7-Hydroxy flavonoids and their preparation methods and applications

Abstract

The present invention discloses a 7-hydroxyflavonoid compound, or an intermediate, prodrug, deuterated compound, pharmaceutically acceptable salt thereof. The structure of the 7-hydroxyflavonoid compound is shown as formula (I); in the formula: R1 is H or C1-C5 alkyl; R2 is H or OH; R3 is halogen; R4 is -X-linker-R'; X is N, O, S or absent, linker is an alkyl chain containing or not containing N, O, S atoms with a length of 0-6 atoms, and R' is alkyl, cycloalkyl, C1-C6 alkoxy, C1-C6 alkylamino, C1-C6 cycloalkylamino or C1-C6 heterocycloalkylamino. The present invention also discloses a preparation method of the 7-hydroxyflavonoid compound, and the use of the 7-hydroxyflavonoid compound or an intermediate, prodrug, deuterated compound, pharmaceutically acceptable salt thereof. The present invention discloses that the 7-hydroxyflavonoid compound has good inhibitory activity against 3βHSD1. #imgabs0#

Description

Technical Field

[0001] The present invention relates to the field of pharmaceutical technologies, and particularly relates to a 7-hydroxy flavonoid compound, a preparation method thereof, and an application thereof. Background Art

[0002] Prostate cancer cells can utilize testosterone from the testis or dehydroepiandrosterone (DHEA) from the adrenal gland to synthesize the androgen dihydrotestosterone (DHT). DHT binds to AR, initiates downstream gene transcription, and promotes disease evolution. Androgen Deprivation Therapy (ADT) can effectively delay the progression of prostate cancer and significantly prolong the survival period of PCa patients by surgically removing both testes or chemically castrating to block the synthesis of testosterone in the testes. However, the vast majority of patients eventually experience disease recurrence and progress to castration-resistant prostate cancer (CRPC). In 2011, the US FDA approved abiraterone and enzalutamide for the treatment of CRPC. Abiraterone treats CRPC by targeting the metabolic enzyme CYP17A and inhibiting the synthesis of DHEA. Enzalutamide treats CRPC by directly competing with the androgen DHT for binding to AR and preventing AR from being activated. Abiraterone and enzalutamide have achieved great success in clinical practice, but drug tolerance is inevitable, and drug-resistant patients are almost in a hopeless situation.

[0003] During the progression of CRPC, the metabolic process of DHEA to a more active androgen is mainly initiated by the metabolic enzyme type I 3β-hydroxysteroid dehydrogenase (3βHSD1). Previous studies have found that the metabolic pathway of DHEA to downstream androgens is more active in the puncture tissues of metastatic cancer patients, suggesting that the activity of the metabolic enzyme 3βHSD1 is related to the metastatic ability of prostate cancer. In addition, the second-generation anti-androgen drugs abiraterone and enzalutamide can both induce an increase in the expression or activity of the metabolic enzyme 3βHSD1. The metabolic enzyme 3βHSD1 regulates the metabolism of the drug abiraterone, accelerates the blood clearance of abiraterone; promotes the progression of prostate cancer by promoting the production of the steroid progesterone; and antagonizes the efficacy of enzalutamide by accelerating the synthesis of androgens in cancer cells. Therefore, as the convergence point of multiple tolerance mechanisms of second-generation anti-androgen drugs, the metabolic enzyme 3βHSD1 is a very important and promising target for the treatment of CRPC.

[0004] The metabolic enzyme 3βHSD1 has gene polymorphisms (A1245C), and different single nucleotide polymorphisms (SNPs) can affect androgen metabolism by altering the protein stability of 3βHSD1. The 1245th base of the wild-type 3βHSD1 gene is adenine A, while in a small number of cells or patients, this site is thymine T, resulting in the 367th amino acid changing from asparagine in the wild type to threonine in the mutant. In cells, the ubiquitin ligase AMFR can normally recognize wild-type 3βHSD1 and promote the degradation of wild-type 3βHSD1. However, the mutant 3βHSD1 cannot be recognized by AMFR, so the stability of the related protein is increased, which can better generate androgens and promote the growth of corresponding cells and tumors. Multiple clinical studies have found that patients with mutant 3βHSD1 have a faster disease progression, including a higher tendency for in situ cancer to metastasize, recurrence after surgical removal, and drug tolerance in metastatic cancer patients, and their overall survival period is also shorter. These clinical studies also indicate that the activity of the metabolic enzyme 3βHSD1 is related to the malignant progression and treatment tolerance of prostate cancer, providing more clinical evidence for targeted 3βHSD1 treatment of prostate cancer.

[0005] Biochanin A (BCA) is the most effective 3βHSD1 inhibitor discovered so far. By inhibiting 3βHSD1-mediated DHEA metabolism and being able to inhibit the conversion of DHEA in a dose-dependent manner, it can antagonize the expression of AR target genes. However, BCA has significant drawbacks such as low activity and poor drug-likeness, which severely limit its clinical application. Therefore, it is crucial to discover 3βHSD1 inhibitors with better activity and drug-likeness for new drug development. Summary of the Invention

[0006] The present invention provides a 7-hydroxyflavonoid compound, which has good inhibitory activity against 3βHSD1.

[0007] The technical solution of the present invention is as follows:

[0008] A 7-hydroxyflavonoid compound, or its intermediate, prodrug, deuterated compound, pharmaceutically acceptable salt, and the structure of the 7-hydroxyflavonoid compound is shown as formula (I),

[0009]

[0010] In the formula:

[0011] R1 is H or C1-C5 alkyl;

[0012] R2 is H or OH;

[0013] R3 is a halogen;

[0014] R4 is -X-linker-R'; X is N, O, S or absent, linker is an alkyl chain with 0-6 atoms containing or not containing N, O, S atoms, and R' is alkyl, cycloalkyl, C1-C6 alkoxy, C1-C6 alkanamine, C1-C6 cycloalkanamine or C1-C6 heterocycloalkanamine.

[0015] The 7-hydroxyflavonoid compound shown by formula (I) in the present invention has 3βHSD1 inhibitory activity equivalent to or better than that of biochanin A (BCA). By inhibiting the DHEA metabolism mediated by 3βHSD1 and inhibiting the conversion of DHEA, it antagonizes the expression of AR target genes, blocks the AR signaling pathway, and inhibits the occurrence and development of prostate cancer. After introducing a water-soluble group into the representative compound of the 7-hydroxyflavonoid compound shown by formula (I), it can be orally absorbed and shows plasma exposure.

[0016] Preferably, in the -X-linker-R', X is N, O or absent, linker is a C0-C5 alkyl chain, and R' is alkyl, cycloalkyl, C1-C6 alkoxy, C1-C6 alkanamine, C1-C6 cycloalkanamine or C1-C6 heterocycloalkanamine.

