Ripk1 kinase target inhibitors and medical uses thereof

Abstract

Disclosed are RIPK1 kinase target inhibitors and medical uses thereof. R1 is selected from a substituted or unsubstituted aromatic ring; R2 is selected from a hydroxyl group or an amino group; R3 is selected from a substituted or unsubstituted aromatic ring; R4 is selected from a C 1‑6 aliphatic hydrocarbon or hydrogen. The present application provides a series of novel kinase targeting inhibitors, and the disclosed compounds can effectively target RIPK1 kinase and can be used as effective therapeutic agents for neurodegenerative diseases and inflammation-related diseases.

Description

[0001] Case Analysis

[0002] This application is a divisional application of the following Chinese invention patent application:

[0003] Parent application number: 2022107034376, Parent invention title: RIPK1 kinase target inhibitor and its pharmaceutical use, Parent application date: June 22, 2022. Technical Field

[0004] This invention relates to the field of medicinal chemistry, specifically to RIPK1 kinase target inhibitors and their pharmaceutical uses. Background Technology

[0005] Alzheimer's disease (AD) is a complex neurodegenerative disease clinically characterized by memory loss, cognitive impairment, and behavioral disturbances. The main pathological features of AD include neuronal loss, β-amyloid (Aβ) deposition, and neurofibrillary tangles (NFTs) induced by hyperphosphorylated Tau protein. Recent studies have demonstrated that these may be triggered by neuronal death and neuroinflammation.

[0006] Receptor-interacting protein kinase 1 (RIPK1) is a serine / threonine family kinase located at the crossroads of cell death and inflammation signaling pathways. Important research has shown that phosphorylated RIPK1 is aberrantly expressed in the brains of Alzheimer's disease (AD) patients. When RIPK1 is inhibited, amyloid protein is reduced, delaying the onset and progression of neuroinflammation and alleviating memory loss. Furthermore, microglia promote the degradation of adenosine beta (Aβ), and studies have found that in Alzheimer's disease, RIPK1 promotes inflammation and Aβ accumulation, inducing microglia into an inflammatory activated state and thus weakening Aβ degradation. These findings suggest that RIPK1 is a crucial therapeutic target for AD.

[0007] The reported RIPK1 inhibitor backbone types are very limited, and small molecules involved in AD treatment are even rarer. Therefore, developing compounds targeting RIPK1 with novel backbones and high kinase selectivity is an urgent and important task with profound significance and potential value. Here, we disclose the development of a novel RIPK1 inhibitor with nanomolar potency for the treatment of neurodegenerative diseases such as AD and its application in the treatment of Alzheimer's disease. Animal behavioral experiments show that our disclosed novel RIPK1 inhibitor has good anti-AD efficacy and demonstrates promising clinical application prospects. Summary of the Invention

[0008] The purpose of this invention is to address the above-mentioned shortcomings of the prior art by providing a novel RIPK1 kinase target inhibitor as an effective therapeutic agent for neurodegenerative diseases and other inflammation-related diseases.

[0009] Another object of the present invention is to provide the application of this novel inhibitor targeting RIPK1 kinase.

[0010] The objective of this invention can be achieved through the following technical solutions:

[0011] A new class of inhibitors targeting RIPK1 kinase, selected from compounds with structures as shown in general formula (I) or general formula (II) or pharmaceutically acceptable salts thereof:

[0012]

[0013] Wherein, R1 is selected from substituted or unsubstituted aromatic rings; R2 is selected from hydroxyl or amino groups; R3 is selected from substituted or unsubstituted aromatic rings; R4 is selected from C... 1-6 Aliphatic hydrocarbons or hydrogen.

[0014] As a preferred embodiment of the present invention, the novel inhibitors targeting RIPK1 kinases are selected from compounds with structures as shown in general formula (I), wherein R3 is selected from substituted or unsubstituted benzene rings; and R4 is hydrogen.

