A c-3 pyrazole substituted indazole derivative and uses thereof

Abstract

The present application belongs to the field of pharmaceutical chemistry, and specifically relates to a C-3 pyrazole-substituted indazole derivative (a compound of a general formula, stereoisomers, pharmaceutically acceptable salts, hydrates, solvates or prodrugs thereof) and use (use in preparation of therapeutic agents, particularly TRK inhibitors). The derivative is a C-3 pyrazole-substituted indazole compound having a structure shown in a general formula (I), or stereoisomers, pharmaceutically acceptable salts, hydrates, solvates or prodrugs thereof; the C-3 pyrazole-substituted indazole derivative having the structure shown in the general formula (I), stereoisomers, pharmaceutically acceptable salts, hydrates, solvates or prodrugs thereof provided by the present application have the activity as protein kinase inhibitors, particularly on TRK kinase.

Description

Technical Field

[0001] This invention belongs to the field of medicinal chemistry, specifically relating to a C-3 pyrazole-substituted indazole derivative (general formula compound, its stereoisomer, pharmaceutically acceptable salt, hydrate, solvate or prodrug) and its use (use in the preparation of therapeutic agents, particularly TRK inhibitors). Background Technology

[0002] TRKs, short for tropomyosin receptor kinases, are members of the transmembrane receptor tyrosine kinase family. Based on their encoding genes, TRKs are divided into three subtypes: TRKA, TRKB, and TRKC, encoded by NTRK1, NTRK2, and NTRK3, respectively. TRK receptor proteins play a crucial role in neuronal development and differentiation, and the types and numbers of neurotrophic factors and their receptors vary depending on the location. TRKA's main endogenous ligand is nerve growth factor (NGF), TRKB's ligands mainly include brain-derived neurotrophic factor (BDNF) and neurotrophin 4 (NT-4), and TRKC's only endogenous ligand is neurotrophin 3 (NT-3). During normal physiological processes, TRKs are primarily activated by their endogenous ligands, thereby activating downstream signaling pathways such as RAS / ERK, PI3K / AKT, and PLC-γ1, regulating cell proliferation, differentiation, migration, and apoptosis. This signal transduction process is regulated by their transmembrane receptor binding domain. Under abnormal conditions, unregulated overactivation of TRK can continuously induce cell differentiation and proliferation, leading to cancer formation and progression.

[0003] To date, TRK overactivation has been observed in various cancer types, characterized by NTRK mutations, TRK overexpression, and NTRK gene fusions. These have been found in both common and rare cancers, including colorectal cancer, lung cancer, large cell neuroendocrine carcinoma, non-small cell lung cancer (NSCLC), melanoma, acute myeloid leukemia, breast cancer, skin cancer (such as basal cell carcinoma), neuroblastoma, cylindrica, and other tumors. NTRK gene fusions have the highest oncogenic probability, similar to the oncogenic fusions of ALK and ROS1 genes. The gene characteristic 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 chaperone gene. Because the extracellular domain of TRK fusion proteins has certain structural or functional deficiencies, antibodies targeting TRKs or their ligands cannot serve as effective anticancer drugs. Blocking abnormal TRK signaling pathways with small molecule inhibitors is an effective approach for treating NTRK gene fusion-type cancers. The successful marketing of larotrectinib demonstrates the feasibility of this strategy. Clinical data from 17 types of NTRK fusion-type cancers, including salivary gland secretory carcinoma (MASC), infantile fibrosarcoma, melanoma, gastrointestinal stromal tumor (GIST), thyroid cancer, colorectal adenocarcinoma, and lung adenocarcinoma, show that larotrectinib has a significant therapeutic effect on patients regardless of gene fusion type (NTRK1, NTRK2, or NTRK3 and partner genes) and histology, and is independent of age (4 months to 76 years). However, long-term use can lead to drug resistance, reducing the efficacy of larotrectinib. Summary of the Invention

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

[0005] To achieve the above objectives, 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 the structure shown in general formula (I), or a stereoisomer thereof, a pharmaceutically acceptable salt, hydrate, solvate, or prodrug;

[0007]

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

[0009] Ring A is selected from unsubstituted aryl or heteroaryl groups or groups that are substituted with 1-3 Ra groups that may be the same or different.

