C-3 pyrazole substituted indazole derivative and use thereof

Abstract

The present invention relates to the field of medicinal chemistry, and in particular to a C-3 pyrazole substituted indazole derivative (a general formula compound, and a stereoisomer, pharmaceutically acceptable salt, hydrate, solvate or prodrug thereof) and a use (a use in the preparation of a therapeutic agent, especially a TRK inhibitor). The derivative is a C-3 pyrazole substituted indazole compound having a structure represented by general formula (I), or a stereoisomer, pharmaceutically acceptable salt, hydrate, solvate or prodrug thereof. According to the present invention, the C-3 pyrazole substituted indazole derivative having a structure represented by general formula (I), or the stereoisomer, pharmaceutically acceptable salt, hydrate, solvate or prodrug thereof have activity as a protein kinase inhibitor, especially against a TRK.

Description

A C-3 pyrazole-substituted indazole derivative and its use Technical Field

[0001] The present invention belongs to the field of medicinal chemistry, and specifically relates to a C-3 pyrazole-substituted indazole derivative (a general compound, its stereoisomers, pharmaceutically acceptable salts, hydrates, solvates or prodrugs) and its use (in the preparation of therapeutic agents, especially TRK inhibitors). Background Art

[0002] TRKs, or tropomyosin receptor kinases, are members of the transmembrane receptor tyrosine kinase family. TRKs are classified into three subtypes based on the genes encoding them: TRKA, TRKB, and TRKC, encoded by NTRK1, NTRK2, and NTRK3, respectively. TRK receptor proteins play a crucial role in neuronal development and differentiation. Furthermore, the type and number of neurotrophic factors and neurotrophic factor receptors vary depending on the location of the neuron. The primary endogenous ligand for TRKA is nerve growth factor (NGF), while the ligands for TRKB primarily include brain-derived neurotrophic factor (BDNF) and neurotrophin 4 (NT-4). The sole endogenous ligand for TRKC is neurotrophin 3 (NT-3). In normal physiology, TRKs are primarily activated by their endogenous ligands, which in turn activate downstream signaling pathways such as RAS / ERK, PI3K / AKT, and PLC-γ1, regulating cell proliferation, differentiation, migration, and apoptosis. This signaling process is regulated by their extracellular transmembrane receptor binding domain. Under abnormal circumstances, unregulated overactivation of TRKs can continuously induce cell differentiation and proliferation, leading to the generation and progression of cancer.

[0003] To date, TRK overactivation has been found in a variety of cancer types, characterized by NTRK mutations, TRK overexpression, and NTRK gene fusions. These findings have been found in both common and rare cancer types, including colorectal cancer, lung cancer, large cell neuroendocrine carcinoma, non-small cell lung cancer (NSCLC), melanoma, acute myeloid leukemia, breast cancer, skin cancers (such as basal cell carcinoma) and lung cancer, neuroblastoma, cylindroma, and other tumors. NTRK gene fusions are the most common oncogenic factors. Similar to oncogenic fusions of ALK and ROS1, these fusions typically involve an in-frame fusion of the catalytic tyrosine kinase domain at the 3' end of the NTRK gene with the 5' end of the partner gene. Because the extracellular domain of TRK fusion proteins lacks structure or function, antibodies targeting TRKs or their ligands are ineffective as anticancer drugs. Blocking the aberrant TRK signaling pathway with small molecule inhibitors is an effective treatment for NTRK fusion-associated cancers. The successful launch of larotrectinib demonstrates the feasibility of this strategy. Clinical data on 17 NTRK fusion cancers, including mammary secretory salivary gland 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). However, long-term use can lead to drug resistance in patients, reducing the efficacy of larotrectinib.

[0004] Summary of the Invention

[0005] The present invention aims to provide a novel C-3 pyrazole-substituted indazole derivative and its use in the preparation of therapeutic agents, especially TRK inhibitors.