[0015] As a preferred embodiment of the present invention, in the compound represented by general formula (I), R4 is hydrogen, and R3 is selected from halogen, cyano, methoxy, trifluoromethylthioalkyl, C1-5 alkyl, C1-3 alkenyl, methylenedioxy, trifluoromethyl, phenoxy monosubstituted or polysubstituted phenyl, or 5,6,7,8-tetrahydro-2-naphthylphenyl, 4-(2-furanyl)phenyl, unsubstituted phenyl. As a preferred embodiment of the present invention, in the compound represented by general formula (I), R4 is hydrogen, and R3 is selected from phenyl, 3,4-dimethylphenyl, 4-cyanophenyl, 3-cyanophenyl, 3,4-(methylenedioxy)-phenyl, 5,6,7,8-tetrahydro-2-naphthylphenyl, 3,5-dimethylphenyl, 2,4-difluorophenyl, 4-methoxyphenyl, 4-trifluoromethylthioalkyl-phenyl, 4-vinylphenyl, 4-(2-furanyl)phenyl, 4-tert-butylphenyl, 4-trifluoromethylphenyl, 2-chlorophenyl, 2-methylphenyl, 4-n-propylphenyl, 4-phenoxyphenyl, 4-methoxy-2-methylphenyl, and 2-fluoro-4-methoxyphenyl.

[0016] As a preferred embodiment of the present invention, R1 is selected from phenyl, halogen, or C1-2 mono- or poly-substituted phenyl, and R2 is selected from hydroxyl or amino; preferably, when R2 is selected from hydroxyl, R1 is selected from phenyl or chlorinated phenyl; when R2 is selected from amino, R1 is selected from phenyl or fluorine, chlorine, or methyl mono- or poly-substituted phenyl.

[0017] As a preferred embodiment of the present invention, when R2 is selected from hydroxyl, R1 is selected from phenyl or 4-chlorophenyl; when R2 is selected from amino, R1 is selected from 3-fluoro-5-methylphenyl, 3-fluoro-4-chlorophenyl, or 2-fluoro-3-chlorophenyl.

[0018] The compounds of this invention are selected from any one of the following:

[0019]

[0020]

[0021] As a preferred embodiment of the present invention, the pharmaceutically acceptable salt of the compound represented by formula I-II is selected from the acid addition salt formed by the compound of formula I-II and an acid, wherein the acid is selected from: hydrogen chloride, hydrogen bromide, sulfuric acid, carbonic acid, oxalic acid, citric acid, succinic acid, tartaric acid, phosphoric acid, lactic acid, pyruvic acid, acetic acid, maleic acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, or ferulic acid.

[0022] A pharmaceutical composition comprising the compound of formula I-II of the present invention or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

[0023] As a preferred embodiment of the present invention, the pharmaceutical composition is prepared into any one of the following dosage forms: tablets, capsules, powders, syrups, liquids, suspensions, lyophilized powder for injection, or injections.

[0024] A method for preparing the RIPK1 kinase target inhibitor according to the present invention:

[0025] Route 1: Synthesis of target compounds 1-17 a .

[0026]

[0027] a Reagents and reaction conditions: (a) substituted aromatic amine, propionic acid, 110℃, 4-6h; (b) thionyl chloride, tetrahydrofuran, reflux, 2h; (c) concentrated ammonia, tetrahydrofuran, 0℃, 30min.

[0028] Route 2: Synthesis of target compounds 18-37 a .

[0029]

[0030] aReagents and reaction conditions: (a) 3-fluoro-5-methylaniline, propionic acid, 110℃, 4h; (b) substituted aromatic amine, tris(dibenzylacetone)dipalladium, 2-dicyclohexylphosphine-2',4',6'-triisopropylbiphenyl, cesium carbonate, nitrogen protection, toluene, 100℃, overnight reaction; (c) ammonia-methanol solution (7M), magnesium methoxy, methanol, 80℃, 24h.

[0031] Route 3: Synthesis of target compounds 38-43 a .

[0032]

[0033] a Reagents and reaction conditions: (a) Lithium hydroxide monohydrate, tetrahydrofuran / methanol / water, reaction at room temperature, 2h; (b) Aliphatic amine, 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate, N,N-diisopropylethylamine, dichloromethane, reaction at room temperature, 4h, 42-89%; (c) Potassium hydroxylamine solution, methanol, reaction at room temperature, 3h, 67%.

