Aminothiazole compound and application thereof as androgen receptor antagonist

Abstract

The invention discloses an aminothiazole compound and an application thereof as an androgen receptor antagonist. The aminothiazole compound comprises pharmaceutically acceptable salts, solvates, prodrugs and isomers of the aminothiazole compound, and pharmaceutical compositions containing the aminothiazole compound and the pharmaceutically acceptable salts, the solvates, the prodrugs and the isomers of the aminothiazole compound. The aminothiazole compound has obvious antagonistic activity to AR, shows better biological activity in in-vivo and in-vitro biological evaluation, and is effective to an enzalutamide drug-resistant ARF876L / T877A two-point mutation model and a comparison caralutamide drug-resistant ARW741C mutant cell model. The percutaneous administration also shows the effect of promoting hair growth on a mouse alopecia model. Therefore, the compound can be used as an AR antagonist to prepare drugs for treating diseases related to androgen receptors, such as prostate cancer, metastatic prostate cancer, castration-resistant prostate cancer, breast cancer, ovarian cancer, androgen-derived alopecia and acne.

Description

Technical Field

[0001] This invention belongs to the pharmaceutical field, specifically relating to aminothiazole compounds and their use as androgen receptor antagonists. It includes aminothiazole compounds or their pharmaceutically acceptable salts, solvates, prodrugs, isomers, metabolites, or pharmaceutical compositions containing such compounds, as well as their use as androgen receptor antagonists in the preparation of drugs for treating prostate cancer and androgenic alopecia. Background Technology

[0002] Prostate cancer is a common malignant tumor of the male urinary system. Traditional androgen deprivation therapy (ADT) can effectively slow disease progression by lowering testosterone levels and inhibiting androgen signaling pathways through surgery or medication. However, after 18–24 months of ADT, most prostate cancer patients develop castration-resistant prostate cancer (CRPC), in which the disease continues to progress even with low testosterone levels, resulting in limited survival and posing a serious threat to human health.

[0003] In recent years, androgen receptor (AR) antagonists have been considered one of the key directions in the treatment of prostate cancer. They are not only of great significance in delaying the progression of hormone-sensitive prostate cancer (HSPC) to CRPC, but also can significantly improve the treatment prognosis of CRPC patients.

[0004] Currently, several small molecule compounds targeting the AR ligand binding pocket (LBP) have been successfully marketed. Among them, the second-generation AR antagonist enzalutamide has significantly improved the treatment efficacy of CRPC and has gained widespread clinical recognition. However, the development of acquired resistance at the LBP binding site has greatly reduced the clinical benefits of these drugs.

[0005] The inventors further analyzed the structure of the AR dimerization interface and discovered a novel binding pocket (DIP) (ACS Cent. Sci. 2023, 9, 675-684). Based on this pocket, a lead molecule with certain AR antagonistic activity was obtained through virtual screening and structural modification (J. Med. Chem. 2024, 67, 17520-17541). Building on this, the inventors systematically optimized the structure of the lead molecule and conducted in-depth analysis of its structure-activity relationship, thereby developing a series of novel aminothiazole derivatives. Summary of the Invention

[0006] This invention provides an aminothiazole compound, a novel compound with androgen receptor antagonistic activity. The aminothiazole compound comprises a pharmaceutically acceptable salt and has the structure of general formula (I): R1 and R2 are selected from hydrogen, halogen, nitro, cyano, hydroxyl, amino, and C, respectively.1-3 Alkyl, C 1-3 Fluoroalkyl, C 1-3 Alkoxy, C 1-3 Fluoroalkoxy groups, C 1-3 alkylamine group, C 1-3 Dialkylamine group, C 3-6 Cycloalkylamine, C 1-3 Alkyl sulfone group; When X is N, Y is selected from S and O; When X is S, Y is selected from N; R3 is selected from hydrogen, halogens, and C. 1-4 Alkyl, C 1-3 Fluoroalkyl, C 3-6 cycloalkyl; R4 is selected from hydrogen, halogen, cyano, and C. 1-3 Alkyl, C 1-3 Fluoroalkyl, C 3-6 cycloalkyl; L is selected from -CH2-, -O-, -CH2CH2-, -CH(CH3)-, -CH(CH3)CH2-, -CH2CH2CH2-, -CH2O-, -CH2CH2O-, -C(=O)-, -CH2C(=O)-, -CH2CH2C(=O)-; G is selected from hydrogen, halogen, hydroxyl, C 1-4 Alkyl, C 1-4 Fluoroalkyl, C 1-3 Alkoxy, C 3-7 cycloalkyl groups, optionally substituted with 1-3 independently selected OH groups. l-6 Alkyl groups, optionally substituted with 1-3 C groups independently selected from OH groups l-6 Alkyl groups, 4-6 membered heterocyclic alkyl groups containing 1-3 heteroatoms independently selected from S and O, -NR5R6, -NH(CH2) n NR5R6, n = 1 - 3; R5 and R6 are independent of each other and can be the same or different, selected from hydrogen, methanesulfonyl, acetyl, 2-hydroxyacetyl, C, etc. 1-4 Alkyl, C 1-4 Fluoroalkyl, C 1-4 Alkoxy, C 3-6 cycloalkyl groups, optionally substituted with 1-3 independently selected OH groups. l-6Alkyl; or NR5R6 is a 3-8 member cyclic amine or substituted cyclic amine selected from morpholine, thiomorpholine, thiomorpholine 1-oxide, thiomorpholine dioxide, piperazine, pyrrolidine, piperidine, 1,4-dioxo-8-azaspiro[4.5]decyl, 2,7-diazaspiro[3.5]nonane, wherein the substituent is selected from hydrogen, halogen, cyano, acetyl, trifluoroacetyl, difluoroacetyl, methoxy, 2-hydroxyacetyl, 2-cyanoacetyl, dimethylaminoacetyl, C 1-6 Alkyl, C 1-6 Fluoroalkyl, C 1-6 Alkoxy, C 1-6 Fluoroalkoxy groups, C 3-6 cycloalkyl, C 1-6 alkylamine group, C 1-6 Dialkylamine group, C-group optionally substituted by 1-3 independently selected OH groups l-6 Alkyl groups, optionally substituted with 1-3 C groups independently selected from OH groups l-6 Alkyl group.

[0007] Preferably, R1 and R2 are selected from hydrogen, fluorine, chlorine, bromine, nitro, cyano, methyl, ethyl, trifluoromethyl, methoxy, difluoromethoxy, trifluoromethoxy, ethoxy, difluoroethoxy, and trifluoroethoxy, respectively. R3 is selected from hydrogen, chlorine, methyl, ethyl, trifluoromethyl, and cyclopropyl. Selected from R4 is selected from hydrogen, fluorine, chlorine, bromine, methyl, ethyl, cyclopropyl, trifluoromethyl, and cyano; L is selected from -CH2-, -CH2O-, -CO-, -CH2C(=O)-, and -CH2CH2-; G is selected from hydrogen, fluorine, chlorine, methyl, ethyl, trifluoromethyl, amino, methylamino, dimethylamino, diethylamino, hydroxyl, methoxy, ethoxy.

[0008] More preferably, the aminothiazole compounds have structural formulas as shown in any of A1-A60, B1-B15, C1-C12, and D1-D12, including pharmaceutically compatible salts or solvates thereof.

[0009] Another object of the present invention is to provide the use of the said aminothiazole compounds, or pharmaceutically acceptable salts or mixtures thereof, tautomers, stereoisomers, solvates, metabolites or prodrug molecules thereof, as androgen receptor antagonists in the preparation of medicaments for the treatment of prostate cancer and androgenic alopecia.

[0010] Its application in the preparation of drugs for treating prostate cancer, metastatic prostate cancer, castration-resistant prostate cancer, breast cancer, ovarian cancer, androgenic alopecia, and acne.

[0011] The drug is made from an aminothiazole compound, or a pharmaceutically acceptable salt thereof, or a mixture thereof, tautomer, stereoisomer, solvate, metabolite, or prodrug molecule, and a pharmaceutically acceptable carrier.

[0012] The term "pharmaceutically acceptable salt" refers to salts prepared by conventional methods, including but not limited to organic acid salts and inorganic acid salts. Organic acid salts include, but are not limited to, oxalates, lactates, p-toluenesulfonates, malates, citrates, fumarates, camphorsulfonates, and methanesulfonates; inorganic acid salts include, but are not limited to, nitrates, sulfates, hydrohalates, and phosphates.

[0013] The present invention also provides a “polymorph” that simultaneously contains a compound of formula (I) and its salts and solvates, that is, a compound of formula (I) or its salts or solvates can also exhibit polymorphism, that is, it can produce different crystal forms and have different physical properties, such as density, stability and solubility.

[0014] This invention demonstrates that the aminothiazole compounds with the above-described structural formula possess excellent AR antagonistic activity. They can inhibit AR activity by blocking AR dimerization, thereby effectively blocking AR-mediated downstream signaling pathways and significantly inhibiting tumor cell proliferation. Furthermore, these compounds exhibit low toxicity and good safety profiles in both in vitro and in vivo experiments, demonstrating high application potential.

[0015] The present invention also provides a pharmaceutical composition for treating tumors associated with aberrant or mutated AR expression, comprising at least one active component selected from compounds with the structural formula shown in Formula (I), pharmaceutically acceptable salts thereof, stereoisomers thereof, prodrug molecules thereof, and pharmaceutically acceptable carriers.

[0016] The pharmaceutical composition also includes a pharmaceutically acceptable carrier. These carriers are a variety of pharmaceutical excipients widely used in the pharmaceutical field, including but not limited to: common diluents and excipients (such as water), fillers (such as starch), binders (such as cellulose derivatives and gelatin), humectants (such as glycerin), disintegrants (such as agar and calcium carbonate), adsorbents (such as kaolin and soap clay), surfactants (such as hexadecyl alcohol), absorption enhancers (such as quaternary ammonium compounds), and lubricants (such as talc). The pharmaceutical formulations of this invention can be prepared according to conventional methods in the pharmaceutical field, including but not limited to capsules, powders, tablets, granules, pills, injections, syrups, oral liquids, inhalers, ointments, suppositories, or patches.

[0017] The compounds provided by this invention can be used in combination with existing antitumor drugs, including but not limited to microtubule degradation inhibitors (such as Taxol), mitotic inhibitors (such as vincristine, vindesine, etc.), antimetabolites (such as 5-fluorouracil, methotrexate, etc.), alkylating agents (cisplatin, carboplatin, etc.), topoisomerase inhibitors (such as etoposide and camptothecin), proteasome inhibitors (such as bortezomib, etc.).

[0018] The beneficial effects of this invention are as follows: This invention provides a novel class of aminothiazole compounds with AR antagonistic activity. One of the key target sites of these compounds is the binding pocket between the AR ligand-binding domain dimer interface, which can block AR dimerization and thus exert its antagonistic effect on androgen receptors. The compounds and their derivatives provided by this invention exhibit significant antagonistic activity against AR, demonstrating good biological activity in both in vitro and in vivo biological evaluations, and are effective against enzalutamide-resistant AR receptors. F876L / T877A Two-point mutation model and AR compared to calutamide resistance W741C The mutant cell model was effective. Furthermore, the compounds provided in this invention, when administered transdermally, also exhibited hair growth-promoting effects in a mouse alopecia model. Therefore, they can be used as AR antagonists in the treatment of androgen receptor-related diseases, including but not limited to the treatment of prostate cancer, metastatic prostate cancer, castration-resistant prostate cancer, breast cancer, ovarian cancer, androgenic alopecia, and acne. Detailed Implementation

[0019] The present invention will be further described below with reference to specific embodiments. These specific embodiments are for illustrative purposes only and should not be construed as limiting the scope of the invention. Furthermore, it should be understood that after reading the teachings of this invention, those skilled in the art can make various alterations or modifications to the invention, and these equivalent forms also fall within the scope defined by the appended claims.

