ASK1 inhibitor and application thereof

Abstract

The invention belongs to the technical field of medicines, and particularly relates to a group of ASK1 inhibitors and application thereof. An ASK1 target spot is related to a plurality of diseases, and in numerous disclosed ASK1 modulators, no marketed medicine exists at present, so that the finding of a more potential ASK1 inhibitor has clinical and scientific significance. The ASK1 small-molecule inhibitor provided by the invention has the advantages of novel structure, good activity and the like, and has the potential of further development.

Description

Technical Field

[0001] The present invention belongs to the field of medical technology, and in particular relates to a group of ASK1 inhibitors and uses thereof. Background Art

[0002] ASK1 is a member of the mitogen-activated protein kinase kinase (MAP3K) family. ASK1 can be activated by a series of stimuli, such as oxidative stress, reactive oxygen species (ROS), LPS, TNF-α, FasL, endoplasmic reticulum stress, and increased intracellular calcium concentration. In response to these stimuli, ASK1 is activated by autophosphorylation at Thr838 and then phosphorylates MAP2K (such as MKK3 / 6 and MKK4 / 7), which then phosphorylates and activates JNK (c-Jun N-terminal kinase) and P38 MAPK (P38 mitogen-activated protein kinase), respectively.

[0003] Phosphorylation of ASK1 may lead to apoptosis or other cellular responses. It has been reported that the activation and signaling of ASK1 play an important role in diseases and conditions caused by chronic fatty and fibrotic degeneration (non-alcoholic fatty liver disease (NAFLD)) and non-alcoholic steatohepatitis (NASH). Neurodegenerative diseases, cardiovascular diseases, diabetes (including diabetic nephropathy and other complications of diabetes), inflammatory diseases, autoimmune diseases, respiratory diseases (including chronic obstructive pulmonary disease (COPD), idiopathic fibrosis (IPF) and acute lung injury) and metabolic-related diseases. In addition, ASK1 is also involved in organ damage after ischemia and reperfusion in the heart, brain, and kidneys. Therefore, ASK1 signaling inhibitors have the potential to remedy or improve the condition of patients with such diseases.

[0004] At present, some literature reports that some ASK1 inhibitors have potential problems such as low molecular activity, poor target selectivity, unclear animal efficacy dose-effect relationship, and poor drug-like properties of the compounds. Therefore, there is still a need to develop a more effective ASK1 kinase small molecule inhibitor in this field. Summary of the invention

[0005] A group of ASK1 inhibitors disclosed in the present invention has the following structure: The group of ASK1 inhibitors, their enantiomers, their diastereomers, their racemates or mixtures thereof or pharmaceutically acceptable salts thereof are characterized in that the acids that can be used for salt formation in the pharmaceutically acceptable salts include inorganic acids and organic acids; the inorganic acids include hydrochloric acid, sulfuric acid, methanesulfonic acid or phosphoric acid; the organic acids include acetic acid, trichloroacetic acid, propionic acid, butyric acid, maleic acid, p-toluenesulfonic acid, malic acid, malonic acid, cinnamic acid, citric acid, fumaric acid, aspartic acid or tartaric acid.

[0006] The use of the group of ASK1 inhibitors, their enantiomers, their diastereomers, their racemates or mixtures thereof or pharmaceutically acceptable salts thereof in the preparation of drugs for treating diseases related to the activity or expression of ASK1 kinase.

[0007] The application is characterized in that the diseases related to the activity or expression of ASK1 kinase include locomotor system diseases, digestive system diseases, respiratory system diseases, circulatory system diseases, nervous system diseases, urinary system diseases, endocrine system diseases or reproductive system diseases.

[0008] The application is characterized in that the diseases include non-alcoholic fatty hepatitis, diabetic nephropathy, pulmonary hypertension, ulcerative colitis, multiple sclerosis, liver cancer, breast cancer, lung cancer, prostate cancer, multiple myeloma, leukemia, gastric cancer, colon cancer, pancreatic cancer, ovarian cancer or human glioma. Beneficial Effects

[0009] The benefit of the present invention is that the ASK1 target is related to many diseases. Among the many ASK1 modulators disclosed, there are currently no drugs on the market, so it is of clinical and scientific significance to find more potential ASK1 inhibitors. The ASK1 small molecule inhibitor provided by the present invention has the advantages of novel structure, good activity, etc., and has the potential for further development. DETAILED DESCRIPTION

[0010] To describe the present invention, the following examples are listed. However, it should be understood that the present invention is not limited to these examples, which are only provided to provide methods for practicing the present invention.

[0011] Generally, the compounds of the present invention can be prepared by the methods described herein, unless otherwise specified. The following reaction schemes and examples are provided to further illustrate the present invention.

[0012] Those skilled in the art will recognize that the chemical reactions described herein can be used to appropriately prepare other compounds of the invention, and other methods for preparing the compounds of the invention are considered to be within the scope of the invention. For example, the synthesis of non-exemplified compounds according to the invention can be successfully accomplished by those skilled in the art through modification methods, such as appropriate protection of interfering groups, by utilizing other known reagents in addition to those described herein, or by making some conventional modifications to the reaction conditions. In addition, the reactions disclosed herein or known reaction conditions are also recognized to be applicable to the preparation of other compounds of the invention.

[0013] In the examples described below, all temperatures are in degrees Celsius unless otherwise indicated. Reagents were purchased from commercial suppliers such as Shanghai Haohong Biopharmaceutical Technology Co., Ltd., Shanghai Aladdin Biochemical Technology Co., Ltd., Shanghai Bid Pharmaceutical Technology Co., Ltd., and Jiangsu Aikang Biopharmaceutical Research and Development Co., Ltd. and were used without further purification. General reagents and anhydrous reagents were purchased from Sinopharm Shanghai Reagent Co., Ltd., Nanjing Chemical Reagent Co., Ltd., Xilong Scientific, and Admas-Beta unless otherwise indicated.

[0014] The following reactions are generally carried out under positive pressure of nitrogen or argon or in a Shrek tube. The reaction bottles are plugged with suitable hollow glass stoppers and the substrate is injected by pipette. The glass dishes are all dried.

[0015] The chromatographic column used was a silica gel column. Silica gel (100-200 mesh) was purchased from Yantai Jiangyou Silica Gel Development Co., Ltd.

