7-aminobenzopyrone or 7-aminoquinolinone compound and application thereof

Abstract

The invention discloses 7-aminobenzopyrone or 7-aminoquinolinone compounds and application thereof, the structural general formula of the compounds is as shown in formula (I), and the compounds are compounds with androgen receptor antagonistic activity. The compound provided by the invention has relatively high activity of antagonizing androgen receptor transcription, shows relatively good biological activity at molecular level, cellular level and animal level, and has relatively good safety. Therefore, the compound can be used as an androgen receptor antagonist in preparation of drugs for treating androgen receptor mediated diseases, including but not limited to prostate cancer, metastatic prostate cancer, castration-resistant prostate cancer, breast cancer, ovarian cancer, androgen-derived alopecia and acne. The side effect of systemic androgen antagonism caused by systemic distribution can be avoided, and a safer and more targeted treatment choice is provided for patients.

Description

Technical Field

[0001] The present invention belongs to the field of medicine, and specifically relates to 7-aminobenzopyrone or 7-aminoquinolinone compounds and their applications. The compounds include pharmaceutically acceptable salts, solvates, prodrugs, isomers or metabolites thereof, pharmaceutical compositions containing such compounds, and these compounds or pharmaceutical compositions. And the application of the compounds in the preparation of drugs for treating androgen receptor-related diseases or conditions, and particularly relates to the application of the compounds as androgen receptor antagonists in the preparation of drugs for treating prostate cancer and androgenic alopecia. Background Art

[0002] According to the 2023 global cancer statistics, prostatic cancer has become the second most common cancer in men worldwide and the fifth most lethal, seriously threatening human health. Castration-resistant prostate cancer (CRPC) is the main cause of the high mortality rate of PCa. The androgen receptor (AR) signaling pathway is closely related to the occurrence and development of prostate cancer. Antagonizing the AR pathway is an important strategy for the treatment of hormone-sensitive prostate cancer and CRPC.

[0003] At present, the clinical use of first- and second-generation AR nonsteroidal antagonists in conjunction with endocrine therapy can effectively control the disease. However, with the widespread use of first-generation AR antagonists in clinical applications, specific site mutations in the AR ligand binding domain (LBD), such as threonine at position 877 to alanine (T877A) and tryptophan at position 741 to leucine or cysteine ​​(W741L / C), have been found to cause flutamide and bicalutamide to switch from antagonists to receptor agonists, and these mutations are one of the main causes of the development of castration-resistant prostate cancer (CRPC). Therefore, second-generation AR antagonists, such as enzalutamide and apalutamide, have become new first-line treatments for PCa by enhancing their ability to bind to mutant androgen receptors. However, enzalutamide has only moderate AR antagonistic activity, and the mutation of phenylalanine at position 876 of the LBD to leucine (F876L) converts enzalutamide and apalutamide into agonists, resulting in poor efficacy. Therefore, there is still a need to develop AR antagonists with stronger AR antagonistic activity and anti-tumor effects to further improve the effectiveness of clinical treatment of AR mutation-resistant CRPC.

[0004] In the early stage, the inventor team paid special attention to the dimerization process of AR. Using the self-constructed AR LBD dimer model, through virtual screening and structural modification, they discovered coumarin derivatives with certain AR transcriptional inhibitory activity and the ability to inhibit AR dimerization. The relevant research results have been published in J.Med.Chem. (J.Med.Chem.2021,64,17221-17238) in 2021. For the relevant structure, see patent CN111533721A. On this basis, we systematically optimized the structure of the above-mentioned lead molecules and conducted in-depth analysis of their structure-activity relationship, thereby developing a series of novel 7-aminobenzopyrone or 7-aminoquinolinone derivatives. This series of compounds showed superior AR antagonism and anti-resistance activity, providing new options for the current drug development for the treatment of advanced prostate cancer. Summary of the invention

[0005] The object of the present invention is to provide a 7-aminobenzopyrone or 7-aminoquinolinone compound, which is a novel compound having androgen receptor antagonist activity. The 7-aminobenzopyrone or 7-aminoquinolinone compound includes a pharmaceutically acceptable salt thereof, having a structure of the general formula (I):

[0006] Wherein, R1 and R2 are independently selected from hydrogen, halogen, nitro, cyano, hydroxyl, amino, methylthio, C 1-6 Alkyl, C 1-6 Fluorinated alkyl, C 1-6 Alkoxy, C 1-6 Fluorinated alkoxy, C 1-6 Alkylamino, C 1-6 Dialkylamino, C 3-6 Cycloalkyl or C 3-8 Cycloalkylamino; or R1 and R2 are connected to form a 3-8 membered saturated or unsaturated carbocyclic or heterocyclic ring; X is selected from O or NR a ; where R a Selected from hydrogen, C 1-6 Alkyl or C 3-8 Cycloalkyl; R3 is selected from hydrogen, halogen, nitro, cyano, hydroxyl, amino, NR b R c , C 1-6 Alkyl or C 1-6 Alkoxy; wherein R b , R c independently selected from hydrogen, C 1-6 Alkyl or C 3-6 Cycloalkyl, or NR b R cis a 3-8 membered cyclic amine or substituted cyclic amine selected from morpholine, thiomorpholine, piperazine, pyrrolidine or piperidine, and the substituent is selected from hydrogen, halogen, acetyl, C 1-6 Alkyl, C 1-6 Alkoxy, C 3-8 Alicyclic group; When R4 is hydrogen, n is selected from 0, 2 or 3; When R4 is selected from C 1-6 Alkyl, C 1-6 Haloalkyl, C 3-8 Cycloalkyl, C 3-8 Saturated carbocyclic or heterocyclic ring, C 1-4 When the hydroxyl group of the lipid chain is saturated, n is selected from 0, 1, 2 or 3; Ring A is selected from an aromatic ring and an aromatic heterocyclic ring; R5 and R6 are independently selected from hydrogen, halogen, cyano, hydroxyl, nitro, amino, acetyl, ester, carbamoyl, C 1-6 Alkyl, C 1-6 Haloalkyl, C 1-6 Alkoxy, C 1-6 Fluorinated alkoxy, C 3-8 Cycloalkyl, substituted or unsubstituted five-membered aromatic heterocycle, wherein the substituent is selected from C 1-4 alkyl.

[0007] Preferably, in the 7-aminobenzopyrone or 7-aminoquinolinone compound or a pharmaceutically acceptable salt thereof, R1 and R2 are respectively selected from H, F, Cl, Br, NO2, CN, CH3, CH2CH3, CF3, CF3CH2, OCH3, OCF3, N(CH3)2, or R1 and R2 are connected to form a saturated or unsaturated five-membered or six-membered carbocyclic ring; X is selected from O, NH, N-CH3, NCH2CH3, NCH(CH3)2 or R3 is selected from H, F, Cl, Br, NO2, CH3, CH2CH3, CF3, NH2, NHCH3, N(CH3)2, N(CH2CH3)2, N(CH2CF3)2, OH, OCH3, OCH2CH3, When R4 is H, n is selected from 0, 2 or 3; When R4 is CH3, CF3, CH2CH3, CH2CF3, CH2CH2CH3, CH(CH3)2, CH2CH2OH, CH2CH2CH2OH, When n is selected from 0, 1, 2 or 3; Ring A is selected from: R5 and R6 are selected from H, F, Cl, Br, NO2, CN, NH2, OH, CH3, CF3, OCH3, OCHF2, OCF3, CH2OH, COCH3,

[0008] More preferably, the structural formula of the 7-aminobenzopyrone or 7-aminoquinolinone compound is shown in any one of A1-A36, B1-B21, C1-C12 in Table 1, or a pharmaceutically acceptable salt thereof.

[0009] Table 1

[0010] The present invention also provides derivatives of the 7-aminobenzopyrone or 7-aminoquinolinone compounds, and pharmaceutically acceptable salts thereof or mixtures, tautomers, stereoisomers, solvates, metabolites, and prodrug molecules thereof.

[0011] The pharmaceutically acceptable salts include, but are not limited to, alkali metal salts, such as sodium salts, potassium salts, magnesium salts, calcium salts, etc.; ammonium salts; inorganic acid salts, such as hydrochlorides, hydrobromides, nitrates, sulfates, phosphates, etc.; organic acid salts, such as formates, acetates, propionates, benzoates, maleates, fumarates, succinates, tartrates, citrates, alkyl sulfonates (such as methyl sulfonates, ethyl sulfonates, etc.), aryl sulfonates (such as benzene sulfonates, p-toluene sulfonates, etc.), etc.

[0012] The solvate refers to a complex with variable stoichiometry formed by a solute (the compound of formula (I) provided by the present invention) and a solvent. The solvent includes but is not limited to pharmaceutically acceptable solvents such as water, methanol, ethanol, acetone and acetic acid.

[0013] The present invention also provides polymorphs of the 7-aminobenzopyrone or 7-aminoquinolinone compound or its salt or its solvate, that is, the compound of formula (I) or its salt or solvate can also show polymorphism, that is, it can produce different crystal forms and have different physical properties, such as density, stability and solubility. Different crystal forms and salt forms of drugs may affect their dissolution and absorption in the body, and thus affect the clinical efficacy and safety of the drug to a certain extent.

[0014] Another object of the present invention is to provide the use of the 7-aminobenzopyrone or 7-aminoquinolinone compound in the preparation of a drug for treating androgen receptor-mediated diseases, including prostate cancer, breast cancer, ovarian cancer, androgenic alopecia and acne.

[0015] The 7-aminobenzopyrone or 7-aminoquinolinone derivatives provided by the present invention can be used as a locally applied androgen receptor antagonist for the treatment of androgenetic alopecia, thereby avoiding the systemic androgen antagonism side effects caused by systemic distribution, and providing patients with a safer and more targeted treatment option.

[0016] The present invention has found that the 7-aminobenzopyrone or 7-aminoquinolinone compound described in the above structural formula has the effect of specifically targeting the androgen receptor and showing a significant inhibitory effect on the proliferation of prostate cancer cells, without obvious cytotoxic side effects, and showing good drugability and anti-tumor activity in in vivo biological evaluation. Therefore, this type of compound can be applied to the treatment of tumors related to abnormal expression or mutation of androgen receptor.