[0020] Unless otherwise specified, the experimental methods used in the following examples are conventional methods.

[0021] Unless otherwise specified, all materials and reagents used in the following examples are commercially available.

[0022] Example 1: Preparation of target molecules A1-A43

[0023] N-(4-((3,3-difluoropyrrolidone-1-yl)methyl)thiazolyl-2-yl)-4-methyl-2-(3-(trifluoromethyl)phenyl)thiazolyl-5-carboxamide (Compound A1)

[0024] (a) Synthesis of 4-methyl-2-(3-(trifluoromethyl)phenyl)thiazol-5-carboxylic acid (compound I-2-1) Under N2 protection, m-trifluoromethylphenylboronic acid (I-1-1, 3.82 g, 20.13 mmol), 2-bromo-4-methylthiazol-5-carboxylic acid (4.47 g, 20.13 mmol), and cesium carbonate (13.12 g, 40.26 mmol) were dissolved in a mixed solvent of 1,4-dioxane (35 mL) and water (10 mL). 1,1'-bis(diphenylphosphine)ferrocene palladium dichloride (731.7 mg, 1 mmol) was added, and the mixture was stirred overnight at 110 °C. After cooling to room temperature, the pH was adjusted to 3-4 by adding 2 mol / L dilute hydrochloric acid. The mixture was extracted with ethyl acetate, and the organic layer was washed with saturated saline solution. The organic layers were combined and dried over anhydrous Na2SO4. The mixture was concentrated under reduced pressure, and the residue was subjected to column chromatography to give a pale yellow solid I-2-1, with a yield of 68%. 1 H NMR (500MHz, DMSO-d6) δ8.24–8.17(m,2H),7.86(d,J=7.9Hz,1H),7.74(t,J=8.1Hz,1H),2.68(s,3H).ESI-MS:m / z=288.0[M+H] + .

[0025] (b) Synthesis of N-(4-(chloromethyl)thiazol-2-yl)-4-methyl-2-(3-(trifluoromethyl)phenyl)thiazol-5-carboxamide (compound I-3-1) Compound I-2-1 (2.87 g, 10 mmol), 2-amino-4-chloromethylthiazolium hydrochloride (1.85 g, 10 mmol), EDCI (3.83 g, 20 mmol), and 4-PPY (592.8 mg, 4 mmol) were dissolved in dichloromethane (50 mL), refluxed and stirred overnight, the solvent was removed under reduced pressure, an appropriate amount of water was added, and the mixture was extracted with ethyl acetate. The organic layer was washed with saturated saline solution, the organic layers were combined, dried over anhydrous Na2SO4, concentrated under reduced pressure, and the residue was subjected to column chromatography to give a white solid I-3-1 in 68% yield. 1 H NMR(500MHz,DMSO-d6)δ12.88(s,1H),8.27–8.24(m,2H),7.94(d,J=7.9Hz,1H),7.8 0(t,J=8.1Hz,1H),7.34(s,1H),4.77(s,2H),2.74(s,3H).ESI-MS:m / z=418.0[M+H] + .

[0026] (c) Synthesis of N-(4-((3,3-difluoropyrrolidone-1-yl)methyl)thiazolyl-2-yl)-4-methyl-2-(3-(trifluoromethyl)phenyl)thiazolyl-5-carboxamide (compound A1) Compound I-3-1 (230 mg, 0.55 mmol) and 3,3-difluoropyrrolidine (89 mg, 0.83 mmol) were dissolved in acetonitrile (10 mL), and DIPEA (192 μL, 1.01 mmol) was added dropwise to the reaction system. The mixture was stirred overnight at room temperature. A suitable amount of water was added, and the mixture was extracted with ethyl acetate. The organic layer was washed with saturated saline solution, and the organic layers were combined. The mixture was then dried over anhydrous Na2SO4 and concentrated under reduced pressure. The residue was subjected to column chromatography to give a yellow solid Al in 88% yield. 1 H NMR(500MHz,DMSO-d6)δ12.96(s,1H),8.28–8.20(m,2H),7.95–7.88(m,1H),7.79(t,J=8.0Hz,1H),7.00(s,1H),3 .66(s,2H),2.95(t,J=13.4Hz,2H),2.76(d,J=2.4Hz,5H),2.26(tt,J=15.0,6.9Hz,2H).ESI-MS:m / z=489.1[M+H] + .

[0027] N-(4-((1,1-dioxothiomorpholino)methyl)thiazo-2-yl)-4-methyl-2-(3-(trifluoromethyl)phenyl)thiazo-5-carboxamide (compound A2) The preparation of target compound A2 was based on the synthesis of compound A1, except that 3,3-difluoropyrrolidine was replaced with 1,1-dioxothiomorpholine, yielding a pale yellow solid in 78% yield. 1 H NMR(500MHz,Chloroform-d)δ8.28(s,1H),8.12(d,J=7.8Hz,1H),7.76(d,J=7.8Hz,1H),7.63(t, J=7.8Hz,1H),6.90(s,1H),3.76(s,2H),3.24–3.05(m,8H),2.89(s,3H).ESI-MS:m / z=517.1[M+H] + .

[0028] N-(4-(morpholinomethyl)thiazo-2-yl)-4-methyl-2-(3-(trifluoromethyl)phenyl)thiazo-5-carboxamide (compound A3) The preparation of target compound A3 was the same as that of compound A1, except that 3,3-difluoropyrrolidine was replaced with morpholine, yielding a yellow solid in 68% yield; 1 H NMR(500MHz,DMSO-d6)δ12.94(s,1H),8.28–8.20(m,2H),7.95–7.88(m,1H),7.79(t,J=8.1Hz,1H) ,6.96(s,1H),3.59(d,J=9.2Hz,4H),2.76(s,3H),2.42(t,J=4.6Hz,4H).ESI-MS:m / z=469.1[M+H] + .

[0029] N-(4-((4,4-difluoropiperidin-1-yl)methyl)thiazolyl-2-yl)-4-methyl-2-(3-(trifluoromethyl)phenyl)thiazolyl-5-carboxamide (compound A4) The preparation of target compound A4 was based on the synthesis of compound A1, except that 3,3-difluoropyrrolidine was replaced with 4,4-difluoropiperidine, yielding a pale yellow solid in 64% yield. 1 H NMR (500MHz, DMSO-d6) δ12.94(s,1H),8.24(dd,J=4.8,2.8Hz,2H),7.92(d,J=7.8Hz,1H),7.79(t,J=8.0Hz,1 H),6.98(s,1H),3.58(s,2H),2.76(s,3H),2.56(t,J=5.6Hz,4H),2.01–1.92(m,4H).ESI-MS:m / z=503.1[M+H] + .

[0030] N-(4-((4-acetylpiperazin-1-yl)methyl)thiazo-2-yl)-4-methyl-2-(3-(trifluoromethyl)phenyl)thiazo-5-carboxamide (compound A5) The preparation of target compound A5 was the same as that of compound A1, except that 3,3-difluoropyrrolidine was replaced with 1-(acetyl)piperazine, yielding a yellow solid in 74% yield; 1 H NMR (500MHz, DMSO-d6) δ12.67(s,1H),8.29–8.20(m,2H),7.92(d,J=7.9Hz,1H),7.80(t,J=8.1Hz,1H),6.97(s,1H),3.5 3(s,2H),3.44(dt,J=10.3,4.9Hz,4H),2.77(s,3H),2.41(dt,J=27.4,5.2Hz,4H),1.98(s,3H).ESI-MS:m / z=510.1[M+H] + .

[0031] N-(4-(thiomorpholinemethyl)thiazo-2-yl)-4-methyl-2-(3-(trifluoromethyl)phenyl)thiazo-5-carboxamide (compound A6) The preparation of target compound A6 was based on the synthesis of compound A1, except that 3,3-difluoropyrrolidine was replaced with thiomorpholine to give a yellow solid in 60% yield. 1 H NMR(500MHz, DMSO-d6)δ12.92(s,1H),8.24(dd,J=4.9,2.8Hz,2H),7.95–7.89(m,1H),7.79(t,J=8.2Hz,1H),6.95( s,1H),3.52(s,2H),2.76(s,3H),2.69(dd,J=6.8,3.0Hz,4H),2.62(dd,J=5.9,3.7Hz,4H).ESI-MS:m / z=485.1[M+H] + .

[0032] N-(4-((4-(methanesulfonyl)piperazin-1-yl)methyl)thiazo-2-yl)-4-methyl-2-(3-(trifluoromethyl)phenyl)thiazo-5-carboxamide (compound A7) The preparation of target compound A7 was based on the synthesis of compound A1, except that 3,3-difluoropyrrolidine was replaced with 1-(methanesulfonyl)piperazine, yielding a pale yellow solid in 76% yield. 1H NMR (500MHz, DMSO-d6) δ12.95(s,1H),8.24(d,J=6.0Hz,2H),7.92(d,J=7.9Hz,1H),7.79(t,J=8.0Hz,1H),6.98(s, 1H),3.56(s,2H),3.12(t,J=4.8Hz,4H),2.88(s,3H),2.76(s,3H),2.54(t,J=4.9Hz,4H).ESI-MS:m / z=546.1[M+H] + .

[0033] N-(4-((4-(cyclopropylcarbonyl)piperazin-1-yl)methyl)thiazo-2-yl)-4-methyl-2-(3-(trifluoromethyl)phenyl)thiazo-5-carboxamide (compound A8) The preparation of target compound A8 was based on the synthesis of compound A1, except that 3,3-difluoropyrrolidine was replaced with 1-(cyclopropylmethylketone)piperazine, yielding a deep yellow solid in 70% yield; 1 H NMR (500MHz, DMSO-d6) δ12.95(s,1H),8.26–8.22(m,2H),7.92(d,J=7.9Hz,1H),7.79(t,J=8.1Hz,1H),6.98(s,1H),3.68( s,2H),2.76(s,3H),2.43(d,J=37.1Hz,4H),1.95(ddd,J=9.5,6.6,3.9Hz,1H),0.73–0.65(m,4H).ESI-MS:m / z=536.1[M+H] + .

[0034] 4-((2-(4-methyl-2-(3-(trifluoromethyl)phenyl)thiazol-5-carboxamido)thiazol-4-yl)methyl)piperazine-1-carboxylic acid ethyl ester (compound A9) The preparation of target compound A9 was based on the synthesis of compound A1, except that 3,3-difluoropyrrolidine was replaced with ethyl piperazine-1-carboxylate to give a deep yellow solid in 69% yield. 1 H NMR (500MHz, DMSO-d6) δ12.95(s,1H),8.30–8.18(m,2H),7.92(d,J=7.9Hz,1H),7.79(t,J=8.0Hz,1H),6.96(s,1H),4.03( q,J=7.1Hz,2H),3.37(t,J=4.9Hz,4H),2.76(s,3H),2.40(t,J=5.1Hz,4H),1.17(t,J=7.1Hz,3H).ESI-MS:m / z=540.1[M+H] + .