[0016] 1 H NMR spectra were recorded using a Bruker 300 MHz or 400 MHz NMR spectrometer. 1 H NMR spectra were measured in CDCl 3， DMSO- d 6 is the solvent (in ppm), TMS (0 ppm) is used as the reference standard, and when multiple peaks are present, the following abbreviations are used: s (singlet), d (doublet), t (triplet), q (quartet), m (multiplet), br (broadened), brs (broadened singlet), dd (doublet of doublets), dt (doublet of triplets). Coupling constants J , expressed in Hertz (Hz).

[0017] The following abbreviations are used throughout this invention: DMF N,N-Dimethylformamide DMSO Dimethyl sulfoxide DMSO- d 6 Deuterated dimethyl sulfoxide THF Tetrahydrofuran g mg milligram M mole per liter mM millimole per liter mol mole mmol millimole mL milliliters μL microliter MPa Megapascal DPPFP 2 1,1-Bis(diphenylphosphine)ferrocenepalladium(II) chloride DMF-DMA N , N -Dimethylformamide dimethyl acetal LiA 4 Lithium aluminum hydride NaCl Sodium chloride K 2 CO 3 Potassium carbonate CuI Cuprous iodide CuBr Cuprous Bromide t - BuONO tert-Butyl nitrite Example 1: Example 1 of the present invention provides a method for preparing an intermediate 1, and its synthetic route is as follows: Step 1: Dissolve compound 1 (50 g, 329 mmol) in hydrazine hydrate (32.9 g, 658 mmol), heat under reflux and stir for 3 hours. TLC detection shows that the reaction is complete, cool the reaction solution to room temperature, filter, and wash the filter cake with ethyl acetate to obtain compound 2.

[0018] 44 g of a white solid was obtained by this method with a yield of 87%.

[0019] Step 2: Compound 2 (44 g, 289 mmol) was dissolved in DMF-DMA (450 mL) and heated under reflux for 18 hours. After TLC detection, the reaction solution was cooled and concentrated, ethyl acetate was added, and the reaction solution system was heated to 50 °C and stirred for 20 minutes, and filtered to obtain compound 3.

[0020] 55 g of white solid was obtained by this method with a yield of 71.20%.

[0021] Step 3: Dissolve compound 3 (20 g, 76 mmol) in acetonitrile / acetic acid (v / v = 5 / 1, 200 mL), add isopropylamine (22 g, 22 mmol) dropwise under ice bath, reflux and heat for 36 hours, cool the reaction solution, evaporate under reduced pressure, alkalize the reaction solution with 1N NaOH solution until pH = 8, filter and dry to obtain intermediate 1.

[0022] 8 g of white solid was obtained by this method with a yield of 28%.

[0023] Example 2: Example 2 of the present invention provides a method for preparing intermediate 2, and its synthetic route is as follows:

[0024] Intermediate 1 (3 g, 14.8 mmol) was dissolved in hydrobromic acid solution (30 mL), cooled to 0 °C and then NaNO was added dropwise. 2 (1 g, 14.8 mmol) in water (10 mL) until the solution turns starch potassium iodide paper blue, then add Br 2 (1 g, 14.8 mmol), stirred at room temperature overnight, adjusted to weak alkalinity with 1N NaOH aqueous solution, extracted with ethyl acetate, washed the organic phase with sodium thiosulfate aqueous solution, dried and column chromatographed (DCM / MeOH = 80 / 1 - 30 / 1) to obtain Intermediate 2.

[0025] 1.17 g of brown solid was obtained by this method with a yield of 27%.

[0026] Example 3: Example 3 of the present invention provides a method for preparing intermediate 3, and its synthetic route is as follows:

[0027] Intermediate 2 (1 g, 3.74 mmol), compound 4 (3.22 g, 37.43 mmol), cuprous iodide (71.30 mg, 374.35 μmol), potassium carbonate (1.03 g, 7.49 mmol) and (1R,2R)-(-)- N , N'-Dimethyl-1,2-cyclohexanediamine (106.50 mg, 748.70 μmol) was placed in a two-necked flask and evacuated with nitrogen three times. A dioxane solution (30 mL) evacuated with nitrogen was added and reacted at 110 °C for 3 hours. The intermediate 3 was obtained by purification by basic alumina column chromatography (DCM / MeOH = 250 / 1).

[0028] 800 mg of a white solid was obtained by this method with a yield of 78.43%.

[0029] The intermediate 3 obtained in this example was identified, and the following results were obtained: MS m / z(ESI ) 273 [M+H] + Example 4: Example 4 of the present invention provides a method for preparing intermediate 4, and its synthetic route is as follows:

[0030] Compound 5 (4.62 g, 29.52 mmol) was added dropwise into a dioxane solution of intermediate 1 (3 g, 14.76 mmol) and potassium carbonate (2.45 g, 17.71 mmol) at 0 °C. The reaction was allowed to react overnight. The solvent was removed by distillation under reduced pressure and column chromatography was used for purification (DCM / MeOH = 80 / 1 - 30 / 1) to obtain intermediate 4.

[0031] The method was used to obtain 3 g of a white solid with a yield of 62.89%.

[0032] Example 5: Example 5 of the present invention provides a method for preparing the final product 1, and its synthetic route is as follows:

[0033] Step 1: Add carbon tetrabromide (2.7 g, 8.14 mmol) to anhydrous DMF of compound 6 (1 g, 7.4 mmol) at 0 °C, and add sodium tert-butoxide (2.56 g, 26.63 mmol). React at room temperature for 30 minutes. Dilute with brine, extract with anhydrous ether, and dry with anhydrous sodium sulfate to obtain compound 7.

[0034] 1.2 g of brown oil was obtained by this method with a yield of 75.95%.

[0035] Step 2: Compound 7 (500 mg, 2.34 mmol) was prepared according to the preparation method of Intermediate 3 and purified by column chromatography (DCM / MeOH = 120 / 1) to obtain Intermediate 8.

[0036] The method was used to obtain 300 mg of a white solid with a yield of 58.59%.

[0037] The compound 8 prepared in this example was identified, and the following results were obtained: 1 H NMR (300 MHz, Chloroform- d ) δ 7.79 (dt, J = 7.7, 1.3 Hz, 2H), 7.42(td, J = 7.6, 1.3 Hz, 1H), 5.12 (s, 1H), 4.51 – 4.30 (m, 2H), 3.85 – 3.67 (m, 2H). Step 3: Intermediate 8 (50.0 mg, 228.04 μmol), intermediate 2 (60.92 mg, 228.04 μmol), cuprous iodide (4.34 mg, 22.80 μmol), potassium carbonate (63.03 mg, 456.08 μmol) and (1R,2R)-(-)- N , N '-Dimethyl-1,2-cyclohexanediamine (6.49 mg, 45.61 μmol) was placed in a two-necked bottle, and after nitrogen was evacuated three times, anhydrous dioxane (3 mL) was added and reacted at 110 °C for 3 hours. The final product 1 was purified by basic alumina column chromatography.