[0034] Beneficial effects:

[0035] This invention provides a series of novel kinase-targeting inhibitors. The compounds disclosed in this invention can effectively target RIPK1 kinase and can be used as effective therapeutic agents for neurodegenerative diseases and inflammation-related diseases.

[0036] Specific implementation cases and methods

[0037] Example 1: Preparation of 2-(phenylamino)nicotinic acid (1):

[0038] Starting material a (1.010 g, 5.0 mmol) and aniline (0.558 g, 6.0 mmol) were dissolved in propionic acid (5 mL), and the mixture was stirred at 110 °C for 4–6 h. After the reaction was complete, the mixture was cooled to room temperature and then cooled to 0 °C. Crystals precipitated and were filtered. The crystals were washed successively with propionic acid and water, and dried to obtain off-white crystals with a yield of 46%. 1 HNMR(600MHz,DMSO-d6)δ10.49(s,1H),8.54–8.10(m,2H),7.67(dd,J=8.5,1.2Hz,2H),7 .36(dd,J=8.5,7.3Hz,2H),7.11–7.05(m,1H),6.91(dd,J=7.7,5.0Hz,1H),5.09(s,1H). 13C NMR(151MHz,DMSO)δ168.68,154.90,151.00,141.73,138.88,131.58,129.01,123.22,121.00,113.84,108.51.HRMS(ESI)calcd for C 12 H 10 N₂O₂[M+H] + :215.0806; found:215.0807.

[0039] Example 2: Preparation of 2-((4-chlorophenyl)amino)nicotinic acid (2):

[0040] Dissolve raw material a (1.010 g, 5.0 mmol) and 4-chloroaniline (0.764 g, 6.0 mmol) in propionic acid (5 mL), and follow the same procedure as compound 1. 1 H NMR(600MHz,DMSO-d6)δ10.51(s,1H),8.38(dd,J=4.9,2.0Hz,1H),8.29(dd,J=7.7,2.0Hz, 1H),7.75(d,J=8.9Hz,2H),7.38(d,J=8.8Hz,2H),6.92(dd,J=7.7,4.9Hz,1H),5.61(s,1H). 13 C NMR(151MHz,DMSO)δ168.82,154.99,151.82,141.17,138.34,128.69,126.11,121.96,114.29,108.36.HRMS(ESI)calcd for C 12 H9ClN2O2[M+H] + :249.0426; found:249.0426.

[0041] Example 3: Preparation of 2-((3-chlorophenyl)amino)nicotinic acid (3):

[0042] Raw material a (1.010 g, 5.0 mmol) and 3-chloroaniline (0.764 g, 6.0 mmol) were dissolved in propionic acid (5 mL), and the remaining procedures were the same as for compound 1. 1H NMR (600MHz, DMSO-d6) δ10.57(s,1H),8.43(dd,J=4.9,2.0Hz,1H),8.30(dd,J=7.7,2.0Hz,1H),8.07(s,1H),7.49(ddd,J =8.2,2.1,1.0Hz,1H),7.34(t,J=8.1Hz,1H),7.07(ddd,J=8.0,2.1,0.9Hz,1H),6.94(dd,J=7.7,4.8Hz,1H),6.20(s,1H). 13 C NMR(151MHz,DMSO)δ168.85,154.99,152.08,141.01,133.19,130.38,121.97,119.35,118.63,114.61,108.49.HRMS(ESI)calcd for C 12 H9ClN2O2[M+H] + :249.0426; found:249.0425.