[0017] The 7-aminobenzopyrone or 7-aminoquinolinone compound provided by the present invention includes a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a prodrug molecule thereof, or a polymorph thereof.

[0018] The present invention also provides a pharmaceutical composition for treating tumors associated with abnormal expression or mutation of androgen receptor, comprising at least one active ingredient, wherein the active ingredient is selected from a compound having a structural formula as shown in formula (I), a pharmaceutically acceptable salt of the compound, an esterification or phosphoesterification product of the compound, a stereoisomer of the compound, a deuterated product of the compound or a prodrug molecule thereof.

[0019] The pharmaceutical composition also includes a pharmaceutically acceptable carrier. These carriers are various pharmaceutical excipients widely used in the pharmaceutical field, including but not limited to: commonly used diluents and excipients (such as water), fillers (such as starch), adhesives (such as cellulose derivatives and gelatin), wetting agents (such as glycerol), disintegrants (such as agar and calcium carbonate), adsorption carriers (such as kaolin and bentonite), surfactants (such as hexadecanol), absorption promoters (such as quaternary ammonium compounds) and lubricants (such as talc). The pharmaceutical preparation of the present 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, inhalants, ointments, suppositories or patches.

[0020] The present invention has the following beneficial effects: The present invention provides a novel 7-aminobenzopyrone or 7-aminoquinolinone AR antagonist. The compounds and their derivatives have significant antagonistic activity against androgen receptors, specifically inhibit the proliferation of prostate cancer cells, and show good safety. They show good biological activity in both in vivo and in vitro biological evaluations, and have good efficacy in AR resistant to enzalutamide. F876L / T877A Double point mutation model and AR resistant to bicalutamide W741CThe mutant cell model is effective. In addition, the compounds provided by the present invention also show a hair growth promoting effect in a mouse hair loss model after percutaneous administration. Therefore, the compounds can be used as androgen receptor antagonists to treat diseases related to androgen receptors, including but not limited to the treatment of prostate cancer, breast cancer, ovarian cancer, androgenic alopecia and acne. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1 This is the result of the affinity test between compound B1 and AR LBD using protein thermal shift assay technology.

[0022] Figure 2 The dual luciferase reporter gene assay compound B1 for AR F876L / T877A , AR W741C Results of the transcriptional inhibitory activity test of the mutants.

[0023] Figure 3 This is the result of oral administration of compound B1 on the proliferation inhibition of LNCaP cell transplant tumor model.

[0024] Figure 4 This is a test of the effects of compounds B1 and B10 on the C57BL / 6 mouse hair growth model. DETAILED DESCRIPTION

[0025] The present invention will be further described below in conjunction with specific embodiments. The specific embodiments are for illustration and should not be construed as limiting the scope of the present invention. In addition, it should be understood that after reading the content taught by the present invention, those skilled in the art may make various changes or modifications to the present invention, and these equivalent forms also fall within the scope limited by the appended claims of the application.

[0026] Unless otherwise specified, the experimental methods used in the following examples are conventional methods.

[0027] Unless otherwise specified, the materials and reagents used in the following examples can be obtained from commercial sources.

[0028] Example 1: Preparation of target molecules A1 to A16

[0029] The synthetic routes of compounds A1 to A16 are shown in the figure below:

[0030] (a) Preparation of intermediate 7-amino-4-trifluoromethyl-2H-benzopyran-2-one (I-2) Dissolve m-aminophenol (10 mmol) and ethyl trifluoroacetoacetate (10 mmol) in ethanol (4 mL), add ZnCl2 (10 mmol), and stir overnight at 80 °C. After the reaction is completed as monitored by TLC, pour the reaction solution into cold water (10 mL). Filter the resulting precipitate and dry it to obtain a yellow solid with a yield of 82%; 1 H NMR (500MHz, Chloroform-d) δ7.48 (d, J = 8.0 Hz, 1H), 6.73 (s, 1H), 6.57 (dd, J1 = 8. 0Hz, J2=2.0Hz, 1H), 6.36 (d, J=2.0Hz, 1H), 4.97 (s, 2H). ESI-MS: m / z=230.1[M+H] + .

[0031] (b) Preparation of 7-((4-bromophenyl)amino)-4-(trifluoromethyl)-2H-benzopyran-2-one (A1) Under N2 protection, compound I-2 (1.2 mmol), p-bromoiodobenzene (1.2 mmol), Pd2(dba)3 (0.12 mmol), Xantphos (0.15 mmol) and Cs2CO3 (2.4 mmol) were dissolved in a dioxane solution, and then the temperature was raised to 110°C for reaction for 12 h. 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 a saturated saline solution, then dried over anhydrous Na2SO4, and concentrated under reduced pressure. The residue was subjected to column chromatography to obtain a yellow solid A1 with a yield of 76%; ESI-MS: m / z=384.0 [M+H] + .

[0032] Preparation of 7-((6-fluoropyridin-3-yl)amino)-4-(trifluoromethyl)-2H-benzopyran-2-one (A2) Referring to the synthesis of compound A1, p-bromoiodobenzene was replaced with 2-fluoro-5-iodopyridine to obtain a yellow solid with a yield of 48%; 1 H NMR(500MHz,DMSO-d6)δ9.26(s,1H),8.13(d,J=2.2Hz,1H),7.90(ddd,J=9.7,7.2,3.0Hz,1H),7.63–7.51(m,1H),7. 21(dd,J=8.7,3.3Hz,1H),7.03(dd,J=8.9,2.4Hz,1H),6.91(d,J=2.3Hz,1H),6.68(s,1H).ESI-MS:m / z=325.1[M+H] + .

[0033] Preparation of 4-((2-oxo-4-(trifluoromethyl)-2H-benzopyran-7-yl)amino)benzonitrile (A3) Referring to the synthesis of compound A1, p-bromoiodobenzene was replaced with 4-iodobenzonitrile to obtain a yellow solid with a yield of 56%; 1 H NMR (500MHz, DMSO-d6) δ9.66 (s, 1H), 7.77–7.71 (m, 2H), 7.63 (dt, J = 9.0, 2.1Hz, 1H), 7.37–7.32 ( m,2H),7.22(dd,J=8.9,2.3Hz,1H),7.17(d,J=2.2Hz,1H),6.78(s,1H).ESI-MS:m / z=331.1[M+H] + .

[0034] Preparation of 7-((4-fluorophenyl)amino)-4-(trifluoromethyl)-2H-benzopyran-2-one (A4) Referring to the synthesis of compound A1, p-bromoiodobenzene was replaced with p-fluoroiodobenzene to obtain a yellow solid with a yield of 73%; 1 H NMR(500MHz,DMSO-d6)δ9.14(s,1H),7.53(d,J=9.0Hz,1H),7.31–7.19(m,4H),6.99 (dd,J=9.0,2.5Hz,1H),6.85(d,J=2.5Hz,1H),6.63(s,1H).ESI-MS:m / z=324.1[M+H] + .

[0035] Preparation of 4-((2-oxo-4-(trifluoromethyl)-2H-benzopyran-7-yl)amino)-2-(trifluoromethyl)benzonitrile (A5) Referring to the synthesis of compound A1, p-bromoiodobenzene was replaced with 2-cyano-5-iodotrifluorotoluene to obtain a yellow solid with a yield of 56%; 1 H NMR(500MHz,DMSO-d6)δ9.99(s,1H),8.01(d,J=8.6Hz,1H),7.69(dd,J=8.7,2.0Hz ,1H),7.64–7.55(m,2H),7.33–7.25(m,2H),6.87(s,1H).ESI-MS:m / z=399.0[M+H] + .

[0036] Preparation of 1-(4-((2-oxo-4-(trifluoromethyl)-2H-benzopyran-7-yl)amino)phenyl)cyclopropane-1-carbonitrile (A6) Referring to the synthesis of compound A1, p-bromoiodobenzene was replaced with 1-(4-bromophenyl)cyclopropanecarbonitrile to obtain a yellow solid with a yield of 71%;1 H NMR(500MHz,DMSO-d6)δ9.25(s,1H),7.54(dt,J=9.0,2.1Hz,1H),7.36–7.30(m,2H),7.28–7.22(m,2H),7.06(dd,J=9 .0,2.3Hz,1H),6.95(d,J=2.3Hz,1H),6.65(s,1H),1.72(q,J=4.8Hz,2H),1.50–1.43(m,2H).ESI-MS:m / z=371.1[M+H] + .

[0037] Preparation of 2-chloro-4-((2-oxo-4-(trifluoromethyl)-2H-benzopyran-7-yl)amino)benzonitrile (A7) Referring to the synthesis of compound A1, 4-bromoiodobenzene was replaced with 4-bromo-2-chlorobenzonitrile to obtain a yellow solid with a yield of 46%; 1 H NMR(500MHz,DMSO-d6)δ9.80(s,1H),7.82(d,J=8.6Hz,1H),7.67(dt,J=7.7,1.9Hz, 1H),7.38(d,J=2.2Hz,1H),7.30–7.21(m,3H),6.84(s,1H).ESI-MS:m / z=365.0[M+H] + .

[0038] Preparation of 5-((2-oxo-4-(trifluoromethyl)-2H-benzopyran-7-yl)amino)-3-(trifluoromethyl)pyridinecarbonitrile (A8) Referring to the synthesis of compound A1, 4-bromoiodobenzene was replaced with 5-bromo-3-(trifluoromethyl)-2-cyanopyridine to obtain a yellow solid with a yield of 54%; 1 H NMR (500MHz, DMSO-d6) δ10.19(s,1H),8.76(d,J=2.5Hz,1H),7.92(d,J=2.5Hz,1H),7.71(d,J =8.8Hz,1H),7.42(d,J=2.2Hz,1H),7.38–7.29(m,1H),6.90(s,1H).ESI-MS:m / z=400.0[M+H] + .

[0039] Preparation of 7-((5-fluoropyridin-2-yl)amino)-4-(trifluoromethyl)-2H-benzopyran-2-one (A9) Referring to the synthesis of compound A1, 4-bromoiodobenzene was replaced with 2-bromo-5-fluoropyridine to obtain a yellow solid with a yield of 42%; 1H NMR (500MHz, DMSO-d6) δ9.97(s,1H),8.30(d,J=3.0Hz,1H),8.20(t,J=1.7Hz,1H),7.68(td,J=8.7,3.3Hz,1H),7.60( dd,J=9.1,2.1Hz,1H),7.43(dt,J=9.0,1.5Hz,1H),7.02(dd,J=9.1,3.7Hz,1H),6.73(s,1H).ESI-MS: m / z=325.0[M+H] + .

