WO2023217028A1 - Hair-growth microneedle patch containing androgen receptor-proteolysis targeting chimeric molecules, preparation method therefor and use thereof - Google Patents

Abstract

Provided is a hair-growth microneedle patch containing androgen receptor-proteolysis targeting chimeric molecules. The hair-growth microneedle patch comprises a sheet-like backing and a soluble microneedle body arranged on the sheet-like backing and containing androgen receptor-proteolysis targeting chimeric molecules. The soluble microneedle body comprises one or more of the androgen receptor-proteolysis targeting chimeric molecules, a pharmaceutically acceptable salt thereof, a stereoisomer thereof, a geometric isomer thereof, a tautomer thereof, an ester thereof, a prodrug thereof, a solvate thereof, a metabolite thereof, a nitrogen oxide thereof, or a deuterated compound thereof. Also provided are a preparation method for the hair-growth microneedle patch containing the androgen receptor-proteolysis targeting chimeric molecules and use thereof in the preparation of a drug for treating androgenetic alopecia. The patch has the advantages of being safe, efficient and convenient to use.

Description

A hair growth microneedle patch containing androgen receptor protein targeting consortium and its preparation method and application Technical field

The invention belongs to the technical field of biomedical materials, and specifically relates to a hair growth microneedle patch containing an androgen receptor protein targeting complex and its preparation method and application.

Background technique

Androgenetic alopecia (AGA) is the most common clinical type of pathological hair loss. Both men and women can suffer from it. It is characterized by progressive miniaturization of hair follicles, manifested by the receding hairline on the forehead, or the gradual thinning and loss of hair on the top, and the progressive reduction of hair density, eventually leading to hair loss, which seriously affects the patient's mental health. The survey shows that the prevalence rate among men and women in my country is about 21.3% and 6.0%, respectively. In recent years, with the rapid development of society, competition has become increasingly fierce, and people's mental stress has increased. The incidence of AGA has been increasing year by year, and is accompanied by a younger trend. Hair loss directly affects personal appearance and social interaction, has a negative impact on the patient's psychology, and seriously affects the patient's quality of life. However, there is still a lack of ideal treatment methods. The existing drug treatments (minoxidil and finasteride) have long courses, many adverse reactions, easy relapse after drug withdrawal, and poor patient compliance. Hair transplantation is also invasive, expensive, and Problems such as unsatisfactory hair density after surgery. Based on the above-mentioned current status of incidence and treatment, starting from the pathogenesis of AGA, exploring safer and more effective treatments is a current research hotspot.

Androgenic alopecia is caused by a combination of factors including androgens. The complex that binds androgens (testosterone, dihydrotestosterone) to androgen receptors inhibits the dephosphorylation of GSK-3β, leading to degradation of β-catenin and downregulation of the Wnt/β-catenin pathway; it also upregulates TGF-β1 and TGF- Paracrine factors such as β2, IL-6 and DKK-1, and changes in the Wnt/β-catenin and TGF-β pathways will lead to prolongation of the hair follicle quiescent phase, delay of the hair follicle cycle, inhibition of epithelial cell proliferation, resulting in hair follicle miniaturization, and ultimately lead to The occurrence of androgenic alopecia. Studies have shown that the expression of androgen receptor (AR) is increased in the hair papilla of hair loss areas in patients with androgenic alopecia, and the affinity of androgens for AR is also higher. In recent years, topical antiandrogen therapy has attracted attention due to its potential efficacy in the treatment of androgenic alopecia and the reduction of side effects compared with systemic drugs. Topical finasteride, ketoconazole shampoo and topical claxoterone have all been used. It has entered clinical trials, but the treatment effects vary among different patients. Androgen Receptor-Proteolysis Targeting Chimeric Molecules (AR-PROTAC) connects the ligand of the androgen receptor target protein and the recruitment ligand of E3 ubiquitin ligase through a linker. After entering the cell Recognizes androgen receptors and tags them with ubiquitin via E3 enzymes, thereby achieving efficient androgen receptor degradation.

Microneedles (MNs) are tiny needles with lengths ranging from 100 μm to 2000 μm made of silicon, metal or other materials through microelectronic manufacturing technology or micro-molding technology. They are usually composed of a large number of microneedles to form an array structure, which is called For microneedle patches. Microneedles can pierce the stratum corneum and form micropores on the skin surface to help drugs enter the skin. The length is not long enough to touch the subcutaneous pain nerves. The micropores formed can also be restored within a few hours, which is minimally invasive and painless. and ease of use. Common microneedles include solid microneedles, hollow microneedles, and soluble microneedles. Among them, solid microneedles lack drug-carrying ability and generally require pretreatment of the skin to form micropores before applying the drug, which is a cumbersome operation. And it is impossible to precisely control the amount of medicine entering the skin. Hollow microneedles have small holes on the axis of the needle that function similarly to traditional injections. They are similar to microinjections, but their preparation process is complicated and the needle tips are easily blocked by dermal tissue and cannot release drugs. Soluble microneedles can carry drugs in precise doses, and the preparation process is relatively simple. Most of their bases are polymer materials with high biocompatibility, which can be completely dissolved or degraded in the skin and are easy to use.

