Biaryl compound with necroptosis resisting activity and derivative and application thereof

Abstract

The invention discloses a biaryl compound with necroptosis resisting activity as well as a derivative and application of the biaryl compound. The biaryl compound is a compound as shown in # imgabs0 # or a pharmaceutically acceptable salt of the biaryl compound. The synthesis method of the compound is simple, and the obtained compound can inhibit in-vitro necrotic apoptosis of HT-29 cells and can relieve hepatic fibrosis progress of a schistosome infected mouse model. The compound can be used for preparing drugs for treating and / or preventing inflammatory, fibrosis, infectious, ischemic, degenerative or tumor diseases caused by necrotic apoptosis, and has a good market prospect.

Description

Technical Field

[0001] The present invention relates to the field of medicinal chemistry, and in particular to biaryl compounds with anti-necroptosis activity and derivatives thereof and applications thereof. Background Art

[0002] The balance between cell proliferation and death is crucial to maintaining the normal physiological function of the body. The imbalance of this process will lead to the occurrence of diseases such as cancer, inflammation, infection, and neurodegenerative diseases. Necroptosis or programmed necrosis is a sophisticated and complex form of cell death in the body, and it is also a regulated cell death mode mediated by receptor-interacting serine / threonine protein kinase 1 (RIPK1 or RIP1). RIPK1 plays a role in cell survival, inflammation, and apoptosis, and has been identified as a major player in necroptosis. RIPK1 has a dual regulatory function. On the one hand, it can act as a scaffold protein to promote the activation of the NF-κB signaling pathway, inhibit apoptosis, and maintain cell survival. On the other hand, abnormally regulated RIPK1 will induce cell necrosis, leading to an inflammatory response. At present, the research on programmed necrosis is mainly based on the TNF-α-mediated cell necrosis signal cascade pathway.

[0003] RIPK1 is considered to be a key factor in regulating necroptosis and is closely related to the occurrence and development of many diseases, such as inflammatory diseases, neurodegenerative diseases, ischemic diseases, viral infections and tumor metastasis. As a potential target in the necroptosis pathway, inhibiting its kinase activity can inhibit the development of related diseases. Since scientists discovered the first RIPK1 inhibitor Nec-1 in 2005, a variety of RIPK1-related inhibitors have entered clinical trials as tool molecules. DNL104 is the first brain-penetrating RIPK1 inhibitor to enter Phase I clinical trials, but it has drug resistance and liver toxicity after administration, and the project has been interrupted. Another brain-penetrating RIPK1 inhibitor, DNL747, was also terminated due to its potential dose-limiting toxicity. At present, most of the clinical trials of RIPK1 inhibitors under development are in the early stages and progress is relatively slow. Potential toxicity issues limit their clinical research. At present, there is still a need to further develop new RIPK1 inhibitors for necroptosis-related diseases. Summary of the invention

[0004] In view of the deficiencies in the prior art, the present invention provides a class of biaryl compounds with anti-necroptosis activity and their derivatives and uses. The compounds and their derivatives are RIPK1 inhibitors with a biaryl skeleton, which can inhibit the in vitro necroptosis of HT-29 cells and alleviate the progression of liver fibrosis in a schistosoma japonicum infected mouse model. They can be used to prepare drugs for treating and / or preventing inflammatory, fibrotic, infectious, ischemic, degenerative or tumor diseases caused by necroptosis, and have good market prospects.

[0005] In order to solve the problems of the prior art, the technical solution adopted by the present invention is:

[0006] A biaryl compound represented by formula (I) or a pharmaceutically acceptable salt thereof,

[0007]

[0008] in:

[0009] R 1 Select from the following structures

[0010] R 4 , R 5 are each independently selected from hydrogen or C1-C2 alkyl; Z 1 is nitrogen or CH; Z 2 is NH or sulfur; Z 3 is nitrogen or CH; R 2 or R 2’ independently selected from hydrogen, C1-C2 alkyl, cyano, halogen, C1-C2 alkoxy;

[0011] R 3 Select from the following structures: R 6 is hydrogen, trifluoromethyl, trifluoromethoxy, difluoromethoxy, C1-C2 alkyl or C1-C2 alkoxy; R 7 is hydrogen, trifluoromethoxy or halogen; R 8 is hydrogen or halogen; X is nitrogen, CF or CH; Y is nitrogen, methylene or NH; n is 0, 2 or 3.

[0012] As an improvement, R 1 for R 4 is selected from hydrogen or C1-C2 alkyl; R 2 or R 2’ R is independently selected from hydrogen, C1-C2 alkyl, cyano, halogen, C1-C2 alkoxy; 3 Select from the following structures: R 6is hydrogen, trifluoromethyl, trifluoromethoxy, difluoromethoxy, C1-C2 alkyl or C1-C2 alkoxy; R 7 is hydrogen, trifluoromethoxy or chlorine; R 8 is hydrogen, fluorine or chlorine; X is nitrogen, CH or CF; Y is nitrogen, methylene or NH; n is 0, 2 or 3. As an improvement, R 1 for R 4 is selected from hydrogen or C1-C2 alkyl; R 2 or R 2’ R is independently selected from hydrogen, C1-C2 alkyl, cyano, halogen, C1-C2 alkoxy; 3 for R 6 is hydrogen, trifluoromethyl, trifluoromethoxy, difluoromethoxy, C1-C2 alkyl or C1-C2 alkoxy; R 7 is hydrogen, trifluoromethoxy or chlorine; R 8 is hydrogen, fluorine or chlorine; X is nitrogen, CH or CF; Y is nitrogen, methylene or NH; n is 0 or 2.

[0013] A further improvement is that R 1 for R 4 is selected from hydrogen or C1-C2 alkyl; R 2 or R 2’ R is independently selected from hydrogen, C1-C2 alkyl, cyano, halogen, C1-C2 alkoxy; 3 for R 6 is hydrogen, trifluoromethyl, trifluoromethoxy, difluoromethoxy, C1-C2 alkyl or C1-C2 alkoxy; R 7 is hydrogen, trifluoromethoxy or chlorine; R 8 is hydrogen, fluorine or chlorine; X is nitrogen, CH or CF; Y is nitrogen; n is 2.

[0014] A further improvement is that R 1 for R 4 is selected from hydrogen or C1-C2 alkyl; R 2 or R 2’ R is independently selected from hydrogen, C1-C2 alkyl, cyano, halogen, C1-C2 alkoxy; 3 for R 6 is hydrogen, trifluoromethyl, trifluoromethoxy, difluoromethoxy, C1-C2 alkyl or C1-C2 alkoxy; R 7 is hydrogen, trifluoromethoxy or chlorine; R 8 is hydrogen, fluorine or chlorine; X is nitrogen, CH or CF; Y is NH radical; n is 0.

[0015] As an improvement, R 1 for R 5 is a C1-C2 alkyl group; R 2 or R 2’ is hydrogen; R 3 for R 6 is trifluoromethoxy; R 7 is hydrogen; R 8 is hydrogen; X is CH; Y is NH radical; n is 0.

[0016] As an improvement, R 1 for Z 1 is nitrogen or CH; Z 2 is NH or sulfur atom; Z 3 is nitrogen or CH;

[0017] R 2 or R 2’ is hydrogen or fluorine;

[0018] R 3 for R 6 is trifluoromethoxy; R 7 is hydrogen; R 8 is hydrogen; X is CH; Y is NH radical; n is 0.

[0019] As an improvement, R 1 for R 2 or R 2’ is a hydrogen atom;

[0020] R 3 for R 6 is trifluoromethoxy; R 7 is hydrogen; R 8 is hydrogen; X is CH; Y is NH atomic group;

[0021] n is 0.

[0022] As an improvement, the biaryl compound or a pharmaceutically acceptable salt thereof is specifically:

[0023] 1-(4-(3-aminobenzo[d]isoxazol-4-yl)-2-methylphenyl)-3-(3-(trifluoromethoxy)phenyl)urea (1),

[0024] 1-(4-(3-aminobenzo[d]isoxazol-4-yl)-2,3-xylyl)-3-(3-(trifluoromethoxy)phenyl)urea (2),

[0025] 1-(4-(3-aminobenzo[d]isoxazol-4-yl)-3-methylphenyl)-3-(3-(trifluoromethoxy)phenyl)urea (3),

[0026] 1-(4-(3-aminobenzo[d]isoxazol-4-yl)-3-fluorophenyl)-3-(3-(trifluoromethoxy)phenyl)urea (4),

[0027] 1-(4-(3-aminobenzo[d]isoxazol-4-yl)-2,3-difluorophenyl)-3-(3-(trifluoromethoxy)phenyl)urea (5),

[0028] 1-(4-(3-aminobenzo[d]isoxazol-4-yl)-2,6-difluorophenyl)-3-(3-(trifluoromethoxy)phenyl)urea (6),

[0029] 1-(4-(3-aminobenzo[d]isoxazol-4-yl)-3,5-difluorophenyl)-3-(3-(trifluoromethoxy)phenyl)urea (7),

[0030] 1-(4-(3-aminobenzo[d]isoxazol-4-yl)-2,5-difluorophenyl)-3-(3-(trifluoromethoxy)phenyl)urea (8),

[0031] 1-(4-(3-aminobenzo[d]isoxazol-4-yl)-3-chloro-2-fluorophenyl)-3-(3-(trifluoromethoxy)phenyl)urea (9),

[0032] 1-(4-(3-aminobenzo[d]isoxazol-4-yl)-2-chlorophenyl)-3-(3-(trifluoromethoxy)phenyl)urea (10),

[0033] 1-(4-(3-aminobenzo[d]isoxazol-4-yl)-3-chlorophenyl)-3-(3-(trifluoromethoxy)phenyl)urea (11),

[0034] 1-(4-(3-aminobenzo[d]isoxazol-4-yl)-2,3-dichlorophenyl)-3-(3-(trifluoromethoxy)phenyl)urea (12),

[0035] 1-(4-(3-aminobenzo[d]isoxazol-4-yl)-3-cyanophenyl)-3-(3-(trifluoromethoxy)phenyl)urea (13),

[0036] 1-(4-(3-aminobenzo[d]isoxazol-4-yl)-2-fluoro-5-methylphenyl)-3-(3-(trifluoromethoxy)phenyl)urea (14),

[0037] 1-(4-(3-aminobenzo[d]isoxazol-4-yl)-5-fluoro-2-methylphenyl)-3-(3-(trifluoromethoxy)phenyl)urea (15),

[0038] 1-(4-(3-aminobenzo[d]isoxazol-4-yl)-2-methoxyphenyl)-3-(3-(trifluoromethoxy)phenyl)urea (16),

[0039] 1-(4-(3-aminobenzo[d]isoxazol-4-yl)phenyl)-3-(3-(trifluoromethoxy)phenyl)urea (17),

[0040] 1-(4-(3-aminobenzo[d]isoxazol-4-yl)-2-fluorophenyl)-3-(3-(trifluoromethoxy)phenyl)urea (18),

[0041] 1-(5-(3-aminobenzo[d]isoxazol-4-yl)pyridin-2-yl)-3-(3-(trifluoromethoxy)phenyl)urea (19),

[0042] 2-(4-(3-aminobenzo[d]isoxazol-4-yl)phenyl)-N-(3-(trifluoromethoxy)phenyl)acetamide (20),

[0043] 1-(4-(3-aminobenzo[d]isoxazol-4-yl)phenyl)-3-(5-(tert-butyl)isoxazol-3-yl)urea (21),

[0044] 1-(4-(3-aminobenzo[d]isoxazol-4-yl)phenyl)-3-(3-isopropoxyphenyl)urea (22),

[0045] 6-(3-aminobenzo[d]isoxazol-4-yl)-N-(3-(trifluoromethoxy)phenyl)-3,4-dihydroquinoline-1(2H)-carboxamide (23), 5-(3-aminobenzo[d]isoxazol-4-yl)-N-(3-(trifluoromethoxy)phenyl)indoline-1-carboxamide (24),

[0046] 1-(4-(3-aminobenzo[d]isoxazol-4-yl)phenyl)-3-(3-(trifluoromethyl)phenyl)urea (25),

[0047] 5-(3-aminobenzo[d]isoxazol-4-yl)-N-(3-chloro-5-(trifluoromethoxy)phenyl)indoline-1-carboxamide (26),

[0048] 5-(3-aminobenzo[d]isoxazol-4-yl)-N-(3-(trifluoromethyl)phenyl)indoline-1-carboxamide (27),

[0049] 5-(3-aminobenzo[d]isoxazol-4-yl)-N-(4-chloro-3-(trifluoromethyl)phenyl)indoline-1-carboxamide (28),

[0050] 5-(3-aminobenzo[d]isoxazol-4-yl)-N-(3-fluoro-5-(trifluoromethyl)phenyl)indoline-1-carboxamide (29),

[0051] 5-(3-aminobenzo[d]isoxazol-4-yl)-N-(3-ethylphenyl)indoline-1-carboxamide (30),

[0052] 5-(3-aminobenzo[d]isoxazol-4-yl)-N-(4-(trifluoromethoxy)phenyl)indoline-1-carboxamide (31),

[0053] 5-(3-aminobenzo[d]isoxazol-4-yl)-N-(3-(difluoromethoxy)phenyl)indoline-1-carboxamide (32),

[0054] 5-(3-aminobenzo[d]isoxazol-4-yl)-N-(3-methoxyphenyl)indoline-1-carboxamide (33),

[0055] 1-(4-(3-amino-6-methylbenzo[d]isoxazol-4-yl)phenyl)-3-(3-(trifluoromethoxy)phenyl)urea (34),

[0056] 1-(4-(3-amino-6-methylbenzo[d]isoxazol-4-yl)-2-fluorophenyl)-3-(3-(trifluoromethoxy)phenyl)urea (35),

[0057] 1-(4-(3-amino-6-methylbenzo[d]isoxazol-4-yl)-2,3-fluorophenyl)-3-(3-(trifluoromethoxy)phenyl)urea (36),

[0058] 1-(4-(4-aminothieno[3,2-c]pyridin-3-yl)phenyl)-3-(3-(trifluoromethoxy)phenyl)urea (37),

[0059] 1-(4-(1-oxoisoindolin-4-yl)phenyl)-3-(3-(trifluoromethoxy)phenyl)urea (38),

[0060] 1-(4-(4-amino-1H-pyrazolo[3,4-d]pyrimidin-3-yl)phenyl)-3-(3-(trifluoromethoxy)phenyl)urea (39),

[0061] 1-(4-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-5-yl)phenyl)-3-(3-(trifluoromethoxy)phenyl)urea (40),

[0062] 1-(4-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)-3-(3-(trifluoromethoxy)phenyl)urea (41),

[0063] 1-(4-(3-amino-6-methylisoxazolo[3,4-b]pyridin-4-yl)phenyl)-3-(3-(trifluoromethoxy)phenyl)urea (42),

[0064] 5-(3-amino-6-methylbenzo[d]isoxazol-4-yl)-N-(3-(trifluoromethoxy)phenyl)indoline-1-carboxamide (43),

[0065] N-(4-(3-amino-6-methylisoxazolo[3,4-b]pyridin-4-yl)phenyl)-2-(3-(trifluoromethoxy)phenyl)acetamide (44), N-(4-(3-amino-6-methylisoxazolo[3,4-b]pyridin-4-yl)phenyl)-6-(trifluoromethoxy)indoline-1-carboxamide (45),

[0066] 1-(7-(3-aminobenzo[d]isoxazol-4-yl)-2,3-dihydro-1H-inden-4-yl)-3-(3-(trifluoromethoxy)phenyl)urea (46), 1-(7-(3-aminobenzo[d]isoxazol-4-yl)-3-oxo-2,3-dihydro-1H-inden-4-yl)-3-(3-(trifluoromethoxy)phenyl)urea (47),

[0067] 1-(3-((3-aminobenzo[d]isoxazol-4-yl)oxy)phenyl)-3-(3-(trifluoromethoxy)phenyl)urea (48),

[0068] 1-(3-((3-Aminobenzo[d]isoxazol-4-yl)oxy)phenyl)-3-(3-(trifluoromethyl)phenyl)urea (49).

[0069] A pharmaceutical composition, characterized in that it contains any of the above-mentioned biaryl compounds or pharmaceutically acceptable salts thereof as an active ingredient and is prepared with a pharmaceutically acceptable carrier. Specifically, the pharmaceutical composition can be prepared by methods well known in the art, such as conventional mixing methods, dissolution methods, granulation methods, emulsification methods, freeze-drying methods, etc.

[0070] Among them, the pharmaceutically acceptable salt mentioned in the present invention is a salt formed by the above-mentioned compound and an acid or a base, and the acid is an organic acid or an inorganic acid; the organic acid is methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, naphthalenesulfonic acid, citric acid, malic acid, tartaric acid, lactic acid, pyruvic acid, acetic acid, maleic acid, succinic acid, fumaric acid, salicylic acid, phenylacetic acid or mandelic acid, and the inorganic acid is hydrochloric acid, hydrobromic acid, sulfuric acid or phosphoric acid; the base is a base containing a basic metal cation, an alkaline earth metal cation or an ammonium cation salt.

[0071] The dosage form of the pharmaceutical composition of the present invention is tablets, capsules, syrups, suspensions, inhalants, gels, powders, granules, suppositories, emulsions, or injections, and common pharmaceutical excipients such as flavors, sweeteners, liquid / solid fillers, diluents, etc. can be added during the preparation process.

[0072] Use of any of the above-mentioned biaryl compounds or pharmaceutically acceptable salts thereof, or the above-mentioned pharmaceutical composition in the preparation of a drug for preventing or treating necroptosis-mediated diseases associated with RIPK1.