[0040] Preparation of 7-(Benzo[d][1,3]dioxol-5-ylamino)-4-(trifluoromethyl)-2H-benzopyran-2-one (A10) Referring to the synthesis of compound A1, 4-bromoiodobenzene was replaced with 5-iodobenzo[d][1,3]dioxolane to obtain a yellow solid with a yield of 67%; ESI-MS: m / z=350.1[M+H] + .

[0041] Preparation of 7-(quinolin-7-ylamino)-4-(trifluoromethyl)-2H-benzopyran-2-one (A11) Referring to the synthesis of compound A1, 4-bromoiodobenzene was replaced with 7-bromoquinoline to obtain a yellow solid with a yield of 69%; ESI-MS: m / z = 357.1 [M+H] + .

[0042] Preparation of 7-((1H-indol-6-yl)amino)-4-(trifluoromethyl)-2H-benzopyran-2-one (A12) Referring to the synthesis of compound A1, p-bromoiodobenzene was replaced with 6-iodo-1H-indole to obtain a yellow solid with a yield of 53%; ESI-MS: m / z=345.1 [M+H] + .

[0043] Preparation of 7-((3,5-dimethyl-1H-pyrazol-4-yl)amino)-4-(trifluoromethyl)-2H-benzopyran-2-one (A13) Referring to the synthesis of compound A1, 4-bromoiodobenzene was replaced with 3,5-dimethyl-4-iodopyrazole to obtain a yellow solid with a yield of 47%; ESI-MS: m / z = 324.1 [M+H] + .

[0044] Preparation of 4-(trifluoromethyl)-7-((1,3,5-trimethyl-1H-pyrazol-4-yl)amino)-2H-benzopyran-2-one (A14) Referring to the synthesis of compound A1, 4-bromoiodobenzene was replaced with 4-bromo-1,3,5-trimethylpyrazole to obtain a yellow solid with a yield of 39%; ESI-MS: m / z = 338.1 [M+H] + .

[0045] Preparation of 7-(isoxazol-4-ylamino)-4-(trifluoromethyl)-2H-benzopyran-2-one (A15) Referring to the synthesis of compound A1, 4-bromoiodobenzene was replaced with 4-bromoisoxazole to obtain a yellow solid with a yield of 62%; ESI-MS: m / z = 297.0 [M+H] + .

[0046] Preparation of 7-((3,5-dimethylisoxazol-4-yl)amino)-4-(trifluoromethyl)-2H-benzopyran-2-one (A16) Referring to the synthesis of compound A1, 4-bromoiodobenzene was replaced with 3,5-dimethyl-4-iodoisoxazole to obtain a yellow solid with a yield of 51%; ESI-MS: m / z=325.1 [M+H] + .

[0047] Example 2: Preparation of target molecule A17

[0048] (a) Preparation of intermediate 7-amino-4-methyl-2H-benzopyran-2-one (I-4) Dissolve m-aminophenol (4.2 mmol) and ethyl acetoacetate (4.2 mmol) in ethanol (2 mL), slowly add concentrated sulfuric acid (1 mL), and then stir at 80°C overnight. After the reaction is completed as monitored by TLC, pour the reaction solution into cold water (10 mL). Filter the resulting precipitate and dry to obtain a brown solid with a yield of 89%; ESI-MS: m / z = 176.2 [M+H] + .

[0049] (b) Preparation of 7-((4-fluorophenyl)amino)-4-methyl-2H-benzopyran-2-one (A17) Under N2 protection, compound I-4 (1.2 mmol), p-fluoroiodobenzene (1.2 mmol), Pd2(dba)3 (0.12 mmol), Xantphos (0.15 mmol) and Cs2CO3 (2.4 mmol) were dissolved in a dioxane solution, and then the temperature was raised to 110°C for reaction for 12 h. 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 a saturated saline solution, then dried over anhydrous Na2SO4, and concentrated under reduced pressure. The residue was subjected to column chromatography to obtain a yellow solid A17 with a yield of 66%; ESI-MS: m / z=270.1 [M+H]+ .

[0050] Example 3: Preparation of target molecules A18, A19

[0051] (a) Preparation of intermediate 7-amino-4-(trifluoromethyl)quinolin-2(1H)-one (I-6-1) Under N2 protection, m-phenylenediamine (10 mmol) and ethyl trifluoroacetoacetate (12 mmol) were dissolved in DMF (20 mL), and then the temperature was raised to 150°C for 24 h. After the reaction was completed by TLC monitoring, the reaction solution was poured into cold water (10 mL). The precipitate was filtered and dried to obtain a brown solid with a yield of 71%; ESI-MS: m / z = 229.1 [M+H] + .

[0052] (b) Preparation of 7-((4-fluorophenyl)amino)-4-(trifluoromethyl)quinolin-2(1H)-one (A18) Referring to the synthesis step (b) of compound A17, compound I-4 was replaced by compound I-6-1 to obtain a brown solid with a yield of 54%; ESI-MS: m / z=323.1 [M+H] + .

[0053] Preparation of 7-((4-fluorophenyl)amino)-4-methylquinolin-2(1H)-one (A19) Referring to the synthesis of compound A18, ethyl trifluoroacetoacetate was replaced with ethyl acetoacetate to obtain a brown solid with a yield of 62%; ESI-MS: m / z=269.1 [M+H] + .

[0054] Example 4: Preparation of target molecules A20-A23

[0055] (a) Preparation of intermediate (3-hydroxyphenyl)carbamic acid isopropyl ester (I-8) Isopropyl chloroformate (10 mmol) was added to a solution of m-aminophenol (I-7, 10 mmol) in ethyl acetate (15 mL), and the reaction mixture was stirred at room temperature for 2 h. The amine hydrochloride precipitate was removed by filtration, and the ethyl acetate was removed under reduced pressure to obtain colorless crystals of I-8 in a yield of 66%; 1H NMR(500MHz,Chloroform-d)δ7.22(s,1H),7.10(t,J=8.1Hz,1H),6.87(s,1H),6.70(ddd,J=8.1,2.1,0.9Hz, 1H), 6.57 (ddd, J=8.2, 2.4, 0.9Hz, 1H), 4.99 (p, J=6.2Hz, 1H), 1.28 (d, J=6.2Hz, 6H). ESI-MS: m / z=196.1[M+H] + .

[0056] (b) Preparation of intermediate (4-oxo-1,2,3,4-tetrahydrocyclopenta[c]chromen-7-yl)carbamic acid isopropyl ester (I-9-1) Compound I-8 (1.0 mmol) and ethyl 2-oxocyclopentanecarboxylate (1.2 mmol) were dissolved in 70% H2SO4 (4 mL) ethanol solution and stirred at room temperature for 4-6 h. After the reaction was completed by TLC detection, the solution was poured into 10 mL of ice water to obtain a large amount of brown crystalline precipitate. A 50% aqueous NaOH solution was added to the system to adjust the pH to 9-10, and then extracted with ethyl acetate, 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 obtain a brown solid I-9-1 with a yield of 78%; ESI-MS: m / z=288.1[M+H] + .

[0057] (c) Preparation of Intermediate 7-amino-2,3-dihydrocyclopenta[c]chromen-4(1H)-one (I-10-1) Compound I-9-1 (2.2 mmol) was refluxed in a mixture of concentrated sulfuric acid and glacial acetic acid (1:1, 5 ml) for 4 h. A yellow precipitate was formed after cooling. The mixture was poured into 100 mL of ice water and allowed to stand overnight. Under low temperature conditions, the pH was adjusted to slightly alkaline with a 50% aqueous solution of NaOH. The brown precipitate formed was filtered and washed with ice water. Ethanol crystallization gave light brown crystals I-10-1 with a yield of 86%; 1 H NMR (500MHz, DMSO-d6) δ7.24(d,J=8.4Hz,1H),6.55(dd,J=8.4,2.1Hz,1H),6.45(d,J=2.1Hz,1H),6.00(s,2H),2. 96(ddt,J=8.4,7.1,1.7Hz,2H),2.67(ddt,J=8.4,6.9,1.7Hz,2H),2.05(p,J=7.6Hz,2H).ESI-MS:m / z=201.1[M+H] + .

[0058] (d) Preparation of 7-((4-fluorophenyl)amino)-2,3-dihydrocyclopenta[c]chromen-4(1H)-one (A20) Referring to the synthesis step (b) of compound A17, compound I-4 was replaced by compound I-10-1 to obtain a brown solid with a yield of 46%; ESI-MS: m / z=296.1 [M+H] + .

[0059] Preparation of 3-((4-fluorophenyl)amino)-7,8,9,10-tetrahydro-6H-benzo[c]chromen-6-one (A21) Referring to the synthetic route of compound A20, ethyl 2-oxocyclopentanecarboxylate was replaced with ethyl 2-cyclohexanonecarboxylate to obtain a brown solid with a yield of 65%; ESI-MS: m / z=310.1 [M+H] + .

[0060] Preparation of 3-chloro-7-((4-fluorophenyl)amino)-4-methyl-2H-benzopyran-2-one (A22) Referring to the synthetic route of compound A20, ethyl 2-oxocyclopentanecarboxylate was replaced with ethyl 2-chloroacetoacetate to obtain a brown solid with a yield of 74%; ESI-MS: m / z=304.0 [M+H] + .

[0061] Preparation of 7-((4-fluorophenyl)amino)-3,4-dimethyl-2H-benzopyran-2-one (A23) Referring to the synthetic route of compound A20, ethyl 2-oxocyclopentanecarboxylate was replaced with ethyl 2-methylacetoacetate to obtain a brown solid with a yield of 47%; ESI-MS: m / z=284.1 [M+H] + .

[0062] Example 5: Preparation of target molecules A24, A25

[0063] (a) Preparation of intermediate 7-amino-1-methyl-4-(trifluoromethyl)quinolin-2(1H)-one (I-13-1) Under N2 protection, compound I-6-1 (1.0 mmol) was dissolved in anhydrous DMF (5 mL), NaH (60%, 1.2 mmol) was added under ice bath and stirred for 15 min, then iodomethane (1.5 mmol) was slowly added dropwise, and the temperature was raised to room temperature and stirred overnight. After TLC monitoring, the reaction was completed and extracted with ethyl acetate. The organic layer was spin-dried and separated by column chromatography to obtain a brown solid with a yield of 65%; ESI-MS: m / z=243.1[M+H]+ .