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

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

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

[0013] Ring A is selected from unsubstituted aryl or heteroaryl groups or groups substituted with 1-3 identical or different Ra groups; Ra is selected from halogens, C1-C4 alkyl groups, C1-C4 haloalkyl groups, C1-C4 alkoxy groups, C1-C4 haloalkoxy groups, amino groups, C3-C6 cycloalkyl groups, 0-2 identical or different Rb groups, phenyl groups, piperazine groups, and morpholino groups; Rb is selected from H, halogens, and C1-C6 alkyl groups.

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

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

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

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

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

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

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

[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)isoxazole-5-yl)urea;

[0038] 1-(3-((3-(1H-pyrazol-4-yl)-1H-indazol-6-yl)amino)phenyl)-3-(5-(tert-butyl)isoxazole-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, its geometric isomers, or pharmaceutically acceptable salts, hydrates, solvates, or prodrugs thereof in the preparation of medicaments for the prevention or treatment of diseases related to the expression or activity of TRK kinase.

[0054] The use of the compound represented by general formula I, its geometric isomers, or pharmaceutically acceptable salts, hydrates, solvates, or prodrugs in the preparation of preventive or antitumor drugs.

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

[0056] The composition is used in the preparation of a medicine for the prevention or treatment of tumors, cancer, or severe pain caused by various reasons.

[0057] The use of the composition in the preparation of preventive or antitumor drugs.

[0058] The following is an explanation of some of the terms used in this invention:

[0059] Halogens: Refers to fluorine, chlorine, bromine, or iodine. Alkyl groups: Straight-chain or branched alkyl groups, such as methyl, ethyl, propyl, isopropyl, n-butyl, or tert-butyl. Cycloalkyl groups: Substituted or unsubstituted cyclic alkyl groups, such as cyclopropyl, cyclopentyl, or cyclohexyl. Substituents include methyl, halogen, etc. Halogenated alkyl groups: Straight-chain or branched alkyl groups where the hydrogen atoms may be partially or completely replaced by halogen atoms, such as chloromethyl, dichloromethyl, trichloromethyl, fluoromethyl, difluoromethyl, trifluoromethyl, etc. Alkoxy groups: Straight-chain or branched alkyl groups where 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 C-3 pyrazole-substituted indazole derivatives or stereoisomers thereof of general formula (I) as described in the first aspect above, pharmaceutically acceptable salts, hydrates, solvates or prodrugs:

[0061]

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

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

[0064] In the above preparation process, R 1 H, methyl, trifluoromethyl, fluorine, chlorine, bromine, R 2 The phenyl or isoxazolyl group 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, unsubstituted or phenyl substituted with one Rb; Rb is H, fluorine, chlorine, or bromine.

[0065] Furthermore, starting with 6-bromo-1H-indazole, an iodination reaction is carried out with iodosuccinimide at high temperature 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, or N,N-dimethylformamide, preferably N,N-dimethylformamide. Intermediate 2 is then reacted with 3,4-dihydro-2H-pyran at low temperature to obtain intermediate 3. The reaction solvent can be acetonitrile, tetrahydrofuran, or dichloromethane, preferably dichloromethane, and the reaction temperature is 0–30°C, preferably 30°C. Intermediate 31-THP-4-pyrazolobionic acid pinacol ester undergoes a 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, or 1,4-dioxane, preferably N,N-dimethylformamide. Starting material 5 undergoes a reduction reaction to obtain intermediate 6. The initial reaction conditions can be Fe / NH4Cl, H2 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℃, preferably 80℃. Intermediate 6 undergoes an acylation reaction with the corresponding aromatic amine under low temperature conditions to obtain intermediate 7. The low temperature reaction temperature is 0-30℃, preferably 30℃. The reaction solvent can be N,N-dimethylformamide, tetrahydrofuran, preferably N,N-dimethylformamide. Intermediate 6 undergoes a condensation reaction with the corresponding aromatic amine to obtain final product 9. The reaction temperature is 0-30℃, preferably 30℃. 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℃, preferably 50℃.