[0017] The cycloalkyl is a C1-C5 cycloalkyl.

[0018] The heterocycloalkanamine is a 3-8 membered heterocycloalkanamine containing or not containing N, O, S atoms.

[0019] More preferably, the -X-linker-R' is selected from Table 1:

[0020] Table 1

[0021]

[0022]

[0023] More preferably, the 7-hydroxyflavonoid compound is selected from Table 2:

[0024] Table 2

[0025]

[0026] The present invention also provides a preparation method of the 7-hydroxyflavonoid compound:

[0027] When R2 is H, the compound of formula (I) is mainly synthesized through the following synthetic route:

[0028] The first method for preparing the compound of formula I-A-1

[0029]

[0030] (1) The compound XA undergoes a Suzuki coupling reaction with different types of boric acids or boronic esters to obtain the intermediate XB;

[0031] (2) The protecting group of the intermediate XB is removed to obtain the compound I-A-1.

[0032] Among them, the definitions of R1, R3, and R4 are as described above.

[0033] In step (1), the Suzuki coupling reaction conditions are that the compound XA reacts with the corresponding boric acid or boronic ester in a solvent under basic conditions and in the presence of a catalyst.

[0034] Preferably, the base used in the basic conditions includes at least one of sodium carbonate, sodium hydroxide, potassium hydroxide, cesium carbonate, and potassium phosphate.

[0035] Preferably, the catalyst includes at least one of palladium acetate, dichlorobis(η5-cyclopentadienyl)iron(II)palladium(II), tetrakis(triphenylphosphine)palladium(0), and dichlorobis(triphenylphosphine)palladium(II).

[0036] Preferably, in step (1), the solvent includes at least one of water, N,N-dimethylformamide, N-methylformamide, and 1,4-dioxane.

[0037] In step (2), the deprotection reaction conditions are that the compound XB reacts in a solvent under acidic conditions.

[0038] Preferably, the acid used in the acidic conditions includes at least one of dilute hydrochloric acid, acetic acid, dilute sulfuric acid, and trifluoroacetic acid.

[0039] Preferably, in step (2), the solvent includes at least one or more of water, methanol, ethanol, acetonitrile, and tetrahydrofuran.

[0040] The second method for preparing the compound of formula I-A-1

[0041]

[0042] (1) The compound XA undergoes a Suzuki coupling reaction with different types of boric acids or boronic esters to obtain the intermediate XB;

[0043] (2) The intermediate XB reacts with a diverse amino compound through a Borch reductive amination reaction to obtain the intermediate XC;

[0044] (3) The protecting group of the intermediate XC is removed to obtain the compound I-A-1.

[0045] Among them, the definitions of R1, R3, and R4 are as described above.

[0046] In step (1), the conditions for the Suzuki coupling reaction are that compound XA reacts with the corresponding boric acid or boronic acid ester in a solvent under basic conditions and in the presence of a catalyst.

[0047] Preferably, the base used for the basic conditions includes at least one of sodium carbonate, sodium hydroxide, potassium hydroxide, cesium carbonate, and potassium phosphate.

[0048] Preferably, the catalyst includes at least one of palladium acetate, dichlorobis(ferrocenyldiphenylphosphine)palladium(II), tetrakis(triphenylphosphine)palladium(0), and dichlorobis(triphenylphosphine)palladium(II).

[0049] Preferably, in step (1), the solvent includes at least one of water, N,N-dimethylformamide, N-methylformamide, and 1,4-dioxane.

[0050] In step (2), the Borch reductive amination reaction is that compound XB and a diverse amino compound react in a solvent under a reducing agent and mild acidic conditions.

[0051] Preferably, the reducing agent includes at least one of sodium cyanoborohydride, sodium triacetoxyborohydride, 2-methylpyridine borane, and sodium borohydride.

[0052] Preferably, in step (2), the acid used for the acidic conditions includes at least one of dilute hydrochloric acid, acetic acid, dilute sulfuric acid, and trifluoroacetic acid.

[0053] Preferably, in step (2), the solvent includes at least one of methanol, ethanol, acetonitrile, dichloromethane, chloroform, dimethyl sulfoxide, and N,N-dimethylformamide.

[0054] In step (3), the conditions for the deprotection reaction are that compound XC reacts in a solvent under acidic conditions.

[0055] Preferably, in step (3), the acid used for the acidic conditions includes at least one of dilute hydrochloric acid, acetic acid, dilute sulfuric acid, and trifluoroacetic acid.

[0056] Preferably, in step (3), the solvent includes at least one of water, methanol, ethanol, acetonitrile, and tetrahydrofuran.

[0057] The third method for preparing the compound of formula I-A-1

[0058]

[0059] (1) Compound XA undergoes a Suzuki coupling reaction with different types of boric acid or boronic acid ester to obtain intermediate XB;

[0060] (2) Intermediate XB undergoes a nucleophilic substitution reaction with different types of halides to obtain intermediate XC;

[0061] (3) The protecting group of intermediate XC is removed to obtain compound I-A-1.

[0062] Among them, the definitions of R1, R3, and R4 are as described above.

[0063] In step (1), the Suzuki coupling reaction conditions are that compound XA reacts with the corresponding boric acid or borate ester in a solvent under alkaline conditions and in the presence of a catalyst.

[0064] Preferably, in step (1), the base used for the alkaline conditions includes at least one of sodium carbonate, sodium hydroxide, potassium hydroxide, cesium carbonate, and potassium phosphate.

[0065] Preferably, the catalyst includes at least one of palladium acetate, dichlorobis(diphenylphosphino)ferrocene palladium(II), tetrakis(triphenylphosphine)palladium(0), and dichlorobis(triphenylphosphine)palladium(II).

[0066] Preferably, in step (1), the solvent includes at least one of water, N,N-dimethylformamide, N-methylformamide, and 1,4-dioxane.

[0067] In step (2), the nucleophilic substitution reaction is that compound XB reacts with different types of halides in a solvent under alkaline conditions.

[0068] Preferably, in step (2), the base used for the alkaline conditions includes at least one of sodium carbonate, sodium hydroxide, potassium hydroxide, cesium carbonate, and potassium phosphate.

[0069] Preferably, in step (2), the solvent includes at least one of water, N,N-dimethylformamide, N-methylformamide, methanol, and ethanol.

[0070] In step (3), the deprotection reaction conditions are that compound XC reacts in a solvent under acidic conditions.

[0071] Preferably, the acid used for the acidic conditions includes at least one of dilute hydrochloric acid, acetic acid, dilute sulfuric acid, and trifluoroacetic acid.