[0035] Preparation of target molecules A10-A24 Referring to the synthesis of compound A1, using m-trifluoromethylphenylboronic acid (I-1-1) and 2-bromo-4-methylthiazol-5-carboxylic acid as starting materials, and employing pyrrolidine, 3-hydroxypyrrolidine, piperidine, 4-hydroxypiperidine, 4-(hydroxymethyl)piperidine, 4-(2-hydroxyethyl)piperazine, cis-2,6-dimethylmorpholine, 4-methylpiperazine, 4-ethylpiperazine, 2-hydroxy-1-(piperazin-1-yl)ethyl ketone, N Compounds A10 to A24 were prepared by substituting 3,3-difluoropyrrolidine with methylpiperazin-1-carboxamide, 2-methoxy-1-(piperazin-1-yl)ethyl ketone, 2,2,2-trifluoro-1-(piperazin-1-yl)ethyl ketone, 3-oxo-3-(piperazin-1-yl)propionitrile, and R-3-(piperazin-1-yl)propane-1,2-diol. The chemical structures and ESI-MS results of the above compounds are shown in Table 1.

[0036] Table 1. Chemical structures and ESI-MS results of target molecules A10–A24

[0037] N-(4-((4-carbamoylpiperazin-1-yl)methyl)thiazo-2-yl)-4-methyl-2-(3-(trifluoromethyl)phenyl)thiazo-5-carboxamide (compound A25) The preparation of target compound A25 was based on the synthesis of compound A1, except that 3,3-difluoropyrrolidine was replaced with piperazine-1-carboxamide to give a yellow solid in 65% yield. 1 H NMR (500MHz, DMSO-d6) δ12.95(s,1H),8.24(d,J=6.6Hz,2H),7.91(d,J=7.8Hz,1H),7.79(t,J=8.0Hz,1H),6.9 7(s,1H),5.95(s,2H),3.51(s,2H),3.30(s,4H),2.76(s,3H),2.38(t,J=5.1Hz,4H).ESI-MS:m / z=511.1[M+H] + .

[0038] N-(4-((1-oxothiomorpholino)methyl)thiazo-2-yl)-4-methyl-2-(3-(trifluoromethyl)phenyl)thiazo-5-carboxamide (compound A26) The preparation of target compound A26 was based on the synthesis of compound A1, except that 3,3-difluoropyrrolidine was replaced with 1-oxothiomorpholine to give a yellow solid in 61% yield. 1H NMR (500MHz, DMSO-d6) δ12.95(s,1H),8.24(d,J=6.3Hz,2H),7.92(d,J=7.8Hz,1H),7.79(t,J=8.0 Hz,1H),7.00(s,1H),3.59(s,2H),2.98–2.83(m,4H),2.81–2.66(m,7H).ESI-MS:m / z=501.1[M+H] + .

[0039] 4-((2-(4-methyl-2-(3-(trifluoromethyl)phenyl)thiazolyl-5-carboxamido)thiazolyl-4-yl)methyl)piperazine-1-carboxylic acid tert-butyl ester (compound A27) The preparation of target compound A27 was based on the synthesis of compound A1, except that 3,3-difluoropyrrolidine was replaced with tert-butyl piperazine-1-carboxylate to give a yellow solid in 55% yield. 1 H NMR(500MHz,DMSO-d6)δ12.95(s,1H),8.27–8.21(m,2H),7.92(d,J=7.8Hz,1H),7.79(t,J=8.1Hz,1H) ,6.96(s,1H),3.51(s,2H),2.76(s,3H),2.38(t,J=5.0Hz,4H),1.39(s,9H).ESI-MS:m / z=568.2[M+H] + .

[0040] N-(4-(piperazin-1-ylmethyl)thiazo-2-yl)4-methyl-2-(3-(trifluoromethyl)phenyl)thiazo-5-carboxamide (compound A28) The preparation of target compound A28 was based on the synthesis of compound A1, except that 3,3-difluoropyrrolidine was replaced with piperazine, yielding a yellow solid in 63% yield. 1 H NMR (500MHz, DMSO-d6) δ8.24–8.17(m,2H),7.86(d,J=7.8Hz,1H),7.76(t,J=7.8Hz,1H ),6.68(s,1H),3.07–2.95(m,4H),2.76(s,3H),2.56(s,4H).ESI-MS:m / z=468.1[M+H] + .

[0041] N-(4-((1,4-dioxa-8-azaspiro[4.5]decane-8-yl)methyl)thiazo-2-yl)-4-methyl-2-(3-(trifluoromethyl)phenyl)thiazo-5-carboxamide (compound A29) The preparation of target compound A29 was based on the synthesis of compound A1, except that 3,3-difluoropyrrolidine was replaced with 1,4-dioxa-8-azaspiro[4.5]decane hydrochloride, yielding a pale yellow solid in 70% yield; 1 H NMR(500MHz,Chloroform-d)δ8.23(s,1H),8.06(d,J=7.8Hz,1H),7.71(d,J=7.8Hz,1H),7.57(t,J=7.9Hz,1H),6.7 5(s,1H),3.91(s,4H),3.48(s,2H),2.82(s,3H),2.58–2.41(m,4H),1.71(t,J=5.7Hz,4H).ESI-MS:m / z=525.1[M+H] + .

[0042] N-(4-((2-acetyl-2,7-diazaspiro[3.5]nonane-7-yl)methyl)thiazolyl-2-yl)-4-methyl-2-(3-(trifluoromethyl)phenyl)thiazolyl-5-carboxamide (compound A30) The preparation of target compound A30 was based on the synthesis of compound A1, except that 3,3-difluoropyrrolidine was replaced with 2-acetyl-2,7-diazaspiro[3.5]nonane, yielding a pale yellow solid in 56% yield; 1 H NMR(500MHz,DMSO-d6)δ8.23(d,J=6.7Hz,2H),7.91(d,J=7.8Hz,1H),7.79(t,J=8.0Hz,1H),6.91 (s,1H),3.77(s,2H),2.76(s,3H),2.40(s,4H),1.72(d,J=18.2Hz,7H).ESI-MS:m / z=550.1[M+H] + .

[0043] N-(4-((4-acetylpiperazin-1-yl)methyl)thiazo-2-yl)-2-(3-cyanophenyl)-4-methylthiazo-5-carboxamide (compound A31) The preparation of target compound A31 was based on the synthesis of compound A1, with 3,3-difluoropyrrolidine replaced by 1-(acetyl)piperazine and m-trifluoromethylphenylboronic acid replaced by m-cyanophenylboronic acid, yielding a pale yellow solid in 72% yield; 1H NMR (500MHz, DMSO-d6) δ12.97(s,1H),8.37(t,J=1.8Hz,1H),8.28(dt,J=8.0,1.4Hz,1H),8.00(dt,J=7.8,1.4Hz,1H),7.75(t,J=7.9Hz,1H),6.97 (s,1H),3.53(s,2H),3.43(q,J=6.1,5.2Hz,4H),2.75(s,3H),2.43(t,J= 5.0Hz,2H),2.38(t,J=5.2Hz,2H),1.98(s,3H).ESI-MS:m / z=467.1[M+H] + .

[0044] N-(4-((4-acetylpiperazin-1-yl)methyl)thiazo-2-yl)-2-(3-methoxyphenyl)-4-methylthiazo-5-carboxamide (compound A32) The preparation of target compound A32 was based on the synthesis of compound A1, with 3,3-difluoropyrrolidine replaced by 1-(acetyl)piperazine and m-trifluoromethylphenylboronic acid replaced by m-methoxyphenylboronic acid, yielding a pale yellow solid in 72% yield; 1 H NMR(500MHz,DMSO-d6)δ12.86(s,1H),7.52(dt,J=7.6,1.3Hz,1H),7.49–7.42(m,2H),7.12(ddd,J=8.2,2.7,1.0Hz,1H),6.96(s,1H),3 .85(s,3H),3.52(s,2H),3.43(q,J=6.4,5.1Hz,4H),2.73(s,3H),2.43(t,J=5.0Hz,2H),2.37(t,J=5.1Hz,2H).ESI-MS:m / z=472.1[M+H] + .

[0045] N-(4-((4-acetylpiperazin-1-yl)methyl)thiazo-2-yl)-4-methyl-2-(3-nitrophenyl)thiazo-5-carboxamide (compound A33) The preparation of target compound A33 was based on the synthesis of compound A1, with 3,3-difluoropyrrolidine replaced by 1-(acetyl)piperazine and m-trifluoromethylphenylboronic acid replaced by m-nitrophenylboronic acid, yielding a yellow solid in 65% yield; 1H NMR (500MHz, DMSO-d6) δ12.98(s,1H),8.70(d,J=2.1Hz,1H),8.37(d,J=8.2Hz,2H),7.83(t,J=8.0Hz,1H),6.97(s,1H),3.53(s ,2H),3.43(q,J=6.8,5.6Hz,4H),2.78(s,3H),2.46–2.41(m,2H),2.38(t,J=5.0Hz,2H),1.98(s,3H).ESI-MS:m / z=487.1[M+H] + .

[0046] N-(4-((4-acetylpiperazin-1-yl)methyl)thiazolyl-2-yl)-4-methyl-2-(3-(trifluoromethoxy)phenyl)thiazolyl-5-carboxamide (compound A34) The preparation of target compound A34 was based on the synthesis of compound A1, with 3,3-difluoropyrrolidine replaced by 1-(acetyl)piperazine and m-trifluoromethylphenylboronic acid replaced by m-trifluoromethoxyphenylboronic acid, yielding a yellow solid in 70% yield; 1 H NMR(500MHz,DMSO-d6)δ12.95(s,1H),7.98(dt,J=7.7,1.3Hz,1H),7.90(dt ,J=2.6,1.2Hz,1H),7.69(t,J=8.0Hz,1H),7.56(ddt,J=8.2,2.3,1.1Hz,1H) ,6.97(s,1H),3.52(s,2H),3.43(p,J=5.3,4.7Hz,4H),2.75(s,3H),2.43(t ,J=5.0Hz,2H),2.38(t,J=5.1Hz,2H),1.98(s,3H).ESI-MS:m / z=526.1[M+H] + .

[0047] N-(4-((4-acetylpiperazin-1-yl)methyl)thiazo-2-yl)-2-(2-fluoro-5-(trifluoromethyl)phenyl)-4-methylthiazo-5-carboxamide (compound A35) The preparation of target compound A35 was based on the synthesis of compound A1, with 3,3-difluoropyrrolidine replaced by 1-(acetyl)piperazine and m-trifluoromethylphenylboronic acid replaced by 2-fluoro-5-(trifluoromethyl)phenylboronic acid, yielding a yellow solid in 60% yield; 1HNMR(500MHz,DMSO-d6)δ13.00(s,1H),8.54(dd,J=6.5,2.4Hz,1H),8.06–7.96(m,1H),7.74(dd,J=10.8,8.7Hz,1H),6. 96(s,1H),3.43(s,4H),2.79(s,3H),2.44(d,J=5.7Hz,2H),2.37(t,J=5.0Hz,2H),1.98(s,3H).ESI-MS:m / z=528.1[M+H] + .