[0043] Example 4: Preparation of 2-(phenylamino)nicotinamide (4):

[0044] Starting material a (1.010 g, 5.0 mmol) and aniline (0.558 g, 6.0 mmol) were dissolved in propionic acid (5 mL) and reacted with stirring at 110 °C for 4–6 h. After the reaction was complete, the mixture was cooled to room temperature and then cooled to 0 °C. Crystals precipitated and were filtered. The crystals were washed successively with propionic acid and water. The corresponding crystals (0.214 g, 1.0 mmol) were dissolved in tetrahydrofuran (1 mL) and thionyl chloride (5 mL) and refluxed for 2 h. After evaporation to dryness, the solution was dissolved in 1 mL of tetrahydrofuran and slowly added dropwise to ammonia water at 0 °C. The reaction was continued for 30 min. After the reaction was complete, the system was poured into water and extracted with ethyl acetate (3 x 5 mL). The organic layer was separated, dried with Na₂SO₄, filtered, concentrated, and purified by preparative TLC (PE:EA = 1:1) to obtain a yellow powder with a yield of 72%. 1 HNMR(600MHz,DMSO-d6)δ11.17(s,1H),8.30(dd,J=4.8,1.8Hz,1H),8.28(s,1H),8.14(dd,J=7.7,1.9Hz,1H),7 .70(s,1H),7.68(d,J=7.4Hz,2H),7.29(dd,J=8.5,7.2Hz,2H),7.02–6.92(m,1H),6.83(dd,J=7.7,4.8Hz,1H). 13C NMR(151MHz,DMSO)δ170.06,154.95,150.89,140.25,137.54,128.74,121.57,119.40,113.34,110.27.HRMS(ESI)calcd for C 12 H 12 N3O[M+H] + :214.0975; found:214.0974.

[0045] Example 5: Preparation of 2-((4-chlorophenyl)amino)nicotinamide (5):

[0046] Raw material a (1.010 g, 5.0 mmol) and 4-chloroaniline (0.764 g, 6.0 mmol) were dissolved in propionic acid (5 mL), and the remaining procedures were the same as for compound 4. 1 H NMR (600MHz, DMSO-d6) δ11.27(s,1H),8.32(dd,J=4.8,1.8Hz,1H),8.30(s,1H),8.16(dd, J=7.7,1.9Hz,1H),7.75–7.71(m,3H),7.33(dd,J=8.9Hz,2H),6.87(dd,J=7.7,4.8Hz,1H). 13 C NMR(151MHz,DMSO)δ169.93,154.63,150.80,139.20,137.60,128.52,124.84,120.78,113.76,110.52.HRMS(ESI)calcd for C 12 H 11 ClN3O[M+H] + :245.0587; found:245.0587.

[0047] Example 6: Preparation of 2-((3-chlorophenyl)amino)nicotinamide (6):

[0048] Raw material a (1.010 g, 5.0 mmol) and 3-chloroaniline (0.764 g, 6.0 mmol) were dissolved in propionic acid (5 mL), and the remaining procedures were the same as for compound 4. 1H NMR (600MHz, DMSO-d6) δ11.36(s,1H),8.37(dd,J=4.8,1.8Hz,1H),8.33(s,1H),8.18(dd,J=7.8,1.9Hz,1H),8.10(t,J=2.1Hz,1H),7 .77(s,1H),7.40(ddd,J=8.3,2.2,0.9Hz,1H),7.30(t,J=8.1Hz,1H),7.00(ddd,J=7.9,2.1,0.9Hz,1H),6.91(dd,J=7.7,4.8Hz,1H). 13 C NMR(151MHz,DMSO)δ169.88,154.54,150.82,141.73,137.67,133.16,130.29,120.99,118.34,117.72,114.10,110.77.HRMS(ESI)calcd forC 12 H 11 ClN3O[M+H] + :247.0586; found:247.0585.

[0049] Example 7: Preparation of 2-((2,4-difluorophenyl)amino)nicotinamide (7):

[0050] Raw material a (1.010 g, 5.0 mmol) and 2,4-difluoroaniline (0.774 g, 6.0 mmol) were dissolved in propionic acid (5 mL), and the remaining procedures were the same as for compound 4. 1 HNMR (600MHz, DMSO-d6) δ11.26(d,J=2.5Hz,1H),8.50(td,J=9.3,6.2Hz,1H),8.31(dd,J=4.8,1.8Hz,2H),8.18(dd,J=7.8,1 .8Hz,1H),7.72(s,1H),7.30(ddd,J=11.7,8.9,2.9Hz,1H),7.05(ddt,J=11.1,8.8,2.2Hz,1H),6.89(dd,J=7.7,4.8Hz,1H). 13C NMR (151MHz, DMSO) δ169.87,157.17,157.09,155.57,155.49,154.68,153.41,153.33,151.79,151.71,150.83,137.59,125.20, 125.18,125.13,125.11,122.24,122.23,122.19,122.17,113.94,110.69,103.78,103.62,103.60,103.44.HRMS(ESI)calcdfor C 12 H 11 F2N3O[M+H] + :250.0786; found:250.0787.

