Use of ml348 in the treatment of androgenetic alopecia

Abstract

The application relates to the technical field of medicines, and discloses application of ML348 in prevention and treatment of androgenetic alopecia. In order to solve the technical problems of preventing and treating androgenetic alopecia, ML348 is applied to intervene in DHT-induced AGA model mice, which shows that ML348 has the effects of promoting hair regeneration, increasing hair length and hair follicle melanin generation, and can effectively resist hair loss caused by DHT. Based on this, a new scheme for treating androgenetic alopecia is provided for preventing and / or treating androgenetic alopecia.

Description

Technical Field

[0001] The present invention relates to the field of medical technology, and in particular to application of ML348 in preventing and treating androgenic alopecia. Background Art

[0002] Androgenetic alopecia (AGA) is one of the most common hair loss disorders worldwide. While it does not affect patients' physical health, it can have a significant negative impact on their psychological well-being and social identity, leading to a decrease in their quality of life. AGA affects both men and women. In men, the condition typically presents with a significantly receding hairline on the forehead and temples, along with thinning hair on the top of the head. In women, the condition presents with thinning and thinning hair between the top of the head and the hairline, with a preserved hairline on the forehead. The pathogenesis of AGA is complex, driven by multiple factors, including genetics, androgen metabolism, psychological factors, diet, and stress. It is generally believed that androgens play a key role in the pathogenesis of AGA. Testosterone is converted to dihydrotestosterone (DHT) catalyzed by 5α-reductase (SRD5A2). Studies have found that 5α-reductase activity in the scalp of AGA patients is significantly higher in areas of hair loss than in areas without hair loss, and DHT levels are significantly elevated in areas of hair loss. Excessive DHT binds to the androgen receptor (AR), initiating signals that regulate hair follicle growth, leading to hair follicle atrophy, smaller hair diameter, and the transformation of hair follicles from terminal hair to vellus hair. This shortens the growth period, affects the normal physiological cycle of hair, and ultimately leads to androgenic alopecia.

[0003] As AGA becomes increasingly common, treatments are under development. The main treatments for AGA include topical minoxidil, oral finasteride or spironolactone, hair transplant surgery, and LDL laser irradiation. The two most effective treatments for AGA are topical minoxidil and oral finasteride. Minoxidil increases vascular endothelial growth factor (VEGF) in dermal papilla cells in a dose-dependent manner and stimulates prostaglandin E2 production, resulting in a prolonged growth phase. Side effects include contact dermatitis, headache, and hirsutism, with a short period of telogen shedding observed during the first eight weeks of treatment. Finasteride is a class of drugs known as 5-α-reductase inhibitors. These drugs block the enzyme 5-α-reductase, thereby preventing the conversion of testosterone to its active form, dihydrotestosterone (DHT), slowing or reversing hair loss. However, finasteride use may be associated with side effects such as sexual dysfunction. Non-pharmacological treatments such as PRP and microneedling are excellent treatment options but are expensive and may not be helpful for patients with severe AGA. Hair transplantation is an excellent option for individuals who have failed medical treatment. Currently, there is insufficient evidence demonstrating the effectiveness of other commonly used medications for this condition, such as topical and hormonal medications, as well as over-the-counter supplements. Furthermore, the multifactorial origin and chronic nature of AGA make it difficult to treat. Therefore, the development of more safe and effective treatment options is urgently needed.

[0004] AGA places a severe psychological burden on patients, leading to long-term mental disorders and, in severe cases, depression. Healthy, beautiful hair enhances social confidence, leading to an increasingly urgent need for hair loss treatments. In this area, there is an urgent need to find more effective and safe drugs to prevent and treat androgenic alopecia to meet these clinical needs. Summary of the Invention

[0005] To address the aforementioned technical issues surrounding androgenic alopecia, the present invention provides the use of ML348 in preventing and treating androgenic alopecia. Researchers applying ML348 to DHT-induced AGA mice found that ML348 promoted hair regeneration, increased hair length, and stimulated melanin production in hair follicles, effectively counteracting DHT-induced hair loss.

[0006] The specific technical solutions of the present invention are:

[0007] In a first aspect, the present invention provides use of ML348 in preparing a product for preventing and / or treating androgenic alopecia.