[0048] N-(4-((4-acetylpiperazin-1-yl)methyl)thiazo-2-yl)-2-(2-chloro-5-(trifluoromethyl)phenyl)-4-methylthiazo-5-carboxamide (compound A36) The preparation of target compound A36 was based on the synthesis of compound A1, with 3,3-difluoropyrrolidine replaced by 1-(acetyl)piperazine and m-trifluoromethylphenylboronic acid replaced by 2-chloro-5-(trifluoromethyl)phenylboronic acid, yielding a yellow solid in 65% yield; 1 HNMR(500MHz,DMSO-d6)δ13.03(s,1H),8.61(d,J=2.1Hz,1H),7.97–7.91(m,2H),6.96(s,1H),3. 42(s,4H),2.79(s,3H),2.43(d,J=5.8Hz,2H),2.37(s,2H),1.98(s,3H).ESI-MS:m / z=544.1[M+H] + .

[0049] N-(4-((4-acetylpiperazin-1-yl)methyl)thiazo-2-yl)-2-(2-fluoro-5-(trifluoromethoxy)phenyl)-4-methylthiazo-5-carboxamide (compound A37) The preparation of target compound A37 was based on the synthesis of compound A1, with 3,3-difluoropyrrolidine replaced by 1-(acetyl)piperazine and m-trifluoromethylphenylboronic acid replaced by 2-fluoro-5-(trifluoromethoxy)phenylboronic acid, yielding a yellow-brown solid in 67% yield; 1 H NMR(500MHz,DMSO-d6)δ13.01(s,1H),8.19–8.09(m,1H),7.69–7.61(m,2H),6.96(s,1H),3.52(s,2H),3.43(q,J= 6.1,5.0Hz,4H),2.78(s,3H),2.43(t,J=4.9Hz,2H),2.38(t,J=5.2Hz,2H),1.98(s,3H).ESI-MS:m / z=544.1[M+H]+ .

[0050] N-(4-((4-acetylpiperazin-1-yl)methyl)thiazo-2-yl)-2-(2-chloro-5-(trifluoromethoxy)phenyl)-4-methylthiazo-5-carboxamide (compound A38) The preparation of target compound A38 was based on the synthesis of compound A1, with 3,3-difluoropyrrolidine replaced by 1-(acetyl)piperazine and m-trifluoromethylphenylboronic acid replaced by 2-chloro-5-(trifluoromethoxy)phenylboronic acid, yielding a yellow-brown solid in 73% yield; 1 H NMR(500MHz,DMSO-d6)δ8.30–8.19(m,1H),7.83(d,J=8.8Hz,1H),7.62–7.54(m,1H),6.95(s,1H),3.52(s,2H),3.4 4(p,J=4.7Hz,4H),2.77(s,3H),2.44(t,J=4.9Hz,2H),2.38(t,J=5.0Hz,2H),1.99(s,3H).ESI-MS:m / z=560.1[M+H] + .

[0051] N-(4-((4-acetylpiperazin-1-yl)methyl)thiazo-2-yl)-2-(3-fluoro-5-(trifluoromethyl)phenyl)-4-methylthiazo-5-carboxamide (compound A39) The preparation of target compound A39 was based on the synthesis of compound A1, with 3,3-difluoropyrrolidine replaced by 1-(acetyl)piperazine and m-trifluoromethylphenylboronic acid replaced by 3-fluoro-5-(trifluoromethyl)phenylboronic acid, yielding a yellow solid in 70% yield; ESI-MS: m / z = 528.1 [M+H] + .

[0052] N-(4-((4-acetylpiperazin-1-yl)methyl)thiazo-2-yl)-2-(3-chloro-5-(trifluoromethyl)phenyl)-4-methylthiazo-5-carboxamide (Compound A40) The preparation of target compound A40 was based on the synthesis of compound A1, with 3,3-difluoropyrrolidine replaced by 1-(acetyl)piperazine and m-trifluoromethylphenylboronic acid replaced by 3-chloro-5-(trifluoromethyl)phenylboronic acid, yielding a yellow solid in 62% yield; ESI-MS: m / z = 544.1 [M+H] + .

[0053] N-(4-((1,1-dioxothiomorpholine)methyl)thiazo-2-yl)-2-(2-fluoro-5-(trifluoromethyl)phenyl)-4-methylthiazo-5-carboxamide (compound A41) The preparation of target compound A41 was based on the synthesis of compound A1, with 3,3-difluoropyrrolidine replaced by 1,1-dioxothiomorpholine and m-trifluoromethylphenylboronic acid replaced by 2-fluoro-5-(trifluoromethyl)phenylboronic acid, yielding a pale yellow solid in 57% yield; ESI-MS: m / z = 535.0 [M+H] + .

[0054] N-(4-(morpholinomethyl)thiazolyl-2-yl)-2-(2-fluoro-5-(trifluoromethyl)phenyl)-4-methylthiazolyl-5-carboxamide (compound A42) The preparation of target compound A42 was based on the synthesis of compound A1, with 3,3-difluoropyrrolidine replaced by morpholine and m-trifluoromethylphenylboronic acid replaced by 2-fluoro-5-(trifluoromethyl)phenylboronic acid, yielding a pale yellow solid in 60% yield; ESI-MS: m / z = 487.1 [M+H] + .

[0055] N-(4-((1,1-dioxothiomorpholine)methyl)thiazolyl-2-yl)-2-(2-fluoro-5-(trifluoromethoxy)phenyl)-4-methylthiazolyl-5-carboxamide (compound A43) The preparation of target compound A43 was based on the synthesis of compound A1, with 3,3-difluoropyrrolidine replaced by 1,1-dioxothiomorpholine and 2-fluoro-5-(trifluoromethyl)phenylboronic acid replaced by 2-fluoro-5-(trifluoromethoxy)phenylboronic acid, yielding a yellow solid in 72% yield; ESI-MS: m / z = 551.0 M + H] + .

[0056] Example 2: Preparation of target molecules A44-A60 N-(4-((4-acetylpiperazin-1-yl)methyl)-5-fluorothiazo-2-yl)-4-methyl-2-(3-(trifluoromethyl)phenyl)thiazo-5-carboxamide (compound A44) Compound A5 (127.4 mg, 0.25 mmol), NFSI (237 mg, 0.75 mmol), and K3PO4 (106 mg, 0.5 mmol) were dissolved in DMF (7 mL), heated to reflux, and stirred for 6 h. A suitable amount of water was added, and the mixture was extracted with ethyl acetate. The organic layer was washed with saturated saline solution, and the two organic layers were combined and dried over anhydrous Na2SO4. The mixture was concentrated under reduced pressure, and the residue was subjected to column chromatography to give a white solid A44, in 25% yield; ESI-MS: m / z = 528.1 [M+H] + .

[0057] N-(4-((4-acetylpiperazin-1-yl)methyl)-5-chlorothiazol-2-yl)-4-methyl-2-(3-(trifluoromethyl)phenyl)thiazol-5-carboxamide (compound A45) Compound A5 (127.4 mg, 0.25 mmol) and NCS (37 mg, 0.275 mmol) were dissolved in DMF (5 mL), heated to reflux, and stirred overnight. A suitable amount of water was added, and the mixture was extracted with ethyl acetate. The organic layer was washed with saturated saline solution, and the two organic layers were combined and dried over anhydrous Na2SO4. The mixture was concentrated under reduced pressure, and the residue was subjected to column chromatography to give a white solid A45 in 62% yield; ESI-MS: m / z = 544.1 [M+H]. + .

[0058] N-(4-((4-acetylpiperazin-1-yl)methyl)-5-bromothiazo-2-yl)-4-methyl-2-(3-(trifluoromethyl)phenyl)thiazo-5-carboxamide (compound A46) The preparation of target compound A46 was performed following the synthesis of compound A45, with NCS replaced by NBS, yielding a pale yellow solid in 55% yield; ESI-MS: m / z = 588.0 [M+H] + .

[0059] N-(4-((4-acetylpiperazin-1-yl)methyl)-5-methylthiazo-2-yl)-4-methyl-2-(3-(trifluoromethyl)phenyl)thiazo-5-carboxamide (compound A47) Under N2 protection, the compound was prepared by dissolving A46 (106 mg, 0.18 mmol), trimethylboroxine (113 mg, 0.9 mmol), Sphos (148 mg, 0.36 mmol), and K3PO4 (115 mg, 0.54 mmol) in toluene (5 mL), followed by the addition of Pd2(dba)3 (83 mg, 0.09 mmol). The mixture was heated to 110 °C and stirred for 6 h. A suitable amount of water was added, and the mixture was extracted with ethyl acetate. The organic layer was washed with a saturated saline solution, and the two organic layers were combined and dried over anhydrous Na2SO4. The mixture was concentrated under reduced pressure, and the residue was subjected to column chromatography to give a white solid A47 in 45% yield; ESI-MS: m / z = 524.1 [M+H]. + .

[0060] N-(4-((4-acetylpiperazin-1-yl)methyl)-5-(trifluoromethyl)thiazo-2-yl)-4-methyl-2-(3-(trifluoromethyl)phenyl)thiazo-5-carboxamide (compound A48) Under N2 protection, compound A46 (150 mg, 0.25 mmol), methyl fluorosulfonyl difluoroacetate (144 mg, 0.75 mmol), and CuI (96 mg, 0.5 mmol) were dissolved in NMP (15 mL). The mixture was heated to 100 °C and stirred for 6 h. The solvent was removed under reduced pressure, and an appropriate amount of water was added. The mixture was extracted with ethyl acetate, and the organic layer was washed with saturated saline solution. The organic layers were combined, dried over anhydrous Na2SO4, and concentrated under reduced pressure. The residue was subjected to column chromatography to give a pale yellow solid A48, with a yield of 33%; ESI-MS: m / z = 578.1 [M+H] + .

[0061] N-(4-((4-acetylpiperazin-1-yl)methyl)-5-cyanothiazol-2-yl)-4-methyl-2-(3-(trifluoromethyl)phenyl)thiazole-5-carboxamide (compound A49) Under controlled conditions, compound A46 (118 mg, 0.2 mmol), Zn(CN)2 (59 mg, 0.5 mmol), and dppf (200 mg, 0.36 mmol) were dissolved in dichloromethane (7 mL), and Pd2(dba)3 (92 mg, 0.1 mmol) was added. The mixture was heated to 85 °C and stirred overnight. The solvent was removed under reduced pressure, and an appropriate amount of water was added. The mixture was extracted with ethyl acetate, and the organic layer was washed with saturated saline solution. The organic layers were combined, dried over anhydrous Na2SO4, and concentrated under reduced pressure. The residue was subjected to column chromatography to give a white solid A49, with a yield of 30%. ESI-MS: m / z = 535.1 [M+H] + .

[0062] N-(4-((1,1-dioxothiomorpholine)methyl)-5-fluorothiazo-2-yl)-4-methyl-2-(3-(trifluoromethyl)phenyl)thiazo-5-carboxamide (Compound A50) The preparation of target compound A50 was based on the synthesis of compound A44, except that acetylpiperazine was replaced with 1,1-dioxothiomorpholine, yielding a white solid in 20% yield; ESI-MS: m / z = 535.0 [M+H] + .

[0063] N-(4-((1,1-dioxothiomorpholine)methyl)-5-methylthiazo-2-yl)-4-methyl-2-(3-(trifluoromethyl)phenyl)thiazo-5-carboxamide (compound A51) The preparation of target compound A51 was based on the synthesis of compound A47, except that acetylpiperazine was replaced with 1,1-dioxothiomorpholine, yielding a white solid in 35% yield; ESI-MS: m / z = 531.1 [M+H]+ .