[0064] (b) Preparation of 7-((4-fluorophenyl)amino)-1-methyl-4-(trifluoromethyl)quinolin-2(1H)-one (A24) Under N2 protection, compound I-13-1 (1.2 mmol), p-fluoroiodobenzene (1.2 mmol), Pd2(dba)3 (0.12 mmol), Xantphos (0.15 mmol) and Cs2CO3 (2.4 mmol) were dissolved in dioxane (10 mL), and then the temperature was raised to 110°C for reaction for 12 h. 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 a saturated saline solution, then dried over anhydrous Na2SO4, and concentrated under reduced pressure. The residue was subjected to column chromatography to obtain brown solid A17 with a yield of 78%; ESI-MS: m / z=337.1 [M+H] + .

[0065] Preparation of 1-cyclopropyl-7-((4-fluorophenyl)amino)-4-(trifluoromethyl)quinolin-2(1H)-one (A25) Referring to the synthetic route of compound A24, methyl iodide was replaced by cyclopropane bromide to obtain a brown solid with a yield of 56%; ESI-MS: m / z=363.1 [M+H] + .

[0066] Example 6: Preparation of target molecules A26-A31

[0067] (a) Preparation of intermediate 4-(4-bromophenyl)-3,5-dimethylisoxazole (I-15-1) Under N2 protection, 4-iodo-3,5-dimethylisoxazole I-14-1 (1.0 mmol), 4-bromophenylboronic acid (2.0 mmol), Pd2(PPh3)Cl2 (0.02 mmol), NaHCO3 (2.0 mmol) were dissolved in a mixed solution of DMF (5 mL) and H2O (2 mL), and then heated to 90°C for 6 h. 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 a saturated saline solution, then dried over anhydrous Na2SO4, and concentrated under reduced pressure. The residue was subjected to column chromatography to obtain a brown solid I-15-1 with a yield of 81%; 1 H NMR (500MHz, DMSO-d6) δ7.69–7.64(m,2H),7.39–7.34(m,2H),2.40(s,3H),2.22(s,3H).ESI-MS:m / z=252.0[M+H] + .

[0068] (b) Preparation of 7-((4-(3,5-dimethylisoxazol-4-yl)phenyl)amino)-4-(trifluoromethyl)-2H-benzopyran-2-one (A26) Under N2 protection, compound I-15-1 (1.2 mmol), compound I-2 (1.2 mmol), Pd2(dba)3 (0.12 mmol), Xantphos (0.15 mmol) and Cs2CO3 (2.4 mmol) were dissolved in dioxane (10 mL), and then the temperature was raised to 110°C to react 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 a saturated saline solution, then dried over anhydrous Na2SO4, and concentrated under reduced pressure. The residue was subjected to column chromatography to obtain a yellow solid A26 with a yield of 78%. 1 HNMR(500MHz,DMSO-d6)δ9.31(s,1H),7.57(dt,J=8.9,2.0Hz,1H),7.40–7.36(m,2H),7.36–7.31(m,2H),7.12 (dd,J=8.9,2.3Hz,1H),7.02(d,J=2.3Hz,1H),6.66(s,1H),2.42(s,3H),2.25(s,3H).ESI-MS:m / z=401.1[M+H] + .

[0069] Preparation of 7-((4-(3,5-dimethyl-1H-pyrazol-4-yl)phenyl)amino)-4-(trifluoromethyl)-2H-benzopyran-2-one (A27) Referring to the synthetic route of compound A26, 4-iodo-3,5-dimethylisoxazole was replaced with 3,5-dimethyl-4-iodopyrazole to obtain a yellow solid with a yield of 51%; ESI-MS: m / z=400.1 [M+H] + .

[0070] Preparation of 4-(trifluoromethyl)-7-((4-(1,3,5-trimethyl-1H-pyrazol-4-yl)phenyl)amino)-2H-benzopyran-2-one (A28) Referring to the synthetic route of compound A26, 4-iodo-3,5-dimethylisoxazole was replaced with 4-iodo-1,3,5-trimethyl-pyrazole to obtain a yellow solid with a yield of 61%; ESI-MS: m / z=414.1 [M+H] + .

[0071] Preparation of 7-((4-(3,5-dimethylisothiazol-4-yl)phenyl)amino)-4-(trifluoromethyl)-2H-benzopyran-2-one (A29) Referring to the synthetic route of compound A26, 4-iodo-3,5-dimethylisoxazole was replaced with 4-iodo-3,5-dimethylisothiazole to obtain a yellow solid with a yield of 74%; ESI-MS: m / z=417.1 [M+H] + .

[0072] Preparation of 7-((4-(4-methyl-1H-pyrazol-5-yl)phenyl)amino)-4-(trifluoromethyl)-2H-benzopyran-2-one (A30) Referring to the synthetic route of compound A26, 4-iodo-3,5-dimethylisoxazole was replaced by 5-iodo-4-methyl 1H-pyrazole to obtain a yellow solid with a yield of 51%; ESI-MS: m / z=386.1 [M+H] + .

[0073] Preparation of 7-((4-(1-ethyl-4-methyl-1H-pyrazol-5-yl)phenyl)amino)-4-(trifluoromethyl)-2H-benzopyran-2-one (A31) Referring to the synthetic route of compound A26, 4-iodo-3,5-dimethylisoxazole was replaced with 1-ethyl-5-iodo-4-methyl-1H-pyrazole to obtain a yellow solid with a yield of 45%; ESI-MS: m / z=414.1 [M+H] + .

[0074] Example 7: Preparation of target molecules A32 to A36

[0075] Preparation of 7-((4-fluorophenyl)(methyl)amino)-4-(trifluoromethyl)-2H-benzopyran-2-one (A32) Under N2 protection, compound A4 (1.0 mmol) was dissolved in anhydrous DMF (5 mL), NaH (60%, 1.2 mmol) was added under ice bath and stirred for 15 min, then iodomethane (1.2 mmol) was slowly added dropwise, the temperature was raised to room temperature and stirred overnight. After TLC monitoring, the reaction was completed, and ethyl acetate was used for extraction. The organic layer was spin-dried and separated by column chromatography to obtain a yellow solid with a yield of 78%. 1 HNMR(500MHz,Chloroform-d)δ7.45(dq,J=7.8,2.0Hz,1H),7.20(ddt,J=8.1,5.5,2.8Hz,2H ),7.17–7.12(m,2H),6.62–6.58(m,2H),6.45(s,1H),3.36(s,3H).ESI-MS: m / z=338.1[M+H] + .

[0076] Preparation of 7-(ethyl(4-fluorophenyl)amino)-4-(trifluoromethyl)-2H-benzopyran-2-one (A33) Referring to the synthetic route of compound A32, methyl iodide was replaced by ethyl iodide to obtain a yellow solid with a yield of 65%; ESI-MS: m / z=352.1 [M+H] + .

[0077] Preparation of 7-((3,5-dimethylisoxazol-4-yl)(methyl)amino)-4-(trifluoromethyl)-2H-benzopyran-2-one (A34) Referring to the synthetic route of compound A32, compound A4 was replaced by compound A16 to obtain a yellow solid with a yield of 83%; ESI-MS: m / z=339.1 [M+H] + .

[0078] Preparation of 7-((3,5-dimethylisoxazol-4-yl)(ethyl)amino)-4-(trifluoromethyl)-2H-benzopyran-2-one (A35) Referring to the synthetic route of compound A32, compound A4 was replaced by compound A16, and iodomethane was replaced by iodoethane to obtain a yellow solid with a yield of 67%; ESI-MS: m / z=353.1 [M+H] + .

[0079] Preparation of 7-((4-(3,5-dimethylisoxazol-4-yl)phenyl)(ethyl)amino)-4-(trifluoromethyl)-2H-benzopyran-2-one (A36) Referring to the synthetic route of compound A32, compound A4 was replaced by compound A26, and iodomethane was replaced by iodoethane to obtain a yellow solid with a yield of 62%. 1 H NMR(500MHz,DMSO-d6)δ7.54–7.45(m,3H),7.41–7.36(m,2H),6.80–6.73(m,2H),6.61(s,1H),3 .88(q,J=7.0Hz,2H),2.46(s,3H),2.28(s,3H),1.21(t,J=7.0Hz,3H).ESI-MS:m / z=429.1[M+H] + .

[0080] Example 8: Preparation of target molecules B1 to B15

[0081] (a) Preparation of intermediate 7-((4-fluorobenzyl)amino)-4-(trifluoromethyl)-2H-benzopyran-2-one (I-16-1) 4-Fluorobenzyl bromide (5.2 mmol) was added to a DMF (10 mL) solution of compound I-2 (4.4 mmol) and anhydrous Cs2CO3 (5.2 mmol), and stirred at room temperature overnight. After the reaction was completed as monitored by TLC, it was extracted with ethyl acetate. The organic layer was dried and separated by column chromatography to obtain a yellow solid with a yield of 73%; 1 H NMR(500MHz,DMSO-d6)δ7.69(t,J=6.0Hz,1H),7.46–7.35(m,3H),7.24–7.13(m,2H),6 .77(dd,J=9.0,2.5Hz,1H),6.54(d,J=2.5Hz,1H),6.46(s,1H),4.41(d,J=6.0Hz,2H). 13 C NMR (126MHz, DMSO-d6) δ162.70,160.78,159.77,157.10,153.32,140.57,135.3 0,129.66,125.86,123.49,115.67,108.30,102.51,97.91,45.45.HRMS(ESI)m / z Calcd for C 17 H 12 F4NO2[M+H] + :338.0799;Found:338.0803.