[0006] In order to achieve the above purpose, the present invention adopts the following technical solutions:

[0007] 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, pharmaceutically acceptable salt, hydrate, solvate or prodrug thereof;

[0008] Wherein, L is selected from amide urea fragment;

[0009] Ring A is selected from aryl or heteroaryl groups which are 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 and substituted by 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, pharmaceutically acceptable salt, hydrate, solvate or 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 with 1-3 identical or different Ra; Ra is selected from halogen, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 alkoxy, C1-C4 haloalkoxy, amino, C3-C6 cycloalkyl, phenyl, piperazinyl, morpholinyl, which are substituted with 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, pharmaceutically acceptable salt, hydrate, solvate or 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 with 1-3 identical or different Ra; Ra is selected from halogen, methyl, ethyl, isopropyl, tert-butyl, cyclohexyl, trifluoromethyl, dimethylamino, methylpiperazinyl, morpholinyl, C6 cycloalkyl, and phenyl substituted with 0-2 identical or different Rb; Rb is selected from H, fluorine, chlorine, and 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, pharmaceutically acceptable salt, hydrate, solvate or 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, and phenyl substituted with 0-2 identical or different Rb; Rb is selected from H, fluorine, chlorine, and bromine;

[0020] More preferably, a C-3 pyrazole-substituted indazole derivative having a structure represented by general formula (I), or a stereoisomer, pharmaceutically acceptable salt, hydrate, solvate or 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 use of the compound represented by general formula I, and its 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.

[0054] Use of the compound represented by general formula I, and its geometric isomers or pharmaceutically acceptable salts, hydrates, solvates or prodrugs 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, 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.

[0056] The composition is used in the preparation of medicines for preventing or treating tumors, cancers or severe pain caused by various 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: a straight-chain or branched alkyl group, such as methyl, ethyl, propyl, isopropyl, n-butyl or tert-butyl. Cycloalkyl: a substituted or unsubstituted cyclic alkyl group, such as cyclopropyl, cyclopentyl or cyclohexyl. Substituents include methyl, halogen, etc. Haloalkyl: a straight-chain or branched alkyl group, the hydrogen atoms on which may be partially or completely replaced by halogen atoms, such as chloromethyl, dichloromethyl, trichloromethyl, fluoromethyl, difluoromethyl, trifluoromethyl, etc. Alkoxy: a straight-chain or branched alkyl group, the hydrogen atoms of the hydroxyl group may be replaced by these straight-chain or branched alkyl groups, such as 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 of the general formula (I) described in the first aspect above, or a stereoisomer, pharmaceutically acceptable salt, hydrate, solvate or prodrug thereof:

[0061] Starting from 6-bromo-1H-indazole, iodination reaction with iodosuccinimide was carried out to obtain intermediate 2. Intermediate 2 was reacted with 3,4-dihydro-2H-pyran to obtain 3. Intermediate 3 was subjected to Suzuki coupling reaction with 1-THP-4-pyrazoleboronic acid pinacol ester to obtain 4. Intermediate 4 was subjected to Buchwald–Hartwig reaction with m-nitroaniline to obtain 5. Intermediate 5 was reduced with hydrazine hydrate and Pd / C to obtain intermediate 6. Intermediate 6 was subjected to condensation reaction with the corresponding aromatic amine to obtain intermediate 7 or 9. Intermediate 7 or 9 was deprotected with p-toluenesulfonic acid to obtain the final product 8 or 10.

[0062] The raw material 11 undergoes an acylation reaction with phenyl chloroformate to obtain intermediate 12, which undergoes a condensation reaction with intermediate 6 to obtain intermediate 13, and intermediate 13 is deprotected with toluenesulfonic acid to obtain the final product 14.

[0063] 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 with one Rb; Rb is H, fluorine, chlorine, or bromine.

[0064] 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 further The original conditions may be Fe / NH4Cl, H2 and Pd / C, hydrazine hydrate and Pd / C, etc., preferably hydrazine hydrate and Pd / C, the reaction solvent may be methanol, ethanol, acetonitrile, etc., preferably ethanol, and the reaction temperature is 45-80°C, preferably 80°C; intermediate 6 undergoes acylation reaction with the 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 may be N,N-dimethylformamide, tetrahydrofuran, preferably N,N-dimethylformamide; intermediate 6 undergoes condensation reaction with the corresponding aromatic amine to obtain final product 9, the reaction temperature is 0-30°C, preferably 30°C, the reaction solvent may 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 may be ethanol:ethyl acetate:water = 3:3:1, and the reaction temperature is 40-80°C, preferably 50°C.

[0065] The raw material 11 undergoes an acylation reaction with phenyl chloroformate to obtain 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 intermediate 6 to obtain 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.

[0066] In a third aspect, the present invention provides the use of C-3 pyrazole-substituted indazole derivatives, the compound represented by general formula I, and its 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.