[0072] Preferably, in step (3), the solvent includes at least one of water, methanol, ethanol, acetonitrile, and tetrahydrofuran.

[0073] When R2 is OH, the compound of formula (1) is mainly synthesized through the synthetic route:

[0074] The first method for preparing compound I-A-2

[0075]

[0076] (1) The compound XA undergoes a Suzuki coupling reaction with different types of boric acids or boronic esters to obtain the intermediate XB;

[0077] (2) The intermediate XB is demethylated to obtain the compound I-A-2.

[0078] Wherein, the definitions of R1, R3, and R4 are as described above.

[0079] In step (1), the conditions of the Suzuki coupling reaction are that the compound XA reacts with the corresponding boric acid or boronic ester in a solvent under basic conditions and in the presence of a catalyst.

[0080] Preferably, the base used under the basic conditions includes at least one of sodium carbonate, sodium hydroxide, potassium hydroxide, cesium carbonate, and potassium phosphate.

[0081] Preferably, the catalyst includes at least one of palladium acetate, dichlorobis(η5-cyclopentadienyl)iron(II) palladium(II), tetrakis(triphenylphosphine)palladium(0), and dichlorobis(triphenylphosphine)palladium(II).

[0082] Preferably, in step (1), the solvent includes at least one of water, N,N-dimethylformamide, N-methylformamide, and 1,4-dioxane.

[0083] In step (2), the conditions of the demethylation reaction are that the compound XB reacts in a solvent under acidic conditions.

[0084] Preferably, the acid used under the acidic conditions includes at least one of dilute hydrochloric acid, acetic acid, dilute sulfuric acid, and trifluoroacetic acid.

[0085] Preferably, in step (2), the solvent includes at least one of water, methanol, ethanol, acetonitrile, and tetrahydrofuran.

[0086] The second method for preparing the compound of formula I-A-2

[0087]

[0088] (1) The compound XA undergoes a Suzuki coupling reaction with different types of boric acids or boronic esters to obtain the intermediate XB;

[0089] (2) The intermediate XB reacts with a diverse amino compound through a Borch reductive amination reaction to obtain the intermediate XC;

[0090] (3) The intermediate XC is demethylated to obtain the compound I-A-2.

[0091] Wherein, the definitions of R1, R3, and R4 are as described above.

[0092] In step (1), the conditions for the Suzuki coupling reaction are that compound XA reacts with the corresponding boric acid or borate ester in a solvent under basic conditions and in the presence of a catalyst.

[0093] Preferably, the base used under the basic conditions includes at least one of sodium carbonate, sodium hydroxide, potassium hydroxide, cesium carbonate, and potassium phosphate.

[0094] Preferably, the catalyst includes at least one of palladium acetate, dichlorobis(η5-cyclopentadienyl)(diphenylphosphine)palladium(II), tetrakis(triphenylphosphine)palladium(0), and dichlorobis(triphenylphosphine)palladium(II).

[0095] Preferably, in step (1), the solvent includes at least one of water, N,N-dimethylformamide, N-methylformamide, and 1,4-dioxane.

[0096] In step (2), the Borch reductive amination reaction is that compound XB and a diverse amino compound react in a solvent under a reducing agent and mild acidic conditions.

[0097] Preferably, the reducing agent includes at least one of sodium cyanoborohydride, sodium triacetoxyborohydride, 2-methylpyridine borane, and sodium borohydride.

[0098] Preferably, the acid used under the acidic conditions includes at least one of dilute hydrochloric acid, acetic acid, dilute sulfuric acid, and trifluoroacetic acid.

[0099] Preferably, in step (2), the solvent includes at least one of methanol, ethanol, acetonitrile, dichloromethane, chloroform, dimethyl sulfoxide, and N,N-dimethylformamide.

[0100] In step (3), the demethylation reaction conditions are that compound XC reacts in a solvent under acidic conditions.

[0101] Preferably, the acid used under the acidic conditions includes at least one of dilute hydrochloric acid, acetic acid, dilute sulfuric acid, and trifluoroacetic acid.

[0102] Preferably, in step (3), the solvent includes at least one of water, methanol, ethanol, acetonitrile, and tetrahydrofuran.

[0103] The third method for preparing the compound of formula I-A-2

[0104]

[0105] (1) Compound XA undergoes a Suzuki coupling reaction with different types of boric acid or borate ester to obtain intermediate XB;

[0106] (2) Intermediate XB undergoes a nucleophilic substitution reaction with different types of halides to obtain intermediate XC;

[0107] (3) The demethylation of intermediate XC gives compound I-A-2.

[0108] Among them, the definitions of R1, R3, and R4 are as described above.

[0109] In step (1), the Suzuki coupling reaction conditions are that compound XA reacts with the corresponding boric acid or borate ester in a solvent under alkaline conditions and in the presence of a catalyst.

[0110] Preferably, in step (1), the base used for the alkaline conditions includes at least one of sodium carbonate, sodium hydroxide, potassium hydroxide, cesium carbonate, and potassium phosphate.

[0111] Preferably, the catalyst includes at least one of palladium acetate, dichlorobis(η5-cyclopentadienyl)iron(II) palladium(II), tetrakis(triphenylphosphine)palladium(0), and dichlorobis(triphenylphosphine)palladium(II).

[0112] Preferably, in step (1), the solvent includes at least one of water, N,N-dimethylformamide, N-methylformamide, and 1,4-dioxane.

[0113] In step (2), the nucleophilic substitution reaction is that compound XB reacts with different types of halides in a solvent under alkaline conditions.

[0114] Preferably, in step (2), the base used for the alkaline conditions includes at least one of sodium carbonate, sodium hydroxide, potassium hydroxide, cesium carbonate, and potassium phosphate.

[0115] Preferably, in step (2), the solvent includes at least one of water, N,N-dimethylformamide, N-methylformamide, methanol, and ethanol.

[0116] The demethylation reaction conditions are that compound XC reacts in a solvent under acidic conditions.

[0117] Preferably, in step (3), the acid used for the acidic conditions includes at least one of dilute hydrochloric acid, acetic acid, dilute sulfuric acid, and trifluoroacetic acid.

[0118] Preferably, in step (3), the solvent includes at least one of water, methanol, ethanol, acetonitrile, and tetrahydrofuran.

[0119] The present invention also provides a pharmaceutical composition comprising the 7-hydroxyflavonoid compound or its intermediate, prodrug, deuterated compound, pharmaceutically acceptable salt, and pharmaceutically acceptable excipients.

[0120] The present invention also provides a 3βHSD1 inhibitor comprising the 7-hydroxyflavonoid compound or its intermediate, prodrug, deuterated compound, pharmaceutically acceptable salt.