[0008] ML348 (CAS: 899713-86-1) is a de-palmitoylation inhibitor that increases palmitoylation, and is currently used in the treatment of renal fibrosis, Huntington's disease. The inventors of the present application applied ML348 to intervene in DHT-induced AGA model mice and found that ML348 has the effects of promoting hair regeneration, increasing hair length, and hair follicle melanin production, and can effectively resist hair loss caused by DHT. Based on this result, the present application can provide a new method for preventing and / or treating androgenetic alopecia.

[0009] The inventors of the present application found in the study of AGA patients that the expression of lysophospholipase 1 (LYPLA1) in the skin lesions of AGA patients was higher than that in non-skin lesions. Through multiple experiments, it was proved that this phenomenon may be related to the progress of AGA. Based on this result, the inventors of the present application further applied LYPLA1 The inventors of the present application applied the inhibitor ML348 to intervene in DHT-induced AGA model mice, and clarified the effect of ML348 on the intervention of AGA, expecting to provide a new scheme for the prevention and treatment of AGA.

[0010] As a preferred embodiment of the above technical scheme of the present application, the product is a pharmaceutical product, a daily chemical product, a food product, or a food additive.

[0011] As a preferred embodiment of the above technical scheme of the present application, the product intervenes and / or treats androgenetic alopecia by resisting the inhibition of hair follicle growth caused by DHT. The product is preferably a pharmaceutical product, a daily chemical product, a food product, or a food additive that resists the inhibition of hair follicle growth caused by DHT.

[0012] The present application discloses the molecular mechanism and application of ML348 in alleviating DHT-induced androgenetic alopecia in mice, and clarifies the effectiveness of ML348 in inhibiting abnormal hair follicle growth caused by DHT in mice, and proves the feasibility of ML348 in promoting hair regeneration, thereby providing a theoretical basis for intervention or / and treatment of androgenetic alopecia, and giving the feasibility of ML348 in intervening and / or treating androgenetic alopecia by resisting the inhibition of hair follicle growth caused by DHT.

[0013] In a second aspect, the present application provides a product for treating androgenetic alopecia, wherein the active ingredient of the product comprises ML348.

[0014] The inventors of the present application applied ML348 to intervene in DHT-induced AGA mice, and confirmed that ML348 has the effects of promoting hair regeneration, increasing hair length, and hair follicle melanin production, and can effectively resist hair loss caused by DHT. Based on this, an effective biological product for preventing and / or treating androgenetic alopecia can be provided, and the active ingredient of the product comprises ML348.

[0015] As a preferred technical scheme of the above-mentioned technical scheme of the present application, the active ingredient of the product is ML348.

[0016] As a preferred technical scheme of the above-mentioned technical scheme of the present application, the product is a pharmaceutical product, a daily chemical product, a food product, or a food additive.

[0017] As a preferred technical scheme of the above-mentioned technical scheme of the present application, the product intervenes and / or treats androgenetic alopecia by resisting the inhibition of hair follicle growth caused by DHT. The product is preferably a pharmaceutical product, a daily chemical product, a food product, or a food additive that resists the inhibition of hair follicle growth caused by DHT. In addition to being used for preparing a pharmaceutical product, ML348 can also be used for preparing a hair loss care product and a skin external additive, etc.

[0018] The present application discloses the application of ML348 in alleviating hair loss of mice under the stimulation of DHT and the related molecular mechanism, determines the effectiveness of ML348 in inhibiting abnormal growth of hair follicles of mice caused by DHT, and proves the feasibility of ML348 in promoting hair regeneration, thereby providing a basis for intervention or treatment of androgenetic alopecia and giving the feasibility of ML348 in intervening and / or treating androgenetic alopecia by resisting the inhibition of hair follicle growth caused by DHT.

[0019] Compared with the prior art, the present application has the following technical effects:

[0020] The researchers of the present application applied ML348 to intervene in DHT-induced AGA model mice, which showed that ML348 has the effects of promoting hair regeneration, increasing hair length, and generating melanin in hair follicles, and can effectively resist hair loss caused by DHT. Based on this, a new scheme for treating androgenetic alopecia is provided for preventing and / or treating androgenetic alopecia. BRIEF DESCRIPTION OF DRAWINGS

[0021] Figure 1 Figure 1 is a diagram of the hair growth conditions of mice at different times in Example 1 of the present application.