[0073] As an improvement, the RIPK1-related necroptosis-mediated diseases include but are not limited to: rheumatoid arthritis, ulcerative colitis, systemic lupus erythematosus, psoriasis, atherosclerosis, liver fibrosis, renal fibrosis, pulmonary fibrosis, metabolic dysfunction-related fatty hepatitis, amyotrophic lateral sclerosis, multiple sclerosis, frontotemporal dementia, Alzheimer's disease, Parkinson's disease, ischemia-reperfusion injury of the brain, retina, heart, kidney and liver, sepsis, abnormally high inflammatory response caused by viral or bacterial infection, acute respiratory distress syndrome, retinal degeneration, breast cancer, glioblastoma, pancreatic cancer, melanoma, and acute kidney, liver and lung injury.

[0074] Beneficial effects:

[0075] Compared with the prior art, the present invention provides biaryl compounds and their derivatives with anti-necroptosis activity and their applications. The biaryl compounds and their derivatives are RIPK1 inhibitors with a biaryl skeleton and are widely used. They can be prepared as drugs for treating and / or preventing inflammatory, fibrotic, infectious, ischemic, degenerative or tumor-related diseases caused by necroptosis. Among them, compound 5 is particularly outstanding, which has EC against necroptosis of HT-29 cells. 50 The concentration of 14.43 nM is 50 mg / kg, and oral administration once a day can significantly inhibit the progression of liver fibrosis caused by Schistosoma japonicum infection in mice. In addition, the synthesis method of the biaryl compound and its derivatives of the present invention is simple, easy to operate, and suitable for industrialization. BRIEF DESCRIPTION OF THE DRAWINGS

[0076] Figure 1In order to conduct a drug administration experiment on mice, the drug is compound 5, and the conditions of different parts of the mice are shown, among which (a) is the mouse liver index, (b) is the mouse spleen index, (c) is the mouse liver, and (d) is the mouse spleen.

[0077] Figure 2 (a) HE staining of mouse liver tissue (left) and the granuloma area index of a single worm egg (right).

[0078] Figure 2 (b) Masson staining of mouse liver tissue (left) and the collagen area index of a single egg granuloma (right).

[0079] Figure 3 These are the results of Western blotting experiments, in which RIP1, pRIP1, RIP3, pRIP3, MLKL, and pMLKL are indicators of necroptosis, IL-6, F4 / 80, and TNF-α are indicators of inflammatory factor levels, and Collagen I and α-SMA are indicators of fibrosis levels.

[0080] Figure 4 (a)-(d) are the contents of TNF-α, IL-17A, IL-4, and IL-1β in the culture supernatant of mouse spleen cells, respectively (indicating the inflammation level of mouse spleen under different drug concentrations). DETAILED DESCRIPTION

[0081] Definitions and explanations of terms

[0082] Necroptosis or programmed necrosis refers to a precisely regulated form of cell death mediated by RIPK1.

[0083] In the present invention Indicates the connection site. m -C n means having an integer number of carbon atoms in the range of mn. For example, "C1-C7" means that the group may have 1 carbon atom, 2 carbon atoms, 3 carbon atoms, 4 carbon atoms, 5 carbon atoms, 6 carbon atoms, or 7 carbon atoms. "Alkyl" means a group having the formula C n H 2n+1 The term "C1-C2 alkyl" is understood to mean a saturated hydrocarbon group having 1 or 2 carbon atoms. "Alkoxy" refers to a group resulting from the loss of a hydrogen atom from a hydroxyl group of a straight or branched alcohol, and can be understood as "alkyloxy" or "alkyl-O-". "C1-C2 alkoxy" can be understood as "C1-C2 alkyloxy" or "C1-C2 alkyl-O-". "Halogen" or "halogen" refers to fluorine, chlorine, bromine or iodine. "Hydroxy" refers to an -OH group. "Cyano" refers to a -CN group. "Amino" refers to an -NH2 group.

[0084] Example 1: Preparation of Compound 1

[0085]

[0086] Intermediate A-1 (4-bromobenzo[d]isoxazol-3-amine)

[0087] Dissolve acetohydroxamic acid (3.731 g, 49.70 mmol) in anhydrous DMF (20 mL), add potassium tert-butoxide (5.577 g, 49.70 mmol), stir at room temperature for 1 h, add 2-fluoro-6-bromobenzonitrile (5.000 g, 24.90 mmol), and continue to react at room temperature for 12 h. After the reaction is completed, add 100 mL of water, extract with EA (50 mL × 10), combine the organic layers, wash twice with saturated brine, dry over anhydrous sodium sulfate, concentrate under reduced pressure, and purify with silica gel column chromatography [PE: EA = 20: 1] to obtain 3.000 g of white solid, with a yield of 57%. mp: 158–160℃. 1 H NMR(300MHz,DMSO-d6)δ7.58–7.51(m,1H),7.49–7.44(m,2H),6.12(s,2H).MS m / z(ESI)[M+H] + :213.1.

[0088] Intermediate A-2 (2-methyl-4-phenylboronic acid pinacol ester)

[0089] Dissolve 2-methyl-4-bromoaniline (500.0 mg, 2.691 mmol), biboronic acid pinacol ester (887.2 mg, 3.492 mmol), Pd(dppf)Cl2 (196.8 mg, 0.2691 mmol), potassium acetate (567.0 mg, 5.381 mmol) in anhydrous dioxane (15 mL), react at 110°C for 5 h under nitrogen protection, and after TLC monitoring the disappearance of the raw material, concentrate under reduced pressure and perform silica gel column chromatography [PE:EA=10:1] to obtain 510.0 mg of a yellow solid with a yield of 82%. MS m / z(ESI)[M+H] + :234.1.

[0090] Intermediate A-3 (4-(4-amino-3-methylphenyl)benzo[d]isoxazol-3-amine)

[0091] Intermediate A-2 (500.0 mg, 2.152 mmol), intermediate A-1 (374.0 mg, 1.651 mmol), Pd(dppf)Cl2 (120.7 mg, 0.1650 mmol) were dissolved in dioxane (10 mL), sodium carbonate (524.0 mg, 4.953 mmol) was dissolved in water (2 mL) and added to the mixture, and reacted at 110 ° C for 5 h under nitrogen protection. After TLC monitoring, the raw material disappeared, the mixture was concentrated under reduced pressure, and purified by silica gel column chromatography [PE: EA = 5: 1] to obtain 265.0 mg of yellow solid, with a yield of 67%. MS m / z (ESI) [M+H] + :240.1.

[0092] 1-(4-(3-aminobenzo[d]isoxazol-4-yl)-2-methylphenyl)-3-(3-(trifluoromethoxy)phenyl)urea (1)

[0093] Under ice bath, 3-trifluoromethoxyaniline (196.1 mg, 1.105 mmol) and triphosgene (328.0 mg, 1.105 mmol) were dissolved in anhydrous DCM (20 mL), stirred for 10 min, and triethylamine (448.1 mg, 4.422 mmol) was added. The reaction was continued for 2 h to form isocyanate, and intermediate A-3 (200.0 mg, 0.8800 mmol) was added to react at room temperature overnight. After the reaction was completed under TLC monitoring, the mixture was concentrated under reduced pressure and purified by silica gel column chromatography [PE:EA=2:1] to obtain 48.00 mg of a white solid with a yield of 10%. mp:180–182℃. 1 H NMR(300MHz,DMSO-d6)δ9.46(s,1H),8.19(s,1H),8.05(d,J=8.3Hz,1H),7.76(s,1H),7.59(m,1 H),7.52–7.41(m,2H),7.38–7.27(m,3H),7.15(m,1H),6.97(m,1H),5.25(s,2H),2.35(s,3H).MS m / z(ESI)[M+H] + :442.9.

[0094] Example 2: Preparation of Compound 2

[0095]

[0096] Intermediate A-4 (2,3-dimethyl-4-phenylboronic acid pinacol ester)

[0097] Using 4-bromo-2,3-dimethylaniline (500.0 mg, 2.511 mmol) and biboronic acid pinacol ester (829.5 mg, 3.271 mmol) as raw materials, the synthesis method was the same as that of intermediate compound A-2 to obtain 513.0 mg of yellow solid with a yield of 83%. MS m / z (ESI) [M+H] + :248.2.

[0098] Intermediate A-5 (4-(4-amino-2,3-xylyl)benzo[d]isoxazol-3-amine)

[0099] Intermediate A-4 (500.0 mg, 2.020 mmol) and intermediate A-1 (417.7 mg, 1.840 mmol) were used as raw materials. The synthesis method was the same as that of intermediate compound A-3 to obtain 300.0 mg of yellow solid with a yield of 64%. MS m / z (ESI) [M+H] + :254.1.

[0100] 1-(4-(3-aminobenzo[d]isoxazol-4-yl)-2,3-xylyl)-3-(3-(trifluoromethoxy)phenyl)urea(2)

[0101] Using 3-trifluoromethoxyaniline (231.9 mg, 1.310 mmol) and intermediate A-5 (265.0 mg, 1.050 mmol) as raw materials, the synthesis method was the same as Example 1 to obtain 43.00 mg of white solid with a yield of 9%. mp: 170–172°C. 1 H NMR (400MHz, DMSO-d6) δ9.44(s,1H),8.27(s,1H),7.75(s,1H),7.71(d,J=8.3Hz,1H),7.60(m,1H),7.51(d,J=8.4Hz,1H),7.42(t,J=8.2Hz ,1H),7.35–7.27(m,1H),7.07(d,J=8.3Hz,1H),7.02(d,J=7.1Hz,1H),6.99–6.93(m,1H),4.93(s,2H),2.25(s,3H),2.02(s,3H).HRMS-ESI m / z[M+H] + Calculate for C 23 H 19 F3N4O3:457.1487,found:457.1488.

[0102] Example 3: Preparation of Compound 3

[0103]

[0104] Intermediate A-6 (4-bromo-3-methylphenylboronic acid pinacol ester)

[0105] Using 4-bromo-3-methylaniline (500.0 mg, 2.690 mmol) and biboronic acid pinacol ester (829.5 mg, 3.490 mmol) as raw materials, the synthesis method was the same as that of intermediate compound A-2 to obtain 520.0 mg of yellow solid with a yield of 83%. MS m / z (ESI) [M+H] + :213.4.

[0106] Intermediate A-7 (4-(4-amino-3-methylphenyl)benzo[d]isoxazol-3-amine)

[0107] Using intermediate A-6 (500.0 mg, 2.150 mmol) and intermediate A-1 (417.7 mg, 1.650 mmol) as raw materials, the synthesis method was the same as that of intermediate compound A-3 to obtain 254.0 mg of yellow solid with a yield of 65%. MS m / z (ESI) [M+H] + :234.1.

[0108] 1-(4-(3-aminobenzo[d]isoxazol-4-yl)-3-methylphenyl)-3-(3-(trifluoromethoxy)phenyl)urea (3)

[0109] Using 3-trifluoromethoxyaniline (235.3 mg, 1.332 mmol) and intermediate A-7 (254.0 mg, 1.061 mmol) as raw materials, the synthesis method was the same as Example 1 to obtain 50.00 mg of white solid with a yield of 11%. mp: 100–102°C. 1 H NMR(300MHz,DMSO-d6)δ9.15(d,J=2.8Hz,1H),8.97(d,J=2.8Hz,1H),7.76(s,1H),7.60(m,1H),7.51(m,2H) ,7.42(m,2H),7.32(d,J=8.3Hz,1H),7.19(m,1H),7.05(m,1H),6.97(d,J=8.0Hz,1H),4.97(s,2H).HRMS-ESI m / z[M+H] + Calculate for C 22 H 17 F3N4O3:443.1331,found:443.1334.

[0110] Example 4: Preparation of Compound 4

[0111]

[0112] Intermediate A-8 (4-bromo-3-fluorophenylboronic acid pinacol ester)

[0113] Using 4-bromo-3-fluoroaniline (500.0 mg, 2.630 mmol) and biboronic acid pinacol ester (868.7 mg, 3.420 mmol) as raw materials, the synthesis method was the same as that of intermediate compound A-2 to obtain 504.0 mg of yellow solid with a yield of 81%. MS m / z (ESI) [M+H] + :237.9.

[0114] Intermediate A-9 (4-(4-amino-3-fluorophenyl)benzo[d]isoxazol-3-amine)

[0115] Intermediate A-8 (500.0 mg, 2.110 mmol) and intermediate A-1 (435.4 mg, 1.921 mmol) were used as raw materials. The synthesis method was the same as that of intermediate compound A-3 to obtain 300.0 mg of yellow solid with a yield of 65%. MS m / z (ESI) [M+H] + :244.1.

[0116] 1-(4-(3-aminobenzo[d]isoxazol-4-yl)-3-fluorophenyl)-3-(3-(trifluoromethoxy)phenyl)urea (4)

[0117] Using 3-trifluoromethoxyaniline (273.3 mg, 1.540 mmol) and intermediate A-9 (300.0 mg, 1.230 mmol) as raw materials, the synthesis method was the same as Example 1 to obtain 70.00 mg of white solid with a yield of 13%. mp: 196–198°C. 1 H NMR(300MHz,DMSO-d6)δ9.22(s,1H),9.20(s,1H),7.72(s,1H),7.69–7.63(m,1H),7.62–7.56(m,1 H),7.54(m,1H),7.49–7.35(m,3H),7.31(m,1H),7.16(d,J=7.0Hz,1H),6.99(m,1H),5.20(s,2H). 13 C NMR (75MHz, DMSO-d6) δ162.99, 158.66, 152.75, 149.19, 142.12 (d, J = 11.3Hz), 141.66, 132.05, 130.91, 130.66, 130. 44,124.86,117.96,117.75,117.50,114.71,114.57,114.49,110.88,109.61,105.94(d,J=27.4Hz),79.64.HRMS-ESI m / z[M+H]+ Calculate for C 21 H 14 F4N4O3:447.1080,found:447.1071.

[0118] Example 5: Preparation of Compound 5

[0119]

[0120] Intermediate A-10 (2,3-difluoro-4-phenylboronic acid pinacol ester)

[0121] Using 4-bromo-2,3-difluoroaniline (500.0 mg, 2.420 mmol) and biboronic acid pinacol ester (797.4 mg, 3.140 mmol) as raw materials, the synthesis method was the same as that of intermediate compound A-2 to obtain 490.0 mg of yellow solid with a yield of 79%. mp: 102–106℃. 1 H NMR(300MHz,DMSO-d6)δ7.08(m,1H),6.60–6.50(m,1H),5.96(s,2H),1.26(s,12H).MS m / z(ESI)[M+H] + :256.1.

[0122] Intermediate A-11 (4-(4-amino-2,3-difluorophenyl)benzo[d]isoxazol-3-amine)

[0123] Using intermediate A-10 (490.0 mg, 1.920 mmol) and intermediate A-1 (396.5 mg, 1.752 mmol) as raw materials, the synthesis method was the same as that of intermediate compound A-3 to obtain 313.0 mg of yellow solid with a yield of 68%. 1 H NMR(300MHz,DMSO-d6)δ7.60–7.48(m,2H),7.12(m,1H),6.92(m,1H),6.68(m,1H),5.79(s,2H),5.24(s,2H).MS m / z(ESI)[M+H] + :262.1.

[0124] 1-(4-(3-aminobenzo[d]isoxazol-4-yl)-2,3-difluorophenyl)-3-(3-(trifluoromethoxy)phenyl)urea(5)

[0125] Using 3-trifluoromethoxyaniline (263.2 mg, 1.482 mmol) and intermediate A-11 (310.0 mg, 1.191 mmol) as raw materials, the synthesis method was the same as Example 1 to obtain 47.00 mg of white solid with a yield of 9%. mp: 227–229℃.1 H NMR(300MHz,DMSO-d6)δ9.47(s,1H),8.99(s,1H),8.10(t,J=7.8Hz,1H),7.75(s,1H),7.66–7.55(m,2H ),7.45(t,J=8.2Hz,1H),7.35–7.28(m,1H),7.21(m,2H),7.01(d,J=8.4Hz,1H),5.35(s,2H).HRMS-ESI m / z[M+H] + Calculate for C 21 H 13 F5N4O3:465.0986,found:465.0994. .

[0126] Example 6: Preparation of Compound 6

[0127]

[0128] Intermediate A-12 (2,6-difluoro-4-phenylboronic acid pinacol ester)

[0129] Using 4-bromo-2,6-difluoroaniline (500.0 mg, 2.423 mmol) and biboronic acid pinacol ester (797.4 mg, 3.142 mmol) as raw materials, the synthesis method was the same as intermediate A-2 to obtain 497.0 mg of yellow solid with a yield of 81%. mp: 63–65℃. 1 H NMR(300MHz,DMSO-d6)δ7.06(m,2H),5.71(s,2H),1.26(s,12H).MS m / z(ESI)[M+H] + :256.3.

[0130] Intermediate A-13 (4-(4-amino-2,6-difluorophenyl)benzo[d]isoxazol-3-amine)

[0131] Intermediate A-12 (497.0 mg, 1.950 mmol) and intermediate A-1 (340.3 mg, 1.500 mmol) were used as raw materials. The synthesis method was the same as that of intermediate A-3 to obtain 311.0 mg of yellow solid with a yield of 79%. MS m / z (ESI) [M+H] + :262.2.

[0132] 1-(4-(3-aminobenzo[d]isoxazol-4-yl)-2,6-difluorophenyl)-3-(3-(trifluoromethoxy)phenyl)urea(6)

[0133] Using 3-trifluoromethoxyaniline (263.2 mg, 1.481 mmol) and intermediate A-13 (311.0 mg, 1.190 mmol) as raw materials, the synthesis method was the same as Example 1 to obtain 46.00 mg of white solid with a yield of 8%. mp: 196–198°C. 1 H NMR (300MHz, DMSO-d6) δ9.31(s,1H),8.43(s,1H),7.72–7.57(m,3H),7.45–7.32(m,4H),7.24(m,1H),6.97(d,J=7.7Hz,1H),5.43(s,2H).HRMS-ESI m / z[M+H] + Calculate for C 21 H 13 F5N4O3:465.0986,found:465.0986.