[0121] The present invention also provides the use of the 7-hydroxyflavonoid compound, or its intermediate, prodrug, deuterated compound, pharmaceutically acceptable salt, or its pharmaceutical composition in the preparation of a drug for treating diseases related to 3βHSD1.

[0122] Preferably, the disease related to 3βHSD1 is prostate cancer.

[0123] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0124] The present invention designs and synthesizes a 3βHSD1 inhibitor of 7-hydroxyflavonoid, which has better activity than the existing inhibitors, and has better oral absorption and more excellent pharmacodynamic effects in vivo. Description of the Drawings

[0125] Figure 1 It is a graph of the in vivo pharmacodynamic results of BCA and the compound of Example 7;

[0126] Figure 2 It is a graph of the change in body weight of mice after administration of BCA and the compound of Example 7. Detailed Embodiments

[0127] The present invention will be further described in detail below with reference to the drawings and embodiments. It should be noted that the following embodiments are intended to facilitate the understanding of the present invention and do not limit it in any way.

[0128] Unless otherwise specified, the reagents, methods, etc. used in the embodiments are all conventional reagents and methods in the art.

[0129] (I) Embodiment of Compound Preparation

[0130] The NMR was measured using a Bruker BioSpin AVANCE NEO (500 MHz room temperature probe) and a Bruker BioSpin AVANCE NEO (400 MHz room temperature probe). The solvents for measurement were deuterated dimethyl sulfoxide (DMSO-d6), deuterated chloroform (CDCl3), and deuterated methanol (CD3OD). The internal standard was tetramethylsilane (TMS). The chemical shift was represented by δ, in ppm. The peak patterns were represented as follows: s (singlet), d (doublet), t (triplet), dd (doublet of doublets), q (quartet), dt (doublet of triplets), m (multiplet). The coupling constant (J) was in Hz. The results were analyzed using MestReNova 14. The MS was measured using an Agilent 6120 liquid chromatography-mass spectrometry (ESI); high performance liquid chromatography (HPLC) analysis was performed using an Agilent Zorbax Eclipse XDB-C18 column; column chromatography was performed using a Biotage Isolera One rapid preparator or a Waters Prep-HPLC preparative high performance liquid chromatography.

[0131] Example 1 Synthesis of 7-Hydroxy-3'-Methylamino-4'-Fluoro Isoflavone (Compound 1)

[0132]

[0133] (1) Synthesis of Intermediate 1B

[0134] Add compound 1A (200 mg, 0.54 mmol), 3-Amino-4-Fluorophenylboronic Acid Pinacol Ester (151 mg, 0.64 mmol), Potassium Carbonate (224 mg, 1.62 mmol), Pd(PPh3)4 (25 mg, 0.022 mmol), Dioxane and H2O (Dioxane:H2O = 4:1) into a round-bottom flask, and heat under reflux at 90 °C for 12 h under argon protection. After the reaction is completed, extract three times with DCM, dry with anhydrous sodium sulfate, combine the organic phases and concentrate, then purify by column chromatography (PE / EA = 79 / 21) to obtain white solid compound 1E (151 mg, yield 80%); MS(ESI): m / z 356.1[(M+H) + .

[0135] (2) Synthesis of Intermediate 1C

[0136] Dissolve compound 1B (60 mg, 0.17 mmol) in 6 mL of DMF, add anhydrous potassium carbonate (94 mg, 0.68 mmol), potassium iodide (42 μL, 0.68 mmol), and heat the reaction in a sealed tube at 90 °C for 24 h. After the reaction is completed, extract three times with ethyl acetate, collect the organic phase, wash it three times with water and three times with saturated brine, and dry with anhydrous sodium sulfate to obtain crude compound 1F. MS(ESI): m / z 370.1[(M+H) + .

[0137] (3) Synthesis of Compound 1

[0138] Dissolve compound 1C (40 mg, 0.11 mmol) in MeOH and THF (MeOH:THF = 1:1), add p-Toluenesulfonic Acid (3 mg, 0.011 mmol), and heat under reflux at 60 °C for 1.5 h. After the reaction is completed, extract three times with ethyl acetate, collect the organic phase, wash it three times with water and three times with saturated brine, and dry with anhydrous sodium sulfate. Concentrate the organic phase and purify by column chromatography (PE / EA = 67 / 33). Obtain white solid compound 1 (30 mg, yield 95%); 11H NMR (500 MHz, DMSO-d6) δ 10.81 (s, 1H), 8.35 (s, 1H), 7.97 (d, J = 8.8 Hz, 1H), 7.04 (dd, J = 12.0, 8.2 Hz, 1H), 6.94 (dd, J = 8.8, 2.2 Hz, 1H), 6.87 (d, J = 2.2 Hz, 1H), 6.83 (dd, J = 8.8, 2.0 Hz, 1H), 6.72 (ddd, J = 8.2, 4.5, 2.1 Hz, 1H), 5.59–5.54 (m, 1H), 2.74 (d, J = 5.0 Hz, 3H). MS (ESI): m / z 286.0 [(M+H) + .

[0139] Example 2 Synthesis of 7-Hydroxy-3′-(dimethylamino)-4′-fluoroisoflavone (Compound 2)

[0140]

[0141] The synthesis method was the same as that in Example 1. Another product was the dimethyl-substituted compound, and Compound 2 was obtained. 1 1H NMR (500 MHz, DMSO-d6) δ 10.81 (s, 1H), 8.39 (s, 1H), 7.97 (d, J = 8.8 Hz, 1H), 7.16 (dd, J = 5.2, 3.1 Hz, 1H), 7.13 (d, J = 8.1 Hz, 1H), 7.06 (ddd, J = 8.3, 4.4, 2.1 Hz, 1H), 6.95 (dd, J = 8.8, 2.2 Hz, 1H), 6.88 (d, J = 2.2 Hz, 1H), 2.80 (s, 6H). MS (ESI): m / z 300.0 [(M+H) + .

[0142] Example 3 Synthesis of 7-Hydroxy-3′-(pyrrolidin-1-yl)-4′-fluoroisoflavone (Compound 3)

[0143]

[0144] Using 3-(Pyrrolidin-1-yl)-4-fluorobenzeneboronic acid pinacol ester instead of 3-Amino-4-fluorobenzeneboronic acid pinacol ester, intermediate Compound 3B was obtained according to the synthesis method of Compound 1B in Example 1. Then, Compound 3 was obtained by deprotecting Compound 3B according to the method in step (3) of Example 1. 11H NMR (500 MHz, DMSO-d6) δ 10.83 (s, 1H), 8.36 (s, 1H), 7.97 (d, J = 8.8 Hz, 1H), 7.09 (dd, J = 13.9, 8.3 Hz, 1H), 6.93 (td, J = 9.1, 2.0 Hz, 2H), 6.87 (d, J = 2.1 Hz, 1H), 6.86–6.82 (m, 1H), 3.35–3.32 (m, 4H), 1.90 (t, J = 6.5 Hz, 4H). MS (ESI): m / z 326.1 [(M+H) + .