[0066] The reactant 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, or tetrahydrofuran, preferably tetrahydrofuran. 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, or tetrahydrofuran, preferably tetrahydrofuran. Intermediate 13 is deprotected by toluenesulfonic acid to obtain 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 a third aspect, the present invention provides the use of C-3 pyrazole-substituted indazole derivatives, compounds of general formula I, their geometric isomers or pharmaceutically acceptable salts, hydrates, solvates or prodrugs thereof, in the preparation of medicaments for the prevention or treatment of diseases related to the expression or activity of TRK kinases.

[0068] In the fourth aspect, the use of the compound represented by general formula I, its geometric isomers, or pharmaceutically acceptable salts, hydrates, solvates, or prodrugs thereof in the preparation of preventive or antitumor 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 a stereoisomer thereof described in the first aspect above, a pharmaceutically acceptable salt, hydrate, solvate or prodrug, and a pharmaceutically acceptable carrier or excipient.

[0070] In a sixth aspect, the present invention provides the use of the composition described in the first aspect above in the preparation of a medicament for the prevention or treatment of 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 various reasons.

[0071] In the seventh aspect, the use of the composition in the preparation of preventive or antitumor drugs.

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

[0073] This invention targets NTRK gene fusion-type cancers by designing a series of novel C-3 pyrazole-substituted indazole derivatives and discovering that compounds with such structures exhibit good inhibitory activity against wild-type TRK kinases and various mutants. These compounds can be used to treat tumors caused by NTRK gene fusions and overcome drug resistance caused by TRK mutations or other diseases related to abnormal TRK expression in clinical practice. Detailed Implementation

[0074] The present invention will be further described below with reference to specific embodiments. It should be understood that the specific embodiments described herein are for illustrative purposes only and are not intended to limit the scope of the invention. The proton NMR spectra of the compounds were determined using a Bruker ARX-400; all reagents used were analytical grade or chemically pure.

[0075] Where specific techniques or conditions are not specified in the examples, they shall be performed in accordance with the techniques or conditions described in the literature in this field, or in accordance with the product instructions. Reagents or instruments whose manufacturers are not specified are all conventional products that can be purchased through legitimate channels.

[0076] Unless otherwise specified, the experimental methods used in the following embodiments are conventional methods. Unless otherwise specified, the experimental materials used in the following embodiments are commercially available products.

[0077] The preparation route of Example 1 is shown below:

[0078]

[0079] The specific synthesis steps are as follows:

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

[0081] 6-Bromo-1H-indazole (4.67 g, 23.63 mmol) was dissolved in N,N-dimethylformamide, and iodosuccinimide (5.32 g, 23.63 mmol) was added at room temperature. The reaction mixture was heated to 65 °C and stirred for 5 hours. After cooling to room temperature, the reaction was monitored by TLC until the starting material was completely reacted. 300 mL of water was slowly added, and a large amount of solid precipitated. The mixture was stirred at room temperature for 30 min, filtered, and 7.59 g of a white solid was obtained, 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. At 0 °C, a DCM solution of DHP (5.93 g, 70.51 mmol) was slowly added dropwise to the reaction mixture. After the addition was complete, the mixture was allowed to stand at room temperature for 5 h. The reaction mixture was monitored by TLC until the starting material was fully reacted. The reaction mixture was then diluted with DCM and extracted with H₂O (50 mL × 3) and saturated sodium chloride. The organic layer was collected and purified by silica gel column chromatography (1%–5% ethyl acetate / petroleum ether) to give 5.39 g of a white solid, in 72% yield.

[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 Cs₂CO₃ (8.6 g, 26.48 mmol) were dissolved in DMF solution. After purging with argon for 10 min, Pd(PPh₃)₄ (0.76 g, 0.66 mmol) was added. The reaction mixture was heated to 80 °C and reacted overnight. The reaction was monitored by TLC until the starting material was completely reacted. The solvent was removed by rotary evaporation, 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 give 3.6 g of a white solid, yield 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-indazole-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 Cs₂CO₃ (2.99 g, 9.19 mmol) were dissolved in 1,4-dioxane. After purging with argon for 10 min, Pd(OAc)₂ (0.05 g, 0.22 mmol) was added. The reaction mixture was heated to 80 °C and reacted for 4 h. The reaction was monitored by TLC until the starting material was completely reacted. The solvent was removed by rotary evaporation, and the mixture was dissolved in ethyl acetate (EA). Extraction was then performed with H₂O (100 mL × 3) and saturated sodium chloride (100 mL × 2). The organic phase was collected and purified by silica gel column chromatography (10%–30% ethyl acetate / petroleum ether) to give 1.4 g of a medium-white solid, yield 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)phenyl-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 with stirring at room temperature. The reaction mixture was heated to 60 °C and stirred for 2 hours. The reaction was monitored by TLC until the starting material was completely reacted. Pd / C was filtered off, the filtrate was collected, the solvent was removed by rotary evaporation, and the filtrate was used directly 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 mixture was stirred at room temperature for 12 hours. After the reaction was complete as monitored by TLC, the reaction mixture was slowly poured into water, resulting in the precipitation of a white solid. The mixture was filtered, the filter cake was collected, and dried to give 0.1 g of a white solid, with a yield of 81.3%.