[0038] This method was used to obtain 48 mg of a white solid with a yield of 51.91%.

[0039] The final product 1 obtained in this example was identified, and the following results were obtained: 1 H NMR (300 MHz, DMSO- d 6 ) d 8.93 (s, 1H), 8.35 (dd, J = 8.5, 0.9 Hz,1H), 8.09 – 7.97 (m, 2H), 7.90 (dd, J = 7.6, 0.9 Hz, 1H), 7.80 (d, J = 8.0Hz, 1H), 7.49 – 7.40 (m, 1H), 7.31 (td, J = 7.6, 1.2 Hz, 1H), 5.48 (p, J=6.7 Hz, 1H), 4.29 (s, 4H), 1.54 (d, J = 6.7 Hz, 6H).

[0040] Example 6: Example 6 of the present invention provides a method for preparing the final product 2, and its synthetic route is as follows:

[0041] Step 1: Intermediate 9 (3 g, 19.72 mmol) was added to H 2 SO 4 / H 2 O mixed solution (v / v = 1 / 3), NaNO was added dropwise at 0 °C. 2 / H 2 O (1.4 g / 5 mL) for diazotization, and KSCN / H 2 O (2.48 g / 5 mL) and CuSCN (2.3 g). Stir overnight and heat to 60 °C for 5 minutes the next day. After cooling, extract with ethyl acetate, dry with anhydrous sodium sulfate, and purify by column chromatography (PE / EA = 50 / 1) to obtain intermediate 10.

[0042] 2.2 g of brown solid was obtained by this method with a yield of 52.22%.

[0043] Step 2: Add intermediate 10 (2.2 g, 11.33 mmol) and metallic tin (10 g, 84.96 mmol) to concentrated hydrochloric acid (10 mL) under shaking, and heat to 65 °C until most of the powdered metallic tin reacts and the remaining metallic tin appears as spherical particles. Tilt the solution to remove the remaining metallic tin, cool and filter to obtain a white salt, which is intermediate 11.

[0044] The method was used to prepare 1 g of white salt with a yield of 53.76%.

[0045] Step 3: Under nitrogen protection, tert-butyl nitrite (609 μl, 5.18 mmol) was added to an acetonitrile solution of copper bromide (742.47 mg, 5.18 mmol), and the mixture was stirred at room temperature for 10 minutes. An acetonitrile solution of intermediate 11 (500 mg, 3.04 mmol) was added to the mixture and reacted at room temperature for 1 hour. The solvent was removed by distillation under reduced pressure, diluted with water, extracted with ethyl acetate, and purified by column chromatography (PE / EA = 30 / 1) to obtain intermediate 12.

[0046] The method was used to obtain 423 mg of brown oil with a yield of 60.95%.

[0047] Step 4: Intermediate 13 (223 mg, 977.61 μmol) was prepared according to the method of intermediate 8 to obtain intermediate 13.

[0048] This method was used to obtain 98 mg of a white solid with a yield of 42.97%.

[0049] Step 5: Compound 13 (70 mg, 262.04 μmol) was prepared according to the method of final product 1 to obtain final product 22.

[0050] This method was used to obtain 27 mg of a white solid with a yield of 21.37%.

[0051] The final product 2 obtained in this example was identified, and the following results were obtained: 1 H NMR (300 MHz, Chloroform- d ) d 8.51 – 8.37 (m, 2H), 8.06 (dd, J =7.6, 0.9 Hz, 1H), 7.98 – 7.84 (m, 1H), 7.67 (dd, J = 7.2, 1.8 Hz, 1H), 7.26 –7.11 (m, 2H), 5.59 (p, J = 6.7 Hz, 1H), 4.53 – 4.27 (m, 4H), 2.68 (s, 3H), 1.63 (d, J = 6.7 Hz, 7H).

[0052] Example 7: Example 7 of the present invention provides a method for preparing the final product 3, and its synthetic route is as follows:

[0053] Final product 3 was prepared according to the method of Example 5.

[0054] The final product 3 obtained in this example was identified, and the following results were obtained: 1 H NMR (300 MHz, Chloroform- d ) d 8.56 (d, J = 1.6 Hz, 1H), 8.47 (d, J = 8.4 Hz, 1H), 8.42 (s, 1H), 8.15 (dd, J= 8.5, 1.7 Hz, 1H), 8.10 (d, J = 7.5Hz, 1H), 7.98 – 7.91 (m, 1H), 7.86 (d, J = 8.5 Hz, 1H), 5.58 (p, J = 6.9 Hz,1H), 4.52 – 4.29 (m, 5H), 3.98 (s, 3H), 1.64 (d, J = 6.7 Hz, 6H).

[0055] Example 8: Example 8 of the present invention provides a method for preparing the final product 4, and its synthetic route is as follows:

[0056] Final product 4 was prepared according to the method of Example 5.

[0057] The final product 4 obtained in this example was identified, and the following results were obtained: 1 H NMR (300 MHz, Chloroform- d ) d 8.53 – 8.38 (m, 2H), 8.07 (d, J =7.6 Hz,1H), 7.92 (t, J = 8.0 Hz, 1H), 7.73 (d, J = 8.2 Hz, 1H), 7.64 (s, 1H), 7.26 (s, 1H), 5.65 – 5.51 (m, 1H), 4.49 – 4.25 (m, 4H), 2.50 (s, 3H), 1.64(d, J = 6.7 Hz, 6H).

[0058] Example 9: Example 9 of the present invention provides a method for preparing the final product 5, and its synthetic route is as follows:

[0059] Step 1: Compound 18 (1 g, 8.25 mmol) and ammonium thiocyanate (12.6 g, 16.50 mmol) were refluxed in 2N HCl solution (10 mL) overnight. After the reaction was complete, the mixture was diluted with water, filtered and dried to obtain compound 19.

[0060] 1.54 g of white solid was obtained by this method with a yield of 100%.

[0061] Step 2: Compound 19 (1.5 g, 8.32 mmol) was dissolved in chloroform solution, and bromine was added dropwise at 0 °C, and the temperature was kept below 5 °C for 4 hours. The obtained product was dried, washed with a saturated sodium thiosulfate solution, and purified by column chromatography (PE / EA = 3 / 1) to obtain compound 20.