[0082] (b) Preparation of 7-((4-fluorobenzyl)(methyl)amino)-4-(trifluoromethyl)-2H-benzopyran-2-one (B1) Under N2 protection, compound I-16-1 (1.0 mmol) was dissolved in anhydrous DMF (5 mL), NaH (60%, 1.2 mmol) was added under ice bath and stirred for 15 min, then iodomethane (1.5 mmol) was slowly added dropwise, and the temperature was raised to room temperature and stirred overnight. After TLC monitoring, the reaction was completed and extracted with ethyl acetate. The organic layer was spin-dried and separated by column chromatography to obtain a yellow solid with a yield of 64%; 1 HNMR(500MHz,Chloroform-d)δ7.50(dq,J=9.0,2.0Hz,1H),7.19–7.11(m,2H),7.07–6.96(m,2 H),6.69(dd,J=9.0,2.5Hz,1H),6.58(d,J=2.5Hz,1H),6.40(s,1H),4.62(s,2H),3.16(s,3H). 13C NMR(126MHz,Chloroform-d)δ163.13,161.18,160.22,156.75,152.63,141.68,132.42,127.96 ,127.90,126.19,115.95,115.78,109.77,108.86,103.46,98.72,55.42,38.94.HRMS(ESI)m / z Calcd for C 18 H 14 F4NO2[M+H] + :352.0955;Found:352.0948.

[0083] Preparation of 7-(ethyl(4-fluorobenzyl)amino)-4-(trifluoromethyl)-2H-benzopyran-2-one (B2) Referring to the synthetic route of compound B1, methyl iodide was replaced by ethyl iodide to obtain a yellow solid with a yield of 56%; 1 HNMR(500MHz,Chloroform-d)δ7.48(dq,J=9.0,2.0Hz,1H),7.18–7.13(m,2H),7.06–7.00(m,2H),6.65(dd,J= 9.0,2.5Hz,1H),6.56(d,J=2.5Hz,1H),6.40(s,1H),4.58(s,2H),3.56(q,J=7.0Hz,2H),1.28(d,J=7.0Hz,3H). 13 C NMR(126MHz,Chloroform-d)δ163.09,161.13,160.32,156.87,151.71,141.56,132.47,127.79,12 7.73,126.29,115.94,115.77,109.75,108.63,103.29,98.65,53.21,45.87,12.10.HRMS(ESI)m / z Calcd for C 19 H 16 F4NO2[M+H] + :366.1112;Found:366.1107.

[0084] Preparation of 7-((4-fluorobenzyl)(isopropyl)amino)-4-(trifluoromethyl)-2H-benzopyran-2-one (B3) Referring to the synthetic route of compound B1, methyl iodide was replaced by 2-iodopropane to obtain a yellow solid with a yield of 62%; ESI-MS: m / z=380.1[M+H] + .

[0085] Preparation of 7-((4-fluorobenzyl)(propyl)amino)-4-(trifluoromethyl)-2H-benzopyran-2-one (B4) Referring to the synthetic route of compound B1, methyl iodide was replaced by 1-iodopropane to obtain a yellow solid with a yield of 55%; ESI-MS: m / z=380.1[M+H] + .

[0086] Preparation of 7-(cyclopropyl(4-fluorobenzyl)amino)-4-(trifluoromethyl)-2H-benzopyran-2-one (B5) Referring to the synthetic route of compound B1, iodomethane was replaced by iodocyclopropane to obtain a yellow solid with a yield of 48%; ESI-MS: m / z=378.1 [M+H] + .

[0087] Preparation of 7-(cyclopentyl(4-fluorobenzyl)amino)-4-(trifluoromethyl)-2H-benzopyran-2-one (B6) Referring to the synthetic route of compound B1, iodomethane was replaced with iodocyclopentane to obtain a yellow solid with a yield of 61%; ESI-MS: m / z=406.1 [M+H] + .

[0088] Preparation of 7-(cyclohexyl(4-fluorobenzyl)amino)-4-(trifluoromethyl)-2H-benzopyran-2-one (B7) Referring to the synthetic route of compound B1, iodomethane was replaced by iodocyclohexane to obtain a yellow solid with a yield of 50%; ESI-MS: m / z=420.1 [M+H] + .

[0089] Preparation of 7-((4-fluorobenzyl)(tetrahydro-2H-pyran-4-yl)amino)-4-(trifluoromethyl)-2H-benzopyran-2-one (B8) Referring to the synthetic route of compound B1, methyl iodide was replaced by 4-iodotetrahydropyran to obtain a yellow solid with a yield of 74%; ESI-MS: m / z=422.1 [M+H] + .

[0090] Preparation of 7-((4-fluorobenzyl)(2-hydroxyethyl)amino)-4-(trifluoromethyl)-2H-benzopyran-2-one (B9) Referring to the synthetic route of compound B1, methyl iodide was replaced with 2-iodoethanol to obtain a yellow solid with a yield of 61%; ESI-MS: m / z=382.1 [M+H] + .

[0091] Preparation of 7-((4-chlorobenzyl)(methyl)amino)-4-(trifluoromethyl)-2H-benzopyran-2-one (B10) Referring to the synthetic route of compound B1, 4-fluorobenzyl bromide was replaced by 4-chlorobenzyl bromide to obtain a yellow solid with a yield of 63%. 1 H NMR(500MHz,Chloroform-d)δ7.50(m,1H),7.36–7.27(m,2H),7.11(d,J=8.0Hz,2H),6. 67(dd,J=9.0,2.5Hz,1H),6.58(d,J=2.5Hz,1H),6.41(s,1H),4.61(s,2H),3.16(s,3H). 13 C NMR(126MHz,Chloroform-d)δ160.19,156.74,152.56,141.68,135.30,133.33,129.13 ,127.69,126.22,122.94,109.76,108.99,103.54,98.76,55.54,39.05.HRMS(ESI)m / z Calcd for C 18 H 14 ClF3NO2[M+H] + :368.0660;Found:368.0655.

[0092] The preparation of 7-((4-chlorobenzyl)(ethyl)amino)-4-(trifluoromethyl)-2H-benzopyran-2-one (B11) was based on the synthetic route of compound B1, except that 4-fluorobenzyl bromide was replaced by 4-chlorobenzyl bromide and iodomethane was replaced by iodoethane to give a yellow solid with a yield of 68%. 1 H NMR(500MHz,Chloroform-d)δ7.48(dq,J=9.0,2.0Hz,1H),7.35–7.27(m,2H),7.12(d,J=8.5Hz,2H),6.63(dd,J =9.0,2.5Hz,1H),6.55(d,J=2.5Hz,1H),6.41(s,1H),4.58(s,2H),3.56(q,J=7.0Hz,2H),1.28(t,J=7.0Hz,3H). 13C NMR(126MHz,Chloroform-d)δ160.26,156.85,151.64,141.64,135.42,133.22,129.10,12 7.56,126.30,122.94,109.75,108.72,103.35,98.68,53.32,45.98,12.13.HRMS(ESI)m / z Calcd forC 19 H 16 ClF3NO2[M+H] + :382.0816;Found:382.0813.

[0093] Preparation of 7-((4-bromobenzyl)(methyl)amino)-4-(trifluoromethyl)-2H-benzopyran-2-one (B12) Referring to the synthetic route of compound B1, 4-fluorobenzyl bromide was replaced by 4-bromobenzyl bromide to obtain a yellow solid with a yield of 80%. 1 H NMR(500MHz,Chloroform-d)δ7.51-7.48(m,1H),7.46(d,J=8.5Hz,2H),7.05(d,J=8.5Hz,2H),6 .67(dd,J1=8.5Hz,J2=2.5Hz,1H),6.57(d,J=2.5Hz,1H),6.42(s,1H),4.60(s,2H),3.17(s,3H). 13 C NMR(126MHz,Chloroform-d)δ160.19,156.72,152.53,141.67,135.83,132.08,12 8.02,126.22,121.35,109.75,109.01,103.55,98.76,55.59,39.07.HRMS(ESI)m / z Calcd for C 18 H 14 BrF3NO2[M+H] + :412.0155;Found:412.0153.

[0094] Preparation of 7-((4-bromobenzyl)(ethyl)amino)-4-(trifluoromethyl)-2H-benzopyran-2-one (B13) Referring to the synthetic route of compound B1, 4-fluorobenzyl bromide was replaced by 4-bromobenzyl bromide, and methyl iodide was replaced by ethyl iodide to obtain a yellow solid with a yield of 63%. 1H NMR(500MHz,Chloroform-d)δ7.49-7.44(m,3H),7.07(d,J=8.5Hz,2H),6.63(dd,J=8.5,2.5Hz,1 H), 6.55 (d, J = 2.5Hz, 1H), 6.40 (s, 1H), 4.56 (s, 2H), 3.56 (q, J = 7.0Hz, 2H), 1.28 (t, J = 7.0Hz, 3H). 13 C NMR(125MHz,Chloroform-d)δ160.23,156.84,151.62,141.66,135.96,132.05,127.92,1 26.30,122.94,121.23,109.76,108.4,103.38,98.70,53.38,46.00,12.13.HRMS(ESI)m / z Calcd for C 19 H 16 BrF3NO2[M+H] + :426.0311;Found:426.0313.

[0095] Preparation of 7-((2,5-difluorobenzyl)(methyl)amino)-4-(trifluoromethyl)-2H-benzopyran-2-one (B14) Referring to the synthetic route of compound B1, 4-fluorobenzyl bromide was replaced by 2,5-difluorobenzyl bromide to obtain a yellow solid with a yield of 56%. 1 H NMR(500MHz,Chloroform-d)δ7.53(dq,J=9.0,2.0Hz,1H),7.09–7.05(m,1H),6.98–6.93(m,1H),6.76– 6.72(m,1H),6.68(dd,J=9.0,2.5Hz,1H),6.59(d,J=2.5Hz,1H),6.45(s,1H),4.66(s,2H),3.20(s,3H). 13 C NMR(126MHz,Chloroform-d)δ160.13,158.02,157.42,156.71,155.49,152.30,141.69,126.33 ,120.73,116.86,115.49,114.41,114.21,109.35,103.82,98.89,50.21,39.15.HRMS(ESI)m / z Calcd for C 18 H 13 F5NO2[M+H] + :370.0861;Found:370.0863.