[0092] Preparation of N-(3-((3-(1H-pyrazol-4-yl)-1H-indazole-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 mixture was heated to 50 °C and stirred for 2 hours. The reaction was monitored by TLC until complete. The mixture was diluted with EA 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%–5% methanol / dichloromethane) to obtain 0.03 g of compound 8 (i.e., the compound of Example 1), in 42.8% yield. 1 HNMR(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.

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

[0096]

[0097] Referring to the method of preparation Example 1, benzoic acid in step f was replaced with m-fluorobenzoic acid in proportion, thus obtaining 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.

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

[0099]

[0100] Referring to the method of preparation Example 1, benzoic acid in step f was replaced with m-chlorobenzoic acid in proportion, thus obtaining 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.

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

[0102]

[0103] Referring to the method of preparation Example 1, benzoic acid in step f was replaced with m-bromobenzoic acid in proportion, thus obtaining 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 CNMR(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.

[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 of Preparation Example 1, benzoic acid in step f was replaced with m-methylbenzoic acid in proportion, thus obtaining 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.

[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 of Preparation Example 1, benzoic acid in step f was replaced with m-trifluoromethylbenzoic acid in proportion, thus obtaining 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.

[0110] The preparation route of Example 7 is shown below:

[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. 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 mixture. The mixture was then cooled to room temperature and stirred at room temperature for 12 hours. TLC analysis revealed a slight residual starting material. The reaction mixture was diluted with DCM and 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%.

[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(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). 13C 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.

[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 of preparation Example 7, aniline in step h was replaced with m-fluoroaniline in an 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).

[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 of preparation Example 7, aniline in step h was replaced with m-chloroaniline in an equal proportion to obtain Example 9. 1H 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). 13 C 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.

[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 of preparation Example 7, aniline in step h was replaced with m-bromoaniline in 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). 13C 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.

[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 of preparation Example 7, the aniline in step h was replaced with m-methylaniline in an equal proportion to obtain Example 11. 1 H 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.

[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 of preparation Example 7, the aniline in step h was replaced with m-trifluoromethylaniline in an equal proportion 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). 13 C 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.

[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 of preparation Example 7, aniline in step h was replaced with 3-isopropylaniline in proportion, thus obtaining Example 13. 1H 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).

[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 of preparation Example 7, the aniline in step h was replaced with 3-tert-butylaniline in an equal proportion to obtain Example 14. 1H 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).

[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 of preparation Example 7, aniline in step h was replaced with 3-(dimethylamino)aniline in proportion, thus obtaining 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).

[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 of preparation Example 7, aniline in step h was replaced with 3-morpholine aniline in proportion, thus obtaining 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).

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

[0146]

[0147] Referring to the method of preparation Example 7, the aniline in step h was replaced with 3-(tert-butyl)isoxazole-5-amine in proportion, thus obtaining 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 CNMR(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).

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

[0149]

[0150] Referring to the method of preparation Example 7, the aniline in step h was replaced with 5-(tert-butyl)isoxazole-3-amine in proportion, thus obtaining 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).