[0062] 1 g of white solid was obtained by this method with a yield of 67.58%.

[0063] Step 3, step 4 and step 5: Compound 21, compound 22 and final product 5 were prepared according to the method of Example 5.

[0064] The final product 5 obtained in this example was identified, and the following results were obtained: 1 H NMR (300 MHz, Chloroform- d ) d 8.55 – 8.35 (m, 2H), 8.07 (dd, J =7.6, 0.9 Hz, 1H), 7.91 (dd, J = 8.5, 7.6 Hz, 1H), 7.50 (s, 1H), 6.96 (s, 1H), 5.59 (p, J = 6.7 Hz, 1H), 4.48 – 4.25 (m, 4H), 2.54 (s, 3H), 2.46 (s, 3H), 1.63 (d, J = 6.7 Hz, 6H).

[0065] Example 10: Example 10 of the present invention provides a method for preparing the final product 6, and its synthetic route is as follows:

[0066] Step 1: Compound 23 (200 mg, 934.12 μmol), compound 24 (126.32 mg, 1.03 mmol), cesium carbonate (669.66 mg, 2.06 mmol) and DPPF palladium dichloride (34.18 mg, 46.71 μmol) were placed in a two-necked bottle, evacuated with nitrogen three times, and dioxane and aqueous solution (v / v = 4 / 1) evacuated with nitrogen were added. The mixture was reacted at 110 °C for 24 hours, diluted with water, extracted with ethyl acetate, dried over anhydrous sodium sulfate, and purified by column chromatography (DCM / MeOH = 80 / 1) to obtain pure compound 25.

[0067] The method was used to obtain 156 mg of a white solid with a yield of 78.79%.

[0068] Step 2: Compound 25 (118 mg, 555.90 μmol) was dissolved in DMF, carbon tetrabromide (202.79 mg, 611.47 μmol) was added under ice bath, and then sodium tert-butoxide particles (192.32 mg, 2.0 mmol) were slowly added. The reaction was carried out at room temperature for 3 hours, quenched with water, extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, and purified by column chromatography (PE / EA = 2 / 1) to obtain compound 26.

[0069] The method was used to obtain 150 mg of brown solid with a yield of 89.28%.

[0070] Step 3: Compound 26 (100 mg, 367.23 μmol), intermediate 3 (128.31 mg, 440.67 μmol), potassium carbonate (111.66 mg, 807.90 μmol), cuprous iodide (34.97 mg, 183.61 μmol) and (1R,2R)-(-)- N , N '-Dimethyl-1,2-cyclohexanediamine (52.24 mg, 367.23 μmol) was placed in a two-necked flask, evacuated with nitrogen three times, and anhydrous and oxygen-free dioxane was added to react at 110 °C for 3 hours. Water was added to quench, extracted with ethyl acetate three times, washed with saturated brine, and dried over anhydrous sodium sulfate. The final product 6 was purified by basic alumina column chromatography (DCM / MeOH = 250 / 1).

[0071] This method was used to obtain 38 mg of a white solid with a yield of 23.03%.

[0072] The final product 6 obtained in this example was identified, and the following results were obtained: 1 H NMR (300 MHz, Chloroform- d ) d 8.95 (d, J = 2.4 Hz, 1H), 8.70 (d, J = 5.5 Hz, 1H), 8.45 (d, J = 8.4 Hz, 1H), 8.41 (s, 1H), 8.09 (d, J = 7.5 Hz,2H), 7.96 – 7.84 (m, 2H), 7.59 (t, J= 7.7 Hz, 1H), 7.47 (dd, J = 7.9, 4.7Hz, 1H), 7.39 (d, J = 7.5 Hz, 1H), 5.61 – 5.51 (m, 1H), 4.53 – 4.28 (m, 4H), 1.64 (d, J = 6.8 Hz, 6H).

[0073] Example 11: Example 11 of the present invention provides a method for preparing the final product 7, and its synthetic route is as follows:

[0074] Final product 7 was prepared according to the method of Example 5.

[0075] The final product 7 obtained in this example was identified, and the following results were obtained: 1 H NMR (300 MHz, Chloroform- d ) d 8.47 (dd, J = 8.4, 0.9 Hz, 1H), 8.41(s, 1H), 8.06 (dd, J = 7.6, 0.9 Hz, 1H), 7.96 – 7.86 (m, 1H), 7.67 (dd, J =7.2, 1.8 Hz, 1H), 7.28 – 7.17 (m, 2H), 5.59 (p, J = 6.7 Hz, 1H), 4.58 – 4.25(m, 4H), 2.68 (s, 3H), 1.63 (d, J = 6.7 Hz, 6H).

[0076] Example 12: Example 12 of the present invention provides a method for preparing the final product 8, and its synthetic route is as follows:

[0077] At 0 °C, sodium hydroxide (68.28 mg, 1.71 mmol) was added to tetrahydrofuran of compound 29 (100 mg, 853.60 μmol) and intermediate 4 (331.22 mg, 1.02 mmol). After reacting at room temperature for 3 hours, water was added to quench the reaction, and the mixture was extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, and purified by column chromatography (DCM / MeOH / NH 3 ·H2 (0 = 240 / 3 / 1) to obtain the final product 8.

[0078] The method was used to prepare 200 mg of a white solid with a yield of 67.78%.

[0079] The final product 8 obtained in this example was identified, and the following results were obtained: 1 H NMR (400 MHz, DMSO- d 6 ) d 10.62 (s, 1H), 8.86 (s, 1H), 8.20 (d, J =8.3 Hz, 1H), 8.09 – 7.97 (m, 2H), 7.93 (d, J = 8.2 Hz, 1H), 7.86 (d, J = 7.5Hz, 1H), 7.64 (d, J = 7.7 Hz, 1H), 7.33 – 7.21 (m, 2H), 6.77 (d, J = 3.7 Hz,1H), 5.66 (p, J = 6.7 Hz, 1H), 1.43 (d, J = 6.7 Hz, 6H).