[0067] In a fourth aspect, the compound represented by the general formula I, and its geometric isomers or pharmaceutically acceptable salts, hydrates, solvates or prodrugs are used in the preparation of preventive or anti-tumor drugs.

[0068] 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.

[0069] 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 diseases 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.

[0070] In the seventh aspect, the composition is used in the preparation of preventive or anti-tumor drugs.

[0071] Compared with the prior art, the present invention has the following beneficial effects:

[0072] 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. They 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

[0073] The present invention is further described below with reference to specific examples. It should be understood that the specific examples described herein are intended only to illustrate the present invention and are not intended to limit its scope. Proton nuclear magnetic resonance spectra of the compounds were measured using a Bruker ARX-400; all reagents used were of analytical or chemical grade.

[0074] If no specific techniques or conditions are specified in the examples, the experiments were carried out according to the techniques or conditions described in the literature in the field or according to the product instructions. If no manufacturer is specified for the reagents or instruments used, they are all conventional products that can be purchased through regular channels.

[0075] 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.

[0076] The preparation route of Example 1 is as follows:

[0077] The specific synthesis steps are as follows:

[0078] Synthesis of 6-bromo-3-iodo-1H-indazole (2)

[0079] Dissolve 6-bromo-1H-indazole (4.67 g, 23.63 mmol) in N,N-dimethylformamide and add iodosuccinimide (5.32 g, 23.63 mmol) at room temperature. Heat the reaction mixture to 65°C and stir for 5 hours. Cool to room temperature and monitor the reaction completion by TLC. Slowly add 300 mL of water, resulting in the precipitation of a large amount of solid. Stir at room temperature for 30 minutes and filter to obtain 7.59 g of a white solid (99% yield).

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

[0081] Intermediate 2 (7.59 g, 23.5 mmol) and p-toluenesulfonic acid (0.2 g, 1.17 mmol) were dissolved in DCM. A DCM solution of DHP (5.93 g, 70.51 mmol) was slowly added dropwise to the reaction mixture at 0°C. After completion of the addition, the mixture was allowed to react at room temperature for 5 h. TLC confirmed the completion of the reaction. The reaction mixture was diluted with DCM and extracted with H2O (50 mL x 3) and saturated sodium chloride. The organic layer was collected and purified by silica gel column chromatography (1%-5% ethyl acetate / petroleum ether) to afford 5.39 g of a white solid in a 72% yield.

[0082] Synthesis of 6-bromo-1-(tetrahydro-2H-pyran-2-yl)-3-(1-(tetrahydro-1H-pyran-2-yl)-1H-pyrazol-4-yl)-1-indazole (4)

[0083] Intermediate 3 (5.39 g, 13.24 mmol), 1-THP-4-pyrazoleboronic acid pinacol ester (4.42 g, 15.89 mmol) and Cs2CO3 (8.6 g, 26.48 mmol) were dissolved in DMF solution and purged with argon for 10 min. Pd(PPh3)4 (0.76 g, 0.66 mmol) was added. The reaction solution was heated to 80°C and allowed to react overnight. TLC monitored the completion of the reaction of the raw materials, the solvent was removed by vortexing, and the mixture was dissolved in ethyl acetate, then extracted with H2O (100 mL × 3) and saturated sodium chloride (100 mL × 2). 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%.

[0084] 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)

[0085] Intermediate 4 (1.88 g, 4.37 mmol), m-nitroaniline (0.6 g, 4.37 mmol), Xantphos (0.5 g, 0.87 mmol), and CsCO (2.99 g, 9.19 mmol) were dissolved in 1,4-dioxane and purged with argon for 10 minutes. Pd(OAc) (0.05 g, 0.22 mmol) was then added. The reaction mixture was heated to 80°C for 4 hours. The reaction was monitored by TLC. The solvent was removed and the mixture was dissolved in EA, followed by extraction with H2O (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 yield 1.4 g of a medium-white solid in a 65.7% yield.

[0086] 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)

[0087] 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 with stirring at room temperature. The reaction solution was heated to 60°C and stirred for 2 hours. After TLC monitoring, the reaction was complete, and the Pd / C was filtered out. The filtrate was collected, the solvent was removed by vortexing, and it was used directly in the next step without further purification.

[0088] 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)

[0089] 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 mixture was stirred at room temperature for 12 hours. TLC monitored the reaction completion of the raw materials. The reaction mixture was slowly poured into water, and a white solid precipitated. The filter cake was collected by filtration and dried to obtain 0.1 g of a white solid with a yield of 81.3%.