[0096] Preparation of 7-((2,5-difluorobenzyl)(ethyl)amino)-4-(trifluoromethyl)-2H-benzopyran-2-one (B15) Referring to the synthetic route of compound B1, 4-fluorobenzyl bromide was replaced by 2,5-difluorobenzyl bromide, and iodomethane was replaced by iodoethane to obtain a yellow solid with a yield of 42%. 1 H NMR(500MHz,Chloroform-d)δ7.51(dq,J=9.0,2.0Hz,1H),7.10–7.05(m,1H),6.97–6.92(m,1H),6.76–6.73(m,1H),6.6 4(dd,J=9.0,2.5Hz,1H),6.55(d,J=2.5Hz,1H),6.43(s,1H),4.62(s,2H),3.58(q,J=7.0Hz,2H),1.30(t,J=7.0Hz,3H). 13 C NMR(126MHz,Chloroform-d)δ160.16,158.06,157.39,156.85,155.47,151.41,141.64,126.42,12 5.85,116.78,115.35,114.18,109.62,109.08,103.64,98.77,47.90,45.99,12.10.HRMS(ESI)m / z Calcd for C 19 H 15 F5NO2[M+H] + :384.1017;Found:384.1013.

[0097] Example 9: Preparation of target molecules B16 to B21

[0098] Preparation of 7-((4-fluorobenzyl)(methyl)amino)-4-methyl-2H-benzopyran-2-one (B16) Referring to the synthetic route of compound B1, compound I-2 was replaced by compound I-4 to obtain a yellow solid with a yield of 75%; ESI-MS: m / z=298.1[M+H] + .

[0099] Preparation of 7-(ethyl(4-fluorobenzyl)amino)-4-methyl-2H-benzopyran-2-one (B17) Referring to the synthetic route of compound B1, compound I-2 was replaced by compound I-4, and iodomethane was replaced by iodoethane to obtain a yellow solid with a yield of 61%; ESI-MS: m / z=312.1 [M+H] + .

[0100] Preparation of 7-((4-fluorobenzyl)(methyl)amino)-1-methyl-4-(trifluoromethyl)quinolin-2(1H)-one (B18) Referring to the synthetic route of compound B1, compound I-2 was replaced by compound I-13-1 to obtain a yellow solid with a yield of 54%; ESI-MS: m / z=365.1 [M+H] + .

[0101] Preparation of 7-((4-fluorobenzyl)(methyl)amino)-2,3-dihydrocyclopenta[c]chromen-4(1H)-one (B19) Referring to the synthetic route of compound B1, compound I-2 was replaced by compound I-10-1 to obtain a yellow solid with a yield of 56%; ESI-MS: m / z=324.1 [M+H] + .

[0102] Preparation of 3-((4-fluorobenzyl)(methyl)amino)-7,8,9,10-tetrahydro-6H-benzo[c]chromen-6-one (B20) Referring to the synthetic route of compound B1, compound I-2 was replaced by compound I-10-2 to obtain a yellow solid with a yield of 42%; ESI-MS: m / z=338.1 [M+H] + .

[0103] Preparation of 7-((4-fluorophenethyl)amino)-4-(trifluoromethyl)-2H-benzopyran-2-one (B21) Referring to the synthetic route (a) of compound B1, 4-fluorobenzyl bromide was replaced with 4-fluorobromoethylbenzene to obtain a yellow solid with a yield of 55%; ESI-MS: m / z=352.1 [M+H] + .

[0104] Example 9: Preparation of target molecule C1

[0105] (a) Preparation of intermediate 7-amino-8-nitro-4-(trifluoromethyl)-2H-benzopyran-2-one (I-18) Concentrated nitric acid (6 mmol) was added to a glacial acetic acid solution (10 mL) of compound I-2 and stirred at room temperature for 5 h. After the reaction was completed as monitored by TLC, the reaction solution was poured into cold water (10 mL). The resulting precipitate was filtered and separated by column chromatography to obtain a yellow solid with a yield of 38%; ESI-MS: m / z = 275.0 [M+H] + .

[0106] (b) Preparation of Intermediate 7-((4-fluorophenyl)amino)-8-nitro-4-(trifluoromethyl)-2H-benzopyran-2-one (I-19) Under N2 protection, compound I-18 (1.2 mmol), 4-fluoroiodobenzene (1.8 mmol), Pd2(dba)3 (0.12 mmol), Xantphos (0.15 mmol) and Cs2CO3 (2.4 mmol) were dissolved in dioxane (10 mL), and then the temperature was raised to 110°C for overnight reaction. 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 a saturated saline solution, then dried over anhydrous Na2SO4, and concentrated under reduced pressure. The residue was subjected to column chromatography to obtain a yellow solid I-19 with a yield of 74%; ESI-MS: m / z=369.0 [M+H] + .

[0107] (c) Preparation of 8-amino-7-((4-fluorophenyl)amino)-4-(trifluoromethyl)-2H-benzopyran-2-one (C1) Compound I-19 (1.0 mmol) was dissolved in ethanol (5 mL), iron powder (8.0 mmol) and ammonium chloride (0.1 mmol) were added, and the mixture was heated under reflux overnight. After the reaction was completed as monitored by TLC, the reaction solution was poured into cold water (10 mL). The mixture was extracted with ethyl acetate, the organic phases were combined, dried over anhydrous Na2SO4, and spin-dried. A yellow solid was obtained by column chromatography with a yield of 62%; ESI-MS: m / z=339.1 [M+H] + .

[0108] Example 10: Preparation of target molecules C2, C3

[0109] (a) Intermediate 7-((4-fluorophenyl)(methyl)amino)-8-nitro-4-(trifluoromethyl)-2H-benzopyran-2-one (I-20-1) preparation Under N2 protection, compound I-19 (1.0 mmol) was dissolved in anhydrous DMF (5 mL), NaH (60%, 1.2 mmol) was added under ice bath and stirred for 15 min, then iodomethane (1.5 mmol) was slowly added dropwise, and the temperature was raised to room temperature and stirred overnight. After TLC monitoring, the reaction was completed and extracted with ethyl acetate. The organic layer was spin-dried and separated by column chromatography to obtain a yellow solid with a yield of 66%; ESI-MS: m / z = 383.1 [M+H] + .

[0110] (b) 8-amino-7-((4-fluorophenyl)(methyl)amino)-4-(trifluoromethyl)-2H-benzopyran-2-one (C2) Compound I-20-1 (1.0 mmol) was dissolved in ethanol (5 mL), iron powder (6.0 mmol) and ammonium chloride (0.2 mmol) were added, and the mixture was heated under reflux overnight. After the reaction was completed as monitored by TLC, the reaction solution was poured into cold water (10 mL). The mixture was extracted with ethyl acetate, the organic phases were combined, dried over anhydrous Na2SO4, and spin-dried. A yellow solid was obtained by column chromatography with a yield of 79%; ESI-MS: m / z=353.1 [M+H] + .

[0112] Preparation of 8-amino-7-((4-fluorophenyl)(ethyl)amino)-4-(trifluoromethyl)-2H-benzopyran-2-one (C3) Referring to the synthetic route of compound C2, methyl iodide was replaced by ethyl iodide to obtain a yellow solid with a yield of 52%; ESI-MS: m / z=367.1 [M+H] + .

[0113] Example 11: Preparation of target molecule C4

[0114] Preparation of 4-(4-fluorophenyl)-7-(trifluoromethyl)-1,2,3,4-tetrahydro-9H-pyrano[2,3-f]quinoxalin-9-one (C4) Add 1,2-dibromoethane (2.0 mmol) to a DMF (5 mL) solution of compound C1 (1.0 mmol) and anhydrous Cs2CO3 (2.0 mmol), and stir overnight at room temperature. After the reaction is complete as monitored by TLC, extract with ethyl acetate. The organic layer is dried and separated by column chromatography to obtain a yellow solid with a yield of 40%; ESI-MS: m / z = 365.1 [M+H] + .

[0115] Example 12: Preparation of target molecule C5

[0116] (a) Preparation of Intermediate 7-((4-fluorobenzyl)amino)-8-nitro-4-(trifluoromethyl)-2H-benzopyran-2-one (I-21) 4-Fluorobenzyl bromide (5.2 mmol) was added to a DMF (10 mL) solution of compound I-18 (4.4 mmol) and anhydrous Cs2CO3 (5.2 mmol), and stirred overnight at room temperature. After the reaction was completed as monitored by TLC, it was extracted with ethyl acetate. The organic layer was dried and separated by column chromatography to obtain a yellow solid with a yield of 61%; ESI-MS: m / z=382.1[M+H] + .

[0117] (b) Preparation of 8-amino-7-((4-fluorobenzyl)amino)-4-(trifluoromethyl)-2H-benzopyran-2-one (C5) Compound I-21 (1.0 mmol) was dissolved in ethanol (5 mL), iron powder (4.0 mmol) and ammonium chloride (0.1 mmol) were added, and the mixture was heated under reflux overnight. After the reaction was completed as monitored by TLC, the reaction solution was poured into cold water (10 mL). The mixture was extracted with ethyl acetate, the organic phases were combined, dried over anhydrous Na2SO4, and dried by spin drying. A yellow solid was obtained by column chromatography with a yield of 59%; ESI-MS: m / z=352.1[M+H] + .

[0118] Example 13: Preparation of target molecules C6-C10

[0119] (a) Preparation of Intermediate 7-((4-fluorobenzyl)(methyl)amino)-8-nitro-4-(trifluoromethyl)-2H-benzopyran-2-one (I-22) Under N2 protection, compound I-21 (1.0 mmol) was dissolved in anhydrous DMF (5 mL), NaH (60%, 1.2 mmol) was added under ice bath and stirred for 15 min, then iodomethane (1.5 mmol) was slowly added dropwise, and the temperature was raised to room temperature and stirred overnight. After TLC monitoring, the reaction was completed and extracted with ethyl acetate. The organic layer was spin-dried and separated by column chromatography to obtain a yellow solid with a yield of 82%; ESI-MS: m / z = 397.1 [M+H] + .

[0120] (b) 8-amino-7-((4-fluorobenzyl)(methyl)amino)-4-(trifluoromethyl)-2H-benzopyran-2-one (C6) Compound I-22 (1.0 mmol) was dissolved in ethanol (5 mL), iron powder (4.0 mmol) and ammonium chloride (0.1 mmol) were added, and the mixture was heated under reflux overnight. After the reaction was completed as monitored by TLC, the reaction solution was poured into cold water (10 mL). The mixture was extracted with ethyl acetate, the organic phases were combined, dried over anhydrous Na2SO4, and spin-dried. A light yellow solid was obtained by column chromatography with a yield of 77%; ESI-MS: m / z=367.1 [M+H] + .