[0080] Example 13: Example 13 of the present invention provides a method for preparing the final product 9, and its synthetic route is as follows:

[0081] Step 1: Compound 30 (200 mg, 1.32 mmol), compound 24 (178.39 mg, 1.45 mmol), potassium sulfate (476.08 mg, 2.24 mmol), tricyclohexylphosphine (11.10 mg, 39.58 μmol) and tris(dibenzylideneacetone)dipalladium (12.08 mg, 13.19 μmol) were placed in a double-necked flask, evacuated with nitrogen three times, and added with oxygen-depleted dioxane aqueous solution (v / v = 4 / 1) (30 mL) at 100 °C, and reacted for 18 hours. Extracted with ethyl acetate and water, washed with saturated brine, dried over anhydrous sodium sulfate, and purified by column chromatography (DCM / MeOH = 150 / 1) to obtain compound 31.

[0082] The method was used to obtain 156 mg of a white solid with a yield of 60.94%.

[0083] Step 2: Final product 9 was prepared according to the method of Example 12.

[0084] The final product 9 obtained in this example was identified, and the following results were obtained: 1 H NMR (300 MHz, DMSO- d 6 ) d 10.79 (s, 1H), 8.89 (s, 2H), 8.67 – 8.45 (m, 2H), 8.16 – 7.98 (m, 4H), 7.90 (dd, J = 7.1, 1.4 Hz, 1H), 7.80 (d, J =8.2 Hz, 1H), 7.62 (dd, J = 8.2, 1.6 Hz, 1H), 7.58 – 7.45 (m, 1H), 6.90 – 6.80(m, 1H), 5.69 (p, J = 6.6 Hz, 1H), 1.44 (d, J = 6.7 Hz, 6H).

[0085] Example 14: Example 14 of the present invention provides a method for preparing the final product 10, and its synthetic route is as follows:

[0086] Step 1: Compound 33 (1.30 g, 6.60 mmol) and 48% hydrogen bromide solution (2 mL) were added dropwise to a solution of compound 32 (1.0 g, 6.53 mmol) in ethanol (10 mL). The mixture was heated at 80 °C for 16 hours, and the remaining filtrate was neutralized with saturated sodium bicarbonate and extracted with ethyl acetate to confirm compound 34.

[0087] 400 mg of a yellow solid was obtained by this method with a yield of 34.48%.

[0088] Step 2: Compound 34 (400 mg, 2.06 mmol) was added to a tetrahydrofuran aqueous solution (v / v = 9 / 1) (4 mL) of lithium hydroxide (98.63 mg, 4.12 mmol), and the mixture was reacted for two days. The organic solution was removed under reduced pressure, and hydrochloric acid was added to precipitate the solid, which was filtered and dried thoroughly. The filtrate was extracted with ethyl acetate, dried over anhydrous sodium sulfate, and then the mixture was combined and directly used for the next step.

[0089] Step 3: Add compound 35, intermediate 1 (418.56 mg, 2.06 mmol), pyridine (325.80 mg, 4.2 mmol) in DMF (4 mL), and add phosphorus oxychloride (947.23 mg, 6.18 mmol) dropwise at 0 °C under nitrogen protection. React at 0 °C for 2 hours, add water to quench, adjust the alkalinity with saturated sodium bicarbonate, extract with ethyl acetate, dry with anhydrous sodium sulfate, and purify by basic alumina column chromatography (DCM / MeOH = 250 / 1) to obtain the final product 10.

[0090] The method was used to obtain 197 mg of a white solid with a yield of 22.93%.

[0091] The final product 10 obtained in this example was identified, and the following results were obtained: 1 H NMR (400 MHz, Chloroform-d) d 9.74 (s, 1H), 8.56 (s,1H), 8.25-8.13(m,2H), 8.09 (dd, J=7.5, 1.3 Hz,1H), 7.93 (dd, J=7.7, 1.3 Hz, 1H), 7.86 (d, J=9.2 Hz, 1H),7.72(d, J=9.2 Hz,1H),7.24(d, J=4.2 Hz,1H),4.85-4.72 (m, 1H), 1.50(s, 6H).

[0092] Example 15: Example 15 of the present invention provides a method for preparing the final product 11, and its synthetic route is as follows:

[0093] Step 1: Compound 36 (2.5 g, 15.43 mmol) and compound 37 (2 g, 16.20 mmol) were added to an acetic acid solution and refluxed at 120 °C. The precipitated solid was filtered out, dried and collected to obtain compound 38.

[0094] 2.48 g of white solid was obtained by this method with a yield of 58.41%.

[0095] Step 2: The final product 11 is prepared according to the methods of steps 2 and 3 of Example 14.

[0096] The final product 11 obtained in this example was identified, and the following results were obtained: 1 H NMR (300 MHz, DMSO- d 6 ) d 10.88 (s, 1H), 9.64 (d, J = 2.2 Hz, 1H),8.89 (d, J = 2.7 Hz, 2H), 8.18 (dd, J = 18.2, 8.3 Hz, 1H), 8.05 (t, J = 8.0Hz, 1H), 7.93 – 7.79 (m, 1H), 6.63 (s, 1H), 5.69 (p, J = 6.7 Hz, 1H), 2.17(q, J = 5.2, 4.4 Hz, 1H), 1.46 (d, J = 6.5 Hz, 6H), 1.09 (dt, J = 8.5, 3.2Hz, 2H), 0.97 – 0.89 (m, 2H).

[0097] Example 16: Example 16 of the present invention provides a method for preparing the final product 12, and its synthetic route is as follows:

[0098] The final product 12 was prepared according to the method of the second and third steps of Example 14.

[0099] The final product 12 obtained in this example was identified, and the following results were obtained: .

[0100] Example 17: Example 17 of the present invention provides a method for preparing the final product 13, and its synthetic route is as follows:

[0101] Step 1: Add compound 41 (1 g, 3.42 mmol), compound 42 (1.30 g, 5.14 mmol), potassium acetate (672.04 mg, 6.85 mmol) and DPPF palladium dichloride (125.26 mg, 171.19 μmol) into a Shrek tube, evacuate nitrogen three times, add anhydrous and oxygen-free DMSO solution, and react at 110 °C for 18 hours. Quench with water, extract with ethyl acetate, wash with saturated brine, dry with anhydrous sodium sulfate, and purify by column chromatography (PE / EA = 10 / 1) to obtain compound 43.

[0102] 1 g of white solid was obtained by this method with a yield of 100%.

[0103] Step 2: Compound 43 (500 mg, 1.71 mmol), cesium carbonate (1.23 g, 3.77 mmol), compound 44 (274.26 mg, 1.54 mmol), and tetrakis(triphenylphosphine)palladium (98.89 mg, 85.58 μmol) were added to a Shrek tube, nitrogen was evacuated three times, and anhydrous and oxygen-free dioxane water (v / v = 5 / 1) solution was added, and the mixture was reacted at 110 °C for 24 hours. The mixture was quenched with water, extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, and purified by column chromatography (PE / EA = 10 / 1) to obtain compound 45.