[0090] Preparation of N-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)benzamide (Example 1)

[0091] Intermediate 7 (0.1 g, 0.18 mmol) was dissolved in a 3:3:1 mixture of ethanol, ethyl acetate, and water, followed by the addition of p-toluenesulfonic acid (0.09 g, 0.53 mmol). The reaction mixture was heated to 50°C and stirred for 2 hours. TLC confirmed the completion of the reaction, followed by dilution with EA and extraction with H2O (100 mL x 3) and saturated sodium chloride (100 mL x 2). The organic phase was collected and purified by silica gel column chromatography (1%-5% methanol / dichloromethane) to yield 0.03 g of compound 8 (i.e., Example 1 compound) in a yield of 42.8%. 1 H NMR(600MHz,DMSO-d6)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). 13C NMR (151MHz, DMSO-d6) 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.

[0092] Preparation of N-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-fluorobenzamide (Example 2)

[0093] Referring to the method of Preparation Example 1, the benzoic acid in step f was replaced with m-fluorobenzoic acid in equal proportion to obtain Example 2. 1 H NMR(600MHz,DMSO-d6)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 (151MHz, 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.

[0094] Preparation of N-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-chlorobenzamide (Example 3)

[0095] Referring to the method for preparing Example 1, the benzoic acid in step f was replaced with m-chlorobenzoic acid in equal proportion to obtain Example 3. 1 H NMR(600MHz,DMSO-d6)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,2H),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(151MHz,DMSO-d6)δ164.50,144.22,142.99,142.71,140.28,138.14,137.48,137.07,133.65,131.83, 130.87,129.75,127.89,127.00,126.14,121.65,115.36,115.00,114.56,113.71,112.75,109.50,94.69.

[0096] Preparation of N-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-bromobenzamide (Example 4)

[0097] Referring to the method for preparing Example 1, the benzoic acid in step f was replaced with m-bromobenzoic acid in equal proportion to obtain Example 4. 1H NMR(600MHz,DMSO-d6)δ13.04(s,1H),12.47(s,1H),10.29(s,1H),8.41(s,1H),8.1 4(t,J=1.9Hz,3H),7.95(d,J=7.8Hz,1H),7.88(d,J=8.7Hz,1H),7.79(dd,J=8.0,2.0 Hz,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 C NMR(151MHz,DMSO-d6)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.

[0098] Preparation of N-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-methylbenzamide (Example 5)

[0099] Referring to the method for preparing Example 1, the benzoic acid in step f was replaced with m-toluic acid in equal proportion to obtain Example 5. 1 H NMR(600MHz,DMSO-d6)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=6.9,2.1H z,1H),7.44–7.37(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). 13C NMR(151MHz,DMSO-d6)δ166.12,144.13,143.01,142.79,140.60,138.14,137.05,135.56,132.56,130.13,129 .69,128.74,128.60,126.24,125.29,121.65,115.32,115.01,114.55,113.48,112.74,109.53,94.53,21.44.

[0100] Preparation of N-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-(trifluoromethyl)benzamide (Example 6)

[0101] Referring to the method for preparing Example 1, the benzoic acid in step f was replaced with m-trifluoromethylbenzoic acid in equal proportion to obtain Example 6. 1 H NMR(600MHz,DMSO-d6)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),7.2 6(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(151MHz,DMSO-d6)δ164.55,144.23,142.98,142.72,140.14,138.15,137.10,136.34,132.30,130.22,129.83,129.74,129.5 3,128.61,128.58,126.20,125.36,124.70(q,J=3.9Hz),123.55,121.68,115.37,114.96,114.65,113.86,112.87,109.60,94.75.

[0102] The preparation route of Example 7 is as follows:

[0103] The specific synthesis steps are as follows:

[0104] 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)

[0105] 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 sequentially at 0°C under 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 brought to room temperature and stirred at room temperature for 12 hours. TLC detection showed that there was a slight residue of starting material. The reaction solution was diluted with DCM and then extracted with H2O (100 mL×3) and saturated sodium chloride (100 mL×2). 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%.

[0106] Preparation of 1-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-phenylurea (Example 7)

[0107] Compound 10 (Example 7) was obtained by referring to step g of Preparation Example 1. 1 H NMR(600MHz,DMSO-d6)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(151MHz,DMSO-d6)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.