[0121] Preparation of 8-(diethylamino)-7-((4-fluorobenzyl)(methyl)amino)-4-(trifluoromethyl)-2H-benzopyran-2-one (C7) Under N2 protection, compound C6 (1.0 mmol) was dissolved in glacial acetic acid (10 mL), and NaBH4 (3.0 mmol) was added in batches, and stirred at room temperature overnight. Then, the mixture was heated to 70°C, and sodium borohydride (3.0 mmol) was added in batches again. After the reaction was completed as monitored by TLC, the reaction solution was poured into cold water (10 mL). The mixture was extracted with ethyl acetate, and the organic phases were combined, dried over anhydrous Na2SO4, spin-dried, and separated by column chromatography to obtain a light yellow solid with a yield of 70%; ESI-MS: m / z=423.2 [M+H] + .

[0122] Preparation of 8-(bis(2,2,2-trifluoroethyl)amino)-7-((4-fluorobenzyl)(methyl)amino)-4-(trifluoromethyl)-2H-benzopyran-2-one (C8) Referring to the synthetic route of compound C7, glacial acetic acid was replaced with trifluoroacetic acid to obtain a light yellow solid with a yield of 63%; ESI-MS: m / z=531.1 [M+H] + .

[0123] Preparation of 7-((4-fluorobenzyl)(methyl)amino)-8-(pyrrolidin-1-yl)-4-(trifluoromethyl)-2H-benzopyran-2-one (C9) 1,4-dibromobutane (1.1 mmol) was added to a solution of compound C6 (1.0 mmol) and anhydrous K2CO3 (2.5 mmol) in acetonitrile (10 mL) and heated under reflux overnight. After the reaction was completed as monitored by TLC, the mixture was extracted with ethyl acetate. The organic layer was dried and separated by column chromatography to obtain a light yellow solid with a yield of 73%; ESI-MS: m / z=421.1[M+H] + .

[0124] Preparation of 7-((4-fluorobenzyl)(methyl)amino)-8-morpholinyl-4-(trifluoromethyl)-2H-benzopyran-2-one (C10) Referring to the synthetic route of compound C9, 1,4-dibromobutane was replaced with 2,2'-dibromodiethyl ether to obtain a light yellow solid with a yield of 74%; ESI-MS: m / z = 437.1 [M+H] + .

[0125] Example 14: Preparation of target molecules C11, C12

[0126] (a) Preparation of intermediate 7-amino-4-methyl-8-nitro 2H-benzopyran-2-one (I-23) Concentrated nitric acid (6 mmol) was added to a glacial acetic acid solution (10 mL) of compound I-4 and stirred at room temperature for 4 h. After the reaction was completed as monitored by TLC, the reaction solution was poured into cold water (10 mL). The resulting precipitate was filtered and separated by column chromatography to obtain a yellow solid with a yield of 32%; ESI-MS: m / z = 221.0 [M+H] + .

[0127] (b) Preparation of Intermediate 7-((4-fluorobenzyl)amino)-4-methyl-8-nitro-2H-benzopyran-2-one (I-24) 4-Fluorobenzyl bromide (5.2 mmol) was added to a DMF (10 mL) solution of compound I-23 (4.4 mmol) and anhydrous Cs2CO3 (5.2 mmol), and stirred overnight at room temperature. After the reaction was completed as monitored by TLC, it was extracted with ethyl acetate. The organic layer was dried and separated by column chromatography to obtain a yellow solid with a yield of 83%; ESI-MS: m / z=329.1 [M+H] + .

[0128] (c) Preparation of Intermediate 7-((4-fluorobenzyl)(methyl)amino)-4-methyl-8-nitro-2H-benzopyran-2-one (I-25) Under N2 protection, compound I-24 (1.0 mmol) was dissolved in anhydrous DMF (5 mL), NaH (60%, 1.2 mmol) was added under ice bath and stirred for 15 min, then iodomethane (1.5 mmol) was slowly added dropwise, and the temperature was raised to room temperature and stirred overnight. After TLC monitoring, the reaction was completed and extracted with ethyl acetate. The organic layer was spin-dried and separated by column chromatography to obtain a yellow solid with a yield of 65%; ESI-MS: m / z=343.1[M+H] + .

[0129] (d) Preparation of 8-amino-7-((4-fluorobenzyl)(methyl)amino)-4-methyl-2H-benzopyran-2-one (C11) Compound I-25 (1.0 mmol) was dissolved in ethanol (5 mL), iron powder (4.0 mmol) and ammonium chloride (0.1 mmol) were added, and the mixture was heated under reflux overnight. After the reaction was completed as monitored by TLC, the reaction solution was poured into cold water (10 mL). The mixture was extracted with ethyl acetate, the organic phases were combined, dried over anhydrous Na2SO4, and dried by spin drying. A light yellow solid was obtained by column chromatography with a yield of 49%; ESI-MS: m / z=313.1 [M+H] + .

[0130] Preparation of 8-amino-7-((4-fluorobenzyl)(methyl)amino)-4-(trifluoromethyl)quinolin-2(1H)-one (C12) Referring to the synthetic route of compound C11, compound I-4 was replaced by compound I-6-1 to obtain a light yellow solid with a yield of 44%; ESI-MS: m / z=366.1 [M+H] + .

[0131] Example 15: Testing of the transcriptional inhibitory activity of compounds on androgen receptor Detection principle: As a transcription factor, the transcriptional activity of the androgen receptor depends on binding to androgen response elements. In the androgen-dependent prostate cancer cell line (LNCaP), we stably transfected a plasmid containing the promoter ARR2PB, which is highly responsive to the androgen receptor (AR), to drive the expression of enhanced green fluorescent protein (eGFP). Therefore, the expression level of eGFP directly reflects the transcriptional activity of AR. In this way, the LNCaP-ARR2PB-eGFP cell line we constructed can quantitatively evaluate the antagonistic effect of compounds on AR activity by measuring the expression of eGFP.

[0132] Detection steps: First, LNCaP-ARR2PB-eGFP cells were cultured in RPMI-1640 medium containing 5% hormone-free fetal bovine serum treated with activated carbon and dextran for 4 to 5 days to consume endogenous hormones in the cells and reduce background interference. When the cells grew to an appropriate density, they were digested and collected by centrifugation, and then the concentration of the cell suspension was adjusted and planted in a 96-well plate at a density of 3.5×10^4 cells per well. After the cells adhered, 5nM dihydrotestosterone (DHT) and different concentrations of the test compound were added to each well on the next day, and a blank control group containing only culture medium was set up. After incubation for another 72 hours, the fluorescence intensity was measured using a microplate reader at an excitation wavelength of 485nm and an emission wavelength of 535nm to calculate the half-inhibitory concentration (IC) of the compound on AR.50 ).

[0133] Test results: As shown in Table 2, most compounds showed strong AR transcriptional inhibitory activity, and the activity of many compounds was higher than that of Enzalutamide (Enz), showing good application prospects.

[0134] Table 2. AR transcriptional inhibitory activity of representative compounds

[0135] Example 16: Testing of the Proliferation Activity of Compounds on AR-Positive Prostate Cancer Cells LNCaP Detection principle: Thiazolyl blue (MTT) detection is a common method for evaluating cell proliferation inhibition. In the mitochondria of living cells, MTT is reduced to water-insoluble blue-purple formazan crystals under the catalysis of succinate dehydrogenase and cytochrome C. There is a linear relationship between the amount of formazan produced and the number of living cells. LNCaP cells are AR-positive and androgen-dependent cell lines, so AR antagonists can reduce the proliferation of LNCaP cells by inhibiting the action of androgens. Formazan is generated by adding MTT solution to treated LNCaP cells, and then the formazan crystals are dissolved with DMSO or triple solution. The optical density value measured at a wavelength of 570nm using an enzyme reader can reflect the number of living cells, thereby evaluating the effect of drugs on LNCaP cell proliferation.

[0136] Detection steps: First, LNCaP cells grown to the logarithmic phase were collected and prepared and planted in a 96-well transparent plate at a density of 3000 cells / well. After 24 hours, after the cells attached, 10 μL of the compound solution diluted with the culture medium was added to the wells. At the same time, a blank control group without cells and a solvent control group with only the culture medium added were set up, and the culture was continued for 72 hours. Subsequently, 10 μL of 5 mg / mL MTT solution was added to each well, incubated for 2 to 4 hours, and then 100 μL of triple solution was added to the wells overnight to dissolve the formazan. Finally, the absorbance (OD) value was measured at 570 nm and 620 nm using an ELISA reader to calculate the proliferation inhibition rate.

[0137] Test results: As shown in Table 3, all tested compounds showed significant proliferation inhibition effects on LNCaP cells, exceeding Enz, demonstrating excellent therapeutic potential at the cellular level.

[0138] Table 3. Proliferation inhibition activity of compounds on LNCaP cells serial number <![CDATA[IC 50 (μM)]]> serial number <![CDATA[IC 50 (μM)]]> serial number <![CDATA[IC 50 (μM)]]> A1 ++++ A2 ++++ A3 ++++ A4 ++++ A5 +++ A6 +++ A7 +++ A8 ++++ A9 +++ B1 +++ B2 +++ B5 +++ B10 +++ B11 +++ B12 +++ C4 ++++ C8 ++++ Enz ++ “++++” represents <5 μM; “+++” represents 5-20 μM; “++” represents 20-50 μM; “+” represents >50 μM.

[0139] Example 17: Protein thermal shift assay to detect the binding of compound B1 to AR LBD Detection principle: Protein thermal shift assay (TSA) is a method for evaluating the binding force between molecules and proteins, based on the determination of protein stability. This technique gradually heats the protein and observes the exposure of autofluorescent groups (such as tryptophan and tyrosine) during the unfolding of its secondary structure. The unfolding process of the protein is tracked by monitoring the changes in these fluorescent groups and the melting point (T m ) changes to assess protein stability. When a protein binds to a small molecule compound, it may induce conformational changes in the protein, making it more stable or unstable. This change is measured by T m The increase or decrease of the value reflects the binding affinity of the compound to the protein.