[0064] N-(5-chloro-4-((1,1-dioxothiomorpholino)methyl)thiazo-2-yl)-4-methyl-2-(3-(trifluoromethyl)phenyl)thiazo-5-carboxamide (compound A52) The preparation of target compound A52 was based on the synthesis of compound A45, except that acetylpiperazine was replaced with 1,1-dioxothiomorpholine, yielding a pale yellow solid in 60% yield; ESI-MS: m / z = 551.0 [M+H] + .

[0065] N-(5-chloro-4-((1,1-dioxothiomorpholino)methyl)thiazo-2-yl)-2-(2-fluoro-5-(trifluoromethyl)phenyl)-4-methylthiazo-5-carboxamide (compound A53) The preparation of target compound A53 was based on the synthesis of compound A45, with acetylpiperazine replaced by 1,1-dioxothiomorpholine and m-trifluoromethylphenylboronic acid replaced by 2-fluoro-5-(trifluoromethyl)phenylboronic acid, yielding a yellow solid in 55% yield: ESI-MS: m / z = 569.0 [M+H] + .

[0066] N-(4-((1,1-dioxothiomorpholine)methyl)-5-methylthiazo-2-yl)-2-(2-fluoro-5-(trifluoromethyl)phenyl)-4-methylthiazo-5-carboxamide (compound A54) The preparation of target compound A54 was based on the synthesis of compound A47, with acetylpiperazine replaced by 1,1-dioxothiomorpholine and m-trifluoromethylphenylboronic acid replaced by 2-fluoro-5-(trifluoromethyl)phenylboronic acid, yielding a pale yellow solid in 30% yield; ESI-MS: m / z = 549.1 [M+H] + .

[0067] N-(5-chloro-4-((1,1-dioxothiomorpholino)methyl)thiazo-2-yl)-2-(2-fluoro-5-(trifluoromethoxy)phenyl)-4-methylthiazo-5-carboxamide (compound A55) The preparation of target compound A55 was based on the synthesis of compound A45, with acetylpiperazine replaced by 1,1-dioxothiomorpholine and m-trifluoromethylphenylboronic acid replaced by 2-fluoro-5-(trifluoromethoxy)phenylboronic acid, yielding a yellow solid in 58% yield; ESI-MS: m / z = 585.0 [M+H] + .

[0068] N-(4-((1,1-dioxothiomorpholine)methyl)-5-methylthiazo-2-yl)-2-(2-fluoro-5-(trifluoromethoxy)phenyl)-4-methylthiazo-5-carboxamide (compound A56) The preparation of target compound A56 was based on the synthesis of compound A47, with acetylpiperazine replaced by 1,1-dioxothiomorpholine and m-trifluoromethylphenylboronic acid replaced by 2-fluoro-5-(trifluoromethoxy)phenylboronic acid, yielding a white solid in 39% yield; ESI-MS: m / z = 565.1 [M+H] + .

[0069] N-(4-((4-acetylpiperazin-1-yl)methyl)-5-chlorothiazol-2-yl)-2-(2-fluoro-5-(trifluoromethyl)phenyl)-4-methylthiazol-5-carboxamide (compound A57) The preparation of target compound A57 was based on the synthesis of compound A45, except that m-trifluoromethylphenylboronic acid was replaced with 2-fluoro-5-(trifluoromethyl)phenylboronic acid, yielding a pale yellow solid in 56% yield; ESI-MS: m / z = 562.1 [M+H] + .

[0070] N-(4-((4-acetylpiperazin-1-yl)methyl)-5-chlorothiazol-2-yl)-2-(2-fluoro-5-(trifluoromethoxy)phenyl)-4-methylthiazol-5-carboxamide (compound A58) The preparation of target compound A58 was based on the synthesis of compound A45, with m-trifluoromethylphenylboronic acid replaced by 2-fluoro-5-(trifluoromethoxy)phenylboronic acid, yielding a pale yellow solid in 60% yield; ESI-MS: m / z = 578.1 [M+H] + .

[0071] N-(4-((4-acetylpiperazin-1-yl)methyl)-5-methylthiazo-2-yl)-2-(2-fluoro-5-(trifluoromethyl)phenyl)-4-methylthiazo-5-carboxamide (compound A59) The preparation of target compound A59 was based on the synthesis of compound A47, except that m-trifluoromethylphenylboronic acid was replaced with 2-fluoro-5-(trifluoromethyl)phenylboronic acid, yielding a white solid in 34% yield; ESI-MS: m / z = 542.1 [M+H] + .

[0072] N-(4-((4-acetylpiperazin-1-yl)methyl)-5-methylthiazo-2-yl)-2-(2-fluoro-5-(trifluoromethoxy)phenyl)-4-methylthiazo-5-carboxamide (compound A60) The preparation of target compound A60 was based on the synthesis of compound A47, with m-trifluoromethylphenylboronic acid replaced by 2-fluoro-5-(trifluoromethoxy)phenylboronic acid, yielding a pale yellow solid in 30% yield; ESI-MS: m / z = 558.1 [M+H] + .

[0073] Example 3: Preparation of target molecules B1-B6 Synthetic routes of target molecules B1-B3

[0074] N-(4-((4-acetylpiperazin-1-yl)methyl)thiazo-2-yl)-5-methyl-2-(3-(trifluoromethyl)phenyl)thiazo-4-carboxamide (compound B1)

[0075] (a) Synthesis of 1-(4-((2-aminothiazol-4-yl)methyl)piperazin-1-yl)acet-1-one (compound I-6-1) Compound I-5 (149 mg, 1 mmol) and acetylpiperazine (193 mg, 1.5 mmol) were dissolved in acetonitrile (13 mL). DIPEA (380 μL, 2 mmol) was added dropwise to the reaction mixture, which was stirred overnight at room temperature. A suitable amount of water was added, and the mixture was extracted with ethyl acetate. The organic layer was washed with a saturated saline solution, and the two organic layers were combined. The mixture was then dried over anhydrous Na₂SO₄ and concentrated under reduced pressure. The residue was subjected to column chromatography to give a yellow solid I-6-1, with a yield of 71%. ESI-MS: m / z = 241.3 [M+H] + .

[0076] (b) Synthesis of N-(4-((4-acetylpiperazin-1-yl)methyl)thiazo-2-yl)-2-bromo-5-methylthiazo-4-carboxamide (compound I-7-1) Compound I-6-1 (121 mg, 0.5 mmol), 2-bromo-5-methylthiazolyl-4-carboxylic acid (134 mg, 0.6 mmol), N-methylimidazolium (124 mg, 1.5 mmol), and N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate (211 mg, 0.75 mmol) were dissolved in acetonitrile (10 mL). The mixture was stirred at room temperature for 6 h. A suitable amount of water was added, and the mixture was extracted with ethyl acetate. The organic layer was washed with saturated saline solution, and the two organic layers were combined. The mixture was then dried over anhydrous Na₂SO₄ and concentrated under reduced pressure. The residue was subjected to column chromatography to give a white solid I-7-1 in 75% yield; ESI-MS: m / z = 445.4 [M+H]. + .

[0077] (c) Synthesis of N-(4-((4-acetylpiperazin-1-yl)methyl)thiazo-2-yl)-5-methyl-2-(3-(trifluoromethyl)phenyl)thiazo-4-carboxamide (compound B1) Under N2 protection, m-trifluoromethylphenylboronic acid (100 mg, 0.53 mmol), compound I-7-1 (156 mg, 0.35 mmol), and cesium carbonate (228 mg, 0.7 mmol) were dissolved in a mixed solvent of 1,4-dioxane (7 mL) and water (2 mL). 1,1'-bis(diphenylphosphine)ferrocene palladium dichloride (13 mg, 0.02 mmol) was added, and the mixture was stirred overnight at 110 °C. After cooling to room temperature, the pH was adjusted to 3-4 with 2 mol / L dilute hydrochloric acid. The mixture was extracted with ethyl acetate, and the organic layer was washed with saturated saline solution. The organic layers were combined, dried over anhydrous Na2SO4, and concentrated under reduced pressure. The residue was subjected to column chromatography to give a pale yellow solid B1, with a yield of 45%. ESI-MS: m / z = 510.1 [M+H] + .

[0078] N-(4-((1,1-dioxothiomorpholine)methyl)thiazo-2-yl)-5-methyl-2-(3-(trifluoromethyl)phenyl)thiazo-4-carboxamide (compound B2) The preparation of target compound B2 followed the synthesis of compound B1, except that acetylpiperazine was replaced with 1,1-dioxothiomorpholine, yielding a white solid in 58% yield; ESI-MS: m / z = 517.1 [M+H] + .

[0079] N-(4-(morpholinomethyl)thiazo-2-yl)-5-methyl-2-(3-(trifluoromethyl)phenyl)thiazo-4-carboxamide (compound B3) The preparation of target compound B3 followed the synthesis of compound B1, except that acetylpiperazine was replaced with morpholine, yielding a pale yellow solid in 52% yield: ESI-MS: m / z = 469.1 [M+H] + .

[0080] Preparation of target molecules B4-B6

[0081] N-(4-((4-acetylpiperazin-1-yl)methyl)thiazo-2-yl)-2-(3-(trifluoromethyl)phenyl)thiazo-5-carboxamide (compound B4) The preparation of target compound B4 was performed following the synthesis of compound B1, except that 2-bromo-5-methylthiazol-4-carboxylic acid was replaced with 2-bromothiazol-5-carboxylic acid, yielding a white solid in 43% yield; ESI-MS: m / z = 496.1 [M+H] + .

[0082] N-(4-(thiomorpholinemethyl)thiazo-2-yl)-2-(3-(trifluoromethyl)phenyl)thiazo-5-carboxamide (compound B5) The preparation of target compound B5 followed the synthesis of compound B1, except that 2-bromo-5-methylthiazol-4-carboxylic acid was replaced with 2-bromothiazol-5-carboxylic acid, and acetylpiperazine was replaced with morpholine, yielding a pale yellow solid in 62% yield: ESI-MS: m / z = 471.1 [M+H] + .

[0083] N-(4-((4-(methanesulfonyl)piperazin-1-yl)methyl)thiazo-2-yl)-2-(3-(trifluoromethyl)phenyl)thiazo-5-carboxamide (compound B6) The preparation of target compound B6 followed the synthesis of compound B1, except that 2-bromo-5-methylthiazol-4-carboxylic acid was replaced with 2-bromothiazol-5-carboxylic acid, and acetylpiperazine was replaced with methanesulfonylpiperazine, yielding a yellow solid in 50% yield; ESI-MS: m / z = 532.1 [M+H] + .

[0084] Example 4: Preparation of target molecules B7-B9

[0085] N-(4-((4-acetylpiperazin-1-yl)methyl)thiazo-2-yl)-4-methyl-2-(3-(trifluoromethyl)phenyl)oxazol-5-carboxamide (compound B7)

[0086] (a) Synthesis of ethyl 4-methyl-2-(3-(trifluoromethyl)phenyl)oxazol-5-carboxylate (compound I-9) Compound I-8 (378 mg, 2 mmol) and ethyl 2-chloro-3-oxobutyrate (329 mg, 2 mmol) were dissolved in toluene (5 mL). The mixture was stirred overnight at 120 °C. A suitable amount of water was added, and the mixture was extracted with ethyl acetate. The organic layer was washed with a saturated saline solution, and the two organic layers were combined. The mixture was then dried over anhydrous Na₂SO₄ and concentrated under reduced pressure. The residue was subjected to column chromatography to give a white solid I-9 in 56% yield; ESI-MS: m / z = 300.3 [M+H]. + .