[0151] The preparation route of Example 19 is shown below:

[0152]

[0153] The specific synthesis steps are as follows:

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

[0155] 3-(tert-butyl)-1-methyl-1H-pyrazole-5-amine (0.2 g, 1.3 mmol) was dissolved in dry tetrahydrofuran, and pyridine (0.3 g, 3.9 mmol) and phenyl chloroformate (0.3 g, 1.95 mmol) were added. The reaction mixture was stirred at room temperature for 3 hours. The reaction was complete as monitored by TLC. The solvent was removed by rotary evaporation, and the mixture was dissolved in ethyl acetate and extracted with 1N HCl (aq) (50 mL × 3) and saturated sodium chloride. The organic layer was collected and evaporated to dryness, and used directly for the next step without further 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 sequentially. The reaction mixture was heated to 50 °C and stirred for 12 hours. TLC monitoring showed that the reaction was essentially complete. The solvent was removed by rotary evaporation, and the mixture was purified by silica gel column chromatography (1%–4% methanol / dichloromethane) to give 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 (i.e., 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).

[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 of Preparation Example 19, 3-(tert-butyl)-1-methyl-1H-pyrazole-5-amine in step i was replaced by 3-(tert-butyl)-1-isopropyl-1H-pyrazole-5-amine in equal proportion to obtain Example 20. 1 H NMR(600MHz,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).

[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 of Preparation Example 19, 3-(tert-butyl)-1-methyl-1H-pyrazole-5-amine in step i was replaced by 1,3-di-tert-butyl-1H-pyrazole-5-amine in equal proportion 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).

[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 of Preparation Example 19, 3-(tert-butyl)-1-methyl-1H-pyrazole-5-amine in step i was replaced by 3-(tert-butyl)-1-cyclohexyl-1H-pyrazole-5-amine in equal proportion 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.

[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 of Preparation Example 19, 3-(tert-butyl)-1-methyl-1H-pyrazole-5-amine in step i was replaced by 3-(tert-butyl)-1-phenyl-1H-pyrazole-5-amine in equal proportion 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).

[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 of Preparation Example 19, 3-(tert-butyl)-1-methyl-1H-pyrazole-5-amine in step i was replaced by 3-(tert-butyl)-1-(4-fluorophenyl)-1H-pyrazole-5-amine in equal proportion 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).

[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 of Preparation Example 19, 3-(tert-butyl)-1-methyl-1H-pyrazole-5-amine in step i was replaced by 3-(tert-butyl)-1-(4-chlorophenyl)-1H-pyrazole-5-amine in equal proportion 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).

[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 of Preparation Example 19, 3-(tert-butyl)-1-methyl-1H-pyrazole-5-amine in step i was replaced by 1-(4-bromophenyl)-3-(tert-butyl)-1H-pyrazole-5-amine in equal proportion 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).

[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 of Preparation Example 19, 3-(tert-butyl)-1-methyl-1H-pyrazole-5-amine in step i was replaced by 3-(tert-butyl)-1-(2-fluorophenyl)-1H-pyrazole-5-amine in equal proportion 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).

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

[0186]

[0187] Referring to the method of Preparation Example 19, 3-(tert-butyl)-1-methyl-1H-pyrazole-5-amine in step i was replaced by 3-(tert-butyl)-1-(3-fluorophenyl)-1H-pyrazole-5-amine in equal proportion 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=8.0,7.1,1.8Hz,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(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).

[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 of Preparation Example 19, 3-(tert-butyl)-1-methyl-1H-pyrazole-5-amine in step i was replaced by 3-(tert-butyl)-1-(3,4-difluorophenyl)-1H-pyrazole-5-amine in equal proportion 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).

[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 of Preparation Example 19, 3-(tert-butyl)-1-methyl-1H-pyrazole-5-amine in step i was replaced by 3-(tert-butyl)-1-(3,5-difluorophenyl)-1H-pyrazole-5-amine in equal proportion 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).

[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 of Preparation Example 19, 3-(tert-butyl)-1-methyl-1H-pyrazole-5-amine in step i was replaced by 3-(tert-butyl)-1-(2,5-difluorophenyl)-1H-pyrazole-5-amine in equal proportion 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).

[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 of Preparation Example 19, 3-(tert-butyl)-1-methyl-1H-pyrazole-5-amine in step i was replaced by 3-(tert-butyl)-1-(2,4-difluorophenyl)-1H-pyrazole-5-amine in equal proportion 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).

[0200] Example 33: In vitro enzyme inhibitory activity study of the C-3 pyrazole-substituted indazole derivative of the present invention. Experimental materials:

[0201] Tecan F500 microplate reader.