[0051] Example 8: Preparation of 2-((3-fluoro-5-methylphenyl)amino)nicotinamide (8):

[0052] Raw material a (1.010 g, 5.0 mmol) and 3-fluoro-5-methylaniline (0.751 g, 6.0 mmol) were dissolved in propionic acid (5 mL), and the remaining procedures were the same as for compound 4. 1 H NMR (600MHz, DMSO-d6) δ11.34(s,1H),8.36(dd,J=4.8,1.8Hz,1H),8.32(s,1H),8.17(dd,J=7.7,1.9Hz,1H),7.84 –7.71(m,2H),7.00(d,J=1.4Hz,1H),6.89(dd,J=7.7,4.8Hz,1H),6.60(ddd,J=9.6,2.6,1.3Hz,1H),2.29(s,3H). 13 C NMR (151MHz, DMSO) δ169.92,163.21,161.63,154.64,150.84,141.70,141.62,140.11,140.05,1 37.61,115.38,113.90,110.66,108.42,108.28,103.03,102.85,21.05,21.04.HRMS(ESI)calcd for C 13 H 13 FN3O[M+H] + :246.1037; found:246.1038.

[0053] Example 9: Preparation of 2-((3,4-dimethylphenyl)amino)nicotinamide (9)

[0054] Raw material a (1.010 g, 5.0 mmol) and 3,4-dimethylaniline (0.727 g, 6.0 mmol) were dissolved in propionic acid (5 mL), and the remaining procedures were the same as for compound 4. 1 H NMR (600MHz, DMSO-d6) δ11.02(s,1H),8.28(dd,J=4.8,1.8Hz,1H),8.24(s,1H),8.11(dd,J=7.7,1.9Hz,1H),7.66(s,1H),7.4 8(dd,J=8.2,2.4Hz,1H),7.35(d,J=2.4Hz,1H),7.04(d,J=8.1Hz,1H),6.78(dd,J=7.7,4.8Hz,1H),2.20(s,3H),2.16(s,3H). 13 C NMR(151MHz,DMSO)δ170.13,155.13,150.98,137.98,137.45,136.27,129.61,129.30,120.81,117.03,112.84,109.94,19.64,18.74.HRMS(ESI)calcdfor C 13 H 13 FN3O[M+H] + :242.1288; found:242.1288.

[0055] Example 10: Preparation of 2-((3-bromophenyl)amino)nicotinamide (10)

[0056] Raw material a (1.010 g, 5.0 mmol) and 3-bromoaniline (1.032 g, 6.0 mmol) were dissolved in propionic acid (5 mL), and the remaining procedures were the same as for compound 4. 1 H NMR (600MHz, DMSO-d6) δ11.35(s,1H),8.37(dd,J=4.8,1.8Hz,1H),8.33(s,1H),8.22(t,J=2.0Hz,1H),8.18(dd,J=7.8,1.8Hz,1H),7 .77(s,1H),7.46(ddd,J=8.2,2.1,1.0Hz,1H),7.24(t,J=8.0Hz,1H),7.13(ddd,J=7.9,2.0,0.9Hz,1H),6.90(dd,J=7.7,4.8Hz,1H). 13C NMR(151MHz,DMSO)δ169.88,154.51,150.82,141.87,137.66,130.62,123.90,121.74,121.15,118.11,114.11,110.77.HRMS(ESI)calcd forC 12 H 11 BrN3O[M+H] + :292.0080; found:292.0081.