[0104] 400 mg of a white solid was obtained by this method with a yield of 88.89%.

[0105] Step 3: Final product 13 was prepared according to the method of step 2 and step 3 of Example 14.

[0106] The final product 13 obtained in this example was identified, and the following results were obtained: 1 H NMR (400 MHz, DMSO- d 6 ) d 10.69 (s, 1H), 8.89 (s, 1H), 8.39 – 8.28 (m, 2H), 8.14 – 8.01 (m, 2H), 7.89 (d, J = 7.6 Hz, 1H), 7.37 (d, J = 8.8 Hz,1H), 7.31 (d, J = 1.3 Hz, 1H), 5.56 (p, J = 6.7 Hz, 1H), 4.02 (s, 3H), 2.42(s, 3H), 1.50 (d, J = 6.7 Hz, 6H).

[0107] Example 18: Example 18 of the present invention provides a method for preparing the final product 14, and its synthetic route is as follows:

[0108] Step 1: Compound 36 (500 mg, 4.06 mmol) and compound 46 (965.65 mg, 4.47 mmol) were suspended in glacial acetic acid, refluxed for 12 hours, cooled and filtered, and washed with ethanol to obtain compound 47.

[0109] 750 mg of white solid was obtained by this method with a yield of 75%.

[0110] Step 2: In an ice-water bath, iodomethane (101.247 mg, 713.50 μmol) was slowly added dropwise to a DMF solution of potassium carbonate (98.61 mg, 713.50 μmol) and compound 47 (150 mg, 648.63 μmol) and reacted at room temperature. After the reaction was completed, the mixture was quenched with water, extracted with ethyl acetate, washed with saturated brine, and dried over anhydrous sodium sulfate to obtain compound 48.

[0111] The method was used to obtain 162 mg of a white solid with a yield of 94.94%.

[0112] Step 3: Final product 14 is prepared according to the method of step 2 and step 3 of Example 14.

[0113] The final product 14 obtained in this example was identified, and the following results were obtained: 1 H NMR (300 MHz, DMSO- d 6 ) d 10.88 (s, 1H), 9.64 (d, J = 2.2 Hz, 1H),8.89 (d, J = 2.7 Hz, 2H), 8.18 (dd, J = 18.2, 8.3 Hz, 1H), 8.05 (t, J = 8.0Hz, 1H), 7.93 – 7.79 (m, 1H), 6.63 (s, 1H), 5.69 (p, J = 6.7 Hz, 1H), 4.13(s, 3H) 2.17 (q, J = 5.2, 4.4 Hz, 1H), 1.46 (d, J = 6.5 Hz, 6H), 1.09 (dt, J = 8.5, 3.2 Hz, 2H), 0.97 – 0.89 (m, 2H).

[0114] Example 19: Example 19 of the present invention provides a method for preparing the final product 15, and its synthetic route is as follows:

[0115] Step 1: Compound 49 (2 g, 11.10 mmol) was added dropwise to thionyl chloride (2.64 g, 22.10 mmol) at 0 °C, and then to a catalytic amount of DMF, and the mixture was refluxed for 3 hours, and the solvent was evaporated under reduced pressure. The mixture was dissolved in tetrahydrofuran, and added dropwise to a tetrahydrofuran solution of compound 50 (1.55 g, 12.21 mmol) and triethylamine (1.35 g, 13.32 mmol) at 0 °C, and the reaction was continued overnight. The solvent was removed by pressure distillation the next day, and the mixture was recrystallized from acetone to obtain compound 51.

[0116] The method was used to obtain 3 g of a yellowish solid with a yield of 90.09%.

[0117] Step 2: Add hydrazine hydrate (6 mL) to a methanol solution of compound 51 (1 g, 3.18 mmol), reflux at 100°C for 3 hours to precipitate compound 52.

[0118] 717 g of yellow-green solid was obtained by this method with a yield of 71.70%.

[0119] Step 3: Compound 52 (371 mg, 1.24 mmol) was dissolved in DMF, and DMF-DMA (736.16 mg, 6.18 mmol) was added. The mixture was refluxed for 5 hours, and the product was diluted with water, extracted with ethyl acetate, dried over anhydrous sodium sulfate, and the solvent was dried by spin drying without purification and directly used for the next step.

[0120] Step 4: The residue containing compound 53 from the previous step was dissolved in a mixed solution of acetonitrile and acetic acid (v / v = 5 / 1), and compound 54 (374.28 mg, 63.3 mmol) was added dropwise, and the mixture was refluxed for 18 hours. The mixture was alkali-adjusted with aqueous sodium carbonate solution, extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, and purified by column chromatography (DCM / MeOH = 80 / 1 - 30 / 1) to obtain compound 55.

[0121] The method was used to obtain 350 mg of a white solid with a yield of 78.83%.

[0122] Step 5: Reduced iron powder (111.26 mg, 1.99 mmol) and ammonium chloride (266.41 mg, 4.98 mmol) were added to a solution of compound 55 (350 mg, 996.11 μmol) in ethanol water (v / v = 10 / 1). The mixture was refluxed for 5 hours and purified by column chromatography (DCM / MeOH = 80 / 1 - 30 / 1) to obtain compound 56.

[0123] 300 mg of off-white solid was obtained by this method with a yield of 93.75%.

[0124] Step 6: Compound 57 (24.49 mg, 152.11 μmol) was added dropwise to a solution of compound 56 (50 mg, 126.76 μmol) and triethylamine (15.39 mg, 152.11 μmol) in dichloromethane. The mixture was reacted at room temperature overnight and purified by column chromatography (DCM / MeOH = 80 / 1 - 30 / 1) to obtain the final product 15.

[0125] The method was used to obtain 30 mg of a white solid with a yield of 48.27%.

[0126] The final product 15 obtained in this example was identified, and the following results were obtained: 1 H NMR (400 MHz, DMSO- d 6 ) d 9.77 (s, 1H), 8.49 (t, J = 2.2 Hz, 1H), 8.44 (s, 1H), 8.23 ​​(s, 1H), 7.96 (dddd, J = 8.3, 5.0, 2.2, 1.1 Hz, 2H), 7.70(t, J = 8.0 Hz, 1H), 7.52 (t, J = 2.2 Hz, 1H), 7.43 (ddd, J = 7.9, 2.3, 1.3Hz, 1H), 7.25 (d, J = 7.8 Hz, 1H), 7.03 (ddd, J = 7.9, 2.3, 1.2 Hz, 1H), 4.79– 4.65 (m, 1H), 2.95 (s, 3H), 1.50 (s, 6H).