[0108] Preparation of 1-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-(3-fluorophenyl)urea (Example 8)

[0109] 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(600MHz,DMSO-d6)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=12.0,2.3H z,1H),7.41(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).

[0110] Preparation of 1-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-(3-chlorophenyl)urea (Example 9)

[0111] 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(600MHz,DMSO-d6)δ13.04(s,1H),12.44(s,1H),8.95(s,1H),8.82(s,1H),8.34( s,1H),8.31(s,1H),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). 13C NMR(151MHz,DMSO-d6)δ152.83,144.32,143.00,142.79,141.79,140.80,138.15,137.10,133.67,130.88, 129.94,126.18,121.86,121.65,117.95,117.06,115.30,114.98,114.59,111.82,110.83,107.64,94.51.

[0112] Preparation of 1-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-(3-bromophenyl)urea (Example 10)

[0113] 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(600MHz,DMSO-d6)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(151MHz,DMSO-d6)δ152.84,144.31,143.00,142.79,142.00,140.86,138.15,131.18,130.13,129 .91,124.71,122.20,121.65,120.76,117.41,115.30,115.00,114.56,111.77,110.80,107.64,94.48.

[0114] Preparation of 1-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-(3-bromophenyl)urea (Example 11)

[0115] 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. 1H NMR(600MHz,DMSO-d6)δ13.03(s,1H),12.44(s,1H),8.63(s,1H),8.55(s,1H),8.33(s,1H),8 .31(s,1H),8.01(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(151MHz,DMSO-d6)δ152.93,144.30,143.00,142.83,141.11,140.10,138.44,138.15,137.10,129.91,129 .12,126.17,123.02,121.65,119.12,115.79,115.29,115.00,114.57,111.57,110.61,107.43,94.46,21.69.

[0116] Preparation of 1-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-(3-(trifluoromethyl)phenyl)urea (Example 12)

[0117] 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(600MHz,DMSO-d6)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). 13C NMR(151MHz,DMSO-d6)δ152.94,144.33,143.01,142.80,141.06,140.73,138.14,130.42,130.10,129.95,129.89,12 5.59,123.78,122.24,121.66,118.51,115.30,114.96,114.61,114.50(d,J=4.4Hz),111.82,110.91,107.80,94.51.

[0118] Preparation of 1-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-(3-isopropylphenyl)urea (Example 13)

[0119] Referring to the method for preparing Example 7, the aniline in step h was replaced with 3-isopropylaniline in equal proportions to obtain Example 13. 1 H NMR (600MHz, DMSO-d6) 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=8.3,1.3 Hz,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(d t,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 (151MHz, DMSO-d6) 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).

[0120] Preparation of 1-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-(3-(tert-butyl)phenyl)urea (Example 14)

[0121] Referring to the method for preparing Example 7, the aniline in step h was replaced with 3-tert-butylaniline in equal proportions to obtain Example 14. 1 H NMR (600MHz, DMSO-d6) δ13.04(s,1H),12.44(s,1H),8.58(d,J=3.7Hz,2H),8.35(s,1H),8.3 0(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 ,2.1,1.0Hz,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 ,1.0Hz,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(151MHz,DMSO-d6)δ153.01,151.76,144.29,143.01,142.86,141.16,139.90,138.15,137.11,129.88,128.91,1 26.19,121.64,119.31,115.93,115.63,115.28,115.01,114.55,111.42,110.67,107.61,94.40,34.85,31.60(3C).

[0122] Preparation of 1-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-(3-(dimethylamino)phenyl)urea (Example 15)

[0123] Referring to the method for preparing Example 7, the aniline in step h was replaced with 3-(dimethylamino)aniline in equal proportions to obtain Example 15. 1H NMR(600MHz,DMSO-d6)δ13.02(s,1H),12.43(s,1H),8.81(s,1H),8.71(s,1H),8.34(s,1H),8.2 7(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).

[0124] Preparation of 1-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-(3-morpholinophenyl)urea (Example 16)

[0125] 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(600MHz,DMSO-d6)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) ,7.13–7.08(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). 13C NMR(151MHz,DMSO-d6)δ153.02,152.09,144.23,143.04,142.91,141.16,140.99,138.09,129.87,129.66,121 .64,115.22,114.96,114.57,111.49,110.68,109.88,109.59,107.63,105.38,94.32,66.57(2C),49.02(2C).