[0057] Example 11: Preparation of 2-((4-(trifluoromethyl)phenyl)amino)nicotinamide (11):

[0058] Raw material a (1.010 g, 5.0 mmol) and 4-trifluoromethylaniline (0.967 g, 6.0 mmol) were dissolved in propionic acid (5 mL), and the remaining procedures were the same as for compound 4. 1 H NMR(600MHz,DMSO-d6)δ11.56(s,1H),8.38(dd,J=4.8,1.8Hz,1H),8.37(s,1H),8.21(dd,J=7.8,1 .9Hz,1H),7.92(d,J=8.5Hz,2H),7.82(s,1H),7.63(d,J=8.5Hz,2H),6.96(dd,J=7.7,4.8Hz,1H). 13 C NMR(151MHz,DMSO)δ169.84,154.35,150.76,143.81,137.74,126.04,126.02,125.58,123.79,121.21,121.00,118.73,114.61,111.16.HRMS(ESI)calcd for C 13 H 11 F3N3O[M+H] + :282.0849; found:282.0850.

[0059] Example 12: Preparation of 2-((4-isopropylphenyl)amino)nicotinamide (12):

[0060] Raw material a (1.010 g, 5.0 mmol) and 4-isopropylaniline (0.811 g, 6.0 mmol) were dissolved in propionic acid (5 mL), and the remaining procedures were the same as for compound 4. 1HNMR(600MHz,DMSO-d6)δ11.05(s,1H),8.27(dd,J=4.8,1.8Hz,1H),8.25(s,1H),8.11(dd,J=7.7,1.9Hz,1H),7.66(s,1H ),7.56(d,J=8.5Hz,2H),7.16(d,J=8.5Hz,2H),6.79(dd,J=7.7,4.8Hz,1H),2.84(p,J=6.9Hz,1H),1.19(d,J=6.9Hz,6H). 13 C NMR(151MHz,DMSO)δ170.09,155.08,150.94,141.72,137.94,137.48,126.41,119.74,112.98,109.99,32.82,24.07.HRMS(ESI)calcd for C 15 H 18 N3O[M+H] + :256.1444; found:256.1446.

[0061] Example 13: Preparation of 2-((4-chloro-3-fluorophenyl)amino)nicotinamide (13):

[0062] Raw material a (1.010 g, 5.0 mmol) and 3-fluoro-4-chloroaniline (0.873 g, 6.0 mmol) were dissolved in propionic acid (5 mL), and the remaining procedures were the same as for compound 4. 1 H NMR (600MHz, DMSO-d6) δ11.45(s,1H),8.38(dd,J=4.8,1.8Hz,1H),8.35(s,1H),8.19(dd,J=7.8,1.8Hz,1H),8.13(dd, J=12.6,2.5Hz,1H),7.79(s,1H),7.44(t,J=8.7Hz,1H),7.32(ddd,J=8.8,2.5,1.0Hz,1H),6.93(dd,J=7.7,4.8Hz,1H). 13 C NMR(151MHz,DMSO)δ169.76,157.93,156.32,154.29,150.73,140.89,140.82,137.69,13 0.19,116.13,116.11,114.38,110.95,110.53,110.41,107.03,106.85.HRMS(ESI)calcd for C 12 H 10 ClFN3O[M+H] +:266.0491; found:266.0492.

[0063] Example 14: Preparation of 2-((3-chloro-2-fluorophenyl)amino)nicotinamide (14):

[0064] Raw material a (1.010 g, 5.0 mmol) and 2-fluoro-3-chloroaniline (0.873 g, 6.0 mmol) were dissolved in propionic acid (5 mL), and the remaining procedures were the same as for compound 4. 1 HNMR(600MHz,DMSO-d6)δ11.57(d,J=2.8Hz,1H),8.65–8.50(m,1H),8.37(dd,J=4.8,1.8Hz,2H),8.22(dd,J=7.8,1 .8Hz,1H),7.80(s,1H),7.17(td,J=8.2,1.3Hz,1H),7.13(td,J=8.2,6.7,1.7Hz,1H),6.95(dd,J=7.8,4.8Hz,1H). 13 C NMR (151MHz, DMSO) δ169.80,154.36,150.81,148.87,147.25,137.67,130.18,130.1 2,124.97,124.94,121.88,119.56,119.24,119.14,114.58,111.23.HRMS(ESI)calcd forC 12 H 10 ClFN3O[M+H] + :266.0491; found:266.0491.