[0127] Example 20: Example 20 of the present invention provides a method for preparing the final product 16, and its synthetic route is as follows:

[0128] The final product 16 was prepared according to the method of Example 19.

[0129] The final product 16 obtained in this example was identified, and the following results were obtained: 1 H NMR (400 MHz, DMSO- d 6 ) d10.69 (s, 1H), 8.91 (s, 1H), 8.38 (q, J =1.4 Hz, 1H), 8.24 – 8.13 (m, 2H), 7.98 (ddd, J = 7.3, 5.8, 3.3 Hz, 1H), 7.88– 7.71 (m, 2H), 7.62 – 7.52 (m, 2H), 7.40 (s, 2H), 4.45 (p, J = 6.6 Hz, 1H),1.44 (d, J = 6.7 Hz, 6H).

[0130] Example 21: Example 21 of the present invention provides a method for preparing the final product 17, and its synthetic route is as follows:

[0131] Step 1: Formamidine acetate (132.78 mg, 1.28 mmol) was added to a mixed solution of compound 62 (150 mg, 1.06 mmol) in DIPEA (3 mL) and n-butanol (3 mL), reacted at 110 °C for 6 hours, cooled to room temperature, the precipitated solid was filtered, and washed with ether to obtain compound 63.

[0132] The method was used to obtain 145 mg of a white solid with a yield of 100%.

[0133] Step 2: Compound 63 (145 mg, 965.76 µmol), intermediate 2 (257.98 mg, 965.76 µmol), potassium carbonate (293.64 mg, 2.12 mmol), cuprous iodide (18.39 mg, 96.58 µmol) and (1R,2R)-(-)- N , N '-Dimethyl-1,2-cyclohexanediamine (27.48 mg, 193.15 μmol) was placed in a two-necked flask and evacuated with nitrogen three times. A dioxane solution (10 mL) evacuated with nitrogen was added and reacted at 110 °C for 3 hours. The final product 17 was obtained by purification using basic alumina column chromatography (DCM / MeOH = 250 / 1).

[0134] The method was used to obtain 71 mg of a white solid with a yield of 21.86%.

[0135] The final product 17 obtained in this example was identified, and the following results were obtained: 1H NMR (400 MHz, DMSO- d 6 ) d 8.97 (s, 1H), 8.71 (s, 1H), 8.26 (d, J =6.8 Hz, 2H), 8.22 (s, 1H), 7.87 (dd, J = 6.8, 2.0 Hz, 1H), 5.32 (p, J = 6.6Hz, 1H), 3.99 (s, 3H), 1.47 (d, J = 6.7 Hz, 6H).

[0136] Example 22: Example 22 of the present invention provides a method for preparing the final product 18, and its synthetic route is as follows:

[0137] The final product 18 was prepared according to the method of Example 21.

[0138] The final product 18 obtained in this example was identified, and the following results were obtained: .

[0139] Example 23: Example 23 of the present invention provides a method for preparing the final product 19, and its synthetic route is as follows:

[0140] Step 1: Add compound 65 (500 mg, 1.8 mmol), compound 24 (244 mg, 1.99 mmol), sodium carbonate (420 mg, 3.97 mmol) and tetrakis(triphenylphosphine)palladium (104.27 mg, 0.09 mmol) into a Shrek tube, evacuate with nitrogen three times, add anhydrous and oxygen-free dioxane aqueous solution (v / v = 5 / 1), and react at 100 °C for 18 hours. Quench with water, extract with ethyl acetate, wash with saturated brine, dry with anhydrous sodium sulfate, and purify with column chromatography (PE / EA = 5 / 1) to obtain compound 66.

[0141] 304 mg of white solid was obtained by this method, and the yield was 73.96%.

[0142] The second and third steps: The final product 20 is prepared according to the method of Example 22.

[0143] Step 4: The obtained compound 68 (120 mg, 293.07 µmol) was dissolved in tetrahydrofuran solution, sodium cyanoborohydride (55.25 mg, 879.22 µmol) was added and reacted for 2 hours, and purified by column chromatography to obtain a pure product.

[0144] This method was used to obtain 29.4 mg of a white solid with a yield of 24.39%.

[0145] The final product 19 obtained in this example was identified, and the following results were obtained: 1 H NMR (300 MHz, DMSO- d 6 ) d 8.86 (dd, J = 2.3, 1.7 Hz, 1H), 8.65 (dt, J = 4.8, 1.9 Hz, 2H), 8.56 (s, 1H), 8.33 (d, J = 2.2 Hz, 1H), 8.11 (dd, J =7.9, 1.1 Hz, 1H), 7.91 (dt, J = 7.7, 2.2 Hz, 1H), 7.73 – 7.46 (m, 2H), 7.33(dd, J = 7.5, 4.8 Hz, 1H), 7.27 (dd, J = 7.1, 1.1 Hz, 1H), 7.08 (s, 1H), 6.53(t, J = 3.6 Hz, 1H), 5.75 (d, J = 3.7 Hz, 2H), 4.91 – 4.71 (m, 1H), 1.50 (s, 6H).

[0146] Example 24: Example 24 of the present invention provides a method for preparing the final product 20, and its synthetic route is as follows:

[0147] The final product 20 was prepared according to the method of Example 23.

[0148] The final product 20 obtained in this example was identified, and the following results were obtained: 1 H NMR (300 MHz, DMSO- d 6 ) d 8.95 (s, 1H), 8.64 (s, 1H), 8.42 – 8.33 (m, 2H), 8.32 – 8.23 ​​(m, 2H), 8.14 (dd, J = 8.5, 2.2 Hz, 1H), 8.05 (d, J =0.8 Hz, 1H), 7.98 (p, J = 4.1 Hz, 1H), 7.77 (d, J = 8.4 Hz, 1H), 5.35 (p, J =6.7 Hz, 1H), 3.90 (s, 3H), 1.49 (d, J = 6.7 Hz, 6H).

[0149] Example 25: Example 25 of the present invention provides a method for preparing the final product 21, and its synthetic route is as follows:

[0150] The final product 21 was prepared according to the method of Example 13.