[0134] Example 7: Preparation of Compound 7

[0135]

[0136] Intermediate A-14 (3,5-difluoro-4-phenylboronic acid pinacol ester)

[0137] Using 4-bromo-3,5-difluoroaniline (500.0 mg, 2.420 mmol) and biboronic acid pinacol ester (797.4 mg, 3.140 mmol) as raw materials, the synthesis method was the same as that of intermediate compound A-2 to obtain 501.0 mg of yellow solid with a yield of 81%. MS m / z (ESI) [M+H] + :256.1.

[0138] Intermediate A-15 (4-(4-amino-3,5-difluorophenyl)benzo[d]isoxazol-3-amine)

[0139] Using intermediate A-14 (501.0 mg, 1.960 mmol) and intermediate A-1 (340.3 mg, 1.510 mmol) as raw materials, the synthesis method was the same as that of intermediate compound A-3 to obtain 211.0 mg of yellow solid with a yield of 54%. 1 H NMR(300MHz,DMSO-d6)δ7.60–7.42(m,2H),7.12–7.02(m,2H),6.57–6.42(m,2H),5.74(s,2H),5.15(s,2H).MS m / z(ESI)[M+H] + :262.2.

[0140] 1-(4-(3-aminobenzo[d]isoxazol-4-yl)-3,5-difluorophenyl)-3-(3-(trifluoromethoxy)phenyl)urea(7)

[0141] Using 3-trifluoromethoxyaniline (179.0 mg, 1.010 mmol) and intermediate A-15 (211.0 mg, 0.8080 mmol) as raw materials, the synthesis method was the same as Example 1 to obtain 50.00 mg of a white solid with a yield of 13%. mp: 98–100°C. 1 H NMR (300MHz, DMSO-d6) δ9.39(s,1H),9.32(s,1H),7.70(s,1H),7.67–7.62(m,1H),7.61–7.59(m,1H),7.45(d,J=8 .1Hz,1H),7.41(s,1H),7.40(s,1H),7.37(d,J=2.3Hz,1H),7.18(m,1H),7.03–6.97(m,1H),5.30(s,2H).HRMS-ESI m / z[M+H] + Calculate for C 21 H 13 F5N4O3:465.0986,found:465.0986.

[0142] Example 8: Preparation of Compound 8

[0143]

[0144] Intermediate A-16 (2,5-difluoro-4-phenylboronic acid pinacol ester)

[0145] Using 4-bromo-3,5-difluoroaniline (500.0 mg, 2.420 mmol) and biboronic acid pinacol ester (797.4 mg, 3.140 mmol) as raw materials, the synthesis method was the same as that of intermediate compound A-2 to obtain 521.0 mg of yellow solid with a yield of 84%. MS m / z (ESI) [M+H] + :234.3.

[0146] Intermediate A-17 (4-(4-amino-2,5-difluorophenyl)benzo[d]isoxazol-3-amine)

[0147] Using intermediate A-16 (521.0 mg, 2.040 mmol) and intermediate A-1 (356.8 mg, 1.570 mmol) as raw materials, the synthesis method was the same as that of intermediate compound A-3 to obtain 304.0 mg of yellow solid with a yield of 74%. 1H NMR(300MHz,DMSO-d6)δ7.63–7.42(m,2H),7.14–7.06(m,2H),6.67(m,1H),5.76(s,2H),5.24(s,2H).MS m / z(ESI)[M+H] + :262.4.

[0148] 1-(4-(3-aminobenzo[d]isoxazol-4-yl)-2,5-difluorophenyl)-3-(3-(trifluoromethoxy)phenyl)urea(8)

[0149] Using 3-trifluoromethoxyaniline (258.0 mg, 1.460 mmol) and intermediate A-17 (304.0 mg, 1.160 mmol) as raw materials, the synthesis method was the same as Example 1 to obtain 53.00 mg of white solid with a yield of 10%. mp: 198–200℃. 1 H NMR(300MHz,DMSO-d6)δ9.52(s,1H),9.01(s,1H),8.19(m,1H),7.74(s,1H),7.64–7.54(m,2H), 7.49–7.39(m,2H),7.33–7.28(m,1H),7.18(d,J=6.8Hz,1H),7.02(d,J=8.2Hz,1H),5.34(s,2H). 13 C NMR(75MHz,DMSO-d6)δ163.04,158.69,156.39,154.02,152.32,149.25,141.26,131.05,130.38,129.5 7,124.96,117.66,117.47,117.36,114.76,114.61,110.72,109.97,107.64,107.61,107.32.HRMS-ESI m / z[M+H] + Calculate for C 21 H 13 F5N4O3:465.0986,found:465.0989.

[0150] Example 9: Preparation of Compound 9

[0151]

[0152] Intermediate A-18 (4-bromo-3-chloro-2-fluorophenylboronic acid pinacol ester)

[0153] Using 4-bromo-3-chloro-2-fluoroaniline (500.0 mg, 2.230 mmol) and biboronic acid pinacol ester (736.9 mg, 2.900 mmol) as raw materials, the synthesis method was the same as that of intermediate compound A-2 to obtain 534.0 mg of yellow solid with a yield of 87%. mp: 76–80℃. 1 H NMR(300MHz,DMSO-d6)δ7.20(d,J=8.2Hz,1H),6.59(d,J=10.7Hz,1H),5.90(s,2H),1.26(s,12H).MS m / z(ESI)[M+H] + :272.2.

[0154] Intermediate A-19 (4-(4-amino-3-chloro-2-fluorophenyl)benzo[d]isoxazol-3-amine)

[0155] Using intermediate A-18 (524.0 mg, 1.930 mmol) and intermediate A-1 (399.0 mg, 1.760 mmol) as raw materials, the synthesis method was the same as that of intermediate compound A-3 to obtain 300.0 mg of yellow solid with a yield of 62%. 1 H NMR (300MHz, DMSO-d6) δ7.69–7.47(m,3H),7.06(m,1H),6.96(m,1H),6.83(t,J=8.5Hz,1H),5.74(s,2H),5.08(s,2H).MS m / z(ESI)[M+H] + :277.4.

[0156] 1-(4-(3-aminobenzo[d]isoxazol-4-yl)-3-chloro-2-fluorophenyl)-3-(3-(trifluoromethoxy)phenyl)urea (9)

[0157] Using 3-trifluoromethoxyaniline (240.0 mg, 1.350 mmol) and intermediate A-19 (300.0 mg, 1.080 mmol) as raw materials, the synthesis method was the same as Example 1 to obtain 56.00 mg of white solid with a yield of 11%. mp: 129–130℃. 1 H NMR(300MHz,DMSO-d6)δ9.54(s,1H),8.98(d,J=2.7Hz,1H),8.35–8.22(m,1H),7.77(m,1H),7.67–7.56( m,2H),7.46(t,J=8.2Hz,1H),7.34–7.24(m,2H),7.13(m,1H),7.07–6.98(m,1H),5.24(s,2H).HRMS-ESI m / z[M+H] +Calculate for C 21 H 13 ClF4N4O3:481.0690,found:481.0680.

[0158] Example 10: Preparation of Compound 10

[0159]

[0160] Intermediate A-20 (4-bromo-2-chlorophenylboronic acid pinacol ester)

[0161] Using 4-bromo-2-chloroaniline (500.0 mg, 2.430 mmol) and biboronic acid pinacol ester (801.0 mg, 3.161 mmol) as raw materials, the synthesis method was the same as that of intermediate compound A-2 to obtain 470.0 mg of yellow solid with a yield of 76%. MS m / z (ESI) [M+H] + :254.4.

[0162] Intermediate A-21 (4-(4-amino-2-chlorophenyl)benzo[d]isoxazol-3-amine)

[0163] Using intermediate A-20 (470.0 mg, 1.860 mmol) and intermediate A-1 (324.0 mg, 1.431 mmol) as raw materials, the synthesis method was the same as that of intermediate compound A-3 to obtain 290.0 mg of yellow solid with a yield of 78%. MS m / z (ESI) [M+H] + :260.2.

[0164] 1-(4-(3-aminobenzo[d]isoxazol-4-yl)-2-chlorophenyl)-3-(3-(trifluoromethoxy)phenyl)urea (10)

[0165] Using 3-trifluoromethoxyaniline (247.9 mg, 1.400 mmol) and intermediate A-21 (290.0 mg, 1.120 mmol) as raw materials, the synthesis method was the same as Example 1 to obtain 65.00 mg of white solid with a yield of 13%. mp: 190–192°C. 1 H NMR (300MHz, DMSO-d6) δ9.90 (s, 1H), 8.61 (s, 1H), 8.33 (d, J = 8.6Hz, 1H), 7.76 (s, 1H), 7.68–7.39(m,5H),7.31(m,1H),7.19(m,1H),7.05–6.95(m,1H),5.34(s,2H).HRMS-ESI m / z[M+H] + Calculate for C 21 H 14ClF3N4O3:463.0785,found:463.0784.

[0166] Example 11: Preparation of Compound 11

[0167]

[0168] Intermediate A-22 (4-bromo-3-chlorophenylboronic acid pinacol ester)

[0169] Using 4-bromo-3-chloroaniline (500.0 mg, 2.430 mmol) and biboronic acid pinacol ester (801.0 mg, 3.162 mmol) as raw materials, the synthesis method was the same as that of intermediate compound A-2 to obtain 470.0 mg of yellow solid with a yield of 76%. MS m / z (ESI) [M+H] + :254.2.

[0170] Intermediate A-23 (4-(4-amino-3-chlorophenyl)benzo[d]isoxazol-3-amine)

[0171] Using intermediate A-22 (470.0 mg, 1.860 mmol) and intermediate A-1 (324.0 mg, 1.430 mmol) as raw materials, the synthesis method was the same as that of intermediate compound A-3 to obtain 295.0 mg of yellow solid with a yield of 80%. 1 H NMR (300MHz, DMSO-d6) δ7.62–7.42(m,2H),7.05(m,2H),6.77(d,J=2.0Hz,1H),6.64(d,J=8.3Hz,1H),5.70(s,2H),5.01(s,2H).MS m / z(ESI)[M+H] + :260.1.

[0172] 1-(4-(3-aminobenzo[d]isoxazol-4-yl)-3-chlorophenyl)-3-(3-(trifluoromethoxy)phenyl)urea (11)

[0173] Using 3-trifluoromethoxyaniline (252.2 mg, 1.420 mmol) and intermediate A-23 (295.0 mg, 1.142 mmol) as raw materials, the synthesis method was the same as Example 1 to obtain 69.00 mg of white solid with a yield of 13%. mp: 115–117°C. 1H NMR (400MHz, DMSO-d6) δ9.23(s,1H),9.21(s,1H),7.90(d,J=2.1Hz,1H),7.72(s,1H),7.61(m,1H),7.54(d,J=8.4Hz,1H) ,7.48–7.45(m,1H),7.42(d,J=8.1Hz,1H),7.36(m,2H),7.10(d,J=7.1Hz,1H),7.01–6.96(m,1H),5.04(s,2H).HRMS-ESI m / z[M+H] + Calculate for C 21 H 14 ClF3N4O3:463.0785,found:463.0789.

[0174] Example 12: Preparation of Compound 12

[0175]

[0176] Intermediate A-24 (2,3-dichloro-4-phenylboronic acid pinacol ester)

[0177] Using 4-bromo-2,3-dichloroaniline (500.0 mg, 2.090 mmol) and biboronic acid pinacol ester (690.6 mg, 2.720 mmol) as raw materials, the synthesis method was the same as that of intermediate compound A-2 to obtain 495.0 mg of yellow solid with a yield of 83%. MS m / z (ESI) [M+H] + :288.2.

[0178] Intermediate A-25 (4-(4-amino-2,3-dichlorophenyl)benzo[d]isoxazol-3-amine)

[0179] Using intermediate A-24 (495.0 mg, 1.722 mmol) and intermediate A-1 (301.0 mg, 1.330 mmol) as raw materials, the synthesis method was the same as that of intermediate compound A-3 to obtain 305.0 mg of yellow solid with a yield of 78%. 1 H NMR (300MHz, DMSO-d6) δ7.63–7.46(m,2H),7.15–7.02(m,2H),6.87(d,J=8.4Hz,1H),5.96(s,2H),5.05(s,2H).MSm / z(ESI)[M+H] + :293.4.

[0180] 1-(4-(3-aminobenzo[d]isoxazol-4-yl)-2,3-dichlorophenyl)-3-(3-(trifluoromethoxy)phenyl)urea(12)

[0181] Using 3-trifluoromethoxyaniline (263.0 mg, 1.280 mmol) and intermediate A-25 (301.0 mg, 1.030 mmol) as raw materials, the synthesis method was the same as Example 1 to obtain 60.00 mg of white solid with a yield of 12%. mp: 110–112°C. 1 H NMR (300MHz, DMSO-d6) δ11.28(s,1H),10.03(s,1H),9.65(d,J=8.6Hz,1H),9.09(s,1H),9.00–8.94(m,1H),8.91(d,J=8.2Hz,1H),8.80(t ,J=8.1Hz,1H),8.74(d,J=8.7Hz,1H),8.67(d,J=8.4Hz,1H),8.46(d,J=6.8Hz,1H),8.35(d,J=8.2Hz,1H),6.50(s,2H).HRMS-ESIm / z[M+H] + Calculate for C 21 H 13 Cl2F3N4O3:497.0395,found:497.0399.

[0182] Example 13: Preparation of Compound 13

[0183]

[0184] Intermediate A-26 (4-amino-3-cyanophenylboronic acid pinacol ester)

[0185] Using 5-amino-2-bromobenzonitrile (500.0 mg, 2.551 mmol) and biboronic acid pinacol ester (842.1 mg, 3.310 mmol) as raw materials, the synthesis method was the same as that of intermediate compound A-2 to obtain 469.0 mg of yellow solid with a yield of 75%. MS m / z (ESI) [M+H] + :245.2.

[0186] Intermediate A-27 (4-(4-amino-3-cyano)benzo[d]isoxazol-3-amine)

[0187] Using intermediate A-26 (469.0 mg, 1.920 mmol) and intermediate A-1 (335.6 mg, 1.482 mmol) as raw materials, the synthesis method was the same as that of intermediate compound A-3 to obtain 260.0 mg of yellow solid with a yield of 70%. 1H NMR (400MHz, DMSO-d6) δ7.60(t,J=7.8Hz,1H),7.53(d,J=8.4Hz,1H),7.24(d,J=8.4Hz,1H ),7.14(d,J=7.0Hz,1H),7.03(d,J=2.3Hz,1H),6.95(m,1H),5.88(s,2H),5.12(s,2H).MS m / z(ESI)[M+H] + :251.2.

[0188] 1-(4-(3-aminobenzo[d]isoxazol-4-yl)-3-cyanophenyl)-3-(3-(trifluoromethoxy)phenyl)urea (13)

[0189] Using 3-trifluoromethoxyaniline (232.3 mg, 1.300 mmol) and intermediate A-27 (260.0 mg, 1.040 mmol) as raw materials, the synthesis method was the same as Example 1 to obtain 65.00 mg of white solid with a yield of 14%. mp: 214–216℃. 1 H NMR(400MHz, DMSO-d6)δ9.29(s,1H),9.27(s,1H),8.14(d,J=2.3Hz,1H),7.80(m,1H),7.72(s,1H),7.68–7.59(m,2H),7.54(d, J=8.5Hz,1H),7.44(t,J=8.1Hz,1H),7.36(d,J=8.7Hz,1H),7.23(d,J=6.9Hz,1H),7.00(d,J=8.7Hz,1H),5.22(s,2H).HRMS-ESI m / z[M+H] + Calculate for C 22 H 14 F3N5O3:454.1127,found:454.1122.

[0190] Example 14: Preparation of Compound 14

[0191]

[0192] Intermediate A-28 (4-bromo-2-fluoro-5-methylphenylboronic acid pinacol ester)

[0193] Using 4-bromo-2-fluoro-5-methylaniline (500.0 mg, 2.450 mmol) and biboronic acid pinacol ester (809.1 mg, 3.190 mmol) as raw materials, the synthesis method was the same as that of intermediate compound A-2 to obtain 504.0 mg of yellow solid with a yield of 82%. MS m / z (ESI) [M+H] +:252.2.

[0194] Intermediate A-29 (4-(4-amino-2-fluoro-5-methyl)benzo[d]isoxazol-3-amine)

[0195] Intermediate A-28 (504.0 mg, 2.000 mmol) and intermediate A-1 (347.8 mg, 1.540 mmol) were used as raw materials. The synthesis method was the same as that of intermediate compound A-3 to obtain 283.0 mg of yellow solid with a yield of 72%. MS m / z (ESI) [M+H] + :257.1.

[0196] 1-(4-(3-aminobenzo[d]isoxazol-4-yl)-2-fluoro-5-methylphenyl)-3-(3-(trifluoromethoxy)phenyl)urea (14)

[0197] Using 3-trifluoromethoxyaniline (241.2 mg, 1.360 mmol) and intermediate A-29 (283.0 mg, 1.090 mmol) as raw materials, the synthesis method was the same as Example 1 to obtain 30.00 mg of white solid with a yield of 6%. mp: 155–157°C. 1 H NMR (300MHz, DMSO-d6) δ9.41(s,1H),8.68(s,1H),8.11(d,J=8.2Hz,1H),7.70(s,1H),7.53(d,J=7.7Hz,1H), 7.47(s,1H),7.36(d,J=8.2Hz,1H),7.25–7.09(m,2H),7.04–6.87(m,2H),4.98(s,2H),1.97(s,3H).HRMS-ESI m / z[M+H] + Calculate for C 22 H 16 F4N4O3:461.1237,found:461.1223.