[0145] Example 4 Synthesis of 7-Hydroxy-3′-((dimethylamino)methyl)-4′-fluoroisoflavone (Compound 4)

[0146]

[0147] (1) Synthesis of Intermediate 4E

[0148] Compound 4E was obtained by the same synthetic method as step (1) in Example 1, except that 4-fluoro-3-formylphenylboronic acid was used instead of 3-amino-4-fluorophenylboronic acid pinacol ester.

[0149] (2) Synthesis of Intermediate 4F

[0150] Intermediate compound 4E (140 mg, 0.38 mmol) and dimethylamine (95 μL, 0.19 mmol) were dissolved in 8 mL of methanol. After reacting at room temperature for 1 h, sodium cyanoborohydride (14 mg, 0.23 mmol) and a drop of acetic acid were added, and the reaction continued for 4 h. After the reaction was completed, the solvent was evaporated under reduced pressure and purified by column chromatography (DCM / MEOH = 95 / 5) to obtain white solid compound 4F (66 mg, yield 44%); MS (ESI): m / z 398.2 [(M+H) + .

[0151] (3) Synthesis of Compound 4

[0152] Compound 4 was obtained from compound 4F according to step (3) in Example 1. 11H NMR (500 MHz, DMSO-d6) δ 10.87 (s, 1H), 8.40 (s, 1H), 7.98 (d, J = 8.8 Hz, 1H), 7.60 (dd, J = 7.2, 2.3 Hz, 1H), 7.52–7.46 (m, 1H), 7.23 (dd, J = 9.9, 8.6 Hz, 1H), 6.95 (dd, J = 8.8, 2.2 Hz, 1H), 6.88 (d, J = 2.2 Hz, 1H), 3.47 (s, 2H), 2.18 (s, 6H). MS (ESI): m / z 314.1 [(M+H) + .

[0153] Example 5 Synthesis of 7-Hydroxy-3′-((dimethylamino)methyl)-4′-chloroisoflavone (Compound 5)

[0154]

[0155] Compound 5 was obtained by the same synthetic method as in Example 4, except that 4-chloro-3-formylphenylboronic acid was used instead of 4-fluoro-3-formylphenylboronic acid. 1 1H NMR (500 MHz, DMSO-d6) δ 10.77 (s, 1H), 8.42 (s, 1H), 7.97 (d, J = 8.8 Hz, 1H), 7.69 (s, 1H), 7.47 (s, 2H), 6.94 (dd, J = 8.8, 2.1 Hz, 1H), 6.86 (d, J = 2.1 Hz, 1H), 3.50 (s, 2H), 2.22 (s, 6H). MS (ESI): m / z 330.0 [(M+H) + .

[0156] Example 6 Synthesis of 3-(4-Chloro-3-(pyrrolidin-1-ylmethyl)phenyl)-7-hydroxy-4H-chromen-4-one (Compound 6)

[0157]

[0158] The synthetic method was the same as in Example 5, except that pyrrolidine was used instead of dimethylamine. 1 1H NMR (500 MHz, DMSO-d6) δ 10.85 (s, 1H), 8.42 (s, 1H), 7.98 (d, J = 8.8 Hz, 1H), 7.70 (s, 1H), 7.47 (t, J = 6.0 Hz, 2H), 6.95 (dd, J = 8.7, 1.9 Hz, 1H), 6.88 (d, J = 1.8 Hz, 1H), 3.71 (s, 2H), 2.53 (s, 4H), 1.71 (s, 4H). MS (ESI): m / z 356.0 [(M+H) + .

[0159] Example 7 Synthesis of 3-(4-chloro-3-(morpholinomethyl)phenyl)-7-hydroxy-4H-chromen-4-one (Compound 7)

[0160]

[0161] The synthesis method was the same as that in Example 5 except that morpholine was used instead of dimethylamine. 1 H NMR (500 MHz, DMSO-d6) δ 10.65 (s, 1H), 8.42 (s, 1H), 7.97 (d, J = 8.8 Hz, 1H), 7.70 (s, 1H), 7.49 (s, 2H), 6.94 (dd, J = 8.8, 2.2 Hz, 1H), 6.87 (d, J = 2.1 Hz, 1H), 3.59 (s, 6H), 2.45 (s, 4H). MS (ESI): m / z 372.0 [(M+H) + .

[0162] Example 8 Synthesis of 7-hydroxy-3′-((diethylamino)methyl)-4′-chloroisoflavone (Compound 8)

[0163]

[0164] The synthesis method was the same as that in Example 5 except that diethylamine was used instead of dimethylamine. 1 H NMR (500 MHz, DMSO-d6) δ 10.66 (s, 1H), 8.39 (s, 1H), 7.97 (d, J = 8.8 Hz, 1H), 7.75 (d, J = 1.4 Hz, 1H), 7.48–7.42 (m, 2H), 6.94 (dd, J = 8.8, 2.1 Hz, 1H), 6.86 (d, J = 2.1 Hz, 1H), 3.63 (s, 2H), 2.56–2.51 (m, 4H), 1.00 (t, J = 7.1 Hz, 6H). MS (ESI): m / z 358.1 [(M+H) + .

[0165] Example 9 Synthesis of 7-hydroxy-3′-((methylamino)methyl)-4′-chloroisoflavone (Compound 9)

[0166]

[0167] The synthesis method was the same as that in Example 5 except that methylamine was used instead of dimethylamine. 11H NMR (500 MHz, DMSO-d6) δ 10.16 (s, 1H), 8.39 (s, 1H), 7.98 (d, J = 8.8 Hz, 1H), 7.67–7.62 (m, 1H), 7.50 (ddd, J = 7.6, 5.0, 2.1 Hz, 1H), 7.26–7.20 (m, 1H), 6.95 (dd, J = 8.8, 1.9 Hz, 1H), 6.88 (d, J = 1.7 Hz, 1H), 3.74 (s, 2H), 2.32 (s, 3H).. MS (ESI): m / z 316.0 [(M+H) + .

