),129.05 ,128.83,120.54,120.49,117.33(d,J=18.1Hz,2C),114.22,113.92,113.45 ,113.27,113.13,110.67,109.39,106.23,95.70,93.48,31.46,29.48(3C).

[0191] Preparation of 1-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-(3-(tert-butyl)-1-(3,5-difluorophenyl)-1H-pyrazol-5-yl)urea (Example 30)

[0192]

[0193] Referring to the method for preparing Example 19, the 3-(tert-butyl)-1-methyl-1H-pyrazole-5-amine in step i was replaced with 3-(tert-butyl)-1-(3,5-difluorophenyl)-1H-pyrazole-5-amine in equal proportions to obtain Example 30. 1 H NMR (600 MHz, DMSO-d 6)δ12.45(s,1H),9.10(s,1H),8.55(s,1H),8.33(s,1H),8.15(s,2H),7.86(d,J=8.7Hz,1H),7.41-7.33(m,3H),7.29(tt,J=9.3,2 .3Hz,1H),7.15(t,J=8.0Hz,1H),7.10(d,J=1.8Hz,1H),6.92-6.86(m,2H),6.78(dd,J=8.0,2.2Hz,1H),6.40(s,1H),1.28(s,9H). 13 C NMR (151 MHz, DMSO-d 6 )δ161.78(dd,J=245.8,14.4Hz,2C),161.12,151.12,143.31,141.91,141.62,140.24(t,J=13.0Hz),139.64,137.18,137.04,128.88,120.5 8,114.25,113.88,113.51,110.81,109.53,106.53,106.49,106.38,1 06.33,106.29,101.70(t,J=26.0Hz),96.81,93.59,31.51,29.38(3C).

[0194] Preparation of 1-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-(3-(tert-butyl)-1-(2,5-difluorophenyl)-1H-pyrazol-5-yl)urea (Example 31)

[0195]

[0196] Referring to the method for preparing Example 19, the 3-(tert-butyl)-1-methyl-1H-pyrazole-5-amine in step i was replaced with 3-(tert-butyl)-1-(2,5-difluorophenyl)-1H-pyrazole-5-amine in equal proportions to obtain Example 31. 1 H NMR (600 MHz, DMSO-d 6)δ13.03(s,1H),12.43(s,1H),8.86(s,1H),8.42(s,1H),8.32(s,2H),8.00(s,1 H),7.86(d,J=8.7Hz,1H),7.58-7.50(m,2H),7.46(ddt,J=9.1,7.2,3.5Hz,1H), 7.36(t,J=2.2Hz,1H),7.15(t,J=8.0Hz,1H),7.10(d,J=1.8Hz,1H),6.90(dd,J= 8.7,1.8Hz,1H),6.86-6.83(m,1H),6.78-6.74(m,1H),6.38(s,1H),1.26(s,9H). 13 C NMR (151 MHz, DMSO-d 6 )δ162.50,158.28(d,J=242.4Hz),153.60(d,J=247.1Hz),151.35,144.42,142. 96,142.65,140.56,139.55,138.15,137.08,130.02,127.25-126.96(m),121.6 8,118.64(dd,J=22.4,9.4Hz),117.84(dd,J=24.0,8.3Hz),116.96,116.79,115 .35,114.98,114.60,111.90,110.38,107.09,94.65,93.60,32.60,30.57(3C).

[0197] Preparation of 1-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-(3-(tert-butyl)-1-(2,4-difluorophenyl)-1H-pyrazol-5-yl)urea (Example 32)

[0198]

[0199] Referring to the method for preparing Example 19, the 3-(tert-butyl)-1-methyl-1H-pyrazole-5-amine in step i was replaced with 3-(tert-butyl)-1-(2,4-difluorophenyl)-1H-pyrazole-5-amine in equal proportions to obtain Example 32. 1 H NMR (600 MHz, DMSO-d 6)δ13.04(s,1H),12.44(s,1H),8.97(d,J=19.6Hz,1H),8.51-8.45(m,1H),8.33(s,1H),8.30(s,1H ),8.00(s,1H),7.86(d,J=8.7Hz,1H),7.63(q,J=8.2,7.7Hz,1H),7.59-7.52(m,1H),7.36(d,J=2. 2Hz, 1H), 7.27 (dt, J = 8.6, 4.7Hz, 1H), 7.14 (t, J = 8.0Hz, 1H), 7.10 (d, J = 1.8Hz, 1H), 6.90 (dd, J = 8. 7,1.9Hz,1H),6.84(dd,J=8.0,2.0Hz,1H),6.76(dd,J=8.2,2.2Hz,1H),6.36(s,1H),1.26(s,9H).

[0200] Example 33: Study on in vitro enzyme inhibition activity of C-3 pyrazole-substituted indazole derivatives of the present invention Experimental materials:

[0201] Tecan F500 microplate reader.

