Application of a locally acting androgen receptor antagonist

Abstract

This invention discloses the application of a locally acting androgen receptor antagonist, namely, 4-(3-(4-cyano-3-(trifluoromethyl)phenyl)-5,5-dimethyl-4-oxo-2-thioimidazolidine-1-yl)-2-fluorobenzoate tert-butyl ester as shown in Formula I, or a pharmaceutical composition thereof, in the preparation of a medicament for the prevention or treatment of diseases related to androgen abnormalities. 4-(3-(4-cyano-3-(trifluoromethyl)phenyl)-5,5-dimethyl-4-oxo-2-thioimidazolidine-1-yl)-2-fluorobenzoate tert-butyl ester exhibits good skin stability and slow metabolism, thus exerting an anti-androgenic alopecia effect. However, it has extremely poor stability in plasma and can be directionally and rapidly metabolized into the inactive metabolite enzalutamide, avoiding the systemic androgen antagonistic side effects caused by systemic distribution.

Description

Technical Field

[0001] This invention relates to the pharmaceutical field, specifically to the use of a locally acting androgen receptor antagonist, 4-(3-(4-cyano-3-(trifluoromethyl)phenyl)-5,5-dimethyl-4-oxo-2-thioimidazolidine-1-yl)-2-fluorobenzoate tert-butyl ester, or a pharmaceutical composition thereof, in the preparation of a medicament for treating androgen receptor-related diseases or conditions. Background Technology

[0002] Androgenetic alopecia (AGA), also known as seborrheic alopecia or premature baldness, is divided into male-pattern and female-pattern alopecia and is the most common non-scarring hair loss in clinical practice. AGA mainly occurs in late adolescence and is characterized by interfollicular atrophy. In male patients, the main manifestations are a receding hairline or thinning and thinning of hair on the crown; in female patients, the main manifestations are thinning and thinning of hair on the crown, and a few patients experience diffuse hair thinning without affecting the hairline. Although AGA does not affect the patient's physical health, it affects their mental health and quality of life, and in severe cases, it can lead to depression, anxiety, strained interpersonal relationships, and sensitivity.

[0003] Androgenetic alopecia (AGA) is an androgen-dependent polygenic hereditary disease. Since Hamilton Norwood first proposed its association with androgens in 1942, research on AGA by scholars both domestically and internationally has primarily focused on androgens and their metabolic enzymes in the scalp. However, the exact cause and pathogenesis of this disease remain unclear. In recent years, the incidence of AGA has been increasing and is showing a trend towards affecting younger people. Therefore, treatment is urgently needed.

[0004] In men, androgens are primarily synthesized in the testes, including testosterone, dihydrotestosterone (DHT), androstenedione (DHEA), and dehydroepiandrosterone (ASD). In women, androgens mainly originate from the adrenal cortex. Testosterone and androstenedione can be reduced to DHT by 5α-reductase, and DHT has a 5+ times greater affinity for the androgen receptor (AR) than testosterone. There are two isoenzymes of 5α-reductase in the human body: 5α-reductase I and 5α-reductase II. 5α-reductase I is mainly distributed in the sebaceous glands and liver; 5α-reductase II is mainly distributed in the hair follicles of the scalp, epididymis, vas deferens, seminal vesicles, prostate, and fetal genital skin. Studies have shown that scalp hair follicles are one of the target organs of androgens. Hair biopsy specimens from male AGA patients show that 5α-reductase activity in the bald area of ​​the crown is significantly higher than in the non-bald area of ​​the occipital region. Other studies have also shown that high AR expression in hair follicles in the bald area of ​​the crown accelerates hair loss in that area. DHT has a strong affinity for AR. After binding with AR, it enters the cell nucleus, inhibits adenylate cyclase in hair follicles, reduces the synthesis of cyclic nucleotides and related proteins, and causes hair to enter the resting phase prematurely. It also inhibits hair follicle growth, shrinks hair follicles, stimulates sebaceous gland proliferation and sebum secretion, further reducing the size of hair follicles and causing them to gradually disappear, thus promoting hair loss.