[0198] Example 15: Preparation of Compound 15

[0199]

[0200] Intermediate A-30 (2-methyl-4-bromo-5-fluorophenylboronic acid pinacol ester)

[0201] Using 2-methyl-4-bromo-5-fluoroaniline (500.0 mg, 2.450 mmol) and biboronic acid pinacol ester (809.1 mg, 3.190 mmol) as raw materials, the synthesis method was the same as that of intermediate compound A-2 to obtain 501.0 mg of yellow solid with a yield of 81%. MS m / z (ESI) [M+H]+ :252.2.

[0202] Intermediate A-31 (4-(4-amino-5-fluoro-2-methyl)benzo[d]isoxazol-3-amine)

[0203] Intermediate A-30 (501.0 mg, 2.000 mmol) and intermediate A-1 (347.8 mg, 1.540 mmol) were used as raw materials. The synthesis method was the same as that of intermediate compound A-3 to obtain 280.0 mg of yellow solid with a yield of 71%. MS m / z (ESI) [M+H] + :257.1.

[0204] 1-(4-(3-aminobenzo[d]isoxazol-4-yl)-5-fluoro-2-methylphenyl)-3-(3-(trifluoromethoxy)phenyl)urea (15)

[0205] Using 3-trifluoromethoxyaniline (241.2 mg, 1.360 mmol) and intermediate A-31 (280.0 mg, 1.090 mmol) as raw materials, the synthesis method was the same as Example 1 to obtain 40.00 mg of white solid with a yield of 8%. mp: 107–109°C. 1 H NMR(300MHz,DMSO-d6)δ9.56(s,1H),8.28(s,1H),8.05(d,J=12.8Hz,1H),7.75(m,1H),7.65–7.51(m ,2H),7.45(t,J=8.2Hz,1H),7.34–7.27(m,2H),7.15(m,1H),7.00(m,1H),5.19(s,2H),2.30(s,3H). 13 C NMR(75MHz,DMSO-d6)δ162.94,159.10,158.65,155.92,152.71,149.25,141.63,139.60,139.45,132.77,132.71,131.02,130.72,1 30.43,124.79,123.02,122.97,122.28,118.88,118.01,117.80,117.24,114.54,114.49,110.65,109.58,107.40,107.03,17.61.MS m / z(ESI)[M+H] + :459.2.

[0206] Example 16: Preparation of Compound 16

[0207]

[0208] Intermediate A-32 (2-methoxy-4-phenylboronic acid pinacol ester)

[0209] Using 2-methoxy-4-bromoaniline (500.0 mg, 2.480 mmol) and biboronic acid pinacol ester (817.1 mg, 3.220 mmol) as raw materials, the synthesis method was the same as that of intermediate compound A-2 to obtain 509.0 mg of yellow solid with a yield of 82%. MS m / z (ESI) [M+H] + :250.2.

[0210] Intermediate A-33 (4-(4-amino-2-methoxyphenyl)benzo[d]isoxazol-3-amine)

[0211] Using intermediate A-32 (509.0 mg, 2.040 mmol) and intermediate A-1 (356.9 mg, 1.570 mmol) as raw materials, the synthesis method was the same as that of intermediate compound A-3 to obtain 350.0 mg of yellow solid with a yield of 87%. MS m / z (ESI) [M+H] + :256.2.

[0212] 1-(4-(3-aminobenzo[d]isoxazol-4-yl)-2-methoxyphenyl)-3-(3-(trifluoromethoxy)phenyl)urea(16)

[0213] Using 3-trifluoromethoxyaniline (303.9 mg, 1.720 mmol) and intermediate A-33 (350.0 mg, 1.370 mmol) as raw materials, the synthesis method was the same as Example 1 to obtain 60.00 mg of white solid with a yield of 10%. mp: 185–187°C. 1 H NMR (300MHz, DMSO-d6) δ9.74(s,1H),8.48(s,1H),8.31(d,J=8.4Hz,1H),7.76(s,1H),7.60(t,J=7.8Hz,1H),7.50(d,J=8.3Hz,1 H),7.43(t,J=8.2Hz,1H),7.31–7.25(m,1H),7.24–7.17(m,2H),7.08(m,1H),6.97(d,J=8.3Hz,1H),5.32(s,2H),3.97(s,3H).MS m / z(ESI)[M+H] + :481.1.

[0214] Example 17: Preparation of Compound 17

[0215]

[0216] Intermediate A-34 (4-(4-aminophenyl)benzo[d]isoxazol-3-amine)

[0217] Using 4-aminophenylboronic acid pinacol ester (480.0 mg, 2.190 mmol) and 4-bromobenzo[d]isoxazole-3-amine (310.0 mg, 1.460 mmol) as raw materials, the synthesis method was the same as that of intermediate compound A-3, and finally 180.0 mg of yellow oil was obtained with a yield of 55%. MS m / z(ESI)[M+H] + :226.3.

[0218] 1-(4-(3-aminobenzo[d]isoxazol-4-yl)phenyl)-3-(3-(trifluoromethoxy)phenyl)urea(17)

[0219] Using intermediate A-34 (180.0 mg, 0.7990 mmol) and 3-trifluoromethoxyaniline (170.0 mg, 0.9560 mmol) as raw materials, the synthesis method was the same as Example 1, and finally 125.0 mg of white solid was obtained with a yield of 37%. mp: 200–202℃. 1 HNMR(300MHz,DMSO-d6)δ9.11(s,1H),9.01(s,1H),7.74(s,1H),7.68–7.57(m,3H),7.53–7 .41(m,4H),7.37–7.31(m,1H),7.16(dd,J=7.2,0.9Hz,1H),7.02–6.94(m,1H),5.25(s,2H). 13 CNMR(75MHz,DMSO-d6)δ163.27,158.56,152.85,149.22,149.20,149.18,141.89,140.10,137.62,1 31.16,130.86,130.59,129.79,123.89,118.98,117.33,114.23,113.36,110.71,108.81.HRMS-ESI m / z[M+H] + Calculate for C 21 H 15 F3N4O3:429.1169,found:429.1178.

[0220] Example 18: Preparation of Compound 18

[0221]

[0222] Intermediate A-35 (4-(4-amino-3-fluorophenyl)benzo[d]isoxazol-3-amine)

[0223] Using 3-fluoro-4-aminophenylboronic acid pinacol ester (334.0 mg, 1.410 mmol) and 4-bromobenzo[d]isoxazole-3-amine (200.0 mg, 0.9390 mmol) as raw materials, the synthesis method was the same as that of intermediate compound A-3, and finally 55.00 mg of yellow oil was obtained with a yield of 24%. MS m / z(ESI)[M+H] + :243.3.

[0224] 1-(4-(3-aminobenzo[d]isoxazol-4-yl)-2-fluorophenyl)-3-(3-(trifluoromethoxy)phenyl)urea(18)

[0225] Using intermediate A-35 (55.00 mg, 0.2270 mmol) and 3-trifluoromethoxyaniline (48.00 mg, 0.2720 mmol) as raw materials, the synthesis method was the same as Example 1, and finally 30.00 mg of white solid was obtained, with a yield of 27%. mp: 198–200℃. 1 HNMR(300MHz, DMSO-d6)δ9.48(s,1H),8.82(d,J=2.7Hz,1H),8.33(t,J=8.5Hz,1H),7.77–7.75(m,1H),7.61(dd,J=8.5,7.1Hz,1H),7.53(d d,J=8.5,1.0Hz,1H),7.47–7.41(m,2H),7.33(dd,J=8.1,1.9Hz,2H),7.19(dd,J=7.1,1.0Hz,1H),7.02–6.98(m,1H),5.34(s,2H).HRMS-ESI m / z[M+H] + Calculate for C 21 H 14 F4N4O3:447.1075,found:447.1076.

[0226] Example 19: Preparation of Compound 19

[0227]

[0228] Intermediate A-36 (2-aminopyridine-5-boronic acid pinacol ester)

[0229] Using 2-amino-5-bromopyridine (500.0 mg, 2.890 mmol) and biboronic acid pinacol ester (954.0 mg, 3.760 mmol) as raw materials, the synthesis method was the same as that of intermediate compound A-2 to obtain 501.0 mg of yellow solid with a yield of 79%. mp: 131-135℃.MS m / z(ESI)[M+H] +:221.2.

[0230] Intermediate A-37 (4-(6-aminopyridin-3-yl)benzo[d]isoxazol-3-amine)

[0231] Using intermediate A-36 (501.0 mg, 2.270 mmol) and intermediate A-1 (400.0 mg, 1.760 mmol) as raw materials, the synthesis method was the same as that of intermediate compound A-3 to obtain 203.0 mg of yellow solid with a yield of 51%. 1 H NMR(300MHz, DMSO-d6)δ8.05(m,1H),7.56–7.50(m,2H),7.44(m,1H),7.09(dd,J=7.2,0.9Hz,1H),6.57(m,1H),6.26(s,2H),5.29(s,2H).MS m / z(ESI)[M+H] + :227.3.

[0232] 1-(5-(3-aminobenzo[d]isoxazol-4-yl)pyridin-2-yl)-3-(3-(trifluoromethoxy)phenyl)urea (19)

[0233] Using 3-trifluoromethoxyaniline (196.0 mg, 1.105 mmol) and intermediate A-37 (200.0 mg, 0.8840 mmol) as raw materials, the synthesis method was the same as Example 1 to obtain 44.00 mg of white solid with a yield of 13%. mp: 190–192°C. 1 H NMR (300MHz, DMSO-d6) δ10.78(s,1H),9.74(s,1H),8.46(s,1H),7.94(d,J=8.6Hz,1H),7.82(s,1H),7.70– 7.62(m,2H),7.56(d,J=7.7Hz,1H),7.48–7.42(m,2H),7.24–7.19(m,1H),7.06–7.00(m,1H),5.41(s,2H). 13 C NMR(75MHz,DMSO-d6)δ163.47,158.82,152.78,152.57,149.22,147.05,141.23,139.52,134.19,1 31.01,130.70,127.27,124.31,118.88,117.92,114.92,113.67,112.10,111.34,109.45.HRMS-ESI m / z[M+H] + Calculate for C 20 H 14F3N5O3:430.1127,found:430.1129.

[0234] Example 20: Preparation of Compound 20

[0235]

[0236] Intermediate A-38 (2-(4-(3-aminobenzo[d]isoxazol-4-yl)phenyl)acetic acid)

[0237] Using 4-(carboxymethyl)phenylboronic acid pinacol ester (500.0 mg, 1.910 mmol) as raw material, the synthesis method was the same as intermediate A-2 to obtain 300.0 mg of yellow solid with a yield of 59%. 1 HNMR (300MHz, DMSO-d6) δ12.33(s,1H),7.59(d,J=7.4Hz,1H),7.54(s,1H),7.45(d,J=3.3Hz,4H),7.16(d,J=7.1Hz,1H),5.18(s,2H),3.69(s,2H).MS m / z(ESI)[M+H] + :267.2.

[0238] 2-(4-(3-aminobenzo[d]isoxazol-4-yl)phenyl)-N-(3-(trifluoromethoxy)phenyl)acetamide (20)

[0239] Intermediate A-38 (111.5 mg, 0.4160 mmol), HATU (174.0 mg, 0.4580 mmol), DIEA (162.0 mg, 1.250 mmol) were dissolved in anhydrous DMF (5 mL), stirred at room temperature for 10 min, and then 3-trifluoromethoxyaniline (73.70 mg, 0.4160 mmol) was added, and the reaction was carried out at room temperature for 12 h. After TLC monitoring of the disappearance of the raw material, 30 mL of water was added to the reaction solution, and ethyl acetate (10 mL×10) was used for extraction. The organic layer was washed twice with saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by silica gel column chromatography [PE:EA=2:1] to obtain 43.00 mg of a white solid with a yield of 24%. mp:78–80℃. 1H NMR (400MHz, DMSO-d6) δ10.53(s,1H),7.83(s,1H),7.63–7.58(m,2H),7.53(d,J=5.3Hz,2H),7.50(d,J=2. 0Hz,3H),7.44(d,J=8.2Hz,1H),7.16(d,J=7.2Hz,1H),7.05(d,J=7.9Hz,1H),5.19(s,2H),3.78(s,2H).MS m / z(ESI)[M+H] + :428.0.

[0240] Example 21: Preparation of Compound 21

[0241] 1-(4-(3-aminobenzo[d]isoxazol-4-yl)phenyl)-3-(5-(tert-butyl)isoxazol-3-yl)urea (21)

[0242] Using intermediate A-34 (400.0 mg, 1.780 mmol) and 3-amino-5-tert-butylisoxazole (311.5 mg, 2.225 mmol) as raw materials, the synthesis method was the same as Example 1 to obtain 20.00 mg of white solid with a yield of 3%. mp: 210–212℃. 1 H NMR (300MHz, DMSO-d6) δ9.64 (s, 1H), 9.19–9.07 (m, 1H), 7.63 (d, J = 8.4Hz, 2H), 7.61–7.55 (m,1H),7.53–7.41(m,3H),7.15(d,J=7.1Hz,1H),6.53(s,1H),5.24(s,2H),1.31(s,9H). 13 CNMR(75MHz,DMSO-d6)δ180.69,163.26,158.79,158.55,151.82,139.65,137.52,131.53, 130.60,129.86,123.91,119.03,113.35,108.88,92.94,32.94,28.81.HRMS-ESIm / z[M+H] + Calculate for C 21 H 21 N5O3:392.1722,found:392.1726.

[0243] Example 22: Preparation of Compound 22

[0244] 1-(4-(3-aminobenzo[d]isoxazol-4-yl)phenyl)-3-(3-isopropoxyphenyl)urea(22)

[0245] Using intermediate A-34 (400.0 mg, 1.780 mmol) and 3-isopropoxyaniline (403.7 mg, 2.670 mmol) as raw materials, the synthesis method was the same as Example 1 to obtain 40.00 mg of a white solid with a yield of 5%. 1 H NMR(300MHz,DMSO-d6)δ8.89(s,1H),8.74(s,1H),7.67–7.59(m,3H),7.46–7.40(m,3H),7.20(t,J=2.4Hz,1H),7.18–7.11 (m,2H),6.92(ddd,J=8.0,2.1,0.9Hz,1H),6.58–6.50(m,1H),5.23(s,2H),4.56(p,J=6.0Hz,1H),1.28(d,J=6.0Hz,6H).MS m / z(ESI)[M+H] + :403.3.

[0246] Example 23: Preparation of Compound 23

[0247]

[0248] Intermediate A-39 (6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,2,3,4-tetrahydroquinoline)

[0249] Using 6-bromo-1,2,3,4-tetrahydroquinoline (1000 mg, 4.710 mmol) and biboronic acid pinacol ester (1430 mg, 5.650 mmol) as raw materials, the synthetic method was the same as that of intermediate A-2 to finally obtain 250.0 mg of yellow solid with a yield of 21%. mp: 75–78°C.MS m / z(ESI)[M+H] + :260.3.

[0250] Intermediate A-40 (4-(1,2,3,4-tetrahydroquinolin-6-yl)benzo[d]isoxazol-3-amine)

[0251] Using intermediate A-39 (250.0 mg, 0.9650 mmol) and A-1 (137.0 mg, 0.6430 mmol) as raw materials, the synthesis method was the same as that of intermediate compound A-3, and finally 90.00 mg of yellow oil was obtained with a yield of 53%. MS m / z (ESI) [M+H] + :266.3.

[0252] 6-(3-Aminobenzo[d]isoxazol-4-yl)-N-(3-(trifluoromethoxy)phenyl)-3,4-dihydroquinoline-1(2H)-carboxamide (23)

[0253] Using intermediate A-40 (90.00 mg, 34.00 mmol) and 3-trifluoromethoxyaniline (72.00 mg, 0.4080 mmol) as raw materials, the synthesis method was the same as Example 1 to obtain 49 mg of white solid with a yield of 31%. mp: 96–100℃. 1 H NMR (300MHz, DMSO-d6) δ9.28 (s, 1H), 7.69 (m, 1H), 7.59 (dd, J = 8.4, 7.1Hz, 1H), 7.52–7.45(m,3H),7.39(t,J=8.1Hz,1H),7.34(d,J=2.1Hz,1H),7.25(dd,J=8.4 ,2.2Hz,1H),7.16(dd,J=7.2,0.9Hz,1H),6.96(ddt,J=8.0,2.4,1.2Hz,1H),5.4 0(s,2H),3.78(dd,J=6.8,5.4Hz,2H),2.85(t,J=6.5Hz,2H),1.99–1.92(m,2H). 13 C NMR(75MHz,DMSO-d6)δ163.24,158.51,154.64,148.99,148.96,142.34,139.37,137.57,132.08,131.19,130.55, 130.52,129.47,127.09,123.81,123.71,118.34,114.48,113.33,111.77,108.89,44.85,27.08,23.79.HRMS-ESI m / z[M+H] + Calculate for C 24 H 20 F3N4O3:469.1482,found:469.1485.

[0254] Example 24: Preparation of Compound 24

[0255]

[0256] Intermediate A-41 (5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)indoline)

[0257] 5-bromoindoline (1000 mg, 5.040 mmol), biboronic acid pinacol ester (1020 mg, 4.320 mmol), and the same synthesis method as intermediate A-2 were used to obtain 764 mg of yellow solid with a yield of 59%. mp: 82-87℃. 1 H NMR(300MHz,Chloroform-d)δ7.83(dt,J=6.7,1.5Hz,1H),7.70-7.59(m,2H),3.99(t,J =8.7Hz,2H),3.09(ddd,J=9.5,8.1,1.1Hz,2H),1.36(d,J=4.4Hz,9H),1.28(s,12H).MS m / z(ESI)[M+H] + :246.3.