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

[0203] TRKA protein concentration 0.111 ng / μl, MgCl2, EDTA, DL-Dithiothreitol (DTT), DMSO.

[0204] Experimental methods:

[0205] Step 1: Kinase Response

[0206] First, the compound samples prepared in the above examples were dissolved in DMSO to form a 20 mM solution. Then, according to the testing requirements, the solutions were diluted with kinase reaction buffer to concentrations of 100 μM, 10 μM, and 1 μM. Next, TRKA kinase (0.111 ng / μL), ATP (4 μM), biotin-labeled peptide substrate TK (1 μM), and the 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, during which the kinase phosphorylated the substrate TK. Then, 10 μL of an EDTA-containing detection reagent (included in the kit) was added to detect the phosphorylation product.

[0207] Step 2: Detection of phosphorylation products:

[0208] rare earth element europium (Eu) 3+ The XL665-labeled antibody recognizes phosphorylated substrates, and streptavidin binds to biotin on the substrate. Eu3 + It is a fluorescent donor, and XL665 is a fluorescent acceptor. When Eu 3+ Similar to XL665, Eu 3+ Energy is transferred to the XL665 to generate an HTRF signal.

[0209] Result evaluation method: Fluorescence signal is derived from Eu 3+ The HTRF absorption signals at 620 nm and 665 nm of XL665 were generated. Therefore, the ratio of the HTRF signal (665 / 620) for each well plate reaction was calculated. The result was characterized as Delta F (DF%):

[0210]

[0211] Calculation of inhibition rate (activity %): The DF% of kinase activity without the addition of the compound sample is defined as 100%. When the compound sample is added, the kinase activity rate is:

[0212]

[0213] Computing IC 50 With the addition of the compound, the DF% of kinase activity was plotted on the Y-axis, and the logarithm of the compound concentration was plotted on the X-axis. IC 50 The values ​​were obtained by fitting the data into an S-type stoichiometric response curve.

[0214] Table 1: IC50 of the indole-2-one derivatives of the present invention 50 value

[0215]

[0216]

[0217] The above test results show that all the above embodiments have a good inhibitory effect on TRKA kinase, and the IC50 of most embodiments is [missing information]. 50 The value is in the range of nM.

[0218] Obviously, those skilled in the art can make various modifications and variations to this invention without departing from its spirit and scope. Therefore, if these modifications and variations fall within the scope of the claims of this invention and their equivalents, this invention also intends to include these modifications and variations.

Claims

1. A C-3 pyrazole-substituted indazole derivative, characterized in that: The derivatives are C-3 pyrazole substituted indazoles having the structure of Formula (I), or pharmaceutically acceptable salts thereof; Formula (I) L is selected from amide, urea fragment; A ring is selected from phenyl, isoxazolyl, pyrazolyl, unsubstituted or substituted with 1-3 Ra, which can be the same or different; Ra is selected from halogen, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkylamino, C3-C6 cycloalkyl, phenyl substituted with 0-2 Rb, which can be the same or different, piperazinyl, morpholinyl; Rb is selected from H, halogen, C1-C6 alkyl.

2. The C-3 pyrazole substituted indazole derivatives according to claim 1, characterized by: The derivatives are C-3 pyrazole substituted indazoles having the structure of Formula (I), or pharmaceutically acceptable salts thereof; L is selected from amide, urea fragment; A ring is selected from phenyl, isoxazolyl, pyrazolyl, unsubstituted or substituted with 1-3 Ra, which can be the same or different; Ra is selected from halogen, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 alkylamino, C3-C6 cycloalkyl, phenyl substituted with 0-2 Rb, which can be the same or different, piperazinyl, morpholinyl; Rb is selected from H, halogen, C1-C6 alkyl.

3. The C-3 pyrazole substituted indazole derivatives according to claim 2, characterized by: The derivatives are C-3 pyrazole substituted indazoles having the structure of Formula (I), or pharmaceutically acceptable salts thereof; L is selected from amide, urea fragment; A ring is selected from phenyl, isoxazolyl, pyrazolyl, unsubstituted or substituted with 1-3 Ra, which can be the same or different; Ra is selected from halogen, methyl, ethyl, isopropyl, tert-butyl, trifluoromethyl, dimethylamino, morpholinyl, C6 cycloalkyl, phenyl substituted with 0-2 Rb, which can be the same or different; Rb is selected from H, fluorine, chlorine, bromine.