[0151] The final product 21 obtained in this example was identified, and the following results were obtained: 1 H NMR (300 MHz, DMSO- d 6 ) d 8.55 (d, J = 14.1 Hz, 2H), 8.32 (d, J =2.2 Hz, 1H), 8.11 (dd, J = 7.9, 1.1 Hz, 1H), 7.98 (s, 1H), 7.73 – 7.42 (m,2H), 7.27 (dd, J = 7.1, 1.1 Hz, 1H), 7.05 (s, 1H), 6.53 (t, J = 3.6 Hz, 1H),5.75 (d, J = 3.7 Hz, 2H), 4.85 – 4.69 (m, 1H), 3.92 (s, 3H), 1.50 (s, 6H).

[0152] Example 26: Example 26 of the present invention provides a method for preparing the final product 22, and its synthetic route is as follows:

[0153] Step 1: Compound 73 (700 mg, 1.36 mmol), compound 24 (183.49 mg, 1.49 mmol), cesium carbonate (972.73 mg, 2.99 mmol) and tetrakis(triphenylphosphine)palladium (47.05 mg, 40.71 μmol) were placed in a double-necked flask, evacuated with nitrogen three times, added with a nitrogen-evacuated dioxane aqueous solution (v / v = 4 / 1, 7 mL), reacted at 110 °C for 18 hours, quenched with water, extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, and purified by column chromatography (DCM / MeOH= 100 / 1) to obtain compound 74.

[0154] The method was used to obtain 217 mg of a white solid with a yield of 73.06%.

[0155] Step 2: The final product 22 is prepared according to the method of the second and third steps of Example 14.

[0156] The final product 22 obtained in this example was identified, and the following results were obtained: 1 H NMR (400 MHz, DMSO-d6) d 10.83 (s, 1H), 9.03 (d, J = 2.5 Hz, 1H), 8.88 (d, J = 9.1 Hz, 1H), 8.60 (d, J = 4.7 Hz, 1H), 8.26 – 8.10 (m, 3H), 8.04(t, J = 7.9 Hz, 1H), 7.82 (dd, J = 18.5, 5.7 Hz, 2H), 7.52 (dd, J = 8.0, 4.8Hz, 1H), 5.62 (p, J = 6.7 Hz, 1H), 1.46 (d, J = 6.5 Hz, 6H).

[0157] Example 27: Biological test ASK1 kinase activity assay Screening method: Apoptosis signal-regulating kinase 1 (ASK1) activity screening Materials: ASK1 was purchased from Eurofins (Cat. No. 14-606M, Lot No. 28412U); Staurosporine was purchased from Selleckchem (Cat. No. S1421, Lot No. S142107); ADP-GloKinase Assay was purchased from Promege (Cat. No. v9102 / 3, Lot No. 0000176563).

[0158] Steps: Prepare 1x Kinase buffer (20 mM HEPES pH 7.5, 0.01% Triton X-100, 25 mM MgCl 2 ).

[0159] Preparation of compound concentration gradient: Dilute compound to 100 times the final desired maximum inhibitor concentration in the reaction with 100 % DMSO. Add 100 μL of this compound dilution to one well of a 96-well plate. For all compounds, transfer the compound from the test tube to one well of a 96-well plate and serially dilute the compound by transferring 15 μL to 30 μL of 100 % DMSO in the next well, and so on, for a total of 10 concentrations. Afterwards, add 100 μL of 100 % DMSO to two empty wells in the same 96-well plate for no compound control and no enzyme control. Label the plate as the source plate. Transfer 40 μL of compound from the source plate to a new 384-well plate as an intermediate plate and transfer 50 μL of compound to the detection plate using Echo.

[0160] Use 1x Kinase buffer to prepare a kinase solution with a 2x final concentration. Add 2.5 μL of the kinase solution with a 2x final concentration to each well of the compound detection plate, and add 2.5 μL of 1x Kinase buffer to the control well without enzyme.

[0161] Use 1x Kinase buffer to prepare a substrate solution of MBP substrate and ATP at 4 times the final concentration, and add 2.5 μL of substrate solution to each well of the detection plate to start the reaction.

[0162] Kinase reaction and stop: Incubate at 37°C for 60 minutes, then add 5 μL ADP-Glo ​​reagent and incubate at 37°C for 180 minutes.

[0163] Add 10 μL of kinase assay reagent, equilibrate at room temperature for 30 minutes, and collect Envision data for ADP detection.

[0164] The RLU value was read with Envision microplate reader, and the RLU value was converted into the percentage inhibition value. The data was fitted with curve to obtain the inhibition rate of the compound. value.

[0165]

[0166] All documents mentioned in the present invention are cited as references in this application, just as each document is cited as reference individually. In addition, it should be understood that after reading the above teachings of the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms also fall within the scope defined by the claims attached to this application.

Claims

1. A group of ASK1 inhibitors, their enantiomers, their diastereomers, their racemates or mixtures thereof or pharmaceutically acceptable salts thereof, characterized in that: Its structure is any of the following: 。 2. A group of ASK1 inhibitors according to claim 1, its enantiomers, its diastereomers, its racemates or mixtures thereof or pharmaceutically acceptable salts thereof, characterized in that: The pharmaceutically acceptable salts may be formed from acids including inorganic acids and organic acids; the inorganic acids include hydrochloric acid, sulfuric acid, methanesulfonic acid or phosphoric acid; the organic acids include acetic acid, trichloroacetic acid, propionic acid, butyric acid, maleic acid, p-toluenesulfonic acid, malic acid, malonic acid, cinnamic acid, citric acid, fumaric acid, aspartic acid or tartaric acid.

3. Use of a group of ASK1 inhibitors according to claim 1 or 2, their enantiomers, their diastereomers, their racemates or mixtures thereof or pharmaceutically acceptable salts thereof in the preparation of drugs for treating diseases related to the activity or expression of ASK1 kinase.

4. The use according to claim 3, characterized in that: The diseases related to the activity or expression of ASK1 kinase include locomotor system diseases, digestive system diseases, respiratory system diseases, circulatory system diseases, nervous system diseases, urinary system diseases, endocrine system diseases or reproductive system diseases.

5. The use according to claim 4, characterized in that: The diseases include non-alcoholic steatohepatitis, diabetic nephropathy, pulmonary hypertension, ulcerative colitis, multiple sclerosis, liver cancer, breast cancer, lung cancer, prostate cancer, multiple myeloma, leukemia, gastric cancer, colon cancer, pancreatic cancer, ovarian cancer or human glioma.

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