Contents of the invention

The object of the present invention is to provide a hair growth microneedle patch containing an androgen receptor protein targeting complex and a preparation method thereof. By loading the androgen receptor protein targeting complex in the microneedle, it provides a method for the treatment of AGA. A safe, efficient, minimally invasive and minimally painful treatment.

In order to achieve the above object, according to one aspect of the present invention, a hair growth microneedle patch containing an androgen receptor protein targeting complex is provided, including: a sheet-like backing and a soluble patch arranged on the sheet-like backing. Microneedle body; wherein, the soluble microneedle body includes androgen receptor protein targeting complex, its pharmaceutically acceptable salts, its stereoisomers, its geometric stereoisomers, and its tautomers One or more of its body, its ester, its prodrug, its solvate, its metabolite, its nitrogen oxide or its deuterated compound.

The soluble microneedle body is made by mixing the androgen receptor protein targeting complex with a soluble polymer solution and then drying it so that the androgen receptor protein targeting complex is loaded in the soluble microneedle; the androgen receptor protein targeting complex is mixed The body protein targeting consortium is obtained through chemical synthesis, and its structural formula is as follows:

Pharmaceutically acceptable salts thereof, specifically acid or base addition salts. Where applicable, the term "pharmaceutically acceptable salt" is used in this specification to describe the salt form of one or more of the compounds described herein. Medication acceptable The salts include those derived from pharmaceutically acceptable inorganic or organic bases and acids, where applicable. Suitable salts include those derived from alkali metals (such as potassium and sodium), alkaline earth metals (such as calcium, magnesium and ammonium salts) and a variety of other acids and bases well known in the pharmaceutical art.

The acids used in the preparation of pharmaceutically acceptable acid addition salts of the above-described base compounds useful in the present invention are those which form non-toxic acid addition salts containing pharmacologically acceptable anions. Salts such as hydrochloride, trifluoroacetate, formate, hydrobromide, hydroiodide, nitrate, sulfate, hydrogen sulfate, phosphate, acid phosphate, acetate, Lactate, citrate, acid citrate, tartrate, bitartrate, succinate, maleate, fumarate, gluconate, sucrate, benzoate, methanesulfonate Acid esters, ethanesulfonate esters, benzenesulfonate esters, p-toluenesulfonate esters, and pamoate [i.e., 1,1-methylene-bis-(2-hydroxy-3-naphthoate)] salts and numerous other salts.

Pharmaceutically acceptable base addition salts may also be used to produce pharmaceutically acceptable salt forms of the compounds or derivatives according to the invention. Chemical bases that may be used as reagents for preparing pharmaceutically acceptable base salts of compounds of the invention that are acidic in nature are those that form nontoxic base salts with these compounds. These non-toxic base salts include, but are not limited to, those derived from pharmacologically acceptable cations such as alkali metal cations (e.g., potassium and sodium) and alkaline earth metal cations (e.g., calcium, zinc, and magnesium), ammonium or water-soluble Amine addition salts, such as those of N-methylglucamine-(meglumine) and lower alkanol ammonium and other bases of pharmaceutically acceptable organic amines;

Among them, the backing and the soluble microneedle body are composed of different polymer materials.

Preferably, the polymer used in the backing of the patch is pullulan, polylactic acid, polyglycolic acid, polyethylene oxide, polyacrylic acid, polyacrylamide, poly(methyl vinyl ether/maleic acid) One or more of half-ester copolymer, polyvinylpyrrolidone, polyethylene glycol, polyvinyl alcohol, hydroxypropyl cellulose, hydroxyethyl cellulose, hydroxypropyl methyl cellulose, and carboxymethyl cellulose sodium kind.

Preferably, the polymer used in the soluble microneedle body is hyaluronic acid, polylactic acid, polyglycolic acid, polyethylene oxide, polyacrylic acid, polyacrylamide, poly(methyl vinyl ether/maleic acid) One or more of half-ester copolymer, polyvinylpyrrolidone, polyethylene glycol, polyvinyl alcohol, hydroxypropyl cellulose, hydroxyethyl cellulose, hydroxypropyl methyl cellulose, and carboxymethyl cellulose sodium kind.