4. The C-3 pyrazole substituted indazole derivatives according to claim 3, characterized by: The C-3 pyrazole substituted indazoles having the structure of Formula (I), or pharmaceutically acceptable salts thereof; N - (3 -((3 -(dimethylamino)pyridin- 1 - yl)methyl)-lH-pyrazol-5 -yl)methyl H - (3 -((3 -(dimethylamino)pyridin- 1 - yl)methyl)-lH-pyrazol-5 -yl)methyl H - (3 -((3 -(dimethylamino)pyridin- 1 - yl)methyl)-lH-pyrazol-5 -yl)m N - (3 -((3 -(dimethylamino)- 1 -piperidinyl)carbonyl)- 1 H - (3 -((3 -(dimethylamino)- 1 -piperidinyl)carbonyl)- 1 H - (3 -((3 -(dimethylamino)- 1 -piperidinyl)carbonyl)- 1 N - (3 -((3 -(dimethylamino)- 1 -piperidinyl)carbonyl)- 1 H - (3 -((3 -(dimethylamino)- 1 -piperidinyl)carbonyl)- 1 H - (3 -((3 -(dimethylamino)- 1 -piperidinyl)carbonyl)- 1 N - (3 -((3 -(dimethylamino)pyridin- 1 - yl)methyl)-lH-pyrazol-5 -yl)methyl H - (3 -((3 -(dimethylamino)pyridin- 1 - yl)methyl)-lH-pyrazol-5 -yl)methyl H - (3 -((3 -(dimethylamino)pyridin- 1 - yl)methyl)-lH-pyrazol-5 -yl)methyl N - (3 -((3 -(dimethylamino)pyridin- 1 - yl)methyl)-lH-pyrazol-5 -yl)methyl H - (3 -((3 -(dimethylamino)pyridin- 1 - yl)methyl)-lH-pyrazol-5 -yl)methyl H - (3 -((3 -(dimethylamino)pyridin- 1 - yl)methyl)-lH-pyrazol-5 -yl)methyl N - (3 -((3 -(dimethylamino)- 1 -piperidinyl)sulfonyl)- 1 -piperidinyl)oxy)-3 - (trifluoromethyl)benzamide; H - (3 -((3 -(dimethylamino)- 1 -piperidinyl)sulfonyl)- 1 -piperidinyl)oxy)-3 - (trifluoromethyl)benzamide; H - (3 -((3 -(dimethylamino)- 1 -piperidinyl)sulfonyl)- 1 -(3-((3-(1 H - pyrazol-4-yl)-1 H - indazol-6-yl)amino)phenyl)-3- phenylurea; 1 -(3-((3-(1 H - pyrazol-4-yl)-1 H - indazol-6-yl)amino)phenyl)-3-(3- fluorophenyl)urea; 1 -(3-((3-(1 H - pyrazol-4-yl)-1 H - indazol-6-yl)amino)phenyl)-3-(3- chlorophenyl)urea; 1 -(3-((3-(1 H - pyrazol-4-yl)-1 H - indazol-6-yl)amino)phenyl)-3-(3- bromophenyl)urea; 1 -(3-((3-(1 H - pyrazol-4-yl)-1 H - indazol-6-yl)amino)phenyl)-3-(m-tolyl)urea; 1 -(3-((3-(1 H - pyrazol-4-yl)-1 H - indazol-6-yl)amino)phenyl)-3-(3-(trifluoromethyl)phenyl)urea; 1 -(3-((3-(1 H - pyrazol-4-yl)-1 H - indazol-6-yl)amino)phenyl)-3-(3- isopropylphenyl)urea; 1 -(3-((3-(1 H - pyrazol-4-yl)-1 H - indazol-6-yl)amino)phenyl)-3-(3-(tert-butyl)phenyl)urea; 1 -(3-((3-(1 H - pyrazol-4-yl)-1 H - indazol-6-yl)amino)phenyl)-3-(3-(dimethylamino)phenyl)urea; 1 -(3-((3-(1 H - pyrazol-4-yl)-1 H - indazol-6-yl)amino)phenyl)-3-(3- morpholinophenyl)urea; 1 -(3-((3-(1 H - pyrazol-4-yl)-1 H - indazol-6-yl)amino)phenyl)-3-(3-(tert-butyl)isoxazol-5-yl)urea; 1 -(3-((3-(1 H - pyrazol-4-yl)-1 H - indazol-6-yl)amino)phenyl)-3-(5-(tert-butyl)isoxazol-3-yl)urea; 