[0005] Currently, there are many patients with acute gastrointestinal anemia (AGA), but there are still very few drugs specifically for AGA. Only finasteride and minoxidil are currently approved for marketing and use in the treatment of AGA; most other drugs are used off-label. Existing AGA therapies are effective, but they also bring many side effects, such as male sexual dysfunction caused by finasteride, and propylene glycol allergy and orthostatic hypotension caused by minoxidil. These side effects limit the clinical use of these drugs. Therefore, there is an urgent clinical need for AGA drugs that are locally effective, avoid systemic distribution, have fewer side effects, and are easy to use. Summary of the Invention

[0006] Purpose of the invention: The technical problem to be solved by the present invention is to address the shortcomings of the prior art by providing the application of tert-butyl 4-(3-(4-cyano-3-(trifluoromethyl)phenyl)-5,5-dimethyl-4-oxo-2-thioimidazolidine-1-yl)-2-fluorobenzoate (Formula I) or a pharmaceutically acceptable salt thereof.

[0007] The technical problem to be further solved by the present invention is to provide a method for preventing diseases or conditions mediated by androgens or androgen receptor activity in mammals, reducing the progression of diseases or conditions mediated by androgens or androgen receptor activity in mammals, and treating diseases or conditions mediated by androgens or androgen receptors in mammals.

[0008] To address the first technical problem mentioned above, the present invention discloses the use of tert-butyl 4-(3-(4-cyano-3-(trifluoromethyl)phenyl)-5,5-dimethyl-4-oxo-2-thioimidazolidine-1-yl)-2-fluorobenzoate, as shown in Formula I, or a pharmaceutical composition thereof, in the preparation of a medicament for the prevention or treatment of diseases related to androgen abnormalities.

[0009] .

[0010] In some embodiments, the pharmaceutical composition comprises (i) tert-butyl 4-(3-(4-cyano-3-(trifluoromethyl)phenyl)-5,5-dimethyl-4-oxo-2-thioimidazolidine-1-yl)-2-fluorobenzoate, and (ii) a pharmaceutically acceptable carrier, diluent, or excipient.

[0011] In some implementations, the prevention or treatment of androgen-related diseases is to prevent androgen or androgen receptor-mediated diseases or conditions, reduce the progression of androgen or androgen receptor-mediated diseases or conditions, or treat androgen or androgen receptor-mediated diseases or conditions.

[0012] In some embodiments, the androgen or androgen receptor-mediated disease or condition is selected from prostate cancer, acne, or hair loss; in some embodiments, the androgen or androgen receptor-mediated disease or condition is acne; in some embodiments, the androgen or androgen receptor-mediated disease or condition is hair loss; in some embodiments, the androgen or androgen receptor-mediated disease or condition is androgenetic alopecia.

[0013] To address the second technical problem mentioned above, the present invention discloses a method for preventing diseases or conditions mediated by androgens or androgen receptor activity in mammals, reducing the progression of diseases or conditions mediated by androgens or androgen receptor activity in mammals, and treating diseases or conditions mediated by androgens or androgen receptors in mammals, comprising administering a therapeutically effective amount of a compound of formula (I) or a pharmaceutical composition thereof to a mammal in need of the treatment.

[0014] In some implementations, the mammal is a human.

[0015] Unless otherwise stated, the following terms as used in this application shall have the following meanings. A particular term should not be considered uncertain or unclear unless specifically defined, but should be understood in accordance with its ordinary meaning in the art. When a trade name appears herein, it is intended to refer to the corresponding product or its active ingredient.

[0016] The term "treatment" means administering the compound or preparation described in this application to improve or eliminate a disease or one or more symptoms related to said disease, and includes:

[0017] i. To suppress a disease or disease state, that is, to curb its development;

[0018] ii. To alleviate a disease or disease state, even if the disease or disease state subsides.

[0019] The term “prevention” means administering the compound or formulation described in this application to prevent a disease or one or more symptoms associated with the disease, and includes: preventing the occurrence of a disease or disease state in mammals, particularly when such mammals are susceptible to the disease state but have not yet been diagnosed with the disease state.

[0020] The term "therapeutic effective amount" means the amount of the compound of this application used to treat or prevent a particular disease, condition, or disorder; (ii) to reduce, improve, or eliminate one or more symptoms of a particular disease, condition, or disorder; or (iii) to prevent or delay the onset of one or more symptoms of a particular disease, condition, or disorder described herein. The amount of the compound of this application constituting a "therapeutic effective amount" varies depending on the compound, the disease state and its severity, the route of administration, and the age of the mammal to be treated, but may routinely be determined by a person skilled in the art based on their own knowledge and the present disclosure.