According to another aspect of the present invention, a method for preparing the hair growth microneedle patch containing the androgen receptor protein targeting complex as described above is provided, and the method includes the following steps:

1) First, use compound 1 and compound 2 as raw materials, stir and react under the action of solvent and alkali until the raw materials react completely to obtain compound 3;

2) Dissolve compound 3 in dichloromethane, add organic acid or inorganic acid, react completely at room temperature under argon protection, adjust the pH to 7-11 with saturated sodium bicarbonate, extract, dry and concentrate, beat and filter to obtain compound 4;

3) Using compound 4 and compound 5 as raw materials, react under the action of solvent, amide linker and alkali to obtain compound 6;

4) Dissolve compound 6 in dichloromethane, add organic acid or inorganic acid, react completely at room temperature under argon protection, adjust the pH to 7-11 with saturated sodium bicarbonate, extract, dry and concentrate with dichloromethane to obtain compound 7;

5) Use compound 7 and compound 8 as raw materials, react under the action of solvent, acetic acid, and sodium triacetoxyborohydride, dry and concentrate to obtain compound 9;

6) Dissolve compound 9 in dichloromethane, add organic acid or inorganic acid, react completely at room temperature under the protection of three argon gases, adjust the pH to 7-11 with saturated sodium bicarbonate, extract, dry and concentrate with dichloromethane to obtain compound 10;

7) Using Compound 10 and Compound 11 as raw materials, the reaction is complete under the action of solvent and alkali, and after concentration and purification, the androgen receptor protein targeting consortium is obtained, which can be in the free state of the compound or the salt form of the compound;

8) Prepare the soluble needle polymer solution: Mix the androgen receptor protein targeting consortium and the soluble needle polymer solution to obtain a mixed solution, inject the mixed solution into the microneedle mold, and fill the entire microneedle with the mixed solution by centrifugation. Needle the mold, scrape off excess polymer solution, and dry;

9) Prepare a backing polymer solution, inject the polymer solution into the microneedle mold, fill the backing part of the mold with the backing polymer solution by centrifugation, peel off the microneedle mold after drying, and obtain a hair-growing microneedle patch.

Preferably, in step (1), the solvent is selected from water, ether (such as THF, glyme, etc.) or chlorinated solvent (such as DCM, 1,2-dichloroethane (DCE) or CHCl _3, etc.), toluene, benzene, etc., DMF, DMSO, MeCN; the base includes (but is not limited to) cesium carbonate, potassium carbonate, sodium hydride, triethylamine, DIPEA, etc.

Preferably, in step (1), the reaction is carried out at a temperature between about -78°C and about 150°C; further preferably, the reaction is carried out at a temperature between about 0°C and about 100°C.

Preferably, in step (2), the organic acid is trifluoroacetic acid or formic acid, etc., and the inorganic acid is dioxane hydrochloride, sulfuric acid, etc. Preferably, after extraction with dichloromethane, drying and concentration, methyl tert-butyl ether is added for pulping and suction filtration to obtain compound 4.

Preferably, in step (3), solvents may include (but are not limited to) water, ethers, such as THF, glyme, etc.; chlorinated solvents, such as DCM, 1,2-dichloroethane (DCE). ) or _CHCl3 , etc., toluene, benzene, etc., DMF, DMSO, MeCN; if necessary, use a mixture of these solvents. Further preferably, the solvent is DMF or DCM.

Preferably, in step (3), suitable amide linkers include (but are not limited to) DCC, EDC, HATU, HBTU, PyBOP, etc.

Preferably, in step (3), the base includes (but is not limited to) TEA, DIPEA, etc.

Preferably, in step (3), the reaction is carried out at a temperature between about 0°C and about 100°C.

Preferably, in step (4), the organic acid is trifluoroacetic acid or formic acid, etc., and the inorganic acid is dioxane hydrochloride, sulfuric acid, etc. Extract with dichloromethane, dry and concentrate to obtain compound 7.

Preferably, in step (5), solvents may include (but are not limited to) water, ethers, such as THF, glyme, etc.; chlorinated solvents, such as DCM, 1,2-dichloroethane (DCE). ) or CHCl ₃ , etc., toluene, benzene, etc., DMF, DMSO, MeCN.

Preferably, in step (6), the organic acid is trifluoroacetic acid or formic acid, etc., and the inorganic acid is dioxane hydrochloride, sulfuric acid, etc. Extract with dichloromethane, dry and concentrate to obtain compound 7.

Preferably, in step (7), solvents may include (but are not limited to) water, ethers, such as THF, glyme, etc.; chlorinated solvents, such as DCM, 1,2-dichloroethane (DCE). ) or CHCl ₃ , etc., toluene, benzene, etc., DMF, DMSO, MeCN, etc., add a base to the reaction. Suitable bases include (but are not limited to) TEA, DIPEA, etc. This can be done at a temperature between about -78°C and about 150°C. Further preferably, the reaction is carried out at a temperature between about 0°C and about 100°C.

Preferably, in step 8), the mass fraction of the prepared soluble needle polymer solution is 10 to 100%, and the androgen receptor protein targeting consortium accounts for 10 to 2000mg/ of the total volume of the mixed solution. ml is added and mixed to obtain, the centrifugation speed is 1000~10000rpm, the centrifugation time is 1~30min, and the drying time is 12~24h.

Preferably, in step 9), the mass fraction of the backing polymer solution obtained is 20 to 80%, and The heart speed is 1000~10000rpm, the centrifugation time is 1~30min, and the drying time is 24~72h.

The present invention also provides the application of the above-mentioned hair growth microneedle patch containing the androgen receptor protein targeting complex in preparing a drug for treating androgen-induced alopecia.