[0202] KinEASETM-STK kit (including biotinylated peptide substrate S2, Eu 3+ Labeled monoclonal antibody targeting specific phosphorylation sites, Sa-XL665-labeled streptavidin, kinase reaction buffer solution (KinEASE enzyme reaction buffer), 384 shallow well plate, TRKA full-length protein.

[0203] TRKA protein concentration: 0.111 ng / μl, MgCl 2 , ethylenediaminetetraacetic acid (EDTA), dithiothreitol (DL-Dithiothreitol, DTT), DMSO.

[0204] Experimental methods:

[0205] Step 1: Kinase reaction:

[0206] First, the compound samples prepared in the above examples were prepared into a 20 mM solution with DMSO, and then diluted with kinase reaction buffer solution to 100 μM, 10 μM, 1 μM and other concentrations according to the test requirements. Then TRKA kinase (concentration of 0.111 ng / μL), ATP (4 μM), biotin-labeled peptide substrate TK (1 μM) and compound samples (4 μL) were added to 10 μL kinase reaction buffer solution (containing MgCl 25mM and DTT 1mM), incubate at room temperature for 30 minutes, and the kinase phosphorylates the substrate TK. Then, add 10 μL of detection reagent containing EDTA (provided in the kit) to detect the phosphorylation product.

[0207] Step 2: Detect phosphorylation products:

[0208] Rare earth element Europium (Eu 3+ )-labeled antibody recognizes the phosphorylated substrate, and XL665-labeled streptavidin binds to the biotin on the substrate. + is the fluorescence donor, XL665 is the fluorescence acceptor, when Eu 3+ Close to XL665, Eu 3+ The energy is transferred to XL665, generating HTRF signal.

[0209] Result evaluation method: The fluorescence signal is generated by Eu 3+ The fluorescence absorption signals of 620nm and 665nm of XL665 are generated. Therefore, the ratio of HTRF signal (665 / 620) of each well plate reaction is calculated. The results are characterized as Delta F (DF%):

[0210]

[0211] Calculation of inhibition rate (activity %): In the absence of compound sample, the DF% of kinase activity is defined as 100%. When compound sample is added, the kinase activity rate is:

[0212]

[0213] Calculation IC 50 : DF% of kinase activity in the presence of added compounds is plotted on the Y-axis and the logarithmic concentration of the compound is plotted on the X-axis. IC 50 The values ​​were obtained by fitting the data to a sigmoidal stoichiometric response curve.

[0214] Table 1: IC of indole-2-one derivatives of the present invention 50 value

[0215]

[0216]

[0217] The above test results show that the above embodiments all have good inhibitory effects on TRKA kinase, and the IC 50 Values ​​are in the nM range.

[0218] Obviously, those skilled in the art can make various changes and modifications to the present invention without departing from the spirit and scope of the present invention. Thus, if these modifications and variations of the present invention fall within the scope of the claims of the present invention and their equivalents, the present invention is also intended to include these modifications and variations.

Claims

1. A C-3 pyrazole-substituted indazole derivative, characterized in that: The derivative is a C-3 pyrazole-substituted indazole compound having a structure represented by the general formula (I), or a stereoisomer, a pharmaceutically acceptable salt, a hydrate, a solvate or a prodrug thereof; wherein L is selected from amide and urea fragments; Ring A is selected from aryl or heteroaryl which is unsubstituted or substituted with 1 to 3 R a which may be the same or different; Ra is selected from halogen, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, C1-C6 haloalkoxy, C1-C6 alkylamino, C3-C6 cycloalkyl, 0-2 aryl or heteroaryl groups which may be the same or different from Rb; Rb is selected from H, halogen, C1-C6 alkyl.

2. The C-3 pyrazole-substituted indazole derivative according to claim 1, characterized in that: The derivative is a C-3 pyrazole-substituted indazole compound having a structure represented by the general formula (I), or a stereoisomer, a pharmaceutically acceptable salt, a hydrate, a solvate or a prodrug thereof; wherein L is selected from amide and urea fragments; Ring A is selected from aryl or heteroaryl which is unsubstituted or substituted by 1-3 Ra which may be the same or different; Ra is selected from halogen, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 alkoxy, C1-C4 haloalkoxy, amine, C3-C6 cycloalkyl, phenyl, piperazinyl, morpholinyl which may be substituted by 0-2 Rb which may be the same or different; Rb is selected from H, halogen, C1-C6 alkyl.

3. The C-3 pyrazole-substituted indazole derivative according to claim 2, characterized in that: The derivative is a C-3 pyrazole-substituted indazole compound having a structure represented by the general formula (I), or a stereoisomer, a pharmaceutically acceptable salt, a hydrate, a solvate or a prodrug thereof; wherein L is selected from amide and urea fragments; Ring A is selected from aryl or heteroaryl which is unsubstituted or substituted by 1-3 Ra which may be the same or different; Ra is selected from halogen, methyl, ethyl, isopropyl, tert-butyl, cyclohexyl, trifluoromethyl, dimethylamino, methylpiperazinyl, morpholinyl, C6 cycloalkyl, phenyl which may be substituted by 0-2 Rb which may be the same or different; Rb is selected from H, fluorine, chlorine, and bromine.