1 -(3-((3-(1 H - pyrazol-4-yl)-1 H - indazol-6-yl)amino)phenyl)-3-(3-(tert-butyl)-1 H - pyrazol-5-yl)urea; 1 -(3-((3-(1 H - pyrazol-4-yl)-1 H - indazol-6-yl)amino)phenyl)-3-(3-(tert-butyl)-1 H - pyrazol-5-yl)urea; 1 -(3-((3-(1 H - pyrazol-4-yl)-1 H - indazol-6-yl)amino)phenyl)-3-(1,3-di-tert-butyl-1 H - pyrazol-5-yl)urea; 1 -(3-((3-(1 H - pyrazol-4-yl)-1 H - indazol-6-yl)amino)phenyl)-3-(3-(tert-butyl)-1 H - pyrazol-5-yl)urea; 1 -(3-((3-(1 H - pyrazol-4-yl)-1 H - indazol-6-yl)amino)phenyl)-3-(3-(tert-butyl)-1 -phenyl-1 H - pyrazol-5-yl)urea; 1 -(3-((3-(1 H - pyrazol-4-yl)-1 H - indazol-6-yl)amino)phenyl)-3-(3-(tert-butyl)-1 -(4-fluorophenyl)-1 H - pyrazol-5-yl)urea; 1 -(3-((3-(1 H - pyrazol-4-yl)-1 H - indazol-6-yl)amino)phenyl)-3-(3-(tert-butyl)-1 -(4-chlorophenyl)-1 H - pyrazol-5-yl)urea; 1 -(3-((3-(1 H - pyrazol-4-yl)-1 H - indazol-6-yl)amino)phenyl)-3-(1 -(4-bromophenyl)-3-(tert-butyl)-1 H - pyrazol-5-yl)urea; 1 -(3-((3-(1 H - pyrazol-4-yl)-1 H - indazol-6-yl)amino)phenyl)-3-(3-(tert-butyl)-1 -(2-fluorophenyl)-1 H - pyrazol-5-yl)urea; 1 -(3-((3-(1 H - pyrazol-4-yl)-1 H - indazol-6-yl)amino)phenyl)-3-(3-(tert-butyl)-1 -(3- H - pyrazol-5-yl)urea; 1 -(3-((3-(1 H - pyrazol-4-yl)-1 H - indazol-6-yl)amino)phenyl)-3-(3-(tert-butyl)-1 -(3,4-difluorophenyl)-1 H - pyrazol-5-yl)urea; 1 -(3-((3-(1 H - pyrazol-4-yl)-1 H - indazol-6-yl)amino)phenyl)-3-(3-(tert-butyl)-1 -(3,5-difluorophenyl)-1 H - pyrazol-5-yl)urea; 1 -(3-((3-(1 H - pyrazol-4-yl)-1 H - indazol-6-yl)amino)phenyl)-3-(3-(tert-butyl)-1 -(2,5-difluorophenyl)-1 H - pyrazol-5-yl)urea; 1 -(3-((3-(1 H - pyrazol-4-yl)-1 H - indazol-6-yl)amino)phenyl)-3-(3-(tert-butyl)-1 -(2,4-difluorophenyl)-1 H - pyrazol-5-yl)urea.

5. Use of a C-3 pyrazole substituted indazole derivative according to any one of claims 1 to 4, characterized in that: Use of the compound of Formula I, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for the prevention or treatment of a disease associated with expression or activity of a TRK kinase.

6. A pharmaceutical composition, characterized by: The pharmaceutical composition comprises a therapeutically effective amount of the C-3 pyrazole substituted indazoles of any one of claims 1-4, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or excipient.

7. Use of a pharmaceutical composition according to claim 6, characterized in that: Use of the composition in the manufacture of a medicament for the prevention or treatment of a disease associated with expression or activity of a TRK kinase.

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