[0087] (b) Synthesis of 4-methyl-2-(3-(trifluoromethyl)phenyl)oxazol-5-carboxylic acid (compound I-10) Compound I-9 (299 mg, 1 mmol) and NaOH (320 mg, 8 mmol) were dissolved in a mixed solvent of methanol (4 mL) and water (4 mL). The mixture was stirred at room temperature for 4 h. After partially removing the solvent, an appropriate amount of water was added, and the mixture was extracted with ethyl acetate. The organic layer was washed with a saturated saline solution, and the organic layers were combined. The mixture was then dried over anhydrous Na₂SO₄ and concentrated under reduced pressure. The residue was subjected to column chromatography to give a white solid I-10 in 90% yield; ESI-MS: m / z = 272.2 [M+H]. + .

[0088] (c) Synthesis of N-(4-(chloromethyl)thiazol-2-yl)-4-methyl-2-(3-(trifluoromethyl)phenyl)oxazol-5-carboxamide (compound I-11) Compounds I-10 (217 mg, 0.8 mmol), I-5 (119 mg, 0.8 mmol), EDCI (307 mg, 1.6 mmol), and 4-PPY (48 mg, 0.32 mmol) were dissolved in dichloromethane (10 mL), refluxed and stirred overnight, the solvent was removed under reduced pressure, an appropriate amount of water was added, and the mixture was extracted with ethyl acetate. The organic layer was washed with saturated saline solution, the organic layers were combined, dried over anhydrous Na₂SO₄, concentrated under reduced pressure, and the residue was subjected to column chromatography to give a yellow solid I-11 in 68% yield; ESI-MS: m / z = 402.8 [M+H] + .

[0089] (d) Synthesis of N-(4-((4-acetylpiperazin-1-yl)methyl)thiazo-2-yl)-4-methyl-2-(3-(trifluoromethyl)phenyl)oxazol-5-carboxamide (compound B7) Compound I-11 (201 mg, 0.5 mmol) and acetylpiperazine (97 mg, 0.75 mmol) were dissolved in acetonitrile (8 mL). DIPEA (285 μL, 1.5 mmol) was added dropwise to the reaction mixture, which was stirred overnight at room temperature. A suitable amount of water was added, and the mixture was extracted with ethyl acetate. The organic layer was washed with a saturated saline solution, and the two organic layers were combined. The mixture was then dried over anhydrous Na₂SO₄ and concentrated under reduced pressure. The residue was subjected to column chromatography to give a white solid B7 in 63% yield; ESI-MS: m / z = 494.1 [M+H]. + .

[0090] N-(4-(morpholinomethyl)thiazolyl-2-yl)-4-methyl-2-(3-(trifluoromethyl)phenyl)oxazol-5-carboxamide (compound B8) The preparation of target compound B8 followed the synthesis of compound B7, except that acetylpiperazine was replaced with morpholine, yielding a white solid in 60% yield; ESI-MS: m / z = 453.1 [M+H] + .

[0091] N-(4-((1,1-dioxothiomorpholine)methyl)thiazo-2-yl)-4-methyl-2-(3-(trifluoromethyl)phenyl)oxazol-5-carboxamide (compound B9) The preparation of target compound B9 followed the synthesis of compound B7, except that acetylpiperazine was replaced with 1,1-dioxothiomorpholine, yielding a pale yellow solid in 76% yield; ESI-MS: m / z = 501.1 [M+H] + .

[0092] Example 5: Preparation of target molecules B10-B15

[0093] 1-(4-cyano-3-(trifluoromethyl)phenyl)-N-(4-(morpholinomethyl)thiazo-2-yl)-1H-pyrazole-4-carboxamide (compound B10)

[0094] (a) Synthesis of methyl 1-(4-cyano-3-(trifluoromethyl)phenyl)-1H-pyrazole-4-carboxylate (compound I-13-1) Under N2 protection, 4-bromo-2-trifluoromethylbenzonitrile I-12-1 (249 mg, 1 mmol), methyl 1H-pyrazole-3-carboxylate (152 mg, 1.2 mmol), CuI (38 mg, 0.2 mmol), L-proline (46 mg, 0.4 mmol), and potassium carbonate (276 mg, 2 mmol) were dissolved in DMSO (2 mL). The mixture was stirred at 100 °C for 3 h. A suitable amount of water was added, and the mixture was extracted with ethyl acetate. The organic layer was washed with saturated saline solution, and the two organic layers were combined. The mixture was then dried over anhydrous Na2SO4 and concentrated under reduced pressure. The residue was subjected to column chromatography to give a white solid I-13-1 in 79% yield; ESI-MS: m / z = 296.2 [M+H]. + .

[0095] (b) Synthesis of 1-(4-cyano-3-(trifluoromethyl)phenyl)-1H-pyrazole-4-carboxylic acid (compound I-14-1) Compound I-13-1 (148 mg, 0.5 mmol) was dissolved in a mixed solvent of methanol (5 mL) and THF (3 mL), and 4 mmol / mL sodium hydroxide solution (5 mL) was added dropwise. The mixture was stirred overnight at room temperature. After partially removing the solvent by evaporation, an appropriate amount of water was added, and the mixture was extracted with ethyl acetate. The organic layer was washed with a saturated saline solution, and the organic layers were combined. The mixture was then dried over anhydrous Na₂SO₄ and concentrated under reduced pressure. The residue was subjected to column chromatography to give a white solid I-14-1 in 90% yield; ESI-MS: m / z = 282.2 [M+H] + .

[0096] (c) Synthesis of N-(4-(chloromethyl)thiazo-2-yl)-1-(4-cyano-3-(trifluoromethyl)phenyl)-1H-pyrazole-4-carboxamide (compound I-15-1) Compounds I-14-1 (85 mg, 0.3 mmol), I-5 (52 mg, 0.35 mmol), EDCI (115 mg, 0.6 mmol), and 4-PPY (18 mg, 0.12 mmol) were dissolved in dichloromethane (5 mL), refluxed and stirred overnight, the solvent was removed under reduced pressure, an appropriate amount of water was added, and the mixture was extracted with ethyl acetate. The organic layer was washed with saturated saline solution, the organic layers were combined, dried over anhydrous Na₂SO₄, concentrated under reduced pressure, and the residue was subjected to column chromatography to give a white solid I-15-1 in 70% yield; ESI-MS: m / z = 412.8 [M+H] + .

[0097] (d) Synthesis of 1-(4-cyano-3-(trifluoromethyl)phenyl)-N-(4-(morpholinomethyl)thiazo-2-yl)-1H-pyrazole-3-carboxamide (compound B10) Compound I-15-1 (83 mg, 0.2 mmol) and morpholine (27 mg, 0.3 mmol) were dissolved in acetonitrile (4 mL). DIPEA (114 μL, 0.6 mmol) was added dropwise to the reaction mixture, which was stirred overnight at room temperature. A suitable amount of water was added, and the mixture was extracted with ethyl acetate. The organic layer was washed with a saturated saline solution, and the two organic layers were combined. The mixture was then dried over anhydrous Na₂SO₄ and concentrated under reduced pressure. The residue was subjected to column chromatography to give a white solid B10 in 67% yield; ESI-MS: m / z = 463.1 [M+H]. + .

[0098] N-(4-((4-acetylpiperazin-1-yl)methyl)thiazo-2-yl)-1-(4-cyano-3-(trifluoromethyl)phenyl)-1H-pyrazole-4-carboxamide (compound B11) The preparation of target compound B11 was based on the synthesis of compound B10, except that morpholine was replaced with acetylpiperazine, yielding a white solid in 70% yield; ESI-MS: m / z = 504.1 [M+H] + .

[0099] 1-(4-cyano-3-(trifluoromethyl)phenyl)-N-(4-((1,1-dioxothiomorpholino)methyl)thiazo-2-yl)-1H-pyrazole-4-carboxamide (compound B12) The preparation of target compound B12 followed the synthesis of compound B10, except that morpholine was replaced with 1,1-dioxothiomorpholine, yielding a white solid in 63% yield; ESI-MS: m / z = 511.1 [M+H]+ .

[0100] N-(4-((4-acetylpiperazin-1-yl)methyl)thiazo-2-yl)-1-(3-(trifluoromethyl)phenyl)-1H-pyrazole-4-carboxamide (compound B13) The preparation of target compound B13 followed the synthesis of compound B10, with morpholine replaced by acetylpiperazine and 4-bromo-2-trifluoromethylbenzonitrile replaced by 1-bromo-3-trifluoromethylbenzene, yielding a white solid in 67% yield; ESI-MS: m / z = 479.1 [M+H] + .

[0101] N-(4-(thiomorpholinemethyl)thiazolyl-2-yl)-1-(3-(trifluoromethyl)phenyl)-1H-pyrazole-4-carboxamide (compound B14) The preparation of target compound B14 was based on the synthesis of compound B10, with morpholine replaced by thiomorpholine and 4-bromo-2-trifluoromethylbenzonitrile replaced by 1-bromo-3-trifluoromethylbenzene, yielding a white solid in 76% yield; ESI-MS: m / z = 454.1 [M+H] + .

[0102] N-(4-((4,4-difluoropiperidin-1-yl)methyl)thiazolyl-2-yl)-1-(3-(trifluoromethyl)phenyl)-1H-pyrazole-4-carboxamide (compound B15) The preparation of target compound B15 was based on the synthesis of compound B10, with morpholine replaced by 4,4-difluoropiperidine and 4-bromo-2-trifluoromethylbenzonitrile replaced by 1-bromo-3-trifluoromethylbenzene, yielding a white solid in 69% yield; ESI-MS: m / z = 472.1 [M+H] + .

[0103] Example 6: Preparation of target molecules C1-C12 Referring to the synthesis of compound A1, using compound m-trifluoromethylphenylboronic acid (I-1-1) as the starting material, 2-bromo-4-ethylthiazol-5-carboxylic acid, 2-bromo-4-trifluoromethylthiazol-5-carboxylic acid, 2-bromo-4-cyclopropylthiazol-5-carboxylic acid, and 2-bromo-4-chlorothiazol-5-carboxylic acid were used to replace 2-bromo-4-methylthiazol-5-carboxylic acid, and morpholine, 4-acetylpiperazine, and thiomorphon dioxide were used to replace 3,3-difluoropyrrolidine to prepare compounds C1 to C12; The chemical structures and ESI-MS results of the above compounds are shown in Table 2.

[0104] Table 2. Chemical structures and ESI-MS results of C-series target molecules

[0105] Example 7: Preparation of target molecules D1-D3

[0106] 4-Methyl-N-(4-(morpholino-4-formyl)thiazolyl-2-yl)-2-(3-(trifluoromethyl)phenyl)thiazolyl-5-carboxamide (Compound D1)

[0107] (a) Synthesis of ethyl 2-(4-methyl-2-(3-(trifluoromethyl)phenyl)thiazol-5-carboxamido)thiazol-4-carboxylate (compound I-16) The preparation of compound I-16 was based on the synthesis of compound I-3-1, except that 2-amino-4-chloromethylthiazole hydrochloride was replaced with ethyl 2-aminothiazole-4-carboxylate, yielding a pale yellow solid in 89% yield; ESI-MS: m / z = 442.0 [M+H] + .