[0168] Example 10 Synthesis of 3-(4-chloro-3-(piperidin-1-ylmethyl)phenyl)-7-hydroxy-4H-chromen-4-one (Compound 10)

[0169]

[0170] The synthesis method was the same as that in Example 5 except that piperidine was used instead of dimethylamine. 1 1H NMR (500 MHz, DMSO-d6) δ 10.29 (s, 1H), 8.30 (s, 1H), 7.87 (d, J = 8.8 Hz, 1H), 7.68 (s, 1H), 7.45 (s, 2H), 6.80 (dd, J = 8.8, 1.9 Hz, 1H), 6.65 (d, J = 1.7 Hz, 1H), 3.53 (s, 2H), 2.41 (s, 4H), 1.51 (p, J = 5.4 Hz, 4H), 1.43–1.36 (m, 2H). MS (ESI): m / z 370.1 [(M+H) + .

[0171] Example 11 Synthesis of 7-hydroxy-3′-((diethylamino)methyl)-4′-flavone (Compound 11)

[0172]

[0173] The synthesis method was the same as that in Example 8 except that 4-fluoro-3-formylphenylboronic acid was used instead of 4-chloro-3-formylphenylboronic acid. 11H NMR (500 MHz, DMSO-d6) δ 10.58 (s, 1H), 8.37 (s, 1H), 7.97 (d, J = 8.8 Hz, 1H), 7.64 (dd, J = 7.2, 1.9 Hz, 1H), 7.46 (ddd, J = 7.6, 4.9, 2.2 Hz, 1H), 7.25–7.18 (m, 1H), 6.94 (dd, J = 8.8, 2.1 Hz, 1H), 6.86 (d, J = 2.0 Hz, 1H), 3.60 (s, 2H), 2.51 (s, 2H), 2.48 (d, J = 7.1 Hz, 2H), 1.00 (t, J = 7.1 Hz, 6H). MS (ESI): m / z 342.1 [(M+H) + .

[0174] Example 12 Synthesis of 7-Hydroxy-3′-((methylamino)methyl)-4′-fluoroisoflavone (Compound 12)

[0175]

[0176] The synthesis method was the same as that in Example 9, except that 4-fluoro-3-formylphenylboronic acid was used instead of 4-chloro-3-formylphenylboronic acid. 1 1H NMR (500 MHz, DMSO-d6) δ 10.16 (s, 1H), 8.39 (s, 1H), 7.98 (d, J = 8.8 Hz, 1H), 7.67–7.62 (m, 1H), 7.50 (ddd, J = 7.6, 5.0, 2.1 Hz, 1H), 7.26–7.20 (m, 1H), 6.95 (dd, J = 8.8, 1.9 Hz, 1H), 6.88 (d, J = 1.7 Hz, 1H), 3.74 (s, 2H), 2.32 (s, 3H). MS (ESI): m / z 300.0 [(M+H) + .

[0177] Example 13 Synthesis of 3-(4-Chloro-3-(2-morpholinoethoxy)phenyl)-7-hydroxy-4H-chromen-4-one (Compound 13)

[0178]

[0179] (1) Synthesis of Intermediate 13E

[0180] The synthesis method of 13E was the same as that in step (1) of Example 1, except that (4-chloro-3-hydroxyphenyl)boronic acid was used instead of 3-amino-4-fluorophenylboronic acid pinacol ester in step (1) of Example 1.

[0181] (2) Synthesis of Intermediate 13F

[0182] Intermediate compound 13E (150 mg, 0.40 mmol), N-(2-chloroethyl)morpholine hydrochloride (90 mg, 0.48 mmol), and cesium carbonate (456 mg, 1.00 mmol) were dissolved in 6 mL of N,N-dimethylformamide and heated under reflux at 80 °C for 12 h. After the reaction was completed, the mixture was extracted three times with ethyl acetate. The organic phase was collected, washed three times with water, three times with saturated brine, dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography (PE / EA = 97 / 3). A white solid compound 13F (138 mg, yield 71%) was obtained; MS (ESI): m / z 486.1 [(M+H) + .

[0183] (3) Synthesis of compound 13

[0184] According to step (3) of Example 1, compound 13 can be obtained using 13F as the starting material. 1 H NMR (500 MHz, DMSO-d6) δ 10.86 (s, 1H), 8.49 (s, 1H), 7.99 (d, J = 8.8 Hz, 1H), 7.47 (d, J = 8.2 Hz, 1H), 7.39 (d, J = 1.9 Hz, 1H), 7.20 (dd, 1H), 6.97 (dd, J = 8.8, 2.2 Hz, 1H), 6.90 (d, J = 2.2 Hz, 1H), 4.22 (t, J = 5.7 Hz, 2H), 3.60–3.55 (m, 4H), 2.77 (t, J = 5.7 Hz, 2H), 2.53 (d, J = 5.5 Hz, 4H). MS (ESI): m / z 402.1 [(M+H) + .

[0185] Example 14 Synthesis of 3-(4-chloro-3-(2-methoxyethoxy)phenyl)-7-hydroxy-4H-chromen-4-one (compound 14)

[0186]

[0187] The synthesis method was the same as that in Example 13, except that 2-chloroethyl methyl ether was used instead of N-(2-chloroethyl)morpholine hydrochloride. 1HNMR(500MHz,DMSO-d6)δ10.86(s,1H),8.48(s,1H),7.99(d,J = 8.7Hz,1H),7.48(d,J = 8.2Hz,1H),7.37(d,J = 1.9Hz,1H),7.20(dd,J = 8.2,1.9Hz,1H),6.97(dd,J = 8.8,2.3Hz,1H),6.90(d,J = 2.2Hz,1H),4.26–4.19(m,2H),3.75–3.69(m,2H),3.35(s,3H).MS(ESI):m / z 347.1[(M+H) + .

[0188] Synthesis of 3-(4-chloro-3-(2-(dimethylamino)ethoxy)phenyl)-7-hydroxy-4H-chromen-4-one (Compound 15) in Example 15

[0189]

[0190] The synthesis method was the same as that in Example 13, except that tert-butyl (2-chloroethyl)(methyl)carbamate was used instead of N-(2-chloroethyl)morpholine hydrochloride.

[0191] The synthesis method was the same as that in Example 14. 1H NMR(500MHz,CDCl3)δ8.33(s,1H),8.01–7.95(m,1H),7.53–7.45(m,2H),7.18(s,0H),6.90(s,1H),6.89(dd,J = 9.8,2.7Hz,1H),4.24(t,J = 6.6Hz,2H),3.00(t,J = 6.6Hz,2H).MS(ESI):m / z300.0[(M+H) + .

