Application of D-mannose in prevention and treatment of androgenetic alopecia

Abstract

The invention relates to the technical field of medicines, and discloses application of D-mannose in prevention and treatment of androgenetic alopecia. In the prior art, D-mannose can be a potential strategy for treating ferroptosis related diseases, can also be a non-prescription medicine supplement with high safety, and is used for treating neuritis diseases such as multiple sclerosis. The androgenic alopecia mouse model is constructed by utilizing dihydrotestosterone induction, the model mouse is subjected to mannose intervention, and the result shows that application of mannose can promote mouse hair growth, increase cortex thickness, promote hair regeneration and melanogenesis and effectively resist alopecia caused by dihydrotestosterone. The invention provides a new effect of mannose in prevention and treatment of androgenetic alopecia, and also provides a new strategy for prevention and treatment of androgenetic alopecia.

Description

Technical Field

[0001] The invention relates to the technical field of medicines, and in particular to application of D-mannose in preventing and treating androgenic alopecia. Background Art

[0002] Androgenetic alopecia (AGA) is a common type of hair loss in dermatology, which is mainly caused by the miniaturization of hair follicles. AGA has a great negative impact on the patient's mental health and social identity, leading to a decline in quality of life. Early diagnosis and treatment can significantly delay hair loss and improve the patient's quality of life. AGA can occur in both men and women, with a prevalence of approximately 21.3% in men and approximately 6.0% in women. Men usually show a significant receding hairline on the forehead and temples, or sparse hair on the top, usually accompanied by increased oil secretion; women show sparse and fine hair between the top of the head and the hairline, and the forehead hairline is retained, accompanied by increased transdermal oil secretion.

[0003] The pathogenesis of AGA is complex and is the result of the combined effects of multiple factors. Factors such as inflammation around the hair follicles, mental state, diet, and stress can aggravate the symptoms of AGA. Current studies have shown that testosterone in the human body is converted into dihydrotestosterone (DHT) under the catalysis of 5α-reductase, and androgens play an important role in the pathogenesis of AGA. Studies have found that the activity of 5α-reductase in the hair loss area of ​​AGA patients is significantly higher than that in the non-hair loss area, and the DHT content is significantly increased in the hair loss area. Excessive DHT binds to the androgen receptor (AR), activates the signal that regulates the growth of hair follicles, causes hair follicles to atrophy, hair diameter becomes smaller, hair follicles change from terminal hair to vellus hair, the growth period is shortened, and the normal physiological cycle of hair is affected, which ultimately leads to AGA.

[0004] Since AGA is a long-term process that worsens until hair loss, the earlier it is treated, the better the effect. Treatments for AGA include topical minoxidil, oral finasteride or spironolactone, hair transplant surgery, and low-energy laser irradiation, etc.). The two most effective drugs for treating AGA are topical minoxidil and oral finasteride. Minoxidil belongs to a class of drugs called vasodilators, which can normalize hair follicles and increase blood flow to the scalp, thereby promoting hair growth. Side effects of using minoxidil may include contact dermatitis, headaches, and hirsutism, and short-term resting phase shedding may occur in the first 8 weeks of treatment. Finasteride is a drug that specifically inhibits 5-alpha reductase, which can block 5-alpha reductase, thereby inhibiting the conversion of testosterone into the active form of DHT and slowing down the damage to hair follicles. However, the use of finasteride may be accompanied by side effects such as male breast development and impaired sexual function. In addition, non-drug treatments such as autologous platelet-rich plasma and microneedling are also good options, but the cost is high. Hair transplantation is an effective treatment option, which mainly involves extracting hair follicles from non-hair loss areas and transplanting them to hair loss areas, but significant results are usually seen after 6 to 9 months. The current treatments for AGA have their disadvantages. In addition, the multifactorial pathogenesis of AGA makes its treatment complicated, so there is an urgent need to develop more safe and effective treatment options. Summary of the invention

[0005] In order to solve the above technical problems of preventing and treating androgenic alopecia, the present invention provides the use of D-mannose in preventing and treating androgenic alopecia.

[0006] The specific technical scheme of the present invention is: The invention provides application of D-mannose in preparing a product for preventing and / or treating androgenic alopecia.

[0007] Mannose is the C-2 epimer of glucose, which is naturally present in many plants and fruits. It has anti-inflammatory and anti-tumor activities and plays an important role in immune regulation. In the prior art, D-mannose can be a potential strategy for treating ferroptosis-related diseases, or a safe over-the-counter supplement for the treatment of neuroinflammatory diseases such as multiple sclerosis.

[0008] The researchers of the present invention found in the study of AGA patients that the expression of phosphomannose isomerase in the lesional parts of AGA patients was significantly lower than that in the non-lesional parts. Through multiple experiments, it was verified that this phenomenon may be closely related to the progression of AGA. Based on this finding, the inventors further used the MPI agonist D-mannose to intervene in DHT-induced AGA model mice to clarify the intervention effect of D-mannose on AGA, hoping to provide a new solution for the treatment of AGA.