[0022] Figure 2 Figure 2 is a diagram of the H&E staining conditions of the back skin of mice in each group after being treated for 21 days in Example 2 of the present application. DETAILED DESCRIPTION

[0023] The present application will be further described below in conjunction with examples. Those skilled in the art will be able to implement the present application based on these descriptions. In addition, the examples of the present application involved in the following descriptions are generally only a part of the examples of the present application, rather than all the examples. Therefore, based on the examples in the present application, all other examples obtained by those skilled in the art without making creative efforts shall fall within the scope of protection of the present application.

[0024] The experimental animals used in the embodiments of the present application are: 20 male C57BL / 6 mice (6 weeks old, 25g ± 0.5g) purchased from the Experimental Animal Center of Hangzhou Medical College, and all the mice are raised under clean level conditions in the center.

[0025] In the embodiments of the present application, hematoxylin-eosin staining kit is purchased from Shanghai Biyun Tian Biotechnology Co., Ltd.; dihydrotestosterone (Masteril) is purchased from Shanghai Maikelin Biotechnology Co., Ltd.; ML348 is purchased from MCE company; dimethyl sulfoxide (DMSO) is purchased from Sigma-Aldrich company; melanin staining solution (ferrous sulfate method) is purchased from Zhuhai Baisuo Biotechnology Co., Ltd.

[0026] Example 1: Establishment and administration of AGA mouse model

[0027] (1) Experimental preparation

[0028] Before the experiment, the newly purchased male C57BL / 6 mice were adaptively fed for 7 days. Subsequently, according to the random principle, these mice were randomly divided into 4 experimental groups, each group of 5.

[0029] The four experimental groups are respectively:

[0030] Control group: only shaving operation, without any treatment.

[0031] DHT group: DHT is applied to the depilated area on the back to induce AGA model.

[0032] DHT / DMSO group: DHT is applied to the depilated area on the back, and DMSO is given subcutaneously.

[0033] DHT / ML348 group: DHT is applied to the depilated area on the back, and ML348 is given subcutaneously.

[0034] (2) Model establishment and administration

[0035] Day 0: On the first day of the experiment, the back of all mice was shaved and depilatory cream was applied to remove 2 square centimeters (2 cm × 2 cm) of hair as the standard.

[0036] Day 1~ Day21: From the first day of the experiment (Day 1), the mice in the DHT group, DHT / DMSO group and DHT / ML348 group were locally applied with 200 microliters of DHT solution on the back every day for 21 consecutive days. At the same time, the mice in the DHT / DMSO group additionally received subcutaneous injection of the same dose (200 microliters) of DMSO solution once a day; the mice in the DHT / ML348 group additionally received subcutaneous injection of the same dose (200 microliters) of ML348 solution once a day.

[0037] (3) Observation and recording

[0038] Photographing record: On the 7th, 14th and 21st day of the experiment, the treated area on the back of the mice was photographed to record the growth of hair, such as Figure 1 As shown. Figure 1 shown. Figure 1 In the table, Control indicates the control group, DHT indicates the DHT group, DHT / DMSO indicates the DHT / DMSO group, and DHT / ML348 indicates the DHT / ML348 group.

[0039] Day 21: At the end of the experiment, dorsal skin samples of all mice were collected for analysis of biological indicators in Examples 2 and 3.

[0040] Example 2 Hematoxylin-eosin (H&E) staining of mouse skin tissue

[0041] Skin samples measuring 1 cm × 1 cm were obtained from the back of mice. These samples were fixed with 4% paraformaldehyde, dehydrated, and embedded in paraffin. 5 μm sections were cut and oven-baked at 60°C. The sections were then treated in xylene for 15 minutes to remove the paraffin and then immersed a second time to ensure thorough dewaxing. The sections were then stained in an aqueous hematoxylin solution for 5 minutes and then rinsed in running water for 15 minutes. After staining, the sections were dehydrated in 75% and 90% ethanol, each for 10 minutes. Finally, the sections were immersed in 0.5% ethanol-eosin staining solution for 2 minutes and rinsed in running water for 1 minute. The stained sections were dehydrated in anhydrous ethanol and then cleared in xylene for 3 minutes. The sections were then mounted with neutral resin. The mounted sections were observed and photographed under a microscope.

[0042] The results of H&E staining of the back skin of mice in each group after 21 days of treatment are as follows: Figure 2 As shown, including H&E staining observation results (a), hair follicle number statistics (b) and skin thickness statistics (c). Figure 2 In the table, Control indicates the control group, DHT indicates the DHT group, DHT / DMSO indicates the DHT / DMSO group, and DHT / ML348 indicates the DHT / ML348 group.

[0043] Example 3 Melanin staining of mouse skin

[0044] Slice preparation: Place the slices of Example 2 in a 55°C constant temperature box and preheat for 30 minutes to complete dewaxing and antigen retrieval. Washing: Wash the slices once with distilled water. Melanin staining: Treat the slices with ferrous sulfate solution for 1 hour. Washing: Wash again with distilled water 3 times. Reaction: Add potassium ferrocyanide acetic acid solution and treat for 30 minutes. Washing: Wash with glacial acetic acid solution for 3 seconds and shake dry. VanGieson staining: Stain for 1 minute. Dehydration and transparency: First treat with 95% ethanol for 30 seconds, then dehydrate with anhydrous ethanol, and finally transparentize with xylene. Sealing: Sealing with neutral gum.

[0045] The melanin staining results of the mouse skin in each group were observed and photographed, and melanin staining was quantitatively analyzed as follows: the color of the positive area was selected using the IHC Toolbox of Image J software, and then the melanin-positive area was automatically outlined. The appropriate color threshold was adjusted, and the image was adjusted to 8-bit format. The threshold was adjusted to select the positive area and the entire stained area, and the area value was obtained. The percentage of the melanin-positive area / the total stained area was calculated to obtain the result.

[0046] The melanin staining results of the mouse skin in each group were grouped and analyzed. Control represents the control group, DHT represents the DHT group, DHT / DMSO represents the DHT / DMSO group, and DHT / ML348 represents the DHT / ML348 group.

[0047] Data Analysis

[0048] Figure 1 The data show the hair growth on the back of mice in the control, DHT, DHT / DMSO, and DHT / ML348 groups on days 0, 7, 14, and 21 of the experiment. On day 14, early in the experiment, mice in the control and DHT / ML348 groups showed significant hair regeneration, while the back skin of mice in the DHT and DHT / DMSO groups remained pink with no hair growth. The hair regeneration in the DHT / ML348 group increased significantly, with abundant hair growth on the back, demonstrating a significant hair restoration effect. In contrast, mice in the DHT and DHT / DMSO groups did not begin to show localized hair regeneration until the 21st day of the experiment, but the hair growth was sparse and limited in area, far less than the hair coverage in the control and DHT / ML348 groups. The results indicate that ML348 has a significant effect in resisting the DHT-induced hair loss model and promoting hair regeneration.

[0049] Figure 2 H&E staining results of each treatment group are shown. Figure 2The staining results of a and 2b showed that the hair in the growth phase of the DHT / ML348 group was longer than that in the DHT and DHT / DMSO groups, and the difference was statistically significant ( P <0.005). Meanwhile, the degree of hair regeneration in the DHT / ML348 group was similar to that in the Control group, indicating the potential of ML348 in promoting hair regeneration. Figure 2 a and 2c) showed that the dermis thickness of the Control group and the DHT / ML348 group was thicker than that of the DHT group and the DHT / DMSO group, and the difference was statistically significant ( P <0.05), but there was no statistically significant difference in dermal thickness between the Control and DHT / ML348 groups. This suggests that ML348 can intervene in DHT-induced hair growth inhibition and dermal thinning, promoting hair regeneration.

[0050] Example 3 used the ferrous sulfate method to assess skin melanin levels in different mouse groups. Melanin staining results for each group showed minimal melanin staining in the DHT and DHT / DMSO groups. Melanin levels in the control and DHT / ML348 groups were significantly higher than in the DHT group (P < 0.05), but there was no significant difference between the control and DHT / ML348 groups. This suggests that ML348 can promote melanin production.

[0051] These results suggest that ML348 promotes hair regeneration, increases hair length, and promotes melanin production in hair follicles. It also effectively combats DHT-induced hair loss, demonstrating the potential of ML348 in the treatment of AGA and providing a new treatment option.

[0052] Unless otherwise specified, the raw materials and equipment used in the present invention are commonly used in the art; the methods used in the present invention are conventional methods in the art unless otherwise specified.

[0053] The above description is only a preferred embodiment of the present invention and does not limit the present invention in any way. Any simple modification, change and equivalent transformation made to the above embodiment based on the technical essence of the present invention still fall within the scope of protection of the technical solution of the present invention.

Claims

1. Use of ML348 in the preparation of a medicament for preventing and / or treating androgenic alopecia.

2. The use according to claim 1, characterized in that: The drug is a drug for resisting the hair follicle growth inhibition caused by DHT.

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