[0021] The term "pharmaceutical acceptable" refers to compounds, materials, compositions, and / or dosage forms that, within the bounds of reliable medical judgment, are suitable for use in contact with human and animal tissues without excessive toxicity, irritation, allergic reactions, or other problems or complications, in proportion to a reasonable benefit / risk ratio.

[0022] The term "pharmaceutical composition" refers to a mixture of one or more compounds of this application or their salts with pharmaceutically acceptable excipients. The purpose of a pharmaceutical composition is to facilitate the administration of the compounds of this application to an organism.

[0023] The term "pharmaceuticalally acceptable excipient" refers to excipients that do not cause significant irritation to the organism and do not impair the biological activity and properties of the active compound. Suitable excipients are well known to those skilled in the art, such as carbohydrates, waxes, water-soluble and / or water-swellable polymers, hydrophilic or hydrophobic materials, gelatin, oils, solvents, water, etc.

[0024] The word “comprise” or “comprise” and its English variants such as comprises or comprising should be understood in an open, non-exclusive sense, meaning “including but not limited to”.

[0025] The term "pharmaceuticalally acceptable excipient" refers to excipients that do not cause significant irritation to the organism and do not impair the biological activity and properties of the active compound. Suitable excipients are well known to those skilled in the art, such as carbohydrates, waxes, water-soluble and / or water-swellable polymers, hydrophilic or hydrophobic materials, gelatin, oils, solvents, water, etc.

[0026] The compounds of this invention may contain non-natural proportions of atomic isotopes on one or more atoms constituting the compound. For example, the compounds may be labeled with radioactive isotopes such as tritium (3H), iodine-125 (125I), or C-14 (14C). As another example, deuterated drugs may be formed by replacing hydrogen with deuterium, such as d3-methyl, which indicates that all three hydrogen atoms on the methyl group are replaced by deuterium atoms. The bond between deuterium and carbon is stronger than that between ordinary hydrogen and carbon. Compared to undeuterated drugs, deuterated drugs have advantages such as reduced toxicity, increased drug stability, enhanced efficacy, and prolonged drug biological half-life. All variations in the isotopic composition of the compounds of this invention, regardless of radioactivity, are included within the scope of this invention. "Optional" or "optionally" refers to events or conditions described subsequently that may occur but are not necessary, and the description includes both the occurrence and non-occurrence of said events or conditions.

[0027] This application also includes compounds of this application that are identical to those described herein, but with one or more atoms replaced by isotopes of atomic weights or mass numbers different from those commonly found in nature. Examples of isotopes that can be incorporated into compounds of this application include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, iodine, and chlorine, such as 2H, 3H, 11C, 13C, 14C, 13N, 15N, 15O, 17O, 18O, 31P, 32P, 35S, 18F, 123I, 125I, and 36Cl, respectively.

[0028] Certain isotopically labeled compounds of this application (e.g., those labeled with 3H and 14C) can be used in the analysis of compound and / or substrate tissue distribution. Tritium (i.e., 3H) and carbon-14 (i.e., 14C) isotopes are particularly preferred due to their ease of preparation and detectability. Positron emission isotopes, such as 15O, 13N, 11C, and 18F, can be used in positron emission tomography (PET) studies to determine substrate occupancy. Isotopically labeled compounds of this application can generally be prepared by replacing unlabeled reagents with isotopically labeled reagents using a procedure similar to those disclosed in the schemes and / or examples below.

[0029] In addition, heavier isotopes (such as deuterium) are used. 2H)) substitution can provide certain therapeutic advantages resulting from higher metabolic stability (e.g., increased in vivo half-life or reduced dose requirement), and may therefore be preferred in certain situations, wherein deuterium substitution can be partial or complete, and partial deuterium substitution means that at least one hydrogen is substituted by at least one deuterium, and all such compounds are included within the scope of this application.

[0030] The pharmaceutical compositions of this application can be prepared by combining the compounds of this application with suitable pharmaceutically acceptable excipients, for example, in solid, semi-solid, liquid or gaseous formulations, such as tablets, pills, capsules, powders, granules, ointments, emulsions, suspensions, suppositories, injections, inhalers, gels, microspheres and aerosols.