[0140] Detection steps: First, the purified AR LBD protein was diluted to 0.8 mg / mL in binding buffer. Then, the test compound was also diluted with the same buffer to twice the desired final concentration. The protein and compound were mixed in a 1:1 ratio and incubated for 30 minutes. After that, the mixture was drawn into a high-precision capillary pipette by capillary action and heated from 25°C to 90°C at a rate of 2°C per minute while collecting the fluorescence signal. T was determined by analyzing the first-order derivative of the fluorescence signal. m value.

[0141] Test results: Figure 1 As shown, the binding of dihydrotestosterone (DHT) to AR LBD significantly enhances the stability of the protein and leads to T m Compound B1 increased the T of AR LBD in a dose-dependent manner. m Value (EC 50 =9.97 μM), indicating that compound B1 can stably bind to AR LBD.

[0142] Example 18: Dual luciferase reporter gene assay for the effect of compound B1 on AR F876L / T877A , AR W741C Activity of the mutant Principle of the assay: PC3 is an AR-negative prostate cancer cell line. By transfecting these cells with pCMV-AR, ARR2PB-Luc, and Renilla plasmids, the activity of compounds against mutant AR can be quantitatively evaluated. 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, which is used to correct the transfection efficiency. By comparing the expression ratio of firefly luciferase to Renilla luciferase, the interaction between transcription factors and target promoters can be quantitatively analyzed.

[0143] Detection steps: First, PC3 prostate cancer cells that do not express endogenous AR were starved for 2 to 3 days without hormone serum. Then the cells were collected and the suspension density was adjusted and seeded into 96-well plates at 1×10 4 cells. Next, 72ngpCMV-AR, 24ng ARR2PB-Luc and 5ng Renilla plasmid were mixed with opti-MEM medium and transfection reagent and incubated at room temperature for 30 minutes. 24 hours after transfection, fresh medium was replaced and different concentrations of compounds and 2nM DHT were added. After 24 hours, the medium was discarded and the cells were washed with 100μL PBS, 20μL 1×PLB was added to each well, and the cells were placed on a shaker to fully lyse the cells. Subsequently, 30μL of firefly luciferase substrate was added to each well, and the first fluorescence value was recorded. After the measurement, 30μL of Renilla luciferase substrate was added to each well, the second fluorescence value was measured after mixing, the ratio of the two was calculated, and the transcriptional activity of AR was analyzed.

[0144] Test results: Figure 2 As shown, in the case of transfection with AR F876L / T877A After that, Enz changed from an antagonist to a partial agonist. W741 After that, bicalutamide (Bic) lost its antagonistic activity. However, in both mutation models, compound B1 and darolutamide (Daro) showed strong antagonistic activity, and the antagonistic effect of B1 was better than that of Daro, indicating that it has potential application value in the treatment of drug-resistant prostate cancer.

[0145] Example 19: Detection of the effect of oral administration of compound B1 on the proliferation of LNCaP cell transplant tumor model Detection steps: After 6-week-old male BALB / c nude mice were adaptively raised for one week, LNCaP tumor cells (1×10 7 ), when the average tumor volume reaches 200 mm 3After about 24 hours, the mice were randomly divided into groups according to the tumor volume. A control group (solvent), an enzalutamide group (40 mg / kg, once a day), and a compound B1 group (60 mg / kg, twice a day) were set up. The mice were given intragastric administration, and the weight changes and tumor volume of the mice were monitored during the administration. The length and width of the tumor were measured, and the tumor volume (mm 3 ) = length (mm) × width (mm) 2 / 2. On the last day of drug administration, the tumor was taken and weighed.

[0146] Test results: Figure 3 As shown, compound B1 effectively inhibited the proliferation of prostate cancer xenograft tumors in oral administration of 60 mg / kg. At the same time, the weight of mice did not significantly decrease during administration, indicating that at this dosage, compound B1 has oral efficacy and high safety.

[0147] Example 20: Effects of Compounds B1 and B10 on the C57BL / 6 Mouse Hair Growth Model Experimental procedures: 6- to 7-week-old male C57BL / 6 mice in the telogen phase of the hair cycle were purchased and housed in group cages under standard conditions. The backs were shaved with an electric shaver under deep anesthesia, and then the back hair was removed with a depilatory cream, leaving about 1 cm × 2 cm of bare skin. The next day, mice with no damage to the back skin and pink skin color were selected and randomly divided into: blank control group, model group, compound B1 group, compound B10 group and minoxidil group, 6 mice in each group.

[0148] Blank control group: Apply normal saline twice a day.

[0149] Model group: 200 μL of 0.75% testosterone solution (solvent: ethanol: saline = 1:1) was applied to the hair removal area once a day.

[0150] Drug administration group: 200 μL of 0.75% testosterone solution (solvent is ethanol: saline = 1:1) was applied to the hair removal area once a day, and 150 μL of 1% test drug solution (solvent is NMP: propylene glycol: ethanol = 1:1.2:2.8) was applied to the hair removal area twice a day.

[0151] Minoxidil group: 200 μL of 0.75% testosterone solution (solvent is ethanol: physiological saline = 1:1) was applied to the hair removal area once a day, and 150 μL of minoxidil solution (commercially available 5% minoxidil tincture, Mandi) was applied to the hair removal area twice a day.

[0152] Test results: The hair growth of mice is as follows Figure 4The results showed that after 14 days of treatment, compound B1 and compound B10 had a significant effect of promoting hair growth, and the hair growth promoting effect was better than minoxidil.

Claims

1. 7-aminobenzopyrone or 7-aminoquinolinone compounds, characterized in that: The general structural formula is shown in formula (I): or a pharmaceutically acceptable salt thereof, in, R1 and R2 are independently selected from hydrogen, halogen, nitro, cyano, hydroxyl, amino, methylthio, C 1-6 Alkyl, C 1-6 Fluorinated alkyl, C 1-6 Alkoxy, C 1-6 Fluorinated alkoxy, C 1-6 Alkylamino, C 1-6 Dialkylamino, C 3-6 Cycloalkyl or C 3-8 Cycloalkylamino, or R1 and R2 are linked to form a 3-8 membered saturated or unsaturated carbocyclic or heterocyclic ring; X is selected from O or NR a ; where R a Selected from hydrogen, C 1-6 Alkyl or C 3-8 Cycloalkyl; R3 is selected from hydrogen, halogen, nitro, cyano, hydroxyl, amino, NR b R c , C 1-6 Alkyl, C 1-6 Alkoxy; wherein R b , R c independently selected from hydrogen, C 1-6 Alkyl or C 3-6 Cycloalkyl, or NR b R c is a 3-8 membered cyclic amine or substituted cyclic amine selected from morpholine, thiomorpholine, piperazine, pyrrolidine or piperidine, and the substituent is selected from hydrogen, halogen, acetyl, C 1-6 Alkyl, C 1-6 Alkoxy, C 3-8 Alicyclic group; When R4 is hydrogen, n is selected from 0, 2 or 3; When R4 is selected from C 1-6 Alkyl, C 1-6 Haloalkyl, C 3-8 Cycloalkyl, C 3-8 Saturated carbocyclic or heterocyclic ring, C 1-4 When the hydroxyl group of the lipid chain is saturated, n is selected from 0, 1, 2 or 3; Ring A is selected from an aromatic ring or an aromatic heterocyclic ring; R5 and R6 are independently selected from hydrogen, halogen, cyano, hydroxyl, nitro, amino, acetyl, ester, carbamoyl, C 1-6 Alkyl, C 1-6 Haloalkyl, C 1-6 Alkoxy, C 1-6 Fluorinated alkoxy, C 3-8 Cycloalkyl, substituted or unsubstituted five-membered aromatic heterocycle, wherein the substituent is selected from C 1-4 alkyl.

2. The 7-aminobenzopyrone or 7-aminoquinolinone compound or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that: R1 and R2 are selected from H, F, Cl, Br, NO2, CN, CH3, CH2CH3, CF3, CF3CH2, OCH3, OCF3, N(CH3)2, or R1 and R2 are connected to form a saturated or unsaturated five-membered or six-membered carbocyclic ring; X is selected from O, NH, N-CH3, NCH2CH3, NCH(CH3)2 or R3 is selected from H, F, Cl, Br, NO2, CH3, CH2CH3, CF3, NH2, NHCH3, N(CH3)2, N(CH2CH3)2, N(CH2CF3)2, OH, OCH3, OCH2CH3, When R4 is H, n is selected from 0, 2 or 3; When R4 is CH3, CF3, CH2CH3, CH2CF3, CH2CH2CH3, CH(CH3)2, CH2CH2OH, CH2CH2CH2OH, When n is selected from 0, 1, 2 or 3; Ring A is selected from: R5 and R6 are selected from H, F, Cl, Br, NO2, CN, NH2, OH, CH3, CF3, OCH3, OCHF2, OCF3, CH2OH, COCH3, 3. The 7-aminobenzopyrone or 7-aminoquinolinone compound according to claim 1-2, characterized in that: The structural formula of the compound is shown in any one of A1-A36, B1-B21, and C1-C12, or a pharmaceutically acceptable salt thereof.

4. The compound according to any one of claims 1 to 3, characterized in that The salt formed by the compound is selected from hydrochloride, hydrobromide, methanesulfonate, sulfate, fumarate, tartrate, maleate, malate or citrate.

5. Use of the compound according to any one of claims 1 to 4 as an androgen receptor antagonist in the preparation of a medicament for treating androgen receptor-mediated diseases, characterized in that: Such diseases include prostate cancer, breast cancer, ovarian cancer, androgenic alopecia and acne.

6. The use according to claim 5, characterized in that The drug contains at least one active ingredient, which is selected from a compound with a structural formula as shown in formula (I), a pharmaceutically acceptable salt of the compound, an esterification or phosphate esterification product of the compound, a stereoisomer of the compound, a deuterated product of the compound or a prodrug molecule thereof, and a pharmaceutically acceptable excipient, diluent, carrier or auxiliary material.

7. The use according to claim 5 or 6, characterized in that The drug is in the form of capsules, powders, tablets, granules, pills, injections, syrups, oral liquids, inhalants, ointments, suppositories or patches.

8. The use according to claim 5, characterized in that The drug also includes "polymorphs" of the compound of formula (I) and its salts and solvates, that is, the compound of formula (I) or its salts or solvates also show polymorphism, produce different crystalline forms, and have different physical properties, such as density, stability and solubility.

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