Generally speaking, compared with the existing technology, the above technical solution conceived by the present invention has the following advantages:

(1) The hair growth microneedle patch provided in the present invention is prepared by a two-step centrifugation method. The needle body contains the androgen receptor protein targeting complex, and the patch backing does not contain the androgen receptor protein targeting complex. It will not cause any waste of medicine. When used, the microneedle body absorbs tissue fluid and dissolves quickly, and the patch backing can be removed. The drug administration process is short, does not affect the appearance, and has high patient compliance.

(2) The hair growth microneedle patch provided by the present invention can directly penetrate the stratum corneum barrier of the skin. As the microneedle body dissolves, the androgen receptor protein targeting complex is gradually released, and the androgen receptor protein is The targeted combination is directly delivered to the hair follicle dermal papilla cells and exerts a therapeutic effect by degrading the androgen receptor. It is safe, efficient and easy to use, and has the application prospect of clinical transformation.

(3) In the preparation method of the present invention, the androgen receptor protein targeting consortium recognizes the androgen receptor through the ligand of the androgen receptor target protein in the structure, and at the same time, the E3 ubiquitin ligase in the structure Recruiting ligands causes ubiquitin labeling of androgen receptors through E3 enzymes, thereby achieving efficient androgen receptor degradation, inhibiting the binding of androgens and androgen receptors during the development of AGA, accelerating the entry of hair follicles into the growth phase, and inducing hair regeneration.

(4) The present invention solves the problem based on the pathogenesis of AGA. The hair growth microneedle patch of the androgen receptor protein-targeting combination is used to treat AGA. It has the application prospect of realizing clinical transformation and is a solution for the current AGA treatment that cannot be cured. Solutions are provided for frequently recurring problems.

Description of drawings

Figure 1 is an SEM image of a hair growth microneedle patch in which the needle body is a quadrangular pyramid provided in Example 1 of the present invention.

Figure 2 shows the hair regeneration of AGA model mice using the hair growth microneedle patch containing the androgen receptor protein targeting consortium prepared in the present invention.

Figure 3 is a graph showing the results of new hair coverage in AGA model mice using the hair growth microneedle patch containing the androgen receptor protein targeting consortium prepared in the present invention.

Figure 4 shows the androgen receptor content in the skin of mice using the hair growth microneedle patch containing the androgen receptor protein targeting complex prepared by the present invention.

Detailed ways

The present invention will be further described below in conjunction with specific embodiments and accompanying drawings.

Example 1

This embodiment provides a method for preparing a soluble hair growth microneedle patch, including:

1. Preparation of androgen receptor protein targeting consortium:

Dissolve compound 2 (2.0g, 1.0eq.) in 20mL N,N-dimethylformamide, ice bath, add sodium hydride (60%, 0.56g, 1.5eq.), stir for 0.5h, and add compound 1 ( 1.45g, 1.0eq.) was dissolved in 10mL N,N-dimethylformamide, and the above reaction solution was added dropwise. React at room temperature until the raw material reaction was complete. The reaction solution was concentrated and purified by column chromatography (V _{petroleum ether} /V _{ethyl acetate)} . _Ester =6/1), 3.1g of white solid was obtained, yield 95%, UPLC-MS: m/z 351.15[M+H] ⁺ ;

Dissolve compound 3 (1.0g, 1.0eq.) in 12mL dichloromethane, add 4mL trifluoroacetic acid, protect with argon, react at room temperature until the raw material reaction is complete, adjust the pH to 7 with saturated sodium bicarbonate, and extract with dichloromethane. After drying and concentration, 20 mL of methyl tert-butyl ether was added to beat for 20 minutes, and 0.69 g of white solid was obtained by suction filtration, yield 96%, UPLC-MS: m/z 251.10 [M+H] ⁺ ;

Dissolve compound 5 (1.0g, 1.0eq.) in 15mL anhydrous N,N-dimethylformamide, add 1.2mL (2.0eq.) N,N-diisopropylethylamine, O-(7- Azabenzotriazol-1-yl)-N,N,N',N'-tetramethylurea hexafluorophosphate (1.82g, 1.2eq.), under nitrogen protection, stirred at room temperature for 15 minutes, added compound 4 ( 0.82g, 1.0eq.), react at room temperature until the reaction is complete, the reaction solution is concentrated and then purified by column chromatography (V _{petroleum ether} /V _{ethyl acetate} = 6/1) to obtain 2.02g of white solid, yield 94%, UPLC-MS : m/z 539.24[M+H] ⁺ ;

Dissolve compound 6 (1.0g, 1.0eq.) in 10mL of dichloromethane, add 4mL of trifluoroacetic acid, protect with argon, and react at room temperature until the reaction is complete. The reaction solution is concentrated, and the pH is adjusted to 7 with saturated sodium bicarbonate. Methane extraction, drying Concentrate and purify by column chromatography (D/M 30:1-15:1) to obtain 0.77g of white solid, yield 95%, UPLC-MS: m/z 439.19[M+H] ⁺ ;