[0258] Intermediate A-42 (4-(Indol-5-yl)benzo[d]isoxazol-3-amine)

[0259] Intermediate A-41 (764 mg, 2.970 mmol) and intermediate A-1 (421 mg, 1.980 mmol) were synthesized in the same manner as intermediate A-3 to obtain 358.0 mg of a yellow oil with a yield of 72%. MS m / z (ESI) [M+H] + :252.3.

[0260] 5-(3-Aminobenzo[d]isoxazol-4-yl)-N-(3-(trifluoromethoxy)phenyl)indoline-1-carboxamide (24)

[0261] Using intermediate A-42 (40.00 mg, 0.1590 mmol) and 3-trifluoromethoxyaniline (34.00 mg, 0.1910 mmol) as raw materials, the synthesis method was the same as Example 1 to obtain 32.00 mg of white solid with a yield of 44%. mp: 89–92°C. 1 H NMR(300MHz,Chloroform-d)δ8.38(d,J=8.8Hz,1H),7.53(dd,J=8.5,7.2Hz,1H),7.45–7.39(m,2H),7.37–7.28(m,4H ),7.24–7.17(m,2H),7.12(dd,J=7.2,0.9Hz,1H),4.21(t,J=8.5Hz,2H),3.89(s,2H),3.32(t,J=8.4Hz,2H).HRMS-ESI m / z[M+H] + Calculate for C 23 H 17 F3N4O3:455.1326,found:455.1331.

[0262] Example 25: Preparation of Compound 25

[0263] 1-(4-(3-aminobenzo[d]isoxazol-4-yl)phenyl)-3-(3-(trifluoromethyl)phenyl)urea(25)

[0264] Intermediate A-34 (90.00 mg, 0.4000 mmol), 3-trifluoromethylphenyl isocyanate (74.80 mg, 0.4000 mmol), triethylamine (121.0 mg, 1.200 mmol) were dissolved in anhydrous DCM (5 mL), reacted at room temperature for 4 h, concentrated under reduced pressure after TLC monitoring of the disappearance of the raw material, and purified by silica gel column chromatography (PE:EA=1:5) to obtain 136.0 mg of a white solid with a yield of 82%. mp: 244–246°C. 1 H NMR(300MHz,DMSO-d6)δ9.14(s,1H),9.02(s,1H),8.05(s,1H),7.68–7.57(m,4H ),7.55–7.43(m,4H),7.33(d,J=7.6Hz,1H),7.15(d,J=7.1Hz,1H),5.24(s,2H). 13 C NMR(75MHz,DMSO-d6)δ163.27,158.57,152.96,140.97,140.11,137.62,131.17,130.6 2,130.41,129.81,123.91,122.36,119.01,118.66,114.67,113.35,108.83.HRMS-ESI m / z[M+H] + Calculate for C 21 H 15 F3N4O2:413.1220,found:413.1221.

[0265] Example 26: Preparation of Compound 26

[0266] 5-(3-Aminobenzo[d]isoxazol-4-yl)-N-(3-chloro-5-(trifluoromethoxy)phenyl)indoline-1-carboxamide (26)

[0267]

[0268] Using intermediate A-42 (100.0 mg, 0.3980 mmol) and 3-chloro-5-(trifluoromethoxy)aniline (101.0 mg, 0.4770 mmol) as raw materials, the synthesis method was the same as Example 1 to obtain 18.00 mg of white solid with a yield of 10%. mp: 186–188°C. 1 H NMR (300MHz, DMSO-d6) δ9.04(s,1H),8.02(d,J=8.3Hz,1H),7.84(q,J=3.0,2.0Hz,1H),7.75(s,1H),7.57(d,J=7.6Hz,1H),7.49(d,J=8.3 HRMS-ESI m / z[M+H] + Calculate for C 23 H 16 ClF3N4O3:489.0936,found:489.0938.

[0269] Example 27: Preparation of Compound 27

[0270] 5-(3-Aminobenzo[d]isoxazol-4-yl)-N-(3-(trifluoromethyl)phenyl)indoline-1-carboxamide (27)

[0271] Using intermediate A-42 (100.0 mg, 0.3980 mmol) and 3-trifluoromethylphenyl isocyanate (89.00 mg, 0.4770 mmol) as raw materials, the synthesis method was the same as Example 25 to obtain 65.00 mg of white solid with a yield of 37%. mp: 210–212°C. 1 H NMR (300MHz, DMSO-d6) δ8.94(s,1H),8.12–8.00(m,2H),7.91(d,J=8.6Hz,1H),7.66–7.46(m,3H),7.38(d,J=5.7Hz ,2H),7.29(d,J=8.4Hz,1H),7.14(dd,J=7.2,0.9Hz,1H),5.25(s,2H),4.24(t,J=8.6Hz,2H),3.28(d,J=8.7Hz,2H). 13C NMR(75MHz,DMSO-d6)δ163.24,158.55,152.86,144.23,141.01,137.92,132.62,131.20,130.56,130.05, 128.23,125.59,124.06,123.84,119.19,116.64,116.58,115.36,113.36,108.72,48.11,27.82.HRMS-ESI m / z[M+H] + Calculate for C 23 H 18 F3N4O2:439.1376,found:439.1379.

[0272] Example 28: Preparation of Compound 28

[0273] 5-(3-Aminobenzo[d]isoxazol-4-yl)-N-(4-chloro-3-(trifluoromethyl)phenyl)indoline-1-carboxamide (28)

[0274]

[0275] Using intermediate A-42 (100.0 mg, 0.3980 mmol) and 4-chloro-3-trifluoromethylphenyl isocyanate (105.0 mg, 0.4770 mmol) as raw materials, the synthesis method was the same as Example 25 to obtain 21.00 mg of white solid with a yield of 11%. mp: 230–232°C. 1 H NMR (300MHz, DMSO-d6) δ9.05 (s, 1H), 8.20 (d, J = 2.5Hz, 1H), 8.06–7.93 (m, 2H ),7.66(d,J=8.8Hz,1H),7.58(dd,J=8.4,7.2Hz,1H),7.48(dd,J=8.4,0.9Hz ,1H),7.37(d,J=1.7Hz,1H),7.29(dd,J=8.3,1.9Hz,1H),7.13(dd,J=7.2,0. 9Hz,1H),5.26(s,2H),4.23(t,J=8.6Hz,2H),3.29(t,J=8.6Hz,2H).HRMS-ESI m / z[M+H] + Calculate for C 23 H 17 ClF3N4O2:473.0987,found:473.0991.

[0276] Example 29: Preparation of Compound 29

[0277] 5-(3-Aminobenzo[d]isoxazol-4-yl)-N-(3-fluoro-5-(trifluoromethyl)phenyl)indoline-1-carboxamide (29)

[0278]

[0279] Using intermediate A-42 (100.0 mg, 0.3980 mmol) and 3-fluoro-5-(trifluoromethyl)aniline (85.00 mg, 0.4770 mmol) as raw materials, the synthesis method was the same as Example 1 to obtain 38.00 mg of brown solid with a yield of 21%. mp: 238–240℃. 1 H NMR(300MHz, DMSO-d6)δ9.15(s,1H),8.05(d,J=8.3Hz,1H),7.95–7.89(m,2H),7.61(dd,J=8.4,7.2Hz,1H),7.51(d,J=8.3Hz,1H),7 .40(d,J=1.8Hz,1H),7.32(dd,J=8.1,2.1Hz,2H),7.16(d,J=7.1Hz,1H),5.29(s,2H),4.26(t,J=8.5Hz,2H),3.29(s,2H).HRMS-ESI m / z[M+H] + Calculate for C 23 H 17 F4N4O2:457.1282,found:457.1286.

[0280] Example 30: Preparation of Compound 30

[0281] 5-(3-Aminobenzo[d]isoxazol-4-yl)-N-(3-ethylphenyl)indoline-1-carboxamide (30)

[0282] Intermediate A-42 (100.0 mg, 0.3980 mmol) and 3-ethylaniline (58.00 mg, 0.4770 mmol) were used as raw materials. The synthesis method was the same as in Example 1 to obtain 65.00 mg of a white solid with a yield of 41%. mp: 201–203°C. 1H NMR (300MHz, DMSO-d6) δ8.55(s,1H),8.01(d,J=8.3Hz,1H),7.58(dd,J=8.4,7.2H z,1H),7.45(ddd,J=16.7,8.1,1.0Hz,3H),7.35(d,J=1.7Hz,1H),7.29–7.17(m,2H ),7.13(dd,J=7.2,0.9Hz,1H),6.92–6.86(m,1H),5.27(s,2H),4.21(t,J=8.7Hz, 2H),3.27(t,J=8.7Hz,2H),2.60(q,J=7.6Hz,2H),1.19(d,J=7.6Hz,3H).HRMS-ESI m / z[M+H] + Calculate for C 24 H 23 N4O2:399.1816,found:399.1819.

[0283] Example 31: Preparation of Compound 31

[0284] 5-(3-Aminobenzo[d]isoxazol-4-yl)-N-(4-(trifluoromethoxy)phenyl)indoline-1-carboxamide (31)

[0285]

[0286] Using intermediate A-42 (100.0 mg, 0.3980 mmol) and 4-trifluoromethoxyphenyl isocyanate (97.00 mg, 0.4770 mmol) as raw materials, the synthesis method was the same as Example 25 to obtain 82.00 mg of white solid with a yield of 45%. mp: 136–138°C. 1 H NMR (300MHz, DMSO-d6) δ8.82(s,1H),8.01(d,J=8.3Hz,1H),7.74–7.68(m,2H),7.58(d,J=1.2Hz,1H),7.48(dd,J=8.4,0.9H z,1H),7.37–7.29(m,4H),7.13(dd,J=7.2,0.9Hz,1H),5.26(s,2H),4.22(t,J=8.6Hz,2H),3.28(t,J=8.6Hz,2H).HRMS-ESI m / z[M+H] + Calculate for C 23 H 18 F3N4O3:455.1326,found:455.1329.

[0287] Example 32: Preparation of Compound 32

[0288] 5-(3-Aminobenzo[d]isoxazol-4-yl)-N-(3-(difluoromethoxy)phenyl)indoline-1-carboxamide (32)

[0289]

[0290] Intermediate A-42 (100.0 mg, 0.3980 mmol) and 3-difluoromethoxyaniline (97.00 mg, 0.4770 mmol) were used as raw materials. The synthesis method was the same as in Example 1 to obtain 81.00 mg of a white solid with a yield of 47%. mp: 182–184°C. 1 H NMR(300MHz, DMSO-d6)δ8.79(s,1H),8.01(d,J=8.3Hz,1H),7.57–7.46(m,4H),7.39–7.21(m,4H),7.14(dd ,J=7.1,0.9Hz,1H),6.86–6.82(m,1H),5.27(s,2H),4.22(t,J=8.6Hz,2H),3.28(t,J=8.6Hz,2H).HRMS-ESI m / z[M+H] + Calculate for C 23 H 19 F2N4O3:437.1420,found:437.1426.

[0291] Example 33: Preparation of Compound 33

[0292] 5-(3-Aminobenzo[d]isoxazol-4-yl)-N-(3-methoxyphenyl)indoline-1-carboxamide (33)

[0293] Using intermediate A-42 (100.0 mg, 0.3980 mmol) and m-aminoanisole (65.00 mg, 0.5290 mmol) as raw materials, the synthesis method was the same as Example 1 to obtain 70.00 mg of a white solid with a yield of 44%. 1 H NMR(300MHz,DMSO-d6)δ8.92(s,1H),7.77–7.91(m,4H),7.54–7.58(m,2H),7.24–7.31( m,3H),6.65–6.68(m,1H),5.26(s,2H),4.22(t,2H),3.76(s,3H)3.27(t,2H).HRMS-ESI m / z[M+H] + Calculate for C 23H 21 N4O3:401.1614,found:401.1619.

[0294] Example 34: Preparation of Compound 34

[0295]

[0296] Intermediate A-43 (4-bromo-6-methylbenzo[d]isoxazol-3-amine)

[0297] The synthesis method was the same as that of intermediate A-1 using 2-fluoro-6-bromo-4-methylbenzonitrile (5000 mg, 23.50 mmol) as the raw material to obtain 3030 mg of a white solid with a yield of 57%. 1H NMR (400 MHz, DMSO-d6) δ7.36–7.31 (m, 2H), 6.02 (s, 2H), 2.42 (s, 3H). MS m / z (ESI) [M+H]+: 225.1.

[0298] Intermediate A-44 (1-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-3-(3-(trifluoromethoxy)phenyl)urea)

[0299] Under ice bath, 3-trifluoromethoxyaniline (577.1 mg, 3.260 mmol) and triphosgene (483.6 mg, 1.630 mmol) were dissolved in anhydrous DCM (20 mL). After stirring for 10 min, triethylamine (1.319 mg, 13.04 mmol) was added and reacted for 2 h to form isocyanate. The intermediate p-aminophenylboronic acid pinacol ester (500.0 mg, 2.280 mmol) was added and reacted at room temperature overnight. After the reaction was completed under reduced pressure and concentrated by TLC monitoring, the mixture was purified by silica gel column chromatography [PE:EA=5:1] to obtain 434.0 mg of a white solid with a yield of 45%. 1 H NMR(300MHz,DMSO-d6)δ9.08(s,1H),8.97(s,1H),7.72(s,1H),7.60(d,J=8.1Hz,2H),7.48(d, J=8.1Hz,2H),7.41(t,J=8.1Hz,1H),7.32–7.25(m,1H),6.96(d,J=8.3Hz,1H),1.28(s,12H).MS m / z(ESI)[M+H] + :423.2.

[0300] 1-(4-(3-amino-6-methylbenzo[d]isoxazol-4-yl)phenyl)-3-(3-(trifluoromethoxy)phenyl)urea (34)

[0301] Intermediate A-43 (201.0 mg, 0.9480 mmol) and intermediate A-44 (400.0 mg, 0.9480 mmol) were used as raw materials. The synthesis method was the same as that of intermediate compound A-3 to obtain 26.00 mg of white solid with a yield of 6%. mp: 224–226℃. 1 H NMR(300MHz,DMSO-d6)δ9.18(s,1H),9.08(s,1H),7.82(s,1H),7.71(d,J=8.5Hz,2H),7.51 (t,J=8.5Hz,3H),7.40(d,J=10.9Hz,2H),7.05(d,J=8.0Hz,2H),5.27(s,2H),2.55(s,3H). 13 CNMR(75MHz,DMSO-d6)δ163.87,158.39,152.84,149.20,141.89,141.06,140.02,136.97,131.26,130.87, 129.71,125.47,122.28,118.93,118.88,117.32,114.24,111.22,110.70,108.66,21.68,21.20.HRMS-ESI m / z[M+H] + Calculate for C 22 H 17 F3N4O3:443.1331,found:443.1318.

[0302] Example 35: Preparation of Compound 35

[0303]

[0304] Intermediate A-45 (4-(4-amino-2-fluorophenyl)-6-methylbenzo[d]isoxazol-3-amine)

[0305] Using 2-fluoro-4-aminophenylboronic acid pinacol ester (400.0 mg, 1.690 mmol) and A-43 (381.9 mg, 1.690 mmol) as raw materials, the synthesis method was the same as that of the intermediate compound A-3 to obtain 230.0 mg of a light yellow solid with a yield of 53%. 1 HNMR(300MHz,DMSO-d6)δ7.23(s,1H),7.15(m,1H),7.05–6.99(m,1H),6.94–6.83(m,2H),5.42(s,2H),5.19(s,2H),2.43(s,3H).MS m / z(ESI)[M+H] + :258.1.

[0306] 1-(4-(3-amino-6-methylbenzo[d]isoxazol-4-yl)-2-fluorophenyl)-3-(3-(trifluoromethoxy)phenyl)urea (35)

[0307] The intermediate A-45 (230.0 mg, 0.8950 mmol) was used as the raw material. The synthesis method was the same as in Example 1 to obtain 19.00 mg of white solid with a yield of 5%. mp: 240–242°C. 1 H NMR(300MHz,DMSO-d6)δ9.69(s,1H),8.97(s,1H),8.30(s,1H),7.76(s,1H),7.43(d,J=1 0.2Hz,2H),7.31(d,J=10.1Hz,3H),7.01(d,J=14.6Hz,2H),5.27(s,2H),2.48(s,3H).MS m / z(ESI)[M+H] + :461.0.

[0308] Example 36: Preparation of Compound 36

[0309]

[0310] Intermediate A-46 (4-(4-amino-2,3-difluorophenyl)-6-methylbenzo[d]isoxazol-3-amine)

[0311] A-10 (400.0 mg, 1.570 mmol) and A-43 (381.9 mg, 1.570 mmol) were used as raw materials. The synthesis method was the same as that of the intermediate compound A-3 to obtain 235.0 mg of a light yellow solid with a yield of 54%. mp: 134–138°C. 1 H NMR (300MHz, DMSO-d6) δ7.30(t,J=1.1Hz,1H),6.95(s,1H),6.90(m,1H),6.67(m,1H),5.77(s,2H),5.17(s,2H),2.50(d,J=1.9Hz,3H).MS m / z(ESI)[M+H] + :476.1.

[0312] 1-(4-(3-amino-6-methylbenzo[d]isoxazol-4-yl)-2,3-fluorophenyl)-3-(3-(trifluoromethoxy)phenyl)urea(36)

[0313] The intermediate A-46 (230.0 mg, 0.8360 mmol) was used as the raw material. The synthesis method was the same as in Example 1 to obtain 14.00 mg of white solid with a yield of 4%. mp: 249–250℃. 1H NMR (300MHz, DMSO-d6) δ9.52(s,1H),9.02(s,1H),8.08(t,J=8.0Hz,1H),7.75(s,1H),7.45(t,J=8.2Hz,1H),7.3 8(s,1H),7.31(d,J=8.6Hz,1H),7.21(d,J=7.8Hz,1H),7.01(d,J=8.7Hz,2H),5.28(s,2H),2.47(s,3H).HRMS-ESI m / z[M+H] + Calculate for C 22 H 15 F5N4O3:479.1142,found:479.1151.