[0108] (b) Synthesis of 2-(4-methyl-2-(3-(trifluoromethyl)phenyl)thiazol-5-carboxamido)thiazol-4-carboxylic acid (compound I-17) I-16 (441 mg, 1 mmol) and NaOH (320 mg, 8 mmol) were dissolved in a mixed solvent of tetrahydrofuran (6 mL) and water (6 mL). The mixture was stirred at room temperature for 6 h. After removing the organic solvent by evaporation, an appropriate amount of water was added, and the mixture was extracted with ethyl acetate. The organic layer was washed with saturated brine, and the organic layers were combined, dried over anhydrous Na2SO4, concentrated under reduced pressure, and subjected to column chromatography to obtain a pale yellow solid I-17 with a yield of 93%; ESI-MS: m / z = 414.0 [M+H]+.

[0109] (c) Synthesis of 4-methyl-N-(4-(morpholino-4-formyl)thiazolyl-2-yl)-2-(3-(trifluoromethyl)phenyl)thiazolyl-5-carboxamide (compound D1) Compound I-17 (413 mg, 1 mmol), morpholine (130 μL, 1.5 mmol), HATU (760 mg, 2 mmol), and DIPEA (520 μL, 3 mmol) were dissolved in DMF (10 mL) and stirred overnight at room temperature. After the reaction was complete, an appropriate amount of water was added, and the mixture was extracted with ethyl acetate. The organic layer was washed with saturated saline solution, and the organic layers were combined. The mixture was then dried over anhydrous Na2SO4 and concentrated under reduced pressure. The residue was subjected to column chromatography to give a yellow solid D1 in 61% yield; ESI-MS: m / z = 483.1 [M+H] + .

[0110] N-(4-(4-acetylpiperazin-1-formyl)thiazolyl-2-yl)-4-methyl-2-(3-(trifluoromethyl)phenyl)thiazolyl-5-carboxamide (compound D2) The preparation of target compound D2 followed the synthesis of compound D1, except that morpholine was replaced with acetylpiperazine, yielding a pale yellow solid in 58% yield; ESI-MS: m / z = 524.1 [M+H] + .

[0111] N-(4-(1,1-dioxothiomorpholine-4-carbonyl)thiazolyl-2-yl)-4-methyl-2-(3-(trifluoromethyl)phenyl)thiazolyl-5-carboxamide (compound D3) The preparation of target compound D3 followed the synthesis of compound D1, except that morpholine was replaced with 1,1-dioxothiomorpholine, yielding a pale yellow solid in 63% yield; ESI-MS: m / z = 531.0 [M+H] + .

[0112] Example 8: Preparation of target molecules D4-D9 Preparation of target molecules D4-D6

[0113] 4-Methyl-N-(4-(((tetrahydro-2H-pyran-4-yl)oxy)methyl)thiazolyl-2-yl)-2-(3-(trifluoromethyl)phenyl)thiazolyl-5-carboxamide (compound D4) At 0 °C, a THF (10 mL) solution of tetrahydropyran-4-ol (380 μL, 4 mmol) was slowly added dropwise to a THF (10 mL) suspension of sodium hydride (60% w / w, 200 mg, 5 mmol). After stirring for 0.5 h, a THF (20 mL) solution of compound I-3-1 (1.33 g, 3.2 mmol) was added dropwise, and the reaction was then allowed to proceed at room temperature for 6 h. The mixture was quenched with water, followed by the addition of ethyl acetate. The mixture was washed successively with water and saturated brine. The organic layer was dried over anhydrous Na₂SO₄ and concentrated under reduced pressure. The residue was subjected to column chromatography to give a pale yellow solid D3, yield 43%; ESI-MS: m / z = 484.1 [M+H]. + .

[0114] N-(4-((cyclohexyloxy)methyl)thiazolyl-2-yl)-4-methyl-2-(3-(trifluoromethyl)phenyl)thiazolyl-5-carboxamide (compound D5) The preparation of target compound D5 was based on the synthesis of compound D4, except that tetrahydropyran-4-ol was replaced with cyclohexanol, yielding a white solid in 51% yield; ESI-MS: m / z = 482.1 [M+H] + .

[0115] 4-Methyl-N-(4-(((tetrahydro-2H-thiaran-4-yl)oxy)methyl)thiazolyl-2-yl)-2-(3-(trifluoromethyl)phenyl)thiazolyl-5-carboxamide (compound D6) The preparation of target compound D6 was based on the synthesis of compound D4, except that tetrahydropyran-4-ol was replaced with tetrahydro-2H-thiopyran-4-ol, yielding a white solid in 71% yield; ESI-MS: m / z = 500.1 [M+H] + . Preparation of target molecules D7-D9

[0116] 4-Methyl-N-(4-(2-morpholino-2-oxoethyl)thiazolyl-2-yl)-2-(3-(trifluoromethyl)phenyl)thiazolyl-5-carboxamide (Compound D7) The preparation of target compound D7 followed the synthesis of compound D1, except that ethyl 2-aminothiazolium-4-carboxylate was replaced with methyl 2-amino-4-thiazolium acetate, yielding a white solid in 42% yield; ESI-MS: m / z = 497.1 [M+H] + .

[0117] N-(4-(2-(4-acetylpiperazin-1-yl)-2-oxoethyl)thiazo-2-yl)-4-methyl-2-(3-(trifluoromethyl)phenyl)thiazo-5-carboxamide (compound D8) The preparation of target compound D8 was based on the synthesis of compound D1, with ethyl 2-aminothiazol-4-carboxylate replaced by methyl 2-amino-4-thiazolate and morpholine replaced by acetylpiperazine, yielding a pale yellow solid in 46% yield; ESI-MS: m / z = 538.1 [M+H] + .

[0118] 4-Methyl-N-(4-(2-(4-methylpiperazin-1-yl)-2-oxoethyl)thiazo-2-yl)-2-(3-(trifluoromethyl)phenyl)thiazo-5-carboxamide (Compound D9) The preparation of target compound D9 was based on the synthesis of compound D1, with ethyl 2-aminothiazol-4-carboxylate replaced by methyl 2-amino-4-thiazolate and morpholine replaced by N-methylpiperazine, yielding a pale yellow solid in 52% yield; ESI-MS: m / z = 510.1 [M+H] + .

[0119] Example 9: Preparation of target molecules D10-D12

[0120] 4-Methyl-N-(4-(2-morpholinylethyl)thiazo-2-yl)-2-(3-(trifluoromethyl)phenyl)thiazo-5-carboxamide (Compound (Compound D10))

[0121] (a) Synthesis of methyl 2-(2-(4-methyl-2-(3-(trifluoromethyl)phenyl)thiazol-5-carboxamido)thiazol-4-yl)acetate (compound I-18) The preparation of compound I-18 was based on the synthesis of compound I-3-1, except that 2-amino-4-chloromethylthiazole hydrochloride was replaced with methyl 2-amino-4-thiazole acetate, yielding a pale yellow solid in 69% yield; ESI-MS: m / z = 442.0 [M+H] + .

[0122] (b) Synthesis of N-(4-(2-hydroxyethyl)thiazol-2-yl)-4-methyl-2-(3-(trifluoromethyl)phenyl)thiazol-5-carboxamide (compound I-19) Compound I-18 (441 mg, 1 mmol) was dissolved in 10 mL of THF at -20 °C, followed by the slow addition of a 1 M, 2 mL, 2 mmol solution of LiAlH4 in THF. The reaction was continued at -20 °C for 30 min. Then, 1 mL of water was slowly added dropwise, followed by 1 mL of a 15% NaOH aqueous solution. Anhydrous Na2SO4 was added, and the mixture was filtered through diatomaceous earth. Part of the solvent was removed under reduced pressure, and an appropriate amount of water was added. The mixture was extracted with ethyl acetate, and the organic layer was washed with a saturated saline solution, dried over anhydrous Na2SO4, and concentrated under reduced pressure. The residue was subjected to column chromatography to give a pale yellow solid I-19, with a yield of 60%. ESI-MS: m / z = 414.0 [M+H] + .

[0123] (c) Synthesis of 4-methyl-N-(4-(2-oxoethyl)thiazo-2-yl)-2-(3-(trifluoromethyl)phenyl)thiazo-5-carboxamide (compound I-20) Compound I-19 (413 mg, 1 mmol) was dissolved in dichloromethane (10 mL), and then Dysmartin oxidant (509 mg, 1.2 mmol) was added in portions. The reaction was carried out at room temperature for 1 h. The mixture was quenched with water, extracted with dichloromethane, and the organic layer was washed with saturated saline solution. The organic layers were combined, dried over anhydrous Na₂SO₄, concentrated under reduced pressure, and the residue was subjected to column chromatography to give a yellow solid I-20 in 77% yield; ESI-MS: m / z = 412.0 [M+H]. + .

[0124] (d) Synthesis of 4-methyl-N-(4-(2-morpholinylethyl)thiazo-2-yl)-2-(3-(trifluoromethyl)phenyl)thiazo-5-carboxamide (compound D10) Compound I-20 (822 mg, 2 mmol) and morpholine (260 μL, 3 mmol) were dissolved in dichloromethane (10 mL), and then 2 drops of AcOH were added. The mixture was stirred at room temperature for 30 min. Subsequently, NaBH(OEt)3 (636 mg, 3 mmol) was added in portions, and the mixture was stirred at room temperature for 3 h. The solution was quenched with an appropriate amount of water, extracted with dichloromethane, and the organic layer was washed with saturated saline solution. The solution was then dried over anhydrous Na2SO4, concentrated under reduced pressure, and the residue was subjected to column chromatography to give a pale yellow solid D10 in 58% yield; ESI-MS: m / z = 483.1 [M+H] + .

[0125] N-(4-(2-(4-acetylpiperazin-1-yl)ethyl)thiazolyl-2-yl)-4-methyl-2-(3-(trifluoromethyl)phenyl)thiazolyl-5-carboxamide (compound D11) The preparation of target compound D11 was based on the synthesis of compound D10, with morpholine replaced by acetylpiperazine, yielding a pale yellow solid in 46% yield; ESI-MS: m / z = 524.1 [M+H] + .

[0126] N-(4-(2-(1,1-dioxothiomorpholine)ethyl)thiazolyl-2-yl)-4-methyl-2-(3-(trifluoromethyl)phenyl)thiazolyl-5-carboxamide (compound D12) The preparation of target compound D12 followed the synthesis of compound D10, except that morpholine was replaced with 1,1-dioxothiomorpholine, yielding a pale yellow solid in 41% yield; ESI-MS: m / z = 531.1 [M+H] + .

[0127] Example 10: Evaluation Experiment of Androgen Receptor Transcriptional Ability

[0128] Detection principle: Androgen receptors, as transcription factors, require binding to androgen response elements to exert their transcriptional activity. In androgen-dependent prostate cancer cells (LNCaP), the promoter ARR2PB, which exhibits a strong response to the androgen receptor (AR), was stably transfected. Since the expression of enhanced green fluorescent protein (eGFP) depends on ARR2PB, a stable LNCaP-ARR2PB-eGFP expression model was constructed. After treatment with compounds at different concentration gradients, the fluorescence intensity values ​​directly reflected the transcriptional activity of AR, thus revealing the strength of the compounds' antagonistic ability against the androgen receptor.