[0031] Typical routes of administration for the compounds of this application or their pharmaceutically acceptable salts or pharmaceutical compositions thereof include, but are not limited to, oral, rectal, topical, inhalation, parenteral, sublingual, vaginal, intranasal, intraocular, intraperitoneal, intramuscular, subcutaneous, and intravenous administration.

[0032] The pharmaceutical composition of this application can be manufactured using methods well known in the art, such as conventional mixing, dissolving, granulation, sugar-coated pill making, grinding, emulsification, freeze drying, etc.

[0033] The therapeutic dose of the compound may be determined based on factors such as the specific therapeutic purpose, the method of administration, the patient's health and condition, and the prescribing physician's judgment. The proportion or concentration of the compound in the pharmaceutical composition may not be fixed and depends on various factors, including dosage, chemical properties (e.g., hydrophobicity), and route of administration. For example, the compound may be provided in a physiologically buffered aqueous solution containing about 0.1–10% w / v of the compound for parenteral administration. Some typical dosage ranges are from about 1 μg / kg to about 1 g / kg body weight / day. In some embodiments, the dosage range is from about 0.01 mg / kg to about 100 mg / kg body weight / day. The dosage is likely to depend on variables such as the type and severity of the disease or condition, the general health status of the specific patient, the relative biological potency of the selected compound, the excipient formulation, and the route of administration. The effective dose can be obtained by extrapolation from dose-response curves derived from in vitro or animal model testing systems.

[0034] The compounds of this application can be prepared by a variety of synthetic methods known to those skilled in the art, including the specific embodiments listed below, embodiments formed by combining them with other chemical synthetic methods, and equivalent substitutions known to those skilled in the art. Preferred embodiments include, but are not limited to, the embodiments of this application.

[0035] Beneficial effects: Compared with the prior art, the present invention has the following advantages:

[0036] The tert-butyl 4-(3-(4-cyano-3-(trifluoromethyl)phenyl)-5,5-dimethyl-4-oxo-2-thioimidazolidine-1-yl)-2-fluorobenzoate provided by this invention is a locally acting androgen antagonist. Simultaneously, this compound exhibits good skin stability and slow metabolism, thus exerting an anti-androgenic alopecia effect. Furthermore, it has extremely poor stability in plasma, allowing for targeted and rapid metabolism into the inactive metabolite enzalutamide (CAS: 1242137-15-0), thereby avoiding the systemic androgen antagonistic side effects caused by systemic distribution. In addition, the compound provided by this invention also showed a hair growth-promoting effect in a mouse alopecia model when administered transdermally. Attached Figure Description

[0037] The present invention will be further described in detail below with reference to the accompanying drawings and specific embodiments, and the advantages of the present invention in the above and / or other aspects will become clearer.

[0038] Figure 1 Hair growth was scored for C57BL / 6 mice. Detailed Implementation

[0039] For clarity, the present invention is further illustrated by examples, but these examples are not intended to limit the scope of this application. It will be apparent to those skilled in the art that various changes and modifications can be made to specific embodiments of the invention without departing from the spirit and scope thereof. All reagents used in this application are commercially available and can be used without further purification.

[0040] Unless otherwise specified, the experimental methods described in the following examples are conventional methods; the reagents and materials described are commercially available unless otherwise specified.

[0041] Example 1: Preparation of tert-butyl 4-(3-(4-cyano-3-(trifluoromethyl)phenyl)-5,5-dimethyl-4-oxo-2-thioimidazolidine-1-yl)-2-fluorobenzoate (Compound 1)

[0042]

[0043] Step 1: Synthesis of Compound 1-1

[0044] In a 250 mL three-necked flask, 150 mL of N,N-dimethylformamide and 15 mL of water were added. Then, 10 g of 2-fluoro-4-bromobenzoic acid, 7.25 g of 2-methylalanine, 15.8 g of potassium carbonate, 1.7 g of cuprous iodide, 1.2 g of 2-acetylcyclohexanone, and 0.8 g of triethylamine were added sequentially. The mixture was heated and stirred at 110 °C for 6 h. The mixture was then diluted with 500 mL of water, and the pH was adjusted to 3-4 with citric acid. The mixture was extracted three times with ethyl acetate, and the combined organic phases were washed three times with saturated brine. The organic phase was dried over anhydrous sodium sulfate and concentrated. 50 mL of dichloromethane was added to the resulting crude solid, and the mixture was stirred at room temperature for 1 h. The mixture was then filtered, and the filter cake was washed with dichloromethane. The filter cake was collected and dried to give 9.3 g of compound 1-1.