[0314] Example 37: Preparation of Compound 37

[0315] 1-(4-(4-aminothieno[3,2-c]pyridin-3-yl)phenyl)-3-(3-(trifluoromethoxy)phenyl)urea(37)

[0316] Using 3-bromo-4-aminothieno[3,2-c]pyridine (217.0 mg, 0.9480 mmol) and intermediate A-44 (400.0 mg, 0.9480 mmol) as raw materials, the synthesis method was the same as intermediate A-3 to obtain 35.00 mg of white solid with a yield of 8%. mp: 195–197℃. 1 H NMR (300MHz, DMSO-d6) δ9.12(s,1H),9.01(s,1H),7.83(d,J=5.6Hz,1H),7.72(s,1H),7.61(d,J=8 .5Hz,2H),7.46–7.36(m,4H),7.32(m,1H),7.26(d,J=5.6Hz,1H),6.99–6.93(m,1H),5.43(s,2H). 13 C NMR(75MHz,DMSO-d6)δ155.03,152.92,149.20,148.76,142.30,141.95,139.98,137.08,130.88,1 30.22,130.15,123.11,121.86,119.32,119.05,118.74,117.30,114.20,110.65,108.23.HRMS-ESI m / z[M+H] + Calculate for C 21 H 15F3N4O2S:445.0946,found:445.0944.

[0317] Example 38: Preparation of Compound 38

[0318] 1-(4-(1-Oxoisoindolin-4-yl)phenyl)-3-(3-(trifluoromethoxy)phenyl)urea(38)

[0319] Using 4-bromoisoindolin-1-one (200.0 mg, 0.9480 mmol) and intermediate A-44 (400.0 mg, 0.9480 mmol) as raw materials, the synthesis method was the same as intermediate A-3 to obtain 33.00 mg of white solid with a yield of 8%. mp: 175–177℃. 1 H NMR(300MHz,DMSO-d6)δ9.09(s,1H),8.96(s,1H),8.69(s,1H),7.72(s,1H),7.66(m,2H),7. 59(m,5H),7.42(t,J=8.1Hz,1H),7.32(d,J=8.4Hz,1H),6.96(d,J=8.1Hz,1H),4.53(s,2H). 13 CNMR(75MHz,DMSO-d6)δ170.34,152.87,149.21,149.19,141.91,141.60,139.66,137.09,133.82,132. 37,131.34,130.87,129.00,121.96,121.85,119.31,119.15,117.31,114.20,110.68,45.48.HRMS-ESI m / z[M+H] + Calculate for C 22 H 16 F3N3O3:428.1222,found:428.1206.

[0320] Example 39: Preparation of Compound 39

[0321] 1-(4-(4-amino-1H-pyrazolo[3,4-d]pyrimidin-3-yl)phenyl)-3-(3-(trifluoromethoxy)phenyl)urea(39)

[0322] Using 3-bromo-1H-pyrazolo[3,4-d]pyrimidin-4-amine (202.0 mg, 0.9480 mmol) and intermediate A-44 (400.0 mg, 0.9480 mmol) as raw materials, the synthesis method was the same as that of intermediate compound A-3 to obtain 25.00 mg of white solid with a yield of 6%. mp:>300℃. 1 H NMR(300MHz,DMSO-d6)δ13.79(s,1H),9.39(s,1H),9.29(s,1H),8.48(s,1H),8.00(s,1H ),7.92–7.84(m,4H),7.68(t,J=8.1Hz,1H),7.58(d,J=8.4Hz,1H),7.23(d,J=7.9Hz,1H). 13 C NMR(75MHz,DMSO-d6)δ170.14,152.88,149.20,146.19,142.01,140.12,139.19,131.85,1 31.59,130.87,130.79,129.81,128.41,122.77,117.90,117.26,114.12,110.62.HRMS-ESI m / z[M+H] + Calculate for C 19 H 14 F3N7O2:430.1239,found:430.1241.

[0323] Example 40: Preparation of Compound 40

[0324] 1-(4-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-5-yl)phenyl)-3-(3-(trifluoromethoxy)phenyl)urea(40)

[0325] Using 5-bromo-7H-pyrrolo[2,3-d]pyrimidin-4-amine (201.0 mg, 0.9480 mmol) and intermediate A-44 (400.0 mg, 0.9480 mmol) as raw materials, the synthesis method was the same as that of intermediate compound A-3 to obtain 23.00 mg of white solid with a yield of 58%. mp: 270–272℃. 1H NMR (300MHz, DMSO-d6) δ11.73(s,1H),9.04(s,1H),8.86(s,1H),8.07(s,1H),7.69(s,1H),7.53(d,J=8.5 Hz,2H),7.38–7.34(m,3H),7.29–7.25(m,1H),7.16(d,J=2.4Hz,1H),6.92(d,J=8.1Hz,1H),5.99(s,2H). 13 C NMR(75MHz,DMSO-d6)δ157.66,152.90,152.10,151.76,149.20,149.18,141.99,138.47,130.85,129.40,1 25.67,122.27,120.36,119.36,118.88,117.25,115.94,114.13,110.61,100.36,56.50.HRMS-ESIm / z[M+H] + Calculate for C 20 H 15 F3N6O2:429.1287,found:429.1266.

[0326] Example 41: Preparation of Compound 41

[0327]

[0328] Intermediate A-47 (5-(4-amino-3-fluorophenyl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine)

[0329] Using 3-fluoro-4-aminophenylboronic acid pinacol ester (334.0 mg, 1.410 mmol) and 5-bromo-7H-pyrrolo[2,3-d]pyrimidin-4-amine (200.0 mg, 0.9480 mmol) as raw materials, the synthesis method was the same as that of intermediate A-3, and finally 55.00 mg of yellow oil was obtained with a yield of 24%. MS m / z(ESI)[M+H] + :244.1.

[0330] 1-(4-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)-3-(3-(trifluoromethoxy)phenyl)urea(41)

[0331] Using intermediate A-47 (55.00 mg, 0.2260 mmol) and 3-trifluoromethoxyaniline (48.00 mg, 0.2720 mmol) as raw materials, the synthesis method was the same as Example 1, and finally 25.00 mg of white solid was obtained, with a yield of 25%. 1H NMR (300MHz, DMSO-d6) δ11.84(s,1H),9.41(s,1H),8.71(d,J=2.5Hz,1H),8.20(t,J=8.6Hz,1H),8.11(s,1H),7.74 (s,1H),7.43(t,J=8.2Hz,1H),7.36(d,J=2.0Hz,1H),7.33–7.25(m,3H),7.03–6.94(m,1H),6.11(s,2H).HRMS-ESI m / z[M+H] + Calculate for C 20 H 14 F4N6O2:446.1121,found:446.1114.

[0332] Example 42: Preparation of Compound 42

[0333]

[0334] Intermediate A-48 (4-(4-nitrophenyl)but-3-en-2-one)

[0335] Add p-nitrobenzaldehyde (15.00 g, 0.1000 mol) and potassium carbonate (2.760 g, 0.02000 mol) aqueous solution (20 mL) to acetone (250 mL), stir at room temperature for 24 h, add concentrated HCl (40 mL) and continue stirring at room temperature for 8 h until the reaction is complete. After adding a large amount of water, the product precipitates and is filtered. The filter cake is the crude product, which is purified by silica gel column chromatography (PE:EA=20:1) to obtain 14.61 g of yellow solid with a yield of 77%. mp:253–256℃.MS m / z(ESI)[M+H] + :192.2.

[0336] Intermediate A-49 (6-methyl-4-(4-nitrophenyl)-2-oxo-1,2-dihydropyridine-3-carbonitrile)

[0337] A-48 (4000 mg, 20.90 mmol), cyanoacetamide (3300 mg, 39.20 mmol), sodium ethoxide (7170 mg, 105.5 mmol) were dissolved in ethanol (40 mL) solution, stirred at 80 ° C for 3 h, concentrated under reduced pressure after the reaction was completed, 4N HCl (150 mL) was slowly added, and the pH of the reaction solution was adjusted to neutral. The product was precipitated and filtered, and after drying, 6200 mg of crude product was obtained as a dark brown solid, which was directly used in the next step. mp>280 ° C.MS m / z (ESI) [M+H] + :256.2.

[0338] Intermediate A-50 (6-methyl-4-(4-nitrophenyl)-2-chloro-1,2-dihydropyridine-3-carbonitrile)

[0339] The crude product A-49 (6200 mg, 24.31 mmol) and phosphorus oxychloride (30 mL) were mixed and refluxed for 2 h. After the reaction was completed, the reaction solution was cooled to room temperature, poured into an appropriate amount of ice water for quenching, extracted with ethyl acetate (150 mL×4), and the organic layers were combined, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by silica gel column chromatography (PE:EA=4:1) to obtain 1200 mg of a white oil with a yield of 18%. 1 H NMR (300MHz, DMSO-d6): δ8.4(d,2H,J=8.7Hz),7.8(d,2H,J=8.7Hz),7.3(s,1H),2.5(s,3H).MS m / z(ESI)[M+H] + :274.7.

[0340] Intermediate A-51 (6-methyl-4-(4-aminophenyl)-2-chloro-1,2-dihydropyridine-3-carbonitrile)

[0341] A-50 (1200 mg, 4.380 mmol) and palladium carbon (100.0 mg) were dissolved in methanol (20 mL) and reacted in hydrogen at room temperature for 20 h. The reaction was completed when the starting material disappeared as monitored by TLC. The reaction was filtered, the filtrate was concentrated under reduced pressure, and purified by silica gel column chromatography (PE:EA=2:1) to obtain 1060 mg of yellow solid with a yield of 99%. mp:221–226℃. 1 H NMR (300MHz, DMSO-d6): δ7.5(s,1H),7.4(d,2H,J=8.5Hz),6.7(d,2H,J=8.5Hz),5.8(s,2H),2.5(s,3H).MS m / z(ESI)[M+H] + :244.7.

[0342] Intermediate A-52 (4-(4-aminophenyl)-6-methylisoxazolo[3,4-b]pyridin-3-amine)

[0343] A-51 (1060 mg, 4.350 mmol) and hydroxylamine hydrochloride (1814 mg, 26.10 mmol) were dissolved in tert-butyl alcohol (50 mL). After reflux for 2 h, potassium tert-butylate (2920 mg, 26.10 mmol) was slowly added. After continuing to reflux for 0.5 h, the mixture was concentrated under reduced pressure and purified by silica gel column chromatography (DCM: MeOH = 60: 1) to obtain 300.0 mg of a yellow solid with a yield of 29%. mp: 141–146°C. 1H NMR (300MHz, DMSO-d6): δ7.4(d,2H,J=8.1Hz),7.0(s,1H),6.7(d,2H,J=8.1Hz),5.8(s,2H),5.4(s,2H),2.5(s,3H).MS m / z(ESI)[M+H] + :241.2.

[0344] 1-(4-(3-amino-6-methylisoxazolo[3,4-b]pyridin-4-yl)phenyl)-3-(3-(trifluoromethoxy)phenyl)urea(42)

[0345] Using 3-trifluoromethoxyaniline (88.50 mg, 0.5000 mmol) and intermediate A-52 (100.0 mg, 0.4160 mmol) as raw materials, the synthesis method was the same as Example 1 to obtain 42.00 mg of white solid with a yield of 19%. mp: 130–132°C. 1 H NMR(300MHz,DMSO-d6)δ9.12(s,1H),9.08(s,1H),7.75–7.65(m,3H),7.57(d,J=8.2Hz,2H), 7.37(dd,J=13.4,8.2Hz,2H),7.19(s,1H),6.97(d,J=8.0Hz,1H),5.48(s,2H),2.60(s,3H). 13 CNMR(75MHz,DMSO-d6)δ169.88,160.67,158.43,152.78,149.19,149.17,147.45,141.79,141. 22,130.90,129.81,129.12,119.45,119.00,117.37,114.34,110.73,102.51,24.51.HRMS-ESI m / z[M+H] + Calculate for C 21 H 16 F3N5O3:444.1278,found:444.1273.

[0346] Example 43: Preparation of Compound 43

[0347]

[0348] Intermediate A-53 (4-(indolin-5-yl)-6-methylbenzo[d]isoxazol-3-amine)

[0349] Intermediate A-41 (500.0 mg, 2.040 mmol) and intermediate A-43 (461.0 mg, 2.040 mmol) were used as raw materials, and the synthesis method was the same as A-3 to obtain 240.0 mg of yellow solid with a yield of 44%. MS m / z (ESI) [M+H] + :466.2.

[0350] 5-(3-Amino-6-methylbenzo[d]isoxazol-4-yl)-N-(3-(trifluoromethoxy)phenyl)indoline-1-carboxamide (43)

[0351] Intermediate A-53 (240.0 mg, 0.9060 mmol) was used as the raw material, and the synthesis method was the same as in Example 1 to obtain 30.00 mg of white solid with a yield of 7%. mp: 128–130°C. 1 HNMR(300MHz,DMSO-d6)δ8.89(s,1H),8.01(d,J=8.3Hz,1H),7.75(s,1H),7.62(m,1H),7.43(t,J=8.2Hz,1H),7.35(d,J=1.8Hz, 1H),7.28(d,J=7.5Hz,2H),7.05–6.96(m,2H),5.20(s,2H),4.23(t,J=8.5Hz,2H),3.26(d,J=8.6Hz,2H),2.46(s,3H).HRMS-ESI m / z[M+H] + Calculate for C 24 H 19 F3N4O3:469.1487,found:469.1472.

[0352] Example 44: Preparation of Compound 44

[0353]

[0354] N-(4-(3-amino-6-methylisoxazolo[3,4-b]pyridin-4-yl)phenyl)-2-(3-(trifluoromethoxy)phenyl)acetamide (44)

[0355] 3-Trifluoromethoxyphenylacetic acid (91.00 mg, 0.4160 mmol), HATU (174.0 mg, 0.4580 mmol), DIEA (162.0 mg, 1.250 mmol) were dissolved in anhydrous DMF (5 mL), stirred at room temperature for 10 min, and then intermediate A-52 (100.0 mg, 0.4160 mmol) was added and reacted at room temperature for 4 h. After TLC monitoring of the disappearance of the raw material, an appropriate amount of water was added to the reaction solution, and EA was extracted. The organic layer was washed with saturated brine 2 to 3 times, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by silica gel column chromatography (PE:EA=1:2) to obtain 43.00 mg of a white solid with a yield of 23%. mp:91–92℃. 1 H NMR (300MHz, DMSO-d6) δ10.53(s,1H),7.83(d,J=8.0Hz,2H),7.60(d,J=8.2Hz,2H),7.50(d,J=8.0Hz, 1H),7.41(d,J=8.9Hz,2H),7.29(d,J=8.1Hz,1H),7.20(s,1H),5.48(s,2H),3.81(s,2H),2.61(s,3H). 13 CNMR(75MHz,DMSO-d6)δ169.84,169.29,160.70,158.40,148.77,147.30,140.75,138.89,130 .63,130.36,129.75,128.87,122.17,119.78,119.62,119.51,102.54,43.16,24.50.HRMS-ESI m / z[M+H] + Calculate for C 22 H 18 F3N4O3:443.1326,found:443.1329.

[0356] Example 45: Preparation of Compound 45

[0357]

[0358] N-(4-(3-amino-6-methylisoxazolo[3,4-b]pyridin-4-yl)phenyl)-6-(trifluoromethoxy)indoline-1-carboxamide (45)

[0359] Using intermediate A-52 (100.0 mg, 0.4160 mmol) and 6-trifluoromethoxyindoline (84.00 mg, 0.4160 mmol) as raw materials, the synthesis method was the same as Example 1, and finally 21.00 mg of white solid was obtained with a yield of 10%. mp: 180–182°C.1 HNMR(300MHz,Chloroform-d)δ7.95(d,J=2.1Hz,1H),7.72–7.64(m,2H),7.59–7.52(m,2H),7.19(d,J=8.1Hz,1H),7.04( s,1H),6.90–6.83(m,1H),6.73(s,1H),5.48(s,2H),4.28(d,J=13.6Hz,2H),3.31(t,J=8.5Hz,2H),2.71(s,3H).HRMS-ESI m / z[M+H] + Calculate for C 24 H 23 F3N5O3:486.1748,found:486.1751.

[0360] Example 46: Preparation of Compound 46

[0361]

[0362] Intermediate A-54 (7-bromo-2,3-dihydro-1H-inden-4-amine)

[0363] Dissolve 4-aminoindane (1000 mg, 10.50 mmol) and NBS (1870 mg, 10.50 mmol) in DCM (15 mL), react overnight at room temperature, monitor the disappearance of the starting material by TLC, concentrate under reduced pressure, and purify by silica gel column chromatography (PE:EA=10:1) to obtain 600 mg of a white solid with a yield of 27%. mp: 64–66°C. 1 HNMR(300MHz,DMSO-d6)δ6.97(d,J=8.4Hz,1H),6.35(d,J=8.4Hz,1H),5.01(s,2H),2.75(dt,J=12.6,7.5Hz,4H),1.99(m,2H).MS m / z(ESI)[M+H] + :213.1.

[0364] Intermediate A-55 (7-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2,3-dihydro-1H-inden-4-amine)

[0365] Using intermediate A-54 (600.0 mg, 2.830 mmol) and biboronic acid pinacol ester (863.0 mg, 3.400 mmol) as raw materials, the synthesis method was the same as A-2, and finally 183.0 mg of yellow solid was obtained with a yield of 25%. mp: 116–118℃. 1H NMR(300MHz,DMSO-d6)δ7.20(d,J=7.9Hz,1H),6.81(t,J=7.6Hz,1H),5.22(s,2H),2.9 3(t,J=7.6Hz,2H),2.77(t,J=7.5Hz,2H),1.93(dd,J=7.4,3.2Hz,2H),1.24(s,12H).MS m / z(ESI)[M+H] + :260.5.