[0192] Synthesis of 3-(4-chloro-3-((2-morpholinoethyl)amino)phenyl)-7-hydroxy-4H-chromen-4-one (Compound 16) in Example 16

[0193]

[0194] The synthesis method was the same as that in Example 13, except that 3-amino-4-chlorophenylboronic acid pinacol ester was used instead of (4-chloro-3-hydroxyphenyl)boronic acid. 11H NMR (500 MHz, CDCl3) δ 8.01–7.95 (m, 1H), 7.52 (d, J = 1.1 Hz, 1H), 6.89 (d, J = 8.7 Hz, 1H), 5.68 (t, J = 4.6 Hz, 1H), 3.66–3.61 (m, 2H), 3.48 (td, J = 5.2, 4.5 Hz, 1H), 2.80 (t, J = 5.2 Hz, 1H), 2.59–2.54 (m, 2H). MS (ESI): m / z 360.0 [(M+H) + .

[0195] Synthesis of Example 17 3-(3-(4-Ethylpiperazin-1-yl)-4-fluorophenyl)-7-hydroxy-4H-chromen-4-one (Compound 17)

[0196]

[0197] The synthesis method was the same as that of Example 3, except that (4-Fluoro-3-morpholinophenyl)boronic acid pinacol ester was used instead of 3-(Pyrrolidin-1-yl)-4-fluorobenzeneboronic acid pinacol ester. 1 1H NMR (500 MHz, CDCl3) δ 8.34 (s, 1H), 8.01–7.95 (m, 1H), 7.59 (ddd, J = 8.8, 4.0, 2.2 Hz, 1H), 7.30 (dd, J = 3.6, 2.2 Hz, 1H), 7.24–7.17 (m, 1H), 6.92–6.86 (m, 2H), 3.43–3.37 (m, 4H), 2.86–2.80 (m, 4H), 2.59 (q, J = 7.0 Hz, 2H), 1.06 (t, J = 7.0 Hz, 3H). MS (ESI): m / z 369.1 [(M+H) + .

[0198] Synthesis of Example 18 3-(4-Chloro-3-morpholinomethyl)phenyl)-7-hydroxy-2-methyl-4H-chromen-4-one (Compound 18)

[0199]

[0200] The synthesis method was the same as that of Example 7, except that 3-Iodo-7-methoxy-2-methyl-4H-chromen-4-one was used instead of 3-Iodo-7-((tetrahydro-2H-pyran-2-yl)oxy)-4H-chromen-4-one. 11H NMR (500 MHz, CDCl3) δ 7.91 (d, J = 9.1 Hz, 1H), 7.54–7.47 (m, 2H), 7.38 (s, 1H), 6.89 (dd, J = 9.1, 2.3 Hz, 1H), 6.85 (d, J = 2.3 Hz, 1H), 3.76 (d, J = 0.9 Hz, 2H), 3.68–3.62 (m, 4H), 2.68–2.63 (m, 4H). MS (ESI): m / z 386.1 [(M+H) + .

[0201] Example 19 Synthesis of 3-(4-chloro-3-morpholinomethyl)phenyl)-5,7-dihydroxy-4H-chromen-4-one (Compound 19)

[0202]

[0203] Intermediate 2C can be obtained according to Example 7. MS (ESI): m / z 416.1 [(M+H) + .

[0204] Synthesis of Compound 19

[0205] Intermediate compound 2C (473 mg, 1.14 mmol) was dissolved in 9 mL of 45% aqueous hydrobromic acid and heated under reflux at 130 °C for 24 h. After completion of the reaction, the reaction mixture was poured into ice water for quenching, adjusted to neutral pH, extracted three times with ethyl acetate, and the combined organic phases were concentrated and purified by column chromatography (PE / EA = 79 / 21) to obtain white solid Compound 19 (274 mg, yield 62%). 1 1H NMR (500 MHz, CDCl3) δ 8.22 (s, 1H), 7.57 (dt, J = 1.8, 1.0 Hz, 1H), 7.51 (dd, J = 8.5, 1.9 Hz, 1H), 7.46 (s, 1H), 6.40 (d, J = 2.2 Hz, 1H), 6.20 (d, J = 2.2 Hz, 1H), 3.76 (d, J = 1.0 Hz, 2H), 3.68–3.62 (m, 4H), 2.68–2.63 (m, 4H). MS (ESI): m / z 388.0 [(M+H) + .

[0206] Example 20 Synthesis of 3-(4-chloro-3-morpholinomethyl)phenyl)-5,7-dihydroxy-2-methyl-4H-chromen-4-one (Compound 20)

[0207]

[0208] The synthesis method was the same as that in Example 19, except that 5,7-dihydroxy-3-iodo-2-methyl-4H-chromen-4-one was used instead of 5,7-dihydroxy-3-iodo-4H-chromen-4-one. 1 H NMR(500MHz,CDCl3)δ7.53–7.48(m,1H),7.41–7.35(m,1H),6.47(d,J=1.8Hz,1H),6.20(d,J=1.8Hz,1H),3.76(d,J=0.9Hz,1H),3.68–3.62(m,3H),2.68–2.63(m,3H).MS(ESI):m / z 402.1[(M+H) + .

[0209] Example 21 Synthesis of 3-(4-chloro-3-((4-methylpiperazin-1-yl)methyl)phenyl)-7-hydroxy-4H-chromen-4-one (Compound 21)

[0210]

[0211] The synthesis method was the same as that in Example 5, except that methylpiperazine was used instead of dimethylamine. 1 H NMR(600MHz,DMSO-d6)δ8.36(s,1H),7.91(d,J=10.9Hz,1H),7.68(s,1H),7.47(s,2H),6.86(d,J=8.9Hz,1H),6.75(s,1H),3.57(s,2H),2.50–2.22(m,8H),2.15(s,3H).MS(ESI):m / z 385.1[(M+H) + .

[0212] Example 21 Synthesis of 3-(4-chloro-3-((4-ethylpiperazin-1-yl)methyl)phenyl)-7-hydroxy-4H-chromen-4-one (Compound 21)

[0213]

[0214] The synthesis method was the same as that in Example 5, except that ethylpiperazine was used instead of dimethylamine. 11H NMR (600 MHz, DMSO-d6) δ 11.02 (s, 1H), 8.42 (s, 1H), 7.97 (d, J = 10.4 Hz, 1H), 7.68 (s, 1H), 7.48 (t, J = 6.5 Hz, 2H), 6.95 (d, J = 8.7 Hz, 1H), 6.87 (s, 1H), 3.58 (s, 2H), 2.50–2.34 (m, 8H), 2.30 (q, J = 6.8 Hz, 3H), 0.98 (t, J = 7.3 Hz, 3H). MS (ESI): m / z 399.1 [(M + H) + .