4. The C-3 pyrazole-substituted indazole derivative according to claim 3, characterized in that: The derivative is a C-3 pyrazole-substituted indazole compound having a structure represented by the general formula (I), or a stereoisomer, a pharmaceutically acceptable salt, a hydrate, a solvate or a prodrug thereof; wherein L is selected from amide and urea fragments; Ring A is selected from phenyl, isoxazolyl, pyrazolyl which are unsubstituted or substituted with 1-3 Ra which may be the same or different; Ra is selected from halogen, methyl, ethyl, isopropyl, tert-butyl, cyclohexyl, trifluoromethyl, dimethylamino, methylpiperazinyl, morpholinyl, C6 cycloalkyl, phenyl which may be substituted with 0-2 Rb which may be the same or different; Rb is selected from H, fluorine, chlorine, bromine.

5. The C-3 pyrazole-substituted indazole derivative according to claim 4, characterized in that: A C-3 pyrazole-substituted indazole derivative having a structure represented by general formula (I), or a stereoisomer, a pharmaceutically acceptable salt, a hydrate, a solvate or a prodrug thereof; N-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)benzamide; N-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-fluorobenzamide; N-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-chlorobenzamide; N-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-bromobenzamide; N-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-methylbenzamide; N-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-(trifluoromethyl)benzamide; 1-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-phenylurea; 1-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-(3-fluorophenyl)urea; 1-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-(3-chlorophenyl)urea; 1-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-(3-bromophenyl)urea; 1-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-(m-tolyl)urea; 1-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-(3-(trifluoromethyl)phenyl)urea; 1-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-(3-isopropylphenyl)urea; 1-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-(3-(tert-butyl)phenyl)urea; 1-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-(3-(dimethylamino)phenyl)urea; 1-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-(3-morpholinophenyl)urea; 1-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-(3-(tert-butyl)isoxazol-5-yl)urea; 1-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-(5-(tert-butyl)isoxazol-3-yl)urea; 1-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-(3-(tert-butyl)-1-methyl-1H-pyrazol-5-yl)urea; 1-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-(3-(tert-butyl)-1-isopropyl-1H-pyrazol-5-yl)urea; 1-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-(1,3-di-tert-butyl-1H-pyrazol-5-yl)urea; 1-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-(3-(tert-butyl)-1-cyclohexyl-1H-pyrazol-5-yl)urea; 1-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-(3-(tert-butyl)-1-phenyl-1H-pyrazol-5-yl)urea; 1-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-(3-(tert-butyl)-1-(4-fluorophenyl)-1H-pyrazol-5-yl)urea; 1-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-(3-(tert-butyl)-1-(4-chlorophenyl)-1H-pyrazol-5-yl)urea; 1-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-(1-(4-bromophenyl)-3-(tert-butyl)-1H-pyrazol-5-yl)urea; 1-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-(3-(tert-butyl)-1-(2-fluorophenyl)-1H-pyrazol-5-yl)urea; 1-((3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-(3-(tert-butyl)-1-(3-fluorophenyl)-1H-pyrazol-5-yl)urea; 1-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-(3-(tert-butyl)-1-(3,4-difluorophenyl)-1H-pyrazol-5-yl)urea; 1-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-(3-(tert-butyl)-1-(3,5-difluorophenyl)-1H-pyrazol-5-yl)urea; 1-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-(3-(tert-butyl)-1-(2,5-difluorophenyl)-1H-pyrazol-5-yl)urea; 1-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-(3-(tert-butyl)-1-(2,4-difluorophenyl)-1H-pyrazol-5-yl)urea.

6. Use of a C-3 pyrazole-substituted indazole derivative according to any one of claims 1 to 5, characterized in that: The compound represented by the general formula I, and its geometric isomers or pharmaceutically acceptable salts, hydrates, solvates or prodrugs thereof are used in the preparation of drugs for preventing or treating diseases related to the expression or activity of TRK kinase.

7. Use of a C-3 pyrazole-substituted indazole derivative according to any one of claims 1 to 5, characterized in that: The compound represented by the general formula I, and its geometric isomers or pharmaceutically acceptable salts, hydrates, solvates or prodrugs thereof are used in the preparation of preventive or anti-tumor drugs.

8. A pharmaceutical composition, characterized in that: The pharmaceutical composition comprises a therapeutically effective amount of a C-3 pyrazole-substituted indazole derivative or a stereoisomer thereof, a pharmaceutically acceptable salt, a hydrate, a solvate or a prodrug thereof according to any one of claims 1 to 5 and a pharmaceutically acceptable carrier or excipient.

9. Use of the pharmaceutical composition according to claim 8, characterized in that: The composition is used in preparing medicine for preventing or treating tumors, cancers or severe pain caused by different reasons.

10. Use of the pharmaceutical composition according to claim 9, characterized in that: The composition is used in preparing preventive or anti-tumor drugs.

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