[0366] Intermediate A-56 (4-(7-amino-2,3-dihydro-1H-inden-4-yl)benzo[d]isoxazol-3-amine)

[0367] Using intermediate A-55 (183.0 mg, 0.7040 mmol) and intermediate A-1 (100.0 mg, 0.4690 mmol) as raw materials, the synthesis method was the same as A-3, and finally 50.00 mg of yellow solid was obtained with a yield of 41%. mp: 100–102℃.MS m / z(ESI)[M+H] + :266.3.

[0368] 1-(7-(3-aminobenzo[d]isoxazol-4-yl)-2,3-dihydro-1H-inden-4-yl)-3-(3-(trifluoromethoxy)phenyl)urea (46)

[0369] Intermediate A-56 (50.00 mg, 0.1880 mmol) and 3-trifluoromethoxyaniline (72.00 mg, 0.4080 mmol) were used as raw materials. The synthesis method was the same as in Example 1 to obtain 20.00 mg of a white solid with a yield of 23%. mp: 190–192°C. 1 H NMR (300MHz, DMSO-d6) δ9.38(s,1H),8.27(s,1H),7.96(d,J=8.2Hz,1H),7.76(s,1H),7.58(dd,J=8.4,7.1Hz,1H),7.53–7.37(m,2H) ,7.34–7.25(m,1H),7.10(t,J=7.0Hz,2H),6.97(dd,J=8.1,2.2Hz,1H),5.06(s,2H),2.92(s,2H),2.73(s,2H),2.05(s,2H).HRMS-ESI m / z[M+H] + Calculate for C 24 H 20 F3N4O3:469.1482,found:469.1486.

[0370] Example 47: Preparation of Compound 47

[0371]

[0372] Intermediate A-57 (7-amino-2,3-dihydro-1H-inden-1-one)

[0373] Under ice bath, 4-aminoindan (1000mg, 10.50mmol) and acetic anhydride (1542mg, 14.50mmol) were dissolved in ethanol (10mL) and reacted at room temperature overnight. After the reaction was completed, the solvent was evaporated and the crude product was dissolved in acetone (30mL), and then potassium permanganate (3400mg, 21.52mmol) and magnesium sulfate (1200mg, 10.00mmol) water (6.75mL) were added and mixed, and the reaction was continued at room temperature overnight. After the reaction was completed, celite was filtered, the filtrate was concentrated under reduced pressure, and 6N HCl (15mL) was added to react at 90℃ for 5h. After the reaction was completed, the pH was adjusted to neutral with saturated sodium carbonate aqueous solution, and the mixture was extracted with ethyl acetate (50mL×3). The organic phase was washed with saturated brine (30mL×3), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain 500mg of white oily crude product with a yield of 32%. MSm / z(ESI)[M+H] + :148.2.

[0374] Intermediate A-58 (7-amino-4-bromo-2,3-dihydro-1H-inden-1-one)

[0375] Using intermediate A-57 (500 mg, 3.400 mmol) and NBS (605.0 mg, 3.400 mmol) as raw materials, the synthesis method was the same as intermediate A-54, and finally 560.0 mg of yellow solid was obtained with a yield of 91%. mp: 121–124℃. 1 H NMR (300MHz, DMSO-d6) δ7.43(d,J=8.7Hz,1H),6.76(s,2H),6.54(d,J=8.7Hz,1H),2.91–2.82(m,2H),2.67–2.58(m,2H).MS m / z(ESI)[M+H] + :182.7.

[0376] Intermediate A-59 (7-amino-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2,3-dihydro-1H-inden-1-one)

[0377] Using intermediate A-58 (560.0 mg, 3.080 mmol) and biboronic acid pinacol ester (1170 mg, 4.620 mmol) as raw materials, the synthesis method was the same as intermediate A-2, and finally 193.0 mg of yellow solid was obtained with a yield of 23%. mp: 110–113℃. 1 H NMR (300MHz, DMSO-d6) δ7.54(d,J=8.2Hz,1H),6.96(s,2H),6.52–6.46(m,1H),3.10–3.03(m,2H),2.57–2.52(m,2H),1.17(s,12H).MS m / z(ESI)[M+H] + :274.5.

[0378] Intermediate A-60 (7-amino-4-(3-aminobenzo[d]isoxazol-4-yl)-2,3-dihydro-1H-inden-1-one)

[0379] Using intermediate A-59 (193.0 mg, 0.7070 mmol) and 4-bromobenzo[d]isoxazole-3-amine (100.0 mg, 0.4690 mmol) as raw materials, the synthesis method was the same as that of intermediate compound A-3, and finally 100.0 mg of yellow oil was obtained with a yield of 76%. 1 H NMR(300MHz, DMSO-d6)δ7.47(dd,J=8.4,0.9Hz,1H),7.28–7.18(m,2H),7.11(dd,J=7.2,0.9H z,1H),6.84(s,2H),6.66(d,J=8.3Hz,1H),5.12(s,2H),2.77(s,2H),2.54(d,J=5.7Hz,2H).MS m / z(ESI)[M+H] + :280.3.

[0380] 1-(7-(3-aminobenzo[d]isoxazol-4-yl)-3-oxo-2,3-dihydro-1H-inden-4-yl)-3-(3-(trifluoromethoxy)phenyl)urea (47)

[0381] Using intermediate A-60 (100.0 mg, 0.3580 mmol) and 3-trifluoromethoxyaniline (76.00 mg, 0.4300 mmol) as raw materials, the synthesis method was the same as Example 1, and finally 33.00 mg of white solid was obtained with a yield of 19%. mp: 142–144°C. 1HNMR (300MHz, DMSO-d6) δ10.45(s,1H),10.00(s,1H),8.37(d,J=8.5Hz,1H),7.75(s,1H),7.57(dt,J=13.8,7.4Hz,3H),7.4 5(d,J=8.1Hz,2H),7.16(d,J=7.0Hz,1H),6.99(d,J=7.4Hz,1H),5.18(s,2H),2.87(s,2H),2.69(t,J=5.6Hz,2H).HRMS-ESI m / z[M+H] + calcdfor C 24 H 18 F3N4O4:483.1275,found:483.1279.

[0382] Example 48: Preparation of Compound 48

[0383]

[0384] Intermediate A-61 (tert-butyl 4-(2-cyano-3-fluorophenoxy)phenyl)carbamate)

[0385] Dissolve 2,6-difluorobenzonitrile (500.0 mg, 3.600 mmol) and tert-butyl-(4-hydroxy)carbamate (750.0 mg, 3.600 mmol) in anhydrous DMF (12 mL), add potassium hydroxide (605.0 mg, 10.80 mmol), and stir at room temperature for 10 h. After the reaction is completed, add 100 mL of water, extract with ethyl acetate (50 mL×6), combine the organic layers, wash twice with saturated brine, dry over anhydrous sodium sulfate, concentrate under reduced pressure, and purify with silica gel column chromatography [PE:EA=10:1] to obtain 520.0 mg of white solid, with a yield of 44%. mp:116–120℃. 1 HNMR(300MHz,Chloroform-d)δ7.49–7.36(m,3H),7.12–7.04(m,2H),6.87(m,1H),6.57(m,2H),1.55(s,9H).MS m / z(ESI)[M+H] + :329.2.

[0386] Intermediate A-62 (tert-butyl 4-((3-aminobenzo[d]isoxazol-4-yl)oxy)phenylcarbamate)

[0387] Using intermediate A-61 (500.0 mg, 1.520 mmol) as raw material, the synthesis method was the same as A-1, and 300.0 mg of white oily liquid was obtained with a yield of 58%. mp: 193–197℃.1 HNMR(400MHz,DMSO-d6)δ9.45(s,1H),7.54(d,J=8.8Hz,2H),7.37(t,J=8.2Hz,1H),7.19 –7.12(m,2H),7.09(d,J=8.6Hz,1H),6.31(d,J=7.9Hz,1H),6.04(s,2H),1.49(s,9H).MS m / z(ESI)[M+H] + :342.2.

[0388] Intermediate A-63 (4-(4-aminophenoxy)benzo[d]isoxazol-3-amine)

[0389] Dissolve intermediate A-62 (300.0 mg, 0.8800 mmol) in dichloromethane (13 mL), add trifluoroacetic acid (200.6 mg, 1.760 mmol), and stir at room temperature for 1 h. After the reaction is completed, concentrate under reduced pressure, add 60 mL of water, adjust the pH to 8 with sodium bicarbonate, extract with ethyl acetate (30 mL×10), combine the organic layers, wash twice with saturated brine, dry over anhydrous sodium sulfate, and concentrate under reduced pressure to obtain 200.0 mg of a ginger-yellow solid with a yield of 94%. mp: 82–85°C. 1 H NMR (300MHz, DMSO-d6) δ: 7.33 (t, J = 8.2 Hz, 1H), 7.02 (d, J = 8.1 Hz, 1H), 6.90 (d, J = 8. 6Hz,2H),6.63(d,J=8.7Hz,2H),6.25(d,J=7.9Hz,1H),6.00(s,2H),5.10(s,2H).MS m / z(ESI)[M+H] + :242.1.

[0390] 1-(3-((3-aminobenzo[d]isoxazol-4-yl)oxy)phenyl)-3-(3-(trifluoromethoxy)phenyl)urea (48)

[0391] Intermediate A-63 (200.0 mg, 0.8300 mmol) was used as the raw material, and the synthesis method was the same as Example 1 to obtain 50.00 mg of white solid, yield: 14%. mp: 193–195℃. 1H NMR(300MHz,DMSO-d6)δ9.05(s,1H),8.90(s,1H),7.72(s,1H),7.60–7.53(m,2H),7.44–7.29(m,3H) ,7.22–7.16(m,2H),7.11(d,J=8.3Hz,1H),6.99–6.93(m,1H),6.35(d,J=8.0Hz,1H),6.08(s,2H).MS m / z(ESI)[M+H] + :445.0.

[0392] Example 49: Preparation of Compound 49

[0393] 1-(3-((3-aminobenzo[d]isoxazol-4-yl)oxy)phenyl)-3-(3-(trifluoromethyl)phenyl)urea (49)

[0394]

[0395] Using intermediate A-62 (200.0 mg, 0.8300 mmol) as raw material, the synthesis method was the same as Example 25 to obtain 45.00 mg of white solid, with a yield of 13%. 1 HNMR(300MHz,DMSO-d6)δ9.05(d,J=21.0Hz,2H),7.99(d,J=2.4Hz,1H),7.56(dd,J=15.3,8.0Hz,2H),7.49(t,J=2.1Hz,1H),7.45–7.37(m,2H),7 .33(d,J=7.5Hz,1H),7.27(dt,J=8.3,1.5Hz,1H),7.17(d,J=8.4Hz,1H),6.86(ddd,J=8.1,2.4,1.0Hz,1H),6.50(d,J=7.9Hz,1H),6.06(s,2H).MS m / z(ESI)[M+H] + :429.1.

[0396] Biological activity and related properties test examples

[0397] The compounds in the following test examples were all prepared according to the methods of the above embodiments of the present disclosure.

[0398] Test Example 1 RIPK1 enzyme inhibitory activity test of compounds

[0399] 1. Experimental Materials

[0400] Basic reaction buffer: 50 mM HEPES, 10 mM MgCl2, 0.01% Brij-35, 1 mM EGTA, 2 mM DTT, water, 2.5 mM MnCl2.

[0401] 2. Experimental methods

[0402] Prepare 2x ATP / substrate solution and 2x kinase solution, add kinase reaction buffer; use Echo 655 to transfer 100nL of compound dilution to 384-well plate; add 5μL 2x kinase solution to 384-well plate, centrifuge at 1000rpm for 1min, and incubate at 25℃ for 10min; add 5μL 2× substrate and ATP solution to 384-well plate, centrifuge at 1000rpm for 1min, and incubate at 25℃ for 180min; transfer 5μL ADP-Glo ​​reagent to 384-well plate, centrifuge at 1000rpm for 1min, and incubate at 25℃ for 40min; take 10μL kinase detection reagent to 384-well plate, centrifuge at 1000rpm for 1min, and incubate at 25℃ for 40min; use BMG multi-function microplate reader to read luminescent signal.

[0403] 3. Experimental results

[0404] Table 1 Inhibitory activity of compounds against RIPK1

[0405]

[0406]

[0407] *Compound test concentration: 500nM

[0408] Test Example 2 Anti-necrosis activity test (TSZ method) and anti-proliferation activity test

[0409] 1. Anti-necrosis activity test

[0410] (1) Experimental Materials

[0411] Cell programmed necrosis induction kit (Biyuntian, C1058S), Cell Counting Kit (CCK-8) (Vazyme, product number: A311-01 / 02), PRMI1640 culture medium (Thermo, product number: C11875500BT), PBS (Thermo, product number: C10010500BT), trypsin (TRYPSIN 0.25% EDTA) (Gibio, product number: 25200072), FBS (fetal bovine serum) (Gibio, product number: 10270-106), penicillin / streptomycin mixture (Keygen, KGY0023), microplate reader (Thermo), 96-well microplate (Corning, USA), CO2 incubator (Thermo).

[0412] (2) Experimental methods

[0413] Healthy HT-29 cells were inoculated in a 96-well plate, and 100 μL of cell suspension was added to each well, with an inoculation density of about 12,000 cells / well. The cells were placed in a constant temperature incubator at 37°C and 5% CO2, and cultured overnight to allow the cells to adhere to the wall; 1000x programmed necrosis induction reagent was diluted to 4x with culture medium, and then added to the 96-well plate, and 50 μL was added to each well, so that each well finally contained 1x concentration of TSZ stimulating factor; the compound to be tested was prepared into a 6mM stock solution with DMSO, and then diluted three times with DMSO to eight concentrations of solution, and then the eight concentrations of solution were diluted 50 times with fresh culture medium to obtain a working solution. After stimulating the cells with 1x TSZ solution for 30 min, 50 μL of the working solution of the compound was added to each well, and 50 μL of the culture medium containing 0.5% DMSO was added to the negative control well. The cells were placed in a 37°C, 5% CO2 incubator for 24 h. 20 μL of CCK-8 detection reagent was added to each well and incubated at room temperature for 60 min. The absorbance at a wavelength of 450 nm was monitored by an ELISA reader, and EC was analyzed using GraphPad Prism software. 50 Curve fitting.

[0414] 2. Antiproliferative Activity Test

[0415] (1) Experimental Materials

[0416] Cell Counting-Lite TM2.0 (Vazyme, product number: DD1101-02), PRMI1640 culture medium (Thermo, product number: C11875500BT), PBS (Thermo, product number: C10010500BT), trypsin (TRYPSIN0.25% EDTA) (Gibio, product number: 25200072), FBS (fetal bovine serum) (Gibio, product number: 10270-106), penicillin / streptomycin mixture (Keygen, KGY0023), microplate reader (Thermo), 384-well microplate (white, transparent bottom) (Corning, USA), CO2 incubator (Thermo).

[0417] (2) Experimental methods

[0418] The compound to be tested was prepared into a 6mM stock solution with DMSO, and then diluted three times with DMSO to form ten solutions. The ten solutions were then diluted 100 times with fresh culture medium to obtain working solutions for use. Healthy cells were inoculated in a 384-well plate, and 20μL of cell suspension was added to each well. The inoculation density was about 4,500 / well. The cells were cultured in a constant temperature incubator at 37°C and 5% CO2 overnight. After the cells adhered, 20μL of working solution was added to each well, and 20μL of culture medium containing 0.5% DMSO was added to the negative control wells, which were cultured in a constant temperature incubator at 37°C and 5% CO2 for 72h. During the detection, 20μL of CCL detection reagent was added to each well, and the chemiluminescence intensity was detected by an enzyme reader. IC was analyzed using GraphPad Prism software. 50 Curve fitting.

[0419] 3. Experimental results

[0420] Table 2 The rescue activity and antiproliferative activity of the compounds on HT-29 tumor cell line induced by TSZ combined stimulation

[0421]

[0422] Note: ND means not tested

[0423] Test Example 3 Pharmacological activity of compound 5 on the mouse liver fibrosis model induced by Schistosoma japonicum infection

[0424] (1) Experimental Materials

[0425] DMEM culture medium (Keygen, KGL1211-500), ALT test kit (Nanjing, Nanjing Jiancheng Biotechnology), AST test kit (Nanjing, Nanjing Jiancheng Biotechnology), HE staining solution kit (Solebo, G1120), Masson trichrome staining kit (Solebo, G1340), cellSens Dimension (Olympus, Japan), FACSVerse flow cytometer (BD Biosciences, USA), CD11b-FITC antibody (BD Biosciences, USA), F4 / 80-PE antibody (BD Biosciences, USA), CD45-FITC antibody (BD Biosciences, USA), RIPA lysis buffer (Cwbiotech, Nanjing), RIP3 primary antibody (Cell Signaling Technology, USA), pRIP3 primary antibody (Cell Signaling Technology, USA), RIP1 primary antibody (Cell Signaling Technology, USA), pRIP1 primary antibody (Cell Signaling Technology, USA), MLKL primary antibody (Cell Signaling Technology, USA). Technology), pMLKL primary antibody (Cell Signaling Technology, USA), IL-6 primary antibody (Cell Signaling Technology, USA), F4 / 80 primary antibody (Cell Signaling Technology, USA), TNF-α primary antibody (Cell Signaling Technology, USA), Collagen I primary antibody (Cell Signaling Technology, USA), α-SMA primary antibody (Cell Signaling Technology, USA), BCA kit (Beyotime, P0012S), SDS-PAGE protein loading buffer (Keygen, Nanjing), PVDF membrane (Millipore, USA), GelDoc XR+ system (Bio-Rad, USA), ELISA detection kit (Multisciences, Hangzhou).