Dissolve compound 7 (1.0g, 1.0eq.) in 15mL of dichloromethane, add compound 8 (0.6g, 1.2eq.), 3 drops of acetic acid, react at room temperature for 1h, add sodium triacetoxyborohydride (0.97g, 2.0eq.), react until the raw material reaction is complete, add water to quench the reaction, extract with dichloromethane, concentrate to dryness, and purify by column chromatography (V _{petroleum ether} /V _{ethyl acetate} = 4/1) to obtain 1.33g of white solid, yield 92%, UPLC-MS: m/z 636.33[M+H] ⁺ ;

Dissolve compound 9 (1.0g, 1.0eq.) in 8mL of dichloromethane, add 3.5mL of dichloromethane, protect with argon, react at room temperature until the reaction is complete, concentrate the reaction solution, and adjust the pH to neutral with saturated sodium bicarbonate. Extract with dichloromethane, dry and concentrate, and purify by column chromatography (V _{dichloromethane} /V _methanol = 20/1) to obtain 0.8 g of product, yield 95%, UPLC-MS: m/z536.28 [M+H] ⁺ ;

Dissolve compound 10 (100 mg, 1.0 eq.), compound 11 (62 mg, 1.0 eq.), and 65 μL of N,N-diisopropylethylamine (2.0 eq.) in 6 mL of anhydrous dichloromethane, and react at 90°C The reaction was allowed to proceed overnight until the reaction was complete. The reaction solution was concentrated and purified by preparative HPLC to obtain 103 mg of fluorescent green solid with a yield of 70%. UPLC-MS calculated for C ₄₃ H ₄₆ ClN ₇ O ₆ [M+H] ⁺ :792.33 found:792.36.UPLC-retention time:6.9min. ¹ H NMR(400MHz, DMSO-d ₆ )δ11.06(s, 1H),8.05(d,J＝7.6Hz,1H),7.80(dd,J＝23.7,8.7Hz,3H),7.65(d,J＝8.5Hz,1H),7.34(dd,J＝6.7,2.3 Hz,2H),7.24(dd,J＝8.7,2.3Hz,1H),7.11(dd,J＝8.8,2.4Hz,1H),7.01(d,J＝8.7Hz,2H),5.04(dd,J ＝12.9,5.4Hz,1H),4.50(dp,J＝9.0,4.3Hz,1H),4.07(d,J＝13.1Hz,2H),3.93(d,J＝9.3Hz,2H),3.81–3.72 (m,1H),3.60(d,J＝8.2Hz,2H),3.19–2.79(m,10H),2.61–2.47(m,1H),2.21–1.76(m,8H),1.58–1.39(m ,4H),1.32–1.17(m,2H).

2. Prepare a premixed solution: Prepare a 70% hyaluronic acid solution, add the androgen receptor protein targeting consortium obtained according to step 1 and 50% of the total volume of the hyaluronic acid solution and mix to obtain the result.

3. Prepare the microneedle patch: Inject the premixed solution obtained in step 2 into the customized polydimethylsiloxane (PDMS, Sylgard 184) microneedle array mold. Next, the PDMS microneedle array mold was centrifuged at 3500 rpm for 10 minutes until it was completely filled and then taken out and the excess premixed solution was scraped off. Place the PDMS microneedle array mold in a silica gel desiccator and dry at room temperature for 2 hours. Take out the PDMS microneedle array mold and inject 40% polyvinyl alcohol solution. Next, the PDMS microneedle array mold was centrifuged at 3500 rpm for 5 minutes to form a backing and then taken out. Place the PDMS microneedle array mold in a silica gel desiccator and dry it at room temperature for 24 hours to obtain a hair growth microneedle patch containing the androgen receptor protein targeting complex.

In this embodiment, the androgen receptor protein targeting consortium can be replaced by or further include the compound, its pharmaceutically acceptable salts, its stereoisomers, its geometric stereoisomers, and its tautomers , one or more of its esters, its prodrugs, its solvates, its metabolites, its nitrogen oxides or its deuterated compounds.

In this embodiment, the hyaluronic acid solution may be replaced by or further include polylactic acid, polyglycolic acid, polyethylene oxide, polyacrylic acid, polyacrylamide, poly(methyl vinyl ether/maleic acid) half-ester copolymer , one or more of polyvinylpyrrolidone, polyethylene glycol, polyvinyl alcohol, hydroxypropylcellulose, hydroxyethylcellulose, hydroxypropylmethylcellulose, and carboxymethylcellulose sodium.

In this embodiment, the polyvinyl alcohol solution may be replaced by or further include pullulan, polylactic acid, polyglycolic acid, polyethylene oxide, polyacrylic acid, polyacrylamide, poly(methyl vinyl ether/maleic acid) One or more of half-ester copolymer, polyvinylpyrrolidone, polyethylene glycol, hydroxypropylcellulose, hydroxyethylcellulose, hydroxypropylmethylcellulose, and carboxymethylcellulose sodium.