[0045] Step 2: Synthesis of Compounds 1-2

[0046] 100 mL of methanol and 13 g of compound 1-1 prepared in step 1 were added to a 250 mL three-necked flask. After stirring and dissolving, the mixture was cooled in an ice bath. 25.6 g of thionyl chloride was slowly added dropwise under ice bath conditions. After the addition was complete, the mixture was heated to reflux. After the reaction was complete, the heating was turned off, and the mixture was allowed to cool to room temperature. 50 mL of toluene was added, and the mixture was concentrated under reduced pressure. The resulting residue was redissolved in 300 mL of ethyl acetate, washed once with saturated brine, and then washed three times with saturated sodium bicarbonate solution. The organic phase was dried over anhydrous sodium sulfate and concentrated to give 11.3 g of compound 1-2.

[0047] Step 3: Synthesis of compounds 1-3

[0048] In a 50 mL three-necked flask, 5 mL of dimethyl sulfoxide, 10 mL of isopropyl acetate, 5 g of compounds 1-2 prepared in step 2 above, and 8.9 g of 4-isothiocyano-2-(trifluoromethyl)benzonitrile were added sequentially. The mixture was heated and stirred at 90 °C for 20 h. Heating was turned off, and the mixture was allowed to cool naturally to room temperature. The reaction solution was diluted with 50 mL of water and extracted three times with ethyl acetate. The combined organic phases were washed three times with saturated brine, dried over anhydrous sodium sulfate, and concentrated. The resulting oily substance was added to 35 mL of methanol and stirred at room temperature for 2 h. The mixture was filtered, the filter cake was collected and dried, yielding 5.8 g of compounds 1-3.

[0049] Step 4: Synthesis of compounds 1-4

[0050] 3.8 g of compounds 1-3 prepared in step 3 above were dissolved in 15 mL of tetrahydrofuran, and 15 mL of an aqueous solution containing 0.68 g of sodium hydroxide was added. The mixture was stirred overnight at room temperature. After the reaction was complete, 50 mL of water was added to dilute the solution, and the pH was adjusted to 3-4 with 1 M dilute hydrochloric acid. The mixture was then extracted three times with ethyl acetate. The combined organic phases were dried over anhydrous sodium sulfate, and the organic phase was concentrated to give 3.0 g of compounds 1-4.

[0051] Step 5: Synthesis of Compound 1

[0052] In a 25 mL round-bottom flask, add 0.3 g of compounds 1-4 prepared in step 4 above, 5 mL of tert-butanol, 0.29 g of di-tert-butyl dicarbonate, and 0.14 g of triethylamine, and stir at 30 °C for 6 h. Concentrate under reduced pressure, redissolve the resulting oil in 30 mL of ethyl acetate, and wash three times with saturated brine. Dry the organic phase with anhydrous sodium sulfate and concentrate. The crude product is purified by column chromatography to give 0.16 g of a white solid.

[0053] 1 H NMR (400 MHz, Chloroform-d) δ 8.07 (t, J = 8.1 Hz, 1H), 8.04 –7.95 (m, 2H), 7.85 (dd, J = 8.3, 1.8 Hz, 1H), 7.22 – 7.11 (m, 2H), 1.64 –1.63 (m, 15H).

[0054] MS (ESI, [M+H) + m / z: 508.4.

[0055] Example 2: Preparation of sec-butyl 4-(3-(4-cyano-3-(trifluoromethyl)phenyl)-5,5-dimethyl-4-oxo-2-thioimidazolin-1-yl)-2-fluorobenzoate (Compound 2)

[0056]

[0057] In a 25 mL round-bottom flask, 0.2 g of compounds 1-4 obtained in step 4 of Example 1, 5 mL of N,N-dimethylformamide, 0.25 g of 2-iodobutane, and 0.15 g of potassium carbonate were added sequentially. The mixture was heated at 60 °C overnight. After the reaction was complete, the heating was removed, and the mixture was cooled to room temperature. 25 mL of water was added to dilute the reaction solution. The mixture was extracted with ethyl acetate (15 mL × 3), and the organic phases were combined and washed with saturated brine (15 mL × 3). The organic phase was dried over anhydrous sodium sulfate and concentrated. The crude product was purified by column chromatography to give 0.34 g of a white solid.