[0065] Example 15: Preparation of 2-((2,3,4-trifluorophenyl)amino)nicotinamide (15):

[0066] Raw material a (1.010 g, 5.0 mmol) and 2,3,4-trifluoroaniline (0.883 g, 6.0 mmol) were dissolved in propionic acid (5 mL), and the remaining procedures were the same as for compound 4. 1 H NMR (600MHz, DMSO-d6) δ11.41(d,J=2.1Hz,1H),8.35(s,1H),8.33(dd,J=4.8,1.8Hz,1H),8.32–8.27(m,1H ),8.21(dd,J=7.8,1.8Hz,1H),7.79(s,1H),7.27(td,J=10.4,8.6,2.2Hz,1H),6.94(dd,J=7.7,4.8Hz,1H). 13C NMR (151MHz, DMSO) δ169.79,154.53,150.82,145.67,145.61,144.08,144.01,142.72,141.16,141.10,139.99,139.88,139.79,138.36,13 8.26,138.16,137.65,126.56,126.54,126.51,126.49,115.58,115.54,114.51,111.49,111.46,111.37,111.35,110.93.HRMS(ESI)calcd forC 12 H9F3N3O[M+H] + :268.0692; found:268.0691.

[0067] Example 16: Preparation of 2-((5-chloro-2-fluorophenyl)amino)nicotinamide (16):

[0068] Raw material a (1.010 g, 5.0 mmol) and 5-chloro-2-fluoroaniline (0.873 g, 6.0 mmol) were dissolved in propionic acid (5 mL), and the remaining procedures were the same as for compound 4. 1 HNMR (600MHz, DMSO-d6) δ11.68(d,J=3.2Hz,1H),8.81(dd,J=7.3,2.7Hz,1H),8.43(dd,J=4.8,1.8Hz,1H),8.37(s,1H),8.23 (dd,J=7.8,1.8Hz,1H),7.80(s,1H),7.29(dd,J=11.3,8.6Hz,1H),7.00(dt,J=4.6,2.3Hz,1H),6.98(dd,J=7.8,4.8Hz,1H). 13 C NMR (151MHz, DMSO) δ169.73,154.16,151.59,150.81,149.99,137.74,129.98,129.90,128.1 4,128.12,120.51,120.46,119.39,119.37,116.08,115.94,114.74,111.37.HRMS(ESI)calcd for C 12 H 11 ClFN3O[M+H] + :266.0491; found:266.0491.

[0069] Example 17: Preparation of 2-(p-Tolueneamino)nicotinamide (17):

[0070] Raw material a (1.010 g, 5.0 mmol) and 4-methylaniline (0.643 g, 6.0 mmol) were dissolved in propionic acid (5 mL), and the remaining procedures were the same as for compound 4. 1 H NMR(600MHz,DMSO-d6)δ11.07(s,1H),8.28(dd,J=4.8,1.8Hz,1H),8.25(s,1H),8.12(dd,J=7.7,1.9Hz, 1H),7.67(s,1H),7.55(d,J=8.4Hz,2H),7.10(d,J=8.4Hz,2H),6.79(dd,J=7.7,4.8Hz,1H),2.25(s,3H). 13 C NMR(151MHz,DMSO)δ170.11,155.07,150.94,137.72,137.48,130.43,129.13,119.56,112.96,109.99,20.41.HRMS(ESI)calcd for C 13 H 14 N3O[M+H] + :228.1131; found:228.1131.