In this embodiment, the androgen receptor protein targeting consortium is mixed with the polymer solution to finally obtain a solid soluble microneedle patch. The solid-state soluble microneedle patch has a gentle preparation process, does not require extreme conditions, is simple to prepare, and can be stored at room temperature in a dry environment, effectively realizing the transdermal delivery of the androgen receptor protein-targeting consortium.

In this embodiment, the hair growth microneedle patch is prepared by a two-step centrifugation method. The needle body contains the androgen receptor protein targeting complex, and the backing of the patch does not contain the androgen receptor protein targeting complex, which will not cause Waste of active substances. When used, the microneedle body absorbs tissue fluid and dissolves quickly, and the patch backing can be removed. The drug administration process is short, does not affect the appearance, and has high patient compliance.

Example 2

This embodiment provides a method for preparing a soluble hair growth microneedle patch, including:

1. Preparation of androgen receptor protein targeting consortium:

Dissolve compound 2 (2.0g, 1.0eq.) in 20mL N,N-dimethylformamide, ice bath, add sodium hydride (60%, 0.56g, 1.5eq.), stir for 0.5h, and add compound 1 ( 1.45g, 1.0eq.) was dissolved in 10mL N,N-dimethylformamide, and the above reaction solution was added dropwise. React at room temperature until the raw material reaction was complete. The reaction solution was concentrated and purified by column chromatography (V _{petroleum ether} /V _{ethyl acetate)} . _Ester = 6/1), 3.1 g of white solid was obtained, the yield was 95%. UPLC-MS: m/z 351.15[M+H] ⁺ ;

Dissolve compound 3 (1.0g, 1.0eq.) in 12mL dichloromethane, add 4mL trifluoroacetic acid, protect with argon, react at room temperature until the raw material reaction is complete, adjust the pH to 7 with saturated sodium bicarbonate, and extract with dichloromethane. After drying and concentration, 20 mL of methyl tert-butyl ether was added to beat for 20 min, and 0.69 g of white solid was obtained by suction filtration with a yield of 96%. UPLC-MS: m/z 251.10[M+H] ⁺ ;

Dissolve compound 5 (1.0g, 1.0eq.) in 15mL anhydrous N,N-dimethylformamide, add 1.2mL (2.0eq.) N,N-diisopropylethylamine, O-(7- Azabenzotriazol-1-yl)-N,N,N',N'-tetramethylurea hexafluorophosphate (1.82g, 1.2eq.), under nitrogen protection, stirred at room temperature for 15 minutes, added compound 4 ( 0.82g, 1.0eq.), reacted at room temperature until the reaction was complete, the reaction solution was concentrated and then purified by column chromatography (V _{petroleum ether} /V _{ethyl acetate} = 6/1) to obtain 2.02g of white solid, with a yield of 94%. UPLC-MS: m/z 539.24[M+H] ⁺ ;

Dissolve compound 6 (1.0g, 1.0eq.) in 10mL of methylene chloride, add 4mL of trifluoroacetic acid, protect with argon, and react at room temperature until the reaction is complete. The reaction solution is concentrated, and the pH is adjusted to neutral with saturated sodium bicarbonate. Extract with methyl chloride, dry and concentrate, and purify by column chromatography (D/M 30:1-15:1) to obtain 0.77g of white solid, with a yield of 95%. UPLC-MS: m/z439.19[M+H] ⁺ ;

Dissolve compound 7 (1.0g, 1.0eq.) in 15mL of dichloromethane, add compound 8 (0.6g, 1.2eq.), 3 drops of acetic acid, react at room temperature for 1h, add sodium triacetoxyborohydride (0.97g, 2.0eq.), react until the raw material reaction is complete, add water to quench the reaction, extract with dichloromethane, concentrate to dryness, and purify by column chromatography (V _{petroleum ether} /V _{ethyl acetate} = 4/1) to obtain 1.33g of white solid, yield 92%. UPLC-MS: m/z 636.33[M+H] ⁺ ;

Dissolve compound 9 (1.0g, 1.0eq.) in 8mL of methylene chloride, add 3.5mL of methylene chloride, protect with argon, react at room temperature until the reaction is complete, concentrate the reaction solution, and adjust the pH to 7 with saturated sodium bicarbonate. Extract with methyl chloride, dry and concentrate, and purify by column chromatography (V _{dichloromethane} /V _methanol = 20/1) to obtain 0.8 g of product with a yield of 95%. UPLC-MS: m/z 536.28[M+H] ⁺ ;