[0058] 1H NMR (400 MHz, Chloroform-d) δ 8.13 (t, J = 8.0 Hz, 1H), 8.02 (d, J= 8.3 Hz, 1H), 7.97 (d, J = 2.0 Hz, 1H), 7.85 (dd, J = 8.3, 2.1 Hz, 1H), 7.25– 7.11 (m, 2H), 5.25 – 5.11 (m, 1H), 1.85 – 1.67 (m, 2H), 1.64 (s, 6H), 1.39 (d, J = 6.3 Hz, 3H), 1.02 (t, J = 7.5 Hz, 3H).

[0059] MS (ESI, [M+H) + m / z: 508.4.

[0060] Application Experiment Example 1: Antagonistic Activity of Compounds on Androgen Receptors

[0061] 1) All test compounds were dissolved in DMSO to prepare 15 mM solutions, and then serially diluted 3-fold 10 times to prepare 15 mM, 5 mM, 1.6667 mM, 0.5576 mM, 0.1852 mM, 0.0617 mM, 0.0206 mM, 0.0069 mM, 0.0023 mM and 0.0008 mM solutions.

[0062] 2) Culture HEK293T cells (ATCC, CRL-3216) according to ATCC recommendations (https: / / www.atcc.org / ). Assay the cells during the exponential growth phase.

[0063] 3) Remove the culture medium from the culture dish, rinse the cells with phosphate buffer, add trypsin solution to the culture dish to separate the cells. Wash the cells once with complete culture medium.

[0064] 4) Wash the cells twice with phosphate buffer to remove phenol red and resuspend them in the culture medium to the appropriate concentration.

[0065] 5) 6*10 6 HEK293T cells were seeded in 100 mm culture dishes (only cells with a survival rate greater than 90% were used for analysis) and cultured at 37°C and 5% CO2 for 16 h.

[0066] 6) Transfect the plasmids (pGL4.36 [luc2P / MMTV / Hygro], Promega, E1360; pBIND-AR Vector, Kanglong Chemical; Lipofectamine® LTX & Plus Reagent, Invitrogen, 15338-100) into HEK293T cells and culture them at 37℃ and 5% CO2 for 5-6 h.

[0067] 7) Use a non-contact nano-level ultrasonic pipetting system (Labcyte, Echo550) to transfer 50 nml of compound dilution to a 384-well analytical plate.

[0068] 8) HEK293T cells were seeded into 384-well analytical plates at a final concentration of 15,000 cells / well and 1 nM DHT (MCE, HY-A0120) (25 μl).

[0069] 9) The cells were cultured at 37°C and 5% CO2 for 18-20 hours.

[0070] 10) Add 25 μl of luciferase assay reagent (britelite plus, PerkinElmer, 6066769) to each well of the 384-well assay plate.

[0071] 11) Record the luminescence value using an ELISA reader (Envision 2105, PerkinElmer) and calculate the IC50. 50 .

[0072] Table 1. Antagonistic activity of compounds against androgen receptors

[0073]

[0074] The results indicate that tert-butyl 4-(3-(4-cyano-3-(trifluoromethyl)phenyl)-5,5-dimethyl-4-oxo-2-thioimidazolidine-1-yl)-2-fluorobenzoate has androgen receptor antagonistic activity.

[0075] Application Experiment Example 2: Chemical Stability

[0076] Preparation of warm incubation solution: Add 10 mL of 0.02 M phosphate buffer (PBS) pH=7.4 to 9.8 mL of DMSO, cool in an ice bath, and vortex for 30 s to prepare the warm incubation solution.

[0077] The test compound was prepared as a 10 mM stock solution using DMSO. The test compound was then diluted to 2 mM with DMSO. 15 μL of the 2 mM test compound solution was added to 1485 μL of warm incubation buffer, vortexed for 30 s, and then allowed to stand at room temperature. Samples of 300 μL were taken at 0 h, 2 h, 4 h, 6 h, and 8 h, centrifuged at 14400 rpm for 3 min, and 200 μL of the supernatant was collected for HPLC analysis. The concentration was calculated and determined according to formula T. 1 / 2 = -0.693 / k to calculate the half-life.

[0078] Table 2 Chemical Stability

[0079]

[0080] a No significant degradation was observed after 8 hours, making it impossible to calculate the half-life.

[0081] The results showed that the compounds of this invention have good stability at pH 7.4.