[0009] The researchers of the present invention applied D-mannose to intervene in the DHT-induced AGA mouse model. The results showed that D-mannose has the effects of promoting hair regeneration, increasing hair length, and promoting the production of melanin in hair follicles, and can effectively resist hair loss caused by DHT. Based on this result, the present invention discloses the application of D-mannose in the prevention and treatment of AGA, providing a new method for the prevention and treatment of AGA.

[0010] As a preferred technical solution of the present invention, the product can be food, medicine, daily chemical or skin external additive.

[0011] As a preferred technical solution of the present invention, the product intervenes and / or treats androgenic alopecia by resisting the inhibition of hair follicle growth caused by dihydrotestosterone. The product is preferably a food, medicine, daily cosmetic or external skin additive that inhibits AGA.

[0012] The present invention explores the molecular mechanism and application of D-mannose in alleviating DHT-induced AGA in mice, and clarifies the effectiveness of D-mannose in inhibiting abnormal hair follicle growth in mice caused by DHT. At the same time, the study also proves the feasibility of D-mannose in promoting hair regeneration, providing a theoretical basis for intervention and / or treatment of AGA. The results show that D-mannose has the potential to intervene and / or treat AGA by resisting the inhibition of hair follicle growth caused by DHT.

[0013] The invention provides a preparation for treating androgenic alopecia, wherein the main active ingredient of the preparation is D-mannose.

[0014] The researchers of the present invention applied D-mannose to the experiment of intervening in DHT-induced AGA mice, and confirmed that D-mannose has the effects of promoting hair regeneration, increasing hair length, and promoting the production of melanin in hair follicles, and can effectively resist hair loss caused by DHT. Therefore, this study provides an effective biological product for preventing and / or treating AGA, and its active ingredient is D-mannose.

[0015] As a preferred technical solution of the present invention, the active ingredient of the product is D-mannose.

[0016] As a preferred technical solution of the present invention, the product includes food, medicine, health care product, hair loss care product or food additive.

[0017] As a preferred technical solution of the present invention, the product intervenes and / or treats androgenic alopecia by resisting the inhibition of hair follicle growth caused by dihydrotestosterone. The product is preferably a food, medicine, health product, hair loss care product or food additive that can resist the inhibition of hair follicle growth caused by DHT.

[0018] The present invention explores the application of D-mannose in alleviating hair loss in mice under DHT stimulation and its related molecular mechanism, and clarifies the effectiveness of D-mannose in inhibiting abnormal hair follicle growth in mice caused by DHT. At the same time, the study also proves the feasibility of D-mannose in promoting hair regeneration, providing a scientific basis for intervening or treating AGA. The results show that D-mannose can effectively intervene and / or treat AGA by resisting the inhibition of hair follicle growth caused by DHT.

[0019] Compared with the prior art, the present invention has the following technical effects: In the prior art, D-mannose can be a potential strategy for treating ferroptosis-related diseases, or a highly safe over-the-counter supplement for treating neuroinflammatory diseases such as multiple sclerosis. The present invention uses DHT to induce an AGA mouse model, and intervenes with mannose in the mice. The results show that the application of mannose can promote hair growth, increase cortical thickness, promote hair regeneration and melanin production in mice, and effectively resist hair loss caused by DHT. Based on this, the present invention provides a new role of mannose in the prevention and treatment of AGA, and also provides a new strategy for the prevention and treatment of AGA. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1 The hair growth of mice at different time points in Example 1 of the present invention is shown; Figure 2 The H&E staining of the back skin of each group of mice after 21 days of treatment in Example 2 of the present invention is shown; Figure 3 The melanin staining results of the mouse skin in each group in Example 3 of the present invention are shown. DETAILED DESCRIPTION

[0021] The present invention will be further described below in conjunction with embodiments. Those of ordinary skill in the art will be able to implement the present invention based on these descriptions. In addition, the embodiments of the present invention involved in the following description are generally only embodiments of a part of the present invention, rather than all embodiments. Therefore, based on the embodiments in the present invention, all other embodiments obtained by those of ordinary skill in the art without making creative work should belong to the scope of protection of the present invention.

[0022] The experimental animals used in the present embodiment are: 20 6-week-old male C57BL / 6 mice (weight 50 g±0.2 g) purchased from the Experimental Animal Center of Hangzhou Medical College. All mice were raised under clean-grade conditions of the center.

[0023] In the examples of the present invention, the hematoxylin-eosin staining kit was purchased from Shanghai Bio-Technology Co., Ltd.; DHT was purchased from Shanghai McLean Biochemical Technology Co., Ltