Application of colistin in the preparation of a medicament for treating androgenetic alopecia

Abstract

The present invention discloses the use of colistin in the preparation of a drug for treating androgenic alopecia. The colistin of the present invention can effectively promote hair growth in androgenic alopecia areas, inhibit androgen receptor expression, and promote the proliferation of hair papilla cells. Compared with existing drugs for treating androgenic alopecia, the colistin has fewer side effects and a faster effect. The colistin selected in the present invention is a drug that has been approved for clinical use, has few side effects, a wide drug source, and a simple and economical acquisition method.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine, and particularly relates to the application of colistin in the preparation of a medicament for treating androgenetic alopecia. Background Art

[0002] Androgenetic alopecia (AGA) is a common chronic progressive alopecia, which is more common in men, and women also have a certain probability of suffering from it. AGA is related to genetics, the level of androgens in the human body, and androgen receptors (AR). In male AGA patients, the clinical manifestations are that the hair on the forehead and the top of the head becomes thinner, softer, and less, and the forehead hairline moves backward. In female patients, the hair on the top of the head mostly becomes thinner and less, and the hairline does not change significantly. Testosterone is the main sex hormone in men, and dihydrotestosterone (DHT) is a metabolite of testosterone. In AGA, DHT accumulates in the hair follicle area of the scalp, causing the hair follicles to shrink, resulting in the hair gradually becoming thinner, shorter, and finally stopping growing. High levels of DHT may also trigger an inflammatory response, affecting the normal function of hair follicles. Currently, the drugs clinically used for the treatment of AGA mainly include oral drugs finasteride and dutasteride, and topical drug minoxidil, all of which have certain side effects and cannot cure AGA completely.

[0003] Colistin is an antibiotic against Gram-negative bacilli, with bactericidal effects, and has a more significant effect on Pseudomonas aeruginosa, and also has good effects on Escherichia coli, Salmonella, Shigella, Haemophilus influenzae, Bordetella pertussis, Aerobacter aerogenes, Klebsiella pneumoniae, etc. Clinically, it is mainly used for the treatment of Gram-negative bacterial infections, especially various infections caused by Pseudomonas aeruginosa and Escherichia coli, such as respiratory tract infections, peritonitis, biliary tract infections, urinary tract infections, burn infections, corneal infections, and septicemia, etc. There is no report on its use for the treatment of AGA. Summary of the Invention

[0004] The present invention aims to solve at least one of the technical problems existing in the above-mentioned prior art. For this reason, the present invention proposes the application of colistin in the preparation of a medicament for treating androgenetic alopecia.

[0005] The present invention also proposes a medicament for treating androgenetic alopecia.

[0006] According to the first aspect of the present invention, there is provided the application of colistin in the preparation of a medicament for treating androgenetic alopecia.

[0007] In some embodiments of the present invention, the androgenetic alopecia is androgenetic alopecia caused by testosterone.

[0008] In some embodiments of the present invention, the medicament for treating androgenetic alopecia is a medicament for promoting hair growth in the androgenetic alopecia area.

[0009] In some embodiments of the present invention, the drug for treating androgenic alopecia is a drug that promotes the proliferation of human dermal papilla cells.

[0010] In some embodiments of the present invention, the drug for treating androgenic alopecia is a drug that inhibits the expression of androgen receptors.

[0011] In some embodiments of the present invention, the content of colistin in the drug for treating androgenic alopecia is the content that reaches the therapeutically effective amount.

[0012] In some embodiments of the present invention, the content of colistin in the drug for treating androgenic alopecia can be 0.1%-100%, such as 0.1, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 96, 97, 98, 99, 99.5, 100%.

[0013] In some embodiments of the present invention, the dosage of colistin is 1-20 mg / Kg.

[0014] In some embodiments of the present invention, the dosage of colistin is 1-10 mg / Kg.

[0015] In some embodiments of the present invention, the subjects treated with the drug for treating androgenic alopecia are mammals, including humans, mice, etc. Those skilled in the art can infer the unit body weight dose of humans based on the equal-dose conversion relationship between humans and experimental animals. According to the unit body weight dose conversion, the equivalent dose of mice is about 9 times that of humans.

[0016] In some embodiments of the present invention, the drug for treating androgenic alopecia further includes a pharmaceutically acceptable carrier.

[0017] In some embodiments of the present invention, the pharmaceutically acceptable excipients include at least one of diluents, excipients, fillers, binders, disintegrants, absorption promoters, surfactants, adsorption carriers, lubricants, sweeteners, and flavoring agents.

[0018] In some embodiments of the present invention, the excipient includes water.

[0019] In some embodiments of the present invention, the filler includes at least one of starch and sucrose.

[0020] In some embodiments of the present invention, the binder includes at least one of cellulose derivatives, alginates, gelatin, and polyvinylpyrrolidone.

[0021] In some embodiments of the present invention, the wetting agent includes glycerol.

[0022] In some embodiments of the present invention, the disintegrant includes at least one of agar, calcium carbonate, and sodium bicarbonate.

[0023] In some embodiments of the present invention, the absorption promoter includes quaternary ammonium compounds.

[0024] In some embodiments of the present invention, the surfactant includes cetyl alcohol.

[0025] In some embodiments of the present invention, the adsorption carrier includes at least one of kaolin and saponite clay.

[0026] In some embodiments of the present invention, the lubricant includes at least one of talc, calcium stearate, magnesium stearate, and polyethylene glycol.

[0027] In some embodiments of the present invention, the administration methods of the drug for treating androgenetic alopecia include, but are not limited to, oral administration, enteral administration, subcutaneous injection, intramuscular injection, intravenous injection, nasal administration, transdermal administration, subconjunctival injection, intraocular injection, orbital injection, retrobulbar injection, retinal injection, choroidal injection, intrathecal injection, and so on.

[0028] In some embodiments of the present invention, the dosage forms of the drug for treating androgenetic alopecia include, but are not limited to, tablets, capsules, pills, injections, inhalants, lozenges, suppositories, emulsions, microemulsions, submicroemulsions, nanoparticles, gels, powders, suspensions, creams, jellies, sprays, and so on. The various dosage forms of the drug can be prepared according to the conventional production methods in the pharmaceutical field.

[0029] In some embodiments of the present invention, the colistin is the sole active ingredient of the drug for treating androgenetic alopecia, or is used as an active ingredient together with other drugs for treating androgenetic alopecia.

[0030] According to the second aspect of the present invention, a drug for treating androgenetic alopecia is provided, and the drug contains colistin.

[0031] In some embodiments of the present invention, the drug further contains one or more drugs for treating androgenetic alopecia.

[0032] According to some embodiments of the present invention, there are at least the following beneficial effects: The colistin of the present invention can effectively promote hair growth in androgenetic alopecia areas, inhibit androgen receptor expression, and promote the proliferation of dermal papilla cells. Compared with existing drugs for treating androgenetic alopecia, it has fewer side effects and faster effects. Moreover, the colistin selected in the present invention is a clinically approved drug with small side effects, wide drug sources, and simple and economical acquisition methods. BRIEF DESCRIPTION OF THE DRAWINGS

[0033] The present invention will be further described below in conjunction with the drawings and embodiments, where:

[0034] Figure 1 It is a result diagram of the effect of colistin in Example 1 of the present invention on AR in DPCs;

[0035] Figure 2 It is a result diagram of the cytotoxicity experiment of the effect of colistin in Example 1 of the present invention on the growth of DPCs;

[0036] Figure 3 It is a result diagram of the therapeutic effect of colistin in Example 1 of the present invention on AGA in C57BL-6 male mice;

[0037] Figure 4 It is a result diagram of the therapeutic effect of colistin in Example 1 of the present invention on AGA in C57BL-6 male mice, where "*" is P < 0.05, "**" is P < 0.01, and "***" is P < 0.001;

[0038] Figure 5 It is a result diagram of the therapeutic effect of colistin in Example 1 of the present invention on AGA in C57BL-6 female mice;

[0039] Figure 6 It is a result diagram of the therapeutic effect of colistin in Example 1 of the present invention on AGA in C57BL-6 female mice, where "**" is P < 0.01 and "***" is P < 0.001;

[0040] Figure 7 It is a hematoxylin-eosin staining diagram of the liver after the treatment of AGA in C57BL-6 male mice with colistin in Example 1 of the present invention, where the scale bar is 200 μm;

[0041] Figure 8 It is a hematoxylin-eosin staining diagram of the liver after the treatment of AGA in C57BL-6 female mice with colistin in Example 1 of the present invention, where the scale bar is 200 μm. DETAILED DESCRIPTION OF THE EMBODIMENTS

[0042] The concept of the present invention and the technical effects produced will be clearly and completely described below in conjunction with embodiments to fully understand the purpose, features, and effects of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all embodiments. Based on the embodiments of the present invention, other embodiments obtained by those skilled in the art without creative efforts shall fall within the scope of protection of the present invention.

[0043] Test samples:

[0044] Colistin was purchased from Shanghai Lanmu Chemical Co., Ltd.

[0045] Example 1

[0046] This example provides the application of colistin in the preparation of a drug for treating androgenetic alopecia. The specific verification process is as follows:

[0047] 1. Effect of colistin on androgen receptor (AR) in human dermal papilla cells (DPCs)

[0048] Experimental method: Human dermal papilla cells DPCs were digested with 1 mL of trypsin and then inoculated into 96-well plates, with 1.5×10 5 cells inoculated in each well. After overnight culture at 37°C for cell attachment, colistin with final concentrations of 0 μM, 10 μM, 20 μM, and 40 μM was added to the corresponding culture dishes respectively. After culturing at 37°C for 24 h, 60 μL of RIPA lysis buffer was used to lyse the cells in each well to extract total protein, and 25 μg of total protein was taken for Western Blot experiment to analyze the expression of androgen receptor, with GAPDH as the reference gene.

[0049] The results are as Figure 1 shown. It can be seen from the figure that colistin has a degradation effect on AR in DPCs.