[0126] 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)

[0127] Referring to the method for preparing Example 7, the aniline in step h was replaced with 3-(tert-butyl)isoxazol-5-amine in equal proportions to obtain Example 17. 1 H NMR(600MHz,DMSO-d6)δ13.04(s,1H),12.46(s,1H),10.01(s,1H),8.82(s,1H), 8.37(s,1H),8.31(s,1H),8.01(s,1H),7.87(d,J=8.7Hz,1H),7.44(t,J=2.1Hz,1 H),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 C NMR (151MHz, DMSO-d6) 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).

[0128] 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)

[0129] Referring to the method for preparing Example 7, the aniline in step h was replaced with 5-(tert-butyl)isoxazol-3-amine in equal proportions to obtain Example 18. 1 H NMR(600MHz,DMSO-d6)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,1H),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 (151MHz, DMSO-d6) 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).

[0130] The preparation route of Example 19 is as follows:

[0131] The specific synthesis steps are as follows:

[0132] Synthesis of Phenyl (3-(tert-butyl)-1-methyl-1H-pyrazol-5-yl)carbamate (12)

[0133] Dissolve 3-(tert-Butyl)-1-methyl-1H-pyrazol-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 at room temperature for 3 hours. TLC monitoring indicates the reaction is complete. Remove the solvent by vortexing, dissolve in ethyl acetate, and extract with 1N HCl(aq) (50 mL x 3) and saturated sodium chloride. Collect the organic layer, spin dry, and use it directly in the next step without further treatment.

[0134] 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)

[0135] 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 sequentially. The reaction solution was heated to 50°C and stirred for 12 hours. TLC monitoring indicated that the reaction was nearly complete. The solvent was removed and the product was purified by silica gel column chromatography (1%-4% methanol / dichloromethane) to obtain 0.12 g of Intermediate 13 in a 44% yield.

[0136] 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)

[0137] Compound 14 (ie, Example 19) was obtained by referring to step g of Preparation Example 1. 1 H NMR(600MHz,DMSO-d6)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=8.6,2.3 Hz,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(151MHz,DMSO-d6)δ159.06,152.21,144.34,142.99,142.75,140.85,138.13,137.59,137.02,129.98, 126.22,121.66,115.30,114.97,114.61,111.84,110.58,107.32,94.56,94.00,35.36,32.26,30.83(3C).

[0138] 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)

[0139] 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-d6)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 .2Hz,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 (151MHz, DMSO-d6) 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).

[0140] 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)

[0141] 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. 1H NMR (600MHz, DMSO-d6) 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,1H),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).

[0142] 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)

[0143] 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(600MHz,DMSO-d6)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,1 H),6.91(dd,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,1H),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). 13C NMR(151MHz,DMSO-d6)δ158.81(2C),152.43,144.35,143.02,142.77,140.94,138.10,136.05,130.13,129.97,121.66, 115.30,114.94,114.60,111.76,110.55,107.33,94.59,94.32,55.58,32.70(2C),32.34,30.90(3C),25.63(2C),25.39.

[0144] 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)

[0145] 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(600MHz,DMSO-d6)δ13.06(s,1H),12.46(s,1H),9.30(s,1H),8.64(s,1H),8.3 5(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,1 H),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(151MHz,DMSO-d6)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).

[0146] 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)

[0147] 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(600MHz,DMSO-d6)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(d d,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).

[0148] 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)

[0149] 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. 1H NMR(600MHz,DMSO-d6)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(dd,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(151MHz,DMSO-d6)δ161.67,152.05,147.83,144.37,142.98,142.70,140.73,138.13,137.90,132.00,129.97,129.72(2 C),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).

[0150] 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)

[0151] 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. 1H NMR(600MHz,DMSO-d6)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 (151MHz, DMSO-d6) 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).

[0152] 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)

[0153] 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. 1H NMR(600MHz,DMSO-d6)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(151MHz,DMSO-d6)δ162.11,157.08(d,J=249.8Hz),151.33,144.41,1 42.98,142.66,140.60,139.41,138.13,131.36(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).

[0154] 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)

[0155] 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. 1H NMR(600MHz,DMSO-d6)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=8.0,7.1,1.8H z,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(151MHz,DMSO-d6)δ163.37,161.75,152.14,147.96,144.37,142.97,14 2.71,140.75,140.54(d,J=10.3Hz),138.14,137.94,131.46(d,J=9.1Hz),1 29.96,121.65,120.42(d,J=3.3Hz),115.33,114.98,114.59,114.41,114.2 7,111.86,111.64,111.48,110.55,107.30,96.92,94.63,32.54,30.56(3C).