[0071] Example 18: Detection of the inhibitory activity of RIPK1 fragment long kinase on programmed necrosis in BV2 cells

[0072] The long human RIPK1 (#VA7591) fragment was purchased from Promega. The assay buffer consisted of 25 mM HEPES (pH 7.2), 20 mM MgCl2, 12.5 mM MnCl2, 5 mM EGTA, 2 mM EDTA, 12.5 mM β-glycerophosphate, and 2 mM DTT (added just before use). 1 μL of RIPK1 was incubated with 2 μL of different concentrations of the compound or DMSO in the assay buffer at 24 °C for 15 min. Then, 2 μL of an ATP / MBP mixture (final concentrations of 25 μM and 5 μM, respectively) was added to trigger the kinase reaction, and the mixture was incubated at 37 °C for 90 min. 5 μL of the kinase reaction buffer was then added to 5 μL of LADP-globulin. TM Reagent quenching. After incubation for another 60 min, add 10 μL of Kinase Detection reagent and incubate at 24 °C for 30 min. Read the values ​​using the Lum module of a Molecular Devices multi-plate reader and calculate the inhibition rate (%).

[0073] Mouse microglia BV2 cells (purchased from Wuhan Pronosei Biotechnology Co., Ltd., China) were grown in a medium containing 4 mM L-glutamine, 4500 mg / L glucose, 1 mM sodium pyruvate (Hyclone), 10% heat-inactivated fetal bovine serum, and 1% GlutaMAX™ (Gibco). The culture environment was 37°C with 5% CO2. During the experiment, BV2 cells were cultured at a rate of 4 × 10⁻⁶ cells / year. 3 Cells were seeded per well in 96-well plates for 24 hours. Next, the culture medium was replaced with fresh, FBS-free DMEM containing 200 ng / mL LPS, or none was added, and the cells were incubated for another 23 hours. BV2 cells were then incubated for 1 hour with the specified concentration of the compound or DMSO, followed by incubation for 24 hours with 25 μM zVAD-fmk. Cells were then analyzed using CellTiter. The Aqueous Non-Radioactive Cell Proliferation (MTS) Assay (Promega) was used to detect cell viability.

[0074] IC was calculated using nonlinear regression with normalized dose-response fitting in Prism GraphPad 8.0 software. 50 and EC 50 Values. All experiments were performed independently at least three times.

[0075] Table 1. Inhibition of programmed necrosis in BV2 cells by compounds 1-17 (EC) 50 value a

[0076]

[0077]

[0078] a Enzyme inhibition rate and EC 50 Values, data are expressed as the mean ± SD of three independent experiments.

[0079] We evaluated the inhibitory activity of the target compounds against BV2 cell necrosis (Table 1). Table 1 shows that the compounds of formula (II) series have good inhibitory efficacy against programmed necrosis in BV2 cells.

Claims

1. A class of inhibitors targeting RIPK1 kinase, characterized in that, Compounds with structures as shown in general formula (II) or pharmaceutically acceptable salts thereof: ， R2 is selected from amino groups, and R1 is selected from 3-fluoro-5-methylphenyl, 3-fluoro-4-chlorophenyl, and 2-fluoro-3-chlorophenyl.

2. The inhibitor targeting RIPK1 kinase according to claim 1, characterized in that, The pharmaceutically acceptable salt of the compound represented by formula (II) is selected from the acid addition salt formed by the compound of formula (II) and an acid selected from: hydrogen chloride, hydrogen bromide, sulfuric acid, carbonic acid, oxalic acid, citric acid, succinic acid, tartaric acid, phosphoric acid, lactic acid, pyruvic acid, acetic acid, maleic acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, or ferulic acid.

3. A pharmaceutical composition comprising an inhibitor of RIPK1 kinase as described in any one of claims 1-2 and a pharmaceutically acceptable carrier.

4. Use of the inhibitor targeting RIPK1 kinase according to any one of claims 1-2 in the preparation of a medicament for treating and / or preventing diseases mediated by RIPK1 kinase.

5. The application according to claim 4, characterized in that... The use of the inhibitor targeting RIPK1 kinase according to any one of claims 1-2 in the preparation of a medicament for treating RIPK1 kinase-mediated neurodegenerative diseases or inflammatory-related diseases.

6. The application according to claim 5, characterized in that... The use of the inhibitor targeting RIPK1 kinase according to any one of claims 1-2 in the preparation of a medicament for treating RIPK1 kinase-mediated Alzheimer's disease.

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