[0426] (2) Experimental methods

[0427] At week 5 after C57BL / 6 wild-type mice were infected with schistosome cercariae (15±1), they were treated with praziquantel for 3 days and then treated with different doses (50 mg / kg / d, I50 group; 100 mg / kg / d, I100 group) of compound 5 or solvent control (Ctr group) by intraperitoneal injection once a day for 4 consecutive weeks. The mice were dissected at week 9 after infection. At the same time, a Blank group (C57BL / 6 wild-type mice not infected with schistosome cercariae) was set up and the mice were weighed every week.

[0428] 1) Irrigate and flush out the worms, and count the number of adults and eggs.

[0429] The worms were washed out by perfusion with sodium citrate saline and the number of adults was counted. The liver was digested with 5% KOH and the number of eggs was counted.

[0430] 2) Liver index and spleen index.

[0431] The mice were dissected at 9 weeks after infection, and the liver and spleen indices were recorded. The mice were weighed every week. Figure 1 shown.

[0432] As can be seen from the figure, after using compound 5, the liver index (indicating the proportional relationship between the liver size and body weight of the mouse) and spleen index (indicating the proportional relationship between the spleen size and body weight of the mouse) of different parts of the mouse decreased compared with the Ctr group, and were concentration-dependent, proving that the compound of the present invention has a significant protective effect on the liver and spleen.

[0433] 3) HE staining and Masson staining of mouse liver tissue

[0434] The right anterior lobe of the mouse liver was taken and fixed in 4% paraformaldehyde. After dehydration, transparency, embedding, and dewaxing, the sections were stained using the HE staining kit and the Masson staining kit. After sealing, 6-8 fields of view were randomly selected under a microscope, photographed, and the cellSens Dimension was used to analyze the fibrosis area of ​​a single egg. The results are shown in Figure 2. Figure 2 shown.

[0435] Specifically, Figure 2 (a) HE staining of mouse liver tissue (left) and the granuloma area index of a single worm egg (right, indicating the degree of liver fibrosis in mice). Figure 2 (b) is the Masson staining of mouse liver tissue (left) and the collagen area index of a single egg granuloma (right, indicating the degree of liver fibrosis in mice). As can be seen from the figure, after treatment, the area of ​​a single egg granuloma and the collagen area of ​​a single egg granuloma decreased in the drug-treated group, and there was a concentration-dependent decrease ( Figure 2 ).

[0436] 4) Western blotting experiment

[0437] Proteins in mouse hepatocytes were extracted using RIPA lysis buffer, and the supernatant was centrifuged at 12,000 × g. The protein concentration was detected using the BCA method. The protein (50-100 μg) was electrophoresed using 12% SDS-PAGE and transferred to a PVDF membrane. The membrane was blocked with 5% skim milk powder for 1 hour at 37°C and incubated overnight at 4°C with primary antibodies RIP3, pRIP3, RIP1, pRIP1, MLKL, pMLKL, IL-6, F4 / 80, TNF-α, Collagen I, α-SMA, GAPDH [all, 1:1000]. The membrane was then incubated with goat anti-mouse or anti-rabbit secondary antibodies at room temperature for 1 hour, developed with luminescent fluid, and exposed using the GelDoc XR+ system to obtain the results. The results are shown in Figure 2. Figure 3 As shown. Among them, RIP1, pRIP1, RIP3, pRIP3, MLKL, and pMLKL are indicators of necroptosis, IL-6, F4 / 80, and TNF-α are indicators of inflammatory factor levels, and Collagen I and α-SMA are indicators of fibrosis levels. Compared with the Ctr group, the fibrosis indexes and programmed necrosis indexes, inflammatory factors, and fibrosis levels in the drug-treated group decreased, and there was a concentration-dependency.

[0438] 5) ELISA experiment

[0439] ELISA kits were used to detect TNF-α, IL-17A, IL-4, and IL-1β in the supernatant of spleen cell culture. Figure 4 As shown, Figure 4 (a)-(d) are the contents of TNF-α, IL-17A, IL-4, and IL-1β in the culture supernatant of mouse spleen cells (indicating the level of inflammation in the mouse spleen at different dosing concentrations). The figure shows that compound 5 reduces the inflammatory response of the spleen, and the level of IL-17A in the dosing group decreases and basically returns to the level of the Blank group.

[0440] The present invention and its embodiments are described schematically above, and the description is not restrictive. The drawings show only one embodiment of the present invention, and the actual structure is not limited thereto. Therefore, if a person skilled in the art is inspired by it and designs a structural method and an embodiment similar to the technical solution without creativity without departing from the purpose of the invention, they shall all fall within the protection scope of the present invention.

Claims

1. A biaryl compound represented by formula (I) or a pharmaceutically acceptable salt thereof, in: R 1 Select from the following structures R 4 , R 5 are each independently selected from hydrogen or C1-C2 alkyl; Z 1 is nitrogen or CH; Z 2 is NH or sulfur; Z 3 is nitrogen or CH; R 2 or R 2’ independently selected from hydrogen, C1-C2 alkyl, cyano, halogen, C1-C2 alkoxy; R 3 Select from the following structures: R 6 is hydrogen, trifluoromethyl, trifluoromethoxy, difluoromethoxy, C1-C2 alkyl or C1-C2 alkoxy; R 7 is hydrogen, trifluoromethoxy or halogen; R 8 is hydrogen or halogen; X is nitrogen, CF or CH; Y is nitrogen, methylene or NH radical; n is 0, 2 or 3.

2. The biaryl compound or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that: R 1 for R 4 Selected from hydrogen or C1-C2 alkyl; R 2 or R 2’ independently selected from hydrogen, C1-C2 alkyl, cyano, halogen, C1-C2 alkoxy; R 3 Select from the following structures: R 6 is hydrogen, trifluoromethyl, trifluoromethoxy, difluoromethoxy, C1-C2 alkyl or C1-C2 alkoxy; R 7 is hydrogen, trifluoromethoxy or chlorine; R 8 is hydrogen, fluorine or chlorine; X is nitrogen, CH or CF; Y is nitrogen, methylene or NH; n is 0, 2 or 3.

3. The biaryl compound or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that: R 1 for R 4 Selected from hydrogen or C1-C2 alkyl; R 2 or R 2’ independently selected from hydrogen, C1-C2 alkyl, cyano, halogen, C1-C2 alkoxy; R 3 for R 6 is hydrogen, trifluoromethyl, trifluoromethoxy, difluoromethoxy, C1-C2 alkyl or C1-C2 alkoxy; R 7 is hydrogen, trifluoromethoxy or chlorine; R 8 is hydrogen, fluorine or chlorine; X is nitrogen, CH or CF; Y is a nitrogen atom, a methylene group or an NH group; n is 0 or 2.

4. The biaryl compound or a pharmaceutically acceptable salt thereof according to claim 3, characterized in that: R 1 for R 4 Selected from hydrogen or C1-C2 alkyl; R 2 or R 2’ R is independently selected from hydrogen, C1-C2 alkyl, cyano, halogen, C1-C2 alkoxy; 3 for R 6 is hydrogen, trifluoromethyl, trifluoromethoxy, difluoromethoxy, C1-C2 alkyl or C1-C2 alkoxy; R 7 is hydrogen, trifluoromethoxy or chlorine; R 8 is hydrogen, fluorine or chlorine; X is nitrogen, CH or CF; Y is nitrogen; n is 2.

5. The biaryl compound or a pharmaceutically acceptable salt thereof according to claim 3, characterized in that: R 1 for R 4 Selected from hydrogen or C1-C2 alkyl; R 2 or R 2’ independently selected from hydrogen, C1-C2 alkyl, cyano, halogen, C1-C2 alkoxy; R 3 for R 6 is hydrogen, trifluoromethyl, trifluoromethoxy, difluoromethoxy, C1-C2 alkyl or C1-C2 alkoxy; R 7 is hydrogen, trifluoromethoxy or chlorine; R 8 is hydrogen, fluorine or chlorine; X is nitrogen, CH or CF; Y is a NH radical; n is 0.

6. The biaryl compound or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that: R 1 for R 5 is a C1-C2 alkyl group; R 2 or R 2’ is hydrogen; R 3 for R 6 is trifluoromethoxy; R 7 is hydrogen; R 8 is hydrogen; X is CH; Y is a NH radical; n is 0.

7. The biaryl compound or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that: R 1 for Z 1 is nitrogen or CH; Z 2 is NH or sulfur; Z 3 is nitrogen or CH; R 2 or R 2’ is hydrogen or fluorine; R 3 for R 6 is trifluoromethoxy; R 7 is hydrogen; R 8 is hydrogen; X is CH; Y is a NH radical; n is 0.

8. The biaryl compound or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that: R 1 for R 2 or R 2’ is a hydrogen atom; R 3 for R 6 is trifluoromethoxy; R 7 is hydrogen; R 8 is hydrogen; X is CH; Y is a NH radical; n is 0.

9. The biaryl compound or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that: The biaryl compound or a pharmaceutically acceptable salt thereof is specifically: 1-(4-(3-aminobenzo[d]isoxazol-4-yl)-2-methylphenyl)-3-(3-(trifluoromethoxy)phenyl)urea (1), 1-(4-(3-aminobenzo[d]isoxazol-4-yl)-2,3-xylyl)-3-(3-(trifluoromethoxy)phenyl)urea (2), 1-(4-(3-aminobenzo[d]isoxazol-4-yl)-3-methylphenyl)-3-(3-(trifluoromethoxy)phenyl)urea (3), 1-(4-(3-aminobenzo[d]isoxazol-4-yl)-3-fluorophenyl)-3-(3-(trifluoromethoxy)phenyl)urea (4), 1-(4-(3-aminobenzo[d]isoxazol-4-yl)-2,3-difluorophenyl)-3-(3-(trifluoromethoxy)phenyl)urea (5), 1-(4-(3-aminobenzo[d]isoxazol-4-yl)-2,6-difluorophenyl)-3-(3-(trifluoromethoxy)phenyl)urea (6), 1-(4-(3-aminobenzo[d]isoxazol-4-yl)-3,5-difluorophenyl)-3-(3-(trifluoromethoxy)phenyl)urea (7), 1-(4-(3-aminobenzo[d]isoxazol-4-yl)-2,5-difluorophenyl)-3-(3-(trifluoromethoxy)phenyl)urea (8), 1-(4-(3-aminobenzo[d]isoxazol-4-yl)-3-chloro-2-fluorophenyl)-3-(3-(trifluoromethoxy)phenyl)urea (9), 1-(4-(3-aminobenzo[d]isoxazol-4-yl)-2-chlorophenyl)-3-(3-(trifluoromethoxy)phenyl)urea (10), 1-(4-(3-aminobenzo[d]isoxazol-4-yl)-3-chlorophenyl)-3-(3-(trifluoromethoxy)phenyl)urea (11), 1-(4-(3-aminobenzo[d]isoxazol-4-yl)-2,3-dichlorophenyl)-3-(3-(trifluoromethoxy)phenyl)urea (12), 1-(4-(3-aminobenzo[d]isoxazol-4-yl)-3-cyanophenyl)-3-(3-(trifluoromethoxy)phenyl)urea (13), 1-(4-(3-aminobenzo[d]isoxazol-4-yl)-2-fluoro-5-methylphenyl)-3-(3-(trifluoromethoxy)phenyl)urea (14), 1-(4-(3-aminobenzo[d]isoxazol-4-yl)-5-fluoro-2-methylphenyl)-3-(3-(trifluoromethoxy)phenyl)urea (15), 1-(4-(3-aminobenzo[d]isoxazol-4-yl)-2-methoxyphenyl)-3-(3-(trifluoromethoxy)phenyl)urea (16), 1-(4-(3-aminobenzo[d]isoxazol-4-yl)phenyl)-3-(3-(trifluoromethoxy)phenyl)urea (17), 1-(4-(3-aminobenzo[d]isoxazol-4-yl)-2-fluorophenyl)-3-(3-(trifluoromethoxy)phenyl)urea (18), 1-(5-(3-aminobenzo[d]isoxazol-4-yl)pyridin-2-yl)-3-(3-(trifluoromethoxy)phenyl)urea (19), 2-(4-(3-aminobenzo[d]isoxazol-4-yl)phenyl)-N-(3-(trifluoromethoxy)phenyl)acetamide (20), 1-(4-(3-aminobenzo[d]isoxazol-4-yl)phenyl)-3-(5-(tert-butyl)isoxazol-3-yl)urea (21), 1-(4-(3-aminobenzo[d]isoxazol-4-yl)phenyl)-3-(3-isopropoxyphenyl)urea (22), 6-(3-aminobenzo[d]isoxazol-4-yl)-N-(3-(trifluoromethoxy)phenyl)-3,4-dihydroquinoline-1(2H)-carboxamide (23), 5-(3-aminobenzo[d]isoxazol-4-yl)-N-(3-(trifluoromethoxy)phenyl)indoline-1-carboxamide (24), 1-(4-(3-aminobenzo[d]isoxazol-4-yl)phenyl)-3-(3-(trifluoromethyl)phenyl)urea (25), 5-(3-aminobenzo[d]isoxazol-4-yl)-N-(3-chloro-5-(trifluoromethoxy)phenyl)indoline-1-carboxamide (26), 5-(3-aminobenzo[d]isoxazol-4-yl)-N-(3-(trifluoromethyl)phenyl)indoline-1-carboxamide (27), 5-(3-aminobenzo[d]isoxazol-4-yl)-N-(4-chloro-3-(trifluoromethyl)phenyl)indoline-1-carboxamide (28), 5-(3-aminobenzo[d]isoxazol-4-yl)-N-(3-fluoro-5-(trifluoromethyl)phenyl)indoline-1-carboxamide (29), 5-(3-aminobenzo[d]isoxazol-4-yl)-N-(3-ethylphenyl)indoline-1-carboxamide (30), 5-(3-aminobenzo[d]isoxazol-4-yl)-N-(4-(trifluoromethoxy)phenyl)indoline-1-carboxamide (31), 5-(3-aminobenzo[d]isoxazol-4-yl)-N-(3-(difluoromethoxy)phenyl)indoline-1-carboxamide (32), 5-(3-aminobenzo[d]isoxazol-4-yl)-N-(3-methoxyphenyl)indoline-1-carboxamide (33), 1-(4-(3-amino-6-methylbenzo[d]isoxazol-4-yl)phenyl)-3-(3-(trifluoromethoxy)phenyl)urea (34), 1-(4-(3-amino-6-methylbenzo[d]isoxazol-4-yl)-2-fluorophenyl)-3-(3-(trifluoromethoxy)phenyl)urea (35), 1-(4-(3-amino-6-methylbenzo[d]isoxazol-4-yl)-2,3-fluorophenyl)-3-(3-(trifluoromethoxy)phenyl)urea (36), 1-(4-(4-aminothieno[3,2-c]pyridin-3-yl)phenyl)-3-(3-(trifluoromethoxy)phenyl)urea (37), 1-(4-(1-oxoisoindolin-4-yl)phenyl)-3-(3-(trifluoromethoxy)phenyl)urea (38), 1-(4-(4-amino-1H-pyrazolo[3,4-d]pyrimidin-3-yl)phenyl)-3-(3-(trifluoromethoxy)phenyl)urea (39), 1-(4-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-5-yl)phenyl)-3-(3-(trifluoromethoxy)phenyl)urea (40), 1-(4-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)-3-(3-(trifluoromethoxy)phenyl)urea (41), 1-(4-(3-amino-6-methylisoxazolo[3,4-b]pyridin-4-yl)phenyl)-3-(3-(trifluoromethoxy)phenyl)urea (42), 5-(3-amino-6-methylbenzo[d]isoxazol-4-yl)-N-(3-(trifluoromethoxy)phenyl)indoline-1-carboxamide (43), N-(4-(3-amino-6-methylisoxazolo[3,4-b]pyridin-4-yl)phenyl)-2-(3-(trifluoromethoxy)phenyl)acetamide (44), N-(4-(3-amino-6-methylisoxazolo[3,4-b]pyridin-4-yl)phenyl)-6-(trifluoromethoxy)indoline-1-carboxamide (45), 1-(7-(3-aminobenzo[d]isoxazol-4-yl)-2,3-dihydro-1H-inden-4-yl)-3-(3-(trifluoromethoxy)phenyl)urea (46), 1-(7-(3-aminobenzo[d]isoxazol-4-yl)-3-oxo-2,3-dihydro-1H-inden-4-yl)-3-(3-(trifluoromethoxy)phenyl)urea (47), 1-(3-((3-aminobenzo[d]isoxazol-4-yl)oxy)phenyl)-3-(3-(trifluoromethoxy)phenyl)urea (48), 1-(3-((3-Aminobenzo[d]isoxazol-4-yl)oxy)phenyl)-3-(3-(trifluoromethyl)phenyl)urea (49).

10. A pharmaceutical composition, characterized in that The invention contains any one of the above-mentioned biaryl compounds or a pharmaceutically acceptable salt thereof as an effective ingredient and a pharmaceutically acceptable carrier.

11. The pharmaceutical composition according to claim 10, characterized in that The pharmaceutically acceptable salt is a salt formed by the above-mentioned compound and an acid or a base, wherein the acid is an organic acid or an inorganic acid; the organic acid is methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, naphthalenesulfonic acid, citric acid, malic acid, tartaric acid, lactic acid, pyruvic acid, acetic acid, maleic acid, succinic acid, fumaric acid, salicylic acid, phenylacetic acid or mandelic acid; the inorganic acid is hydrochloric acid, hydrobromic acid, sulfuric acid or phosphoric acid; the base is a base containing a basic metal cation, an alkaline earth metal cation or an ammonium cation salt.

12. Use of the biaryl compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 9, or the pharmaceutical composition according to claim 10, in the preparation of a drug for preventing or treating necroptosis-mediated diseases associated with RIPK1.

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