[0156] 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)

[0157] 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. 1H NMR(600MHz,DMSO-d6)δ13.04(s,1H),12.46(s,1H),9.31(s,1H),8.74(s,1H),8.35(s,1H),8.27(s,1 H),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,1 H),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 (151MHz, DMSO-d6) δ160.63, 151.19, 148.55 (dd, J = 247.0, 14.1Hz), 147.76 (d d,J=245.8,12.8Hz),143.28,141.90,141.65,139.74,137.05,134.74(d,J=7.9H z),129.05,128.83,120.54,120.49,117.33(d,J=18.1Hz,2C),114.22,113.92,1 13.45,113.27,113.13,110.67,109.39,106.23,95.70,93.48,31.46,29.48(3C).

[0158] 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)

[0159] 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. 1H NMR (600MHz, DMSO-d6) 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(t t,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 (151MHz, DMSO-d6) 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 .88,120.58,114.25,113.88,113.51,110.81,109.53,106.53,106.49,106. 38,106.33,106.29,101.70(t,J=26.0Hz),96.81,93.59,31.51,29.38(3C).

[0160] 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)

[0161] 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. 1H NMR(600MHz,DMSO-d6)δ13.03(s,1H),12.43(s,1H),8.86(s,1H),8.42(s,1H),8.32( s,2H),8.00(s,1H),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(d d,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 (151MHz, DMSO-d6) 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),121.68,118.64(dd,J=22.4,9.4Hz),117.84(dd,J=24.0,8.3Hz),116.96,116.7 9,115.35,114.98,114.60,111.90,110.38,107.09,94.65,93.60,32.60,30.57(3C).

[0162] 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)

[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-(2,4-difluorophenyl)-1H-pyrazole-5-amine in equal proportions to obtain Example 32. 1H NMR (600MHz, DMSO-d6) 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).

[0164] Example 33: Study on in vitro enzyme inhibitory activity of C-3 pyrazole-substituted indazole derivatives of the present invention

[0165] Experimental Materials:

[0166] Tecan Microplate reader.

[0167] Kit (including biotinylated peptide substrate S2, Eu 3+ Labeled monoclonal antibody targeting specific phosphorylation sites, Sa-XL665-labeled streptavidin, kinase reaction buffer (KinEASE enzyme reaction buffer), 384 shallow well plate, TRKA full-length protein.

[0168] TRKA protein concentration: 0.111 ng / μl, MgCl2, ethylenediaminetetraacetic acid (EDTA), dithiothreitol (DL-Dithiothreitol, DTT), DMSO.

[0169] Experimental methods:

[0170] Step 1: Kinase reaction:

[0171] First, the compound samples prepared in the above examples were diluted to a 20 mM solution in DMSO. Then, depending on the test requirements, they were diluted to concentrations of 100 μM, 10 μM, or 1 μM using kinase reaction buffer. TRKA kinase (at a concentration of 0.111 ng / μL), ATP (4 μM), biotinylated peptide substrate TK (1 μM), and compound sample (4 μL) were added to 10 μL of kinase reaction buffer (containing 5 mM MgCl2 and 1 mM DTT). The mixture was incubated at room temperature for 30 minutes to allow the kinase to phosphorylate the TK substrate. Then, 10 μL of detection reagent containing EDTA (provided with the kit) was added to detect the phosphorylated product.

[0172] Step 2: Detection of phosphorylation products:

[0173] 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.

[0174] 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 the HTRF signal (665 / 620) for each well reaction is calculated. The results are expressed as Delta F (DF%):

[0175] 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:

[0176] Computing 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. 50 The values ​​were obtained by fitting the data to a sigmoidal stoichiometric response curve.

[0177] Table 1: IC values ​​of indole-2-one derivatives of the present invention 50 value

[0178] The above test results show that the above examples all have good inhibitory effects on TRKA kinase, and the IC 50 Values ​​are in the nM range.

[0179] Obviously, those skilled in the art may make various changes and modifications to the present invention without departing from the spirit and scope of the present invention. Thus, if such changes and modifications fall within the scope of the claims and their equivalents, the present invention is intended to include such changes and modifications.

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 groups 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 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; 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 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; 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 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; 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 various 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.

Images