Dissolve compound 10 (100 mg, 1.0 eq.), compound 11 (62 mg, 1.0 eq.), and 65 μL of N,N-diisopropylethylamine (2.0 eq.) in 6 mL of anhydrous dichloromethane, and react at 90°C overnight until the reaction is complete, the reaction solution is concentrated, and the Prepare HPLC purification to obtain 103 mg of fluorescent green solid, with a yield of 70%. UPLC-MS calculated for C ₄₃ H ₄₆ ClN ₇ O ₆ [M+H] ⁺ :792.33 found:792.36.UPLC-retention time:6.9min. ¹ H NMR(400MHz, DMSO-d ₆ )δ11.06(s, 1H),8.05(d,J＝7.6Hz,1H),7.80(dd,J＝23.7,8.7Hz,3H),7.65(d,J＝8.5Hz,1H),7.34(dd,J＝6.7,2.3 Hz,2H),7.24(dd,J＝8.7,2.3Hz,1H),7.11(dd,J＝8.8,2.4Hz,1H),7.01(d,J＝8.7Hz,2H),5.04(dd,J ＝12.9,5.4Hz,1H),4.50(dp,J＝9.0,4.3Hz,1H),4.07(d,J＝13.1Hz,2H),3.93(d,J＝9.3Hz,2H),3.81–3.72 (m,1H),3.60(d,J＝8.2Hz,2H),3.19–2.79(m,10H),2.61–2.47(m,1H),2.21–1.76(m,8H),1.58–1.39(m ,4H),1.32–1.17(m,2H).

2. Prepare a premixed solution: Prepare a 40% hyaluronic acid solution, add the androgen receptor protein targeting complex obtained according to step 1 and the hyaluronic acid solution at 10% of the total volume and mix to obtain the result.

3. Prepare the microneedle patch: Inject the premixed solution obtained in step 2 into the customized polydimethylsiloxane (PDMS, Sylgard 184) microneedle array mold. Next, the PDMS microneedle array mold was centrifuged at 3000 rpm for 10 minutes until it was completely filled and then taken out, and the excess premixed solution was scraped off. Place the PDMS microneedle array mold in a silica gel desiccator and dry at room temperature for 1 hour. Take out the PDMS microneedle array mold and inject 50% polyvinyl alcohol solution. Next, the PDMS microneedle array mold was centrifuged at 3000 rpm for 5 minutes to form a backing and then taken out. Place the PDMS microneedle array mold in a silica gel desiccator and dry it at room temperature for 24 hours to obtain a hair growth microneedle patch containing the androgen receptor protein targeting complex.

In this embodiment, the androgen receptor protein targeting consortium is mixed with the polymer solution to finally obtain a solid soluble microneedle patch. The solid-state soluble microneedle patch has a gentle preparation process, does not require extreme conditions, is simple to prepare, and can be stored at room temperature in a dry environment, effectively realizing the transdermal delivery of the androgen receptor protein-targeting consortium.

In this embodiment, the hair growth microneedle patch is prepared by a two-step centrifugation method. The needle body contains the androgen receptor protein targeting complex, and the backing of the patch does not contain the androgen receptor protein targeting complex, which will not cause Waste of active substances. When used, the microneedle body absorbs tissue fluid and dissolves quickly, and the patch backing can be removed. The drug administration process is short, does not affect the appearance, and has high patient compliance.

Example 3

This embodiment provides a method for preparing a soluble microneedle patch. The difference between this embodiment and Example 1 is that step 1 in this embodiment involves synthesizing a salt of the androgen receptor protein targeting complex.

In this example, the androgen receptor protein targeting consortium is mixed with the polymer solution to finally obtain to solid dissolvable microneedle patches. The solid-state soluble microneedle patch has a gentle preparation process, does not require extreme conditions, is simple to prepare, and can be stored at room temperature in a dry environment, effectively realizing the transdermal delivery of the androgen receptor protein-targeting consortium.

Example 4

This example provides a method for preparing a soluble microneedle patch. The difference between this example and Example 1 is that step 1 in this example involves synthesizing stereoisomers of the androgen receptor protein targeting consortium. .

In this embodiment, the androgen receptor protein targeting consortium is mixed with the polymer solution to finally obtain a solid soluble microneedle patch. The solid-state soluble microneedle patch has a gentle preparation process, does not require extreme conditions, is simple to prepare, and can be stored at room temperature in a dry environment, effectively realizing the transdermal delivery of the androgen receptor protein-targeting consortium.

Evaluation of efficacy in the treatment of androgenic alopecia

Select the hair growth microneedle patch prepared in Example 1 (Figure 1), apply one microneedle patch on the AGA model mouse by pressing with fingertip strength, and after the 28-day hair cycle is completed, record the hair regeneration of the mouse. and detect new hair coverage. The hair growth microneedle patch prepared in Example 1 was selected, and one microneedle patch was applied to normal mice by pressing with fingertip force. Six days later, the androgen receptor content in the skin was detected.

As shown in Figure 2, the AGA model mice treated with the hair growth microneedle patch containing the androgen receptor protein-targeting consortium experienced significant hair regeneration, while the AGA model mice that did not receive any treatment did not experience hair regeneration.

As shown in Figure 3, the AGA model mice treated with the hair growth microneedle patch containing the androgen receptor protein-targeting consortium had a new hair coverage rate of more than 60%, while the AGA model mice that did not receive any treatment had new hair. Hair coverage is close to 0.

As shown in Figure 4, the androgen receptor content in the skin of mice after application of the hair growth microneedle patch containing the androgen receptor protein-targeting complex was significantly lower than that of mice that did not receive any treatment.