[0215] (II) Examples of Biological Activity Detection

[0216] Evaluation of the Druggability of Preferred Active Compounds

[0217] 1. 3βHSD1 Inhibition Test at the Cellular Level

[0218] Main Experimental Materials:

[0219] LNCaP cells were purchased from the American Type Culture Collection (Manassas, VA). VCaP was generously provided by Dr. Qin Jun of the Shanghai Institute of Health Sciences, Chinese Academy of Sciences. The cells were cultured at 37 °C in a medium containing 10% fetal bovine serum. The cell lines were genotyped by Hybribio (Guangzhou, China) and tested for mycoplasma contamination and found to be free of mycoplasma before being used in the experiment.

[0220] 3βHSD1 Inhibition Activity Test:

[0221] Prostate cancer cells were seeded in 24-well plates; after 24 hours, the medium was changed and the specified concentration of the active compound and 3 H]-labeled DHEA were added; after culturing the cells in a 37 °C, 5% CO2 incubator for a period of time, the cell supernatant was collected; and DHEA and its metabolites were detected by HPLC and compared with the inhibitory effect of the existing inhibitor (biochanin A).

[0222] Results:

[0223] In this invention, the positive drug biochanin A (BCA) was used as a reference and purchased from MCE. The inhibitory results of the representative active compound on 3βHSD1 are shown in Table 3-6. (DHEA is a substrate of 3βHSD1; the higher the percentage of residual DHEA, the stronger the inhibitory effect of the compound on 3βHSD1 activity)

[0224] Table 3

[0225]

[0226] Table 4

[0227]

[0228] Table 5

[0229]

[0230] Table 6

[0231]

[0232] Note: All data are from three independent experiments and the average values are taken.

[0233] Conclusion: As can be seen from the results in Tables 3 - 6, the compounds of Examples 1 - 22 (especially Example 7) of the present invention have good inhibitory activity against 3βHSD1, and are highly efficient 3βHSD1 inhibitors, equivalent to or superior to BCA in activity.

[0234] 2. Pharmacokinetic analysis

[0235] The oral administration dose for rats is 10 mg / kg, and the administration volume is 10 mL / kg. The dissolution condition is a mixed solution of 5% DMSO / 95% HPMC (0.5%).

[0236] The intravenous administration dose for rats is 2 mg / kg, and the administration volume is 5 mL / kg. The dissolution condition is a mixed solution of 5% DMSO, 5% EtOH, 40% PEG300, 50% NaCl (0.9%).

[0237] The body weight of male rats is between 180 - 220 grams. All animals are fasted for 12 hours before dosing. The temperature of the animal room is controlled between 18 - 29 °C, and the relative humidity is 30 - 70%. The temperature and relative humidity are monitored daily. An electronic time-controlled lighting system is used to provide a 12-hour light / 12-hour dark cycle.

[0238] Results:

[0239] Table 7

[0240]

[0241] Note: All data are from three independent experiments and the average values are taken.

[0242] Explanation of the results in Table 7:

[0243] (1) When administered by intravenous injection, the clearance rates of Compound 7 and BCA are comparable.

[0244] (2) When administered orally, the compound 7 has a high plasma exposure, with its C max being 532 ng / mL and AUC being 458 h*ng / mL, while the oral plasma exposure of BCA is below the limit of quantification (LLOQ = 10 ng / mL), and relevant pharmacokinetic parameters cannot be calculated.

[0245] 3. In vivo efficacy evaluation

[0246] Main experimental materials:

[0247] This experiment was conducted with the approval of the Institutional Committee for Animal Protection and Use of the Center for Excellence in Molecular Cell Science, Chinese Academy of Sciences. Six-week-old male B-NDG mice (B: Biocytogen; N: NOD background; D: DNAPK[Prkdc]null; G: IL2rgknockout) were purchased from Beijing Biocytogen. The dissolution condition was a mixed solution of 5% DMSO, 5% DMA, 12% castor oil, and 78% corn oil. The dosing doses were 10 mg / kg and 30 mg / kg, respectively.

[0248] First, mix PBS containing 1×10 7 cells with Matrigel to form a mixed solution. Then, inject 100 μL of the mixed solution into the right axilla of each mouse. Next, regularly observe the volume of the ectopic tumor in the mice until the volume reaches 100 mm 3 (the calculation method is length × width × width × 0.5). When the volume of the ectopic tumor reaches the specified size, perform castration surgery on the mice and implant DHEA sustained-release tablets at the back of the necks of the mice. Continue to regularly observe the mice after the surgery until the volume of the ectopic tumor reaches 100 mm 3 again.

[0249] Before grouping the mice, group them according to the random grouping method. After the mice are grouped, start the dosing treatment. Every two days, measure and record the tumor volume and body weight of the mice by the same person, and administer the drug to the mice every day. After the experiment, euthanize the animals with carbon dioxide and collect the ectopic tumors for further analysis. Finally, perform a statistical significance analysis on the inter-group differences between the control group and the experimental group by T-test.

[0250] Results:

[0251] Figure 1 and Figure 2 show that at the same dose, oral administration of Example 7 has a better ability to inhibit the growth of tumors in mice than oral and intraperitoneal injection of BCA, and there is no obvious toxicity.

[0252] Conclusion: Example 7 has better oral absorption, superior metabolic stability and superior in vivo efficacy compared to BCA.

[0253] The above-described embodiments have detailed the technical solutions and beneficial effects of the present invention. It should be understood that the above are only specific embodiments of the present invention and are not used to limit the present invention. Any modifications, supplements, equivalent replacements, etc. made within the scope of the principles of the present invention should be included within the protection scope of the present invention.

Claims

1. A 7-hydroxyflavonoid compound, or a pharmaceutically acceptable salt thereof, characterized in that: The structure of the 7-hydroxyflavonoid compound is shown in formula (I), Where: R1 is H or C1-C5 alkyl; R2 is H or OH; R3 is halogen; R4 is -X-linker-R'; the -X-linker-R' is selected from Table 1: Table 1 2. The 7-hydroxyflavonoid compound according to claim 1, or a pharmaceutically acceptable salt thereof, characterized in that: The 7-hydroxyflavonoid compound is selected from Table 2: Table 2 3. A pharmaceutical composition, characterized in that The invention comprises the 7-hydroxyflavonoid compound or a pharmaceutically acceptable salt thereof as claimed in claim 1 or 2, and a pharmaceutically acceptable excipient.

4. A 3βHSD1 inhibitor, characterized in that Contains the 7-hydroxyflavonoid compound or a pharmaceutically acceptable salt thereof as claimed in claim 1 or 2.

5. Use of the 7-hydroxyflavonoid compound according to claim 1 or 2, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof in the preparation of a drug for treating diseases associated with 3βHSD1.

6. The use according to claim 5, characterized in that: The disease associated with 3βHSD1 is prostate cancer.

Images