[0129] Testing steps: LNCaP-ARR2PB-eGFP cells were cultured in RPMA-1640 medium containing 10% fetal bovine serum until they reached 80% confluence. The cells were then passaged and cultured for another 4-5 days in RPMA-1640 medium containing 5% fetal bovine serum (without androgens). The cells were then fed at 3.5 × 10⁻⁶ cells / day. 4 Cells were seeded at a density of 10 cells / well into 96-well plates. After cell adhesion the following day, each well was treated with 5 nM dihydrotestosterone (DHT) and different concentration gradients of the test compound. After incubation for 72 hours, fluorescence intensity near 530 nm was detected using a multi-mode microplate reader under 485 nm excitation light. The average value of the 5 nM DHT wells was taken as 0%, and the blank culture group was taken as 100% for normalization of the transcriptional repressive activity of the compound. IC50 was then calculated. 50 The values ​​were obtained by fitting using GraphpadPrism 8.0.2 software, with enzalutamide as a positive control.

[0130] Detection results: As shown in Table 3, most compounds exhibited strong AR transcriptional repression activity.

[0131] Table 3. AR transcriptional repressive activity of relevant compounds "++++" represents <100nM; "+++" represents 100-500nM; "++" represents 0.5-1μM; "+" represents >1μM.

[0132] Example 11: Test of the inhibitory activity of representative compounds on the proliferation of AR-positive prostate cancer cells LNCaP

[0133] Detection principle: MTT assay is a commonly used method for assessing cell proliferation inhibition. In live cells, succinate dehydrogenase and cytochrome C in mitochondria reduce MTT to water-insoluble blue-purple formazan crystals, and the amount produced is positively correlated with the number of live cells. LNCaP cells are an AR-positive androgen-dependent prostate cancer cell line; therefore, AR antagonists can effectively inhibit LNCaP cell proliferation by blocking androgen receptor signaling. In this experiment, MTT solution was added to treated LNCaP cells to generate formazan crystals, which were then dissolved using DMSO or a triple solution. The optical density (OD value) of the solution was measured at 570 nm using a microplate reader, which directly reflects the number of live cells, thus assessing the inhibitory effect of the drug on LNCaP cell proliferation.

[0134] Testing steps: First, LNCaP cells grown to the logarithmic growth phase were collected and diluted, and seeded at a density of 3000 cells per well in 96-well clear plates. After 24 hours of culture, once cell adhesion was complete, 10 μL of the diluted compound solution was added to each well. A cell-free control group and a solvent control group containing only culture medium were also included. After 72 hours of further cell culture, 10 μL of 5 mg / mL MTT solution was added to each well, and the cells were incubated for 2 to 4 hours. Then, 100 μL of a triple solution was added overnight to dissolve the formazan crystals. Finally, the OD values ​​were measured at 570 nm and 620 nm using a microplate reader, and the cell proliferation inhibition rate was calculated based on the results.

[0135] Test results: As shown in Table 4, all tested compounds exhibited significant inhibitory effects on the proliferation of LNCaP cells, which were superior to enzalutamide, demonstrating excellent therapeutic potential at the cellular level.

[0136] Table 4. Inhibitory activity of compounds against the proliferation of LNCaP cells "++++" represents <5μM; "+++" represents 5-20μM; "++" represents 20-50μM; "+" represents >50μM

[0137] Example 12: Dual-luciferase reporter gene assay for the effect of compounds on AR F876L / T877A activity of mutants

[0138] Detection principle: PC3 is an AR-negative prostate cancer cell line. Transfecting these cells with the pCMV-AR, ARR2PB-Luc, and Renilla plasmids allowed for the quantitative assessment of the activity of compounds against mutant AR. Luc is a firefly luciferase, whose expression is regulated by the transcriptional activity of AR downstream of the ARR2PB promoter. The Renilla plasmid encodes Renilla luciferase and was used to correct for transfection efficiency. By comparing the expression ratios of firefly luciferase and Renilla luciferase, the interaction between transcription factors and target promoters could be quantitatively analyzed.

[0139] Testing steps: After digesting and centrifuging cells in the logarithmic growth phase, resuspend them in 10 mL of phenol red-free RPMI 1640 medium. Count the cells and adjust the cell suspension concentration. Seed the cells into 96-well plates with a white background, 10,000 cells per well (75 μL total volume). Incubate at 37°C for 24 hours. Transfect according to the transfection reagent instructions. Add 0.1 μg of total DNA and 0.25 μL of transfection reagent to each well, pre-mixed with 25 μL of Opti-MEM serum-depleted medium and incubated for 30 min before adding to the 96-well plates. The co-transfected DNA was divided into three categories: plasmids expressing the nuclear receptor, plasmids containing the corresponding promoter expressing firefly luciferase, and Renilla plasmids expressing Renilla luciferase. The Renilla transfection ratio was 1 / 20 of the total DNA, and the ratio of the nuclear receptor to its corresponding promoter plasmid was 3:1. Place in an incubator for static culture (pGL4.18-ARR2PB-Luc is AR) F876L / T877A The promoter, pGL4.18-mmTV-Luc, is used for both MR and PRDRAM. After 24 hours, the culture medium containing the transfection reagent was aspirated, and 90 μL of fresh phenol red-free 1640 medium was added to each well. A concentration gradient of the drug and the corresponding hormone was added to each well, with a hormone control group and a blank control group included. 24 hours after administration, fluorescence values ​​were measured according to the kit instructions. The ratio of firefly luciferase to Renilla luciferase in each well was calculated, and the activator / inhibitor ratio of the corresponding compound was calculated. IC50 50 The values ​​were obtained by fitting using Graphpad Prism 8.0.2 software.

[0140] Test results: As shown in Table 5, after transfection with AR F876L / T877A Subsequently, enzalutamide changed from an antagonist to a partial agonist, and several of our compounds exhibited strong antagonistic activity, suggesting its potential application value in the treatment of drug-resistant prostate cancer.

[0141] Table 5. Representative compounds for AR F876L / T877A antagonistic activity serial number <![CDATA[IC 50 ]]> serial number <![CDATA[IC 50 ]]> serial number <![CDATA[IC 50 ]]> A1 +++ A2 ++++ A3 +++ A4 +++ A5 ++++ A6 ++++ A7 ++ A8 +++ A9 +++ A26 +++ A32 ++++ A44 +++ A54 ++ C1 ++++ C12 ++ D1 +++ D5 ++ Enzalutamide Excited "++++" represents <200nM; "+++" represents 200-500nM; "++" represents 0.5-1μM; "+" represents >1μM.

Claims

1. An aminothiazole compound, characterized by, The aminothiazole compounds include pharmaceutically acceptable salts thereof, R1, R2are independently selected from the group consisting of hydrogen, halogen, nitro, cyano, hydroxy, amino, C 1-3 alkyl, C 1-3 fluorinated alkyl, C 1-3 alkoxy, C 1-3 fluorinated alkoxy, C 1-3 alkylamino, C 1-3 dialkylamino, C 3-6 cycloalkylamino, C 1-3 alkylsulfonyl; When X is N, Y is selected from S, O; When X is S, Y is selected from N; R3is selected from hydrogen, halogen, C 1-4 alkyl, C 1-3 fluorinated alkyl, C 3-6 cycloalkyl; R4is selected from hydrogen, halogen, cyano, C 1-3 alkyl, C 1-3 fluorinated alkyl, C 3-6 cycloalkyl; L is selected from -CH2-, -O-, -CH2CH2-, -CH(CH3)-, -CH(CH3)CH2-, -CH2CH2CH2-, -CH2O-, -CH2CH2O-, -C(=O)-, -CH2C(=O)-, -CH2CH2C(=O)-; G is selected from hydrogen, halogen, hydroxyl, C 1-4 alkyl, C 1-4 fluorine-containing alkyl, C 1-3 alkoxy, C 3-7 cycloalkyl, C l-6 alkyl, C l-6 alkoxy, 4-6 membered heterocycloalkyl containing 1-3 heteroatoms independently selected from S and O, -NR5R6, -NH(CH2) n NR5R6, n = 1-3; R5and R6are independently, the same or different, selected from the group consisting of hydrogen, methylsulfonyl, acetyl, 2-hydroxyacetyl, C 1-4 alkyl, C 1-4 fluorine-containing alkyl, C 1-4 alkoxy, C 3-6 cycloalkyl, C l-6 alkyl; or NR4R5is a 3-8 membered cyclic amine and substituted cyclic amine selected from the group consisting of morpholine, thiomorpholine, thiomorpholine 1-oxide, thiomorpholine dioxide, piperazine, pyrrolidine, piperidine, 1,4-dioxy-8-azaspiro[4.5]decyl, 2,7-diazaspiro[3.5]nonane, substituted with a group selected from hydrogen, halogen, cyano, acetyl, trifluoroacetyl, difluoroacetyl, methoxy, 2-hydroxyacetyl, 2-cyanoacetyl, dimethylaminoacetyl, C 1-6 alkyl, C 1-6 fluorine-containing alkyl, C 1-6 alkoxy, C 1-6 fluorine-containing alkoxy, C 3-6 cycloalkyl, C 1-6 alkylamino, C 1-6 dialkylamino, C l-6 alkyl, C l-6 alkoxy.

2. The aminothiazole compound according to claim 1, characterized by R1, R2are independently selected from hydrogen, fluorine, chlorine, bromine, nitro, cyano, methyl, ethyl, trifluoromethyl, methoxy, difluoromethoxy, trifluoromethoxy, ethoxy, difluoroethoxy, trifluoroethoxy; R3is selected from hydrogen, chlorine, methyl, ethyl, trifluoromethyl, cyclopropyl; selected from R4is selected from hydrogen, fluorine, chlorine, bromine, methyl, ethyl, cyclopropyl, trifluoromethyl, cyano; L is selected from -CH2-, -CH2O-, -CO-, -CH2C(=O)-, -CH2CH2-; G is selected from hydrogen, fluorine, chlorine, methyl, ethyl, trifluoromethyl, amino, methylamino, dimethylamino, diethylamino, hydroxy, methoxy, ethoxy, 3. The aminothiazole compound according to claim 1 or 2, characterized by The compounds have the structural formula as shown in any one of A1-A60, B1-B15, C1-C12 and D1-D12:

4. The aminothiazole compound according to any one of claims 1 to 3, characterized in that, The salts formed by the compounds are selected from organic acid salts or inorganic acid salts; the organic acid salts include but are not limited to oxalate, methanesulfonate, maleate, p-toluenesulfonate, malate, citrate, fumarate, camphorsulfonate, citrate, succinate, tartrate, and the inorganic acid salts include but are not limited to nitrate, sulfate, hydrogen halide salt, phosphate.

5. Use of the aminothiazole compound according to claim 1 in the preparation of a medicament for treating prostate cancer, metastatic prostate cancer, castration-resistant prostate cancer, breast cancer, ovarian cancer, androgenic alopecia and acne.

6. A pharmaceutical composition, characterized by, It contains a therapeutically effective amount of one or more aminothiazole compounds according to any one of claims 1-3, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a prodrug molecule thereof, and a pharmaceutically acceptable excipient, diluent, carrier or adjuvant.

7. The pharmaceutical composition of claim 6, wherein: The preparation forms of the pharmaceutical composition include capsules, powders, tablets, granules, pills, injections, syrups, oral liquids, inhalants, ointments, suppositories or patches.

8. The aminothiazole compound according to claim 1, characterized by The "polymorphs" of the compounds of formula (I) and their salts and solvates, i.e. the compounds of formula (I) or their salts or solvates show polymorphism, can produce different crystalline forms, and have different physical properties, such as density, stability and solubility.

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