[0050] 2. Effect of colistin on the growth of DPCs for 24 hours

[0051] Experimental method:

[0052] (1) Prepare a 10 mM colistin stock solution: Weigh colistin powder with an electronic balance and use dimethyl sulfoxide (DMSO) to prepare a stock solution with a concentration of 10 mM, and then dilute it with DMSO to different concentrations.

[0053] (2) Digest DPCs with trypsin and inoculate them into 96-well plates, with 5.5×10 3, After overnight culture at 37°C until adherent, colistin with final concentrations of 0 μM, 10 μM, 20 μM, 40 μM, 80 μM, and 160 μM was added to the corresponding culture dishes. After culturing at 37°C for 24 hours, 10 μL of MTT was added, and the incubation was carried out at 37°C for 4 hours. After 4 hours, the culture medium was aspirated, DMSO was added, and the reaction was carried out at room temperature on a shaker for 30 min. The absorbance was measured at 570 nm, and then a fitting curve of drug concentration corresponding to cell viability was plotted using GraphPad Prism 8.

[0054] The results are as Figure 2 shown. It can be seen from the figure that colistin has less toxicity to DPCs and has a certain effect on promoting the proliferation of dermal papilla cells.

[0055] 3. Animal experiments

[0056] In order to verify the effect of colistin in the treatment of AGA in vivo in this invention, C57BL-6 mice were selected to establish an AGA model for in vivo efficacy verification. The specific experimental method is as follows:

[0057] Experimental animals: 18 male C57BL-6 mice and 18 female C57BL-6 mice at 4 - 5 weeks of age were taken and raised stably in an SPF environment for one week.

[0058] Grouping: The C57BL-6 mice were randomly divided into a blank group (no drug administration) of 6 mice (DHT not injected subcutaneously), among which there were 3 males and 3 females; a modeling group of 30 mice. In the modeling group, DHT (5 mg / Kg) was injected subcutaneously into the back daily / time for 10 days to obtain an AGA mouse model.

[0059] Ten days after the establishment of the AGA mouse model, all mice were anesthetized by intraperitoneal injection of sodium pentobarbital at 10 mg / mL (0.4 mL / 100 g). The back skin of the mice was shaved short with an electric clipper, and a depilatory cream was applied to the back of the mice for depilation. The shaved mice were grouped: the blank group was not treated; in the model group, there were 5 males and 5 females, and only DHT at 10 mg / Kg was injected subcutaneously daily; in the low-dose drug group, there were 5 males and 5 females, and DHT at 10 mg / Kg was given daily, and at the same time, colistin at 2.5 mg / Kg was given daily for treatment; in the high-dose drug group, there were 5 males and 5 females, DHT at 10 mg / Kg was continuously given daily, and colistin at 5 mg / Kg was given daily for treatment.

[0060] Taking the day of hair shaving as the 0th day of the experiment, photos were taken on the 7th, 14th, and 17th days of the experiment to record the hair growth situation, and the relative long hair area of mice in different groups was calculated on the 17th day. After 17 days, the experimental mice were sacrificed by cervical dislocation, blood was collected by eye enucleation, and the blood was centrifuged at 3500 rpm at 4°C for 20 minutes to obtain serum. The back skin of the mice was shaved short with an electric clipper, and depilatory cream was applied to the back of the mice for depilation treatment. The back skin tissue and liver of the mice were dissected, and the liver was photographed and weighed for recording.

[0061] The results are as Figures 3 - 6 shown. It can be seen from Figures 3 - 4 that colistin has a medicinal effect on androgenetic alopecia, and the medicinal effect of high-dose colistin on male mice is better than that of low-dose; it can be seen from Figures 5 - 6 that the medicinal effect of low-dose colistin on female mice is better than that of high-dose; it can be seen from Figures 7 - 8 that colistin has less side effects on the treatment of mice.

[0062] The embodiments of the present invention have been described in detail above in conjunction with the accompanying drawings. However, the present invention is not limited to the above embodiments. Various changes can be made without departing from the spirit of the present invention within the knowledge scope of those of ordinary skill in the art. In addition, the embodiments of the present invention and the features in the embodiments can be combined with each other without conflict.

Claims

1. The use of colistin in the preparation of a drug for treating androgenic alopecia, characterized in that: The colistin is the only active ingredient of the drug for treating androgenic alopecia.

2. The use according to claim 1, characterized in that: The androgenic alopecia is androgenic alopecia caused by testosterone.

3. The use according to claim 1, characterized in that: The drug for treating androgenic alopecia is a drug for promoting hair growth in androgenic alopecia areas.

4. The use according to claim 1, characterized in that: The drug for treating androgenic alopecia is a drug for promoting the proliferation of human hair papilla cells.

5. The use according to claim 1, characterized in that: The drug for treating androgenic alopecia is a drug that inhibits the expression of androgen receptors.

6. The use according to claim 1, characterized in that: The content of colistin in the drug for treating androgenic alopecia is a content that reaches a therapeutically effective amount.

7. The use according to claim 1, characterized in that: The drug for treating androgenic alopecia further comprises a pharmaceutically acceptable carrier.

8. The use according to claim 1, characterized in that: The dosage form of the drug for treating androgenic alopecia is an injection; And / or, the drug for treating androgenic alopecia is administered by subcutaneous injection.

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