[0082] Application Experiment Example 3: Rat Plasma Stability

[0083] The test compound was prepared as a 10 mM stock solution using DMSO. The test compound was diluted to a 2 mM working solution using DMSO. 10 μL of the 2 mM working solution was added to 990 μL of rat plasma, vortexed for 5 s to prepare a reaction system with an initial concentration of 20 μM. The system was incubated at 37°C, and 80 μL of plasma samples were collected at 0 min, 10 min, 20 min, 30 min, 1 h, 2 h, 4 h, and 6 h. 240 μL of methanol was added to the plasma sample, vortexed for 3 min, centrifuged at 14400 rpm for 3 min, and 200 μL of the supernatant was collected for HPLC analysis. The concentration was calculated and determined according to formula T. 1 / 2 The half-life was calculated using -0.693 / k, and the results are shown in Table 3.

[0084] Table 3. Plasma half-life and degradation products in rats

[0085]

[0086] a No significant degradation was observed after 6 hours, making it impossible to calculate the half-life.

[0087] The results showed that tert-butyl 4-(3-(4-cyano-3-(trifluoromethyl)phenyl)-5,5-dimethyl-4-oxo-2-thioxoimidazolidin-1-yl)-2-fluorobenzoate was extremely unstable in plasma, with a plasma half-life of less than 5 min, and the metabolite was enzalutamide acid. Therefore, the compound of the present invention was rapidly inactivated after entering the blood and could not be distributed throughout the body, thus unable to cause systemic androgen antagonism, and would not cause side effects such as decreased libido, reduced sperm production, and ED. It could effectively solve the problem that existing drugs such as finasteride and other drugs were systemically distributed, resulting in systemic androgen inhibition and causing side effects such as decreased libido, reduced sperm production, and ED.

[0088] Application Experimental Example 4: Stability of 10% Skin Homogenate of Mice

[0089] 1) The test compound was prepared into a 10 mM stock solution with DMSO. Take 20 μL of the test compound stock solution and add it to 80 μL of DMSO, vortex for 10 s to prepare a 2 mM working solution;

[0090] 2) The back of C57BL / 6 male mice (license number SCXK (Yu) 2020-0005) was depilated with depilatory cream;

[0091] 3) Under anesthesia, the back skin was dissected, blood vessels, subcutaneous tissues and fats were removed, and rinsed clean with pre-cooled physiological saline;

[0092] 4) Take 2.3 g of skin, add it to 23 mL of pre-cooled physiological saline, and homogenize it充分 (homogenize for 15 s each time and pause for 10 s) under ice bath conditions, centrifuge at 5000 rpm for 5 min, aspirate the supernatant, which is the skin tissue homogenate, and place it on ice for use;

[0093] 5) Take 990 μL of the skin tissue homogenate, add 10 μL of the 2 mM test compound working solution, vortex for 5 s, and place it in a 37 °C water bath for incubation. Samples of 80 μL were taken at 0 min, 5 min, 10 min, 15 min, 30 min, 1 h and 2 h respectively, and 240 μL of methanol was added;

[0094] 6) After vortexing for 3 min, centrifuge at 13000 rpm for 5 min;

[0095] 7) Aspirate 200 μL of the supernatant, analyze it by HPLC injection, calculate the concentration, and calculate the half-life according to the formula T 1 / 2 = -0.693 / k. The results are shown in Table 4.

[0096] Table 4 Stability and Degradation Products of 10% Skin Homogenate of Mice

[0097]

[0098] a No significant degradation was observed after 2 hours, making it impossible to calculate the half-life.

[0099] 4-(3-(4-cyano-3-(trifluoromethyl)phenyl)-5,5-dimethyl-4-oxo-2-thioimidazolidine-1-yl)-2-fluorobenzoate tert-butyl ester has a long half-life and good stability in skin homogenates. It is slowly degraded into the inactive metabolite enzalutamide.

[0100] Application Experiment Example 5: Hair Growth Model in C57BL / 6 Mice

[0101] Male C57BL / 6 mice aged 6 to 7 weeks were purchased and housed in groups under standard conditions for one week. The lower back was shaved with an electric shaver under mild anesthesia, followed by hair removal cream, exposing approximately 2 cm × 2 cm of skin. The next day, mice with intact back skin and pinkish skin were randomly assigned to four groups: a blank control group, a model group, a treatment group (Example 1 group), and a positive control group (minoxidil group), with 6 mice in each group.