Claims (10)

A hair growth microneedle patch containing an androgen receptor protein targeting complex, characterized in that the hair growth microneedle patch includes a sheet-like backing and androgen receptor-containing hair patches arranged on the sheet-like backing A soluble microneedle body of a protein-targeting complex; the soluble microneedle body includes an androgen receptor protein-targeting complex, its pharmaceutically acceptable salts, its stereoisomers, and its geometric stereoisomers, One or more of its tautomers, its esters, its prodrugs, its solvates, its metabolites, its nitrogen oxides or its deuterated compounds. The hair growth microneedle patch containing the androgen receptor protein targeting complex according to claim 1, wherein the backing and the soluble microneedle body are composed of different polymer materials. The hair growth microneedle patch containing androgen receptor protein targeting complex according to claim 1 or 2, characterized in that the polymer used in the backing is pullulan, polylactic acid, polyglycolic acid, Polyethylene oxide, polyacrylic acid, polyacrylamide, poly(methyl vinyl ether/maleic acid) half ester copolymer, polyvinylpyrrolidone, polyethylene glycol, polyvinyl alcohol, hydroxypropyl cellulose, hydroxyethyl One or more of cellulose, hydroxypropyl methylcellulose, and sodium carboxymethylcellulose. The hair growth microneedle patch containing androgen receptor protein targeting combination according to claim 1 or 2, characterized in that the polymer used in the soluble microneedle body is hyaluronic acid, polylactic acid, polylactic acid, Glycolic acid, polyethylene oxide, polyacrylic acid, polyacrylamide, poly(methyl vinyl ether/maleic acid) half ester copolymer, polyvinylpyrrolidone, polyethylene glycol, polyvinyl alcohol, hydroxypropyl cellulose, One or more of hydroxyethyl cellulose, hydroxypropyl methyl cellulose, and carboxymethyl cellulose sodium. A method for preparing a hair growth microneedle patch containing an androgen receptor protein targeting complex according to any one of claims 1 to 4, characterized in that the preparation method includes the following steps:
1) First, use compound 1 and compound 2 as raw materials, stir and react under the action of solvent and alkali until the raw materials react completely to obtain compound 3; 2) Dissolve compound 3 in dichloromethane, add organic acid or inorganic acid, react completely at room temperature under argon protection, adjust the pH to 7-11 with saturated sodium bicarbonate, extract, dry and concentrate, beat and filter to obtain compound 4; 3) Using compound 4 and compound 5 as raw materials, react under the action of solvent, amide linker and alkali to obtain compound 6; 4) Dissolve compound 6 in dichloromethane, add organic acid or inorganic acid, react completely at room temperature under argon protection, adjust the pH to 7-11 with saturated sodium bicarbonate, extract, dry and concentrate with dichloromethane to obtain compound 7; 5) Use compound 7 and compound 8 as raw materials, react under the action of solvent, acetic acid, and sodium triacetoxyborohydride, dry and concentrate to obtain compound 9; 6) Dissolve compound 9 in dichloromethane, add organic acid or inorganic acid, react completely at room temperature under the protection of three argon gases, adjust the pH to 7-11 with saturated sodium bicarbonate, extract, dry and concentrate with dichloromethane to obtain compound 10; 7) Using Compound 10 and Compound 11 as raw materials, the reaction is complete under the action of solvent and alkali, and after concentration and purification, the androgen receptor protein targeting consortium is obtained, which can be in the free state of the compound or the salt form of the compound; 8) Prepare the soluble needle polymer solution: Mix the androgen receptor protein targeting consortium and the soluble needle polymer solution to obtain a mixed solution, inject the mixed solution into the microneedle mold, and fill the entire microneedle with the mixed solution by centrifugation. Needle the mold, scrape off excess polymer solution, and dry; 9) Prepare a backing polymer solution, inject the polymer solution into the microneedle mold, fill the backing part of the mold with the backing polymer solution by centrifugation, peel off the microneedle mold after drying, and obtain a hair-growing microneedle patch. The method for preparing a hair growth microneedle patch containing an androgen receptor protein targeting complex according to claim 5, wherein the mass fraction of the needle polymer solution in step 8) is 10 to 100%. , the androgen receptor protein targeting complex accounts for 10 to 70% of the total volume of the mixed solution. The method for preparing a hair growth microneedle patch containing an androgen receptor protein targeting complex according to claim 5, characterized in that the centrifugation speed of step 8) is 1000-10000 rpm, and the centrifugation time is 1-30 min ; Drying time is 12 to 24 hours. The method for preparing a hair growth microneedle patch containing an androgen receptor protein targeting complex according to claim 5, wherein the mass fraction of the backing polymer solution in step 9) is 20 to 80%. The method for preparing a hair growth microneedle patch containing an androgen receptor protein targeting complex according to claim 5, characterized in that the centrifugation speed of step 9) is 1000-10000 rpm, and the centrifugation time is 1-30 min ; Drying time is 24~72h. The application of a hair growth microneedle patch containing an androgen receptor protein targeting complex according to any one of claims 1 to 4 in the preparation of a drug for the treatment of androgenic alopecia.