[0102] Blank control group: daily subcutaneous injection of a DMSO:glycerin = 1:1 mixture, and application of 20 µL of acetone to the hair removal area twice daily;

[0103] Model group: Daily subcutaneous injection of dihydrotestosterone solution (6 mg / mL, solvent is a mixture of DMSO:glycerol=1:1), dose is 30 mg / kg, and acetone 20 µL is applied to the hair removal site twice daily;

[0104] Example 1 group: Daily subcutaneous injection of dihydrotestosterone solution (6 mg / mL, solvent is a mixture of DMSO:glycerol=1:1), dose is 30 mg / kg, and 20 µL of 5% (g / mL) acetone solution of Example 1 is applied twice daily to the hair removal site.

[0105] Positive control group (minoxidil group): Daily subcutaneous injection of dihydrotestosterone solution (6 mg / mL, solvent is a mixture of DMSO:glycerol=1:1), dose is 30 mg / kg, and 20 µL of 5% (g / mL) minoxidil in acetone solution is applied twice daily to the hair removal site.

[0106] Record hair growth scores every two days.

[0107] The scoring system used for mouse hair growth is as follows:

[0108] a) 0: No hair growth, pink skin tone;

[0109] b)1: The skin color in the shaved area changes from pink to gray, and there is no visible hair growth, indicating that the hair growth phase has started.

[0110] c)2: The skin in the shaved area is black with tiny hairs;

[0111] d)3: Short black hair grows in the shaved area;

[0112] e)4: The hair in the shaved area is almost indistinguishable from the surrounding area.

[0113] Hair scores are shown in Table 5 and Figure 1 As shown. The results showed that after 20 days of treatment, Example 1 had the effect of promoting hair growth.

[0114] Table 5 Hair Scoring

[0115]

[0116] This invention provides a concept and method for the application of a locally acting androgen receptor antagonist. Many methods and approaches exist for implementing this technical solution; the above description is merely a preferred embodiment of the invention. It should be noted that those skilled in the art can make various improvements and modifications without departing from the principles of this invention, and these improvements and modifications should also be considered within the scope of protection of this invention. All components not explicitly stated in this embodiment can be implemented using existing technologies.

Claims

1. The use of tert-butyl 4-(3-(4-cyano-3-(trifluoromethyl)phenyl)-5,5-dimethyl-4-oxo-2-thioimidazolidine-1-yl)-2-fluorobenzoate or a pharmaceutical composition thereof, as shown in Formula I, in the preparation of a medicament for the prevention or treatment of diseases related to androgen abnormalities; 。 2. The application according to claim 1, characterized in that, The pharmaceutical composition comprises (i) tert-butyl 4-(3-(4-cyano-3-(trifluoromethyl)phenyl)-5,5-dimethyl-4-oxo-2-thioimidazolidine-1-yl)-2-fluorobenzoate, and (ii) a pharmaceutically acceptable excipient.

3. The application according to claim 1, characterized in that, The pharmaceutical composition comprises (i) tert-butyl 4-(3-(4-cyano-3-(trifluoromethyl)phenyl)-5,5-dimethyl-4-oxo-2-thioimidazolidine-1-yl)-2-fluorobenzoate, and (ii) a pharmaceutically acceptable carrier.

4. The application according to claim 1, characterized in that, The pharmaceutical composition comprises (i) tert-butyl 4-(3-(4-cyano-3-(trifluoromethyl)phenyl)-5,5-dimethyl-4-oxo-2-thioimidazolidine-1-yl)-2-fluorobenzoate, and (ii) a pharmaceutically acceptable diluent.

5. The application according to claim 1, characterized in that, The prevention or treatment of diseases related to androgen abnormalities refers to the prevention or treatment of diseases or conditions mediated by androgens or androgen receptor activity.

6. The application according to claim 5, characterized in that, The diseases or conditions mediated by androgens or androgen receptor activity are selected from prostate cancer, acne, or hair loss.

7. The application according to claim 5, characterized in that, The disease or condition mediated by androgens or androgen receptor activity is acne.

8. The application according to claim 5, characterized in that, The disease or condition mediated by androgens or androgen receptor activity is hair loss.

9. The application according to claim 5, characterized in that, The disease or condition mediated by androgens or androgen receptor activity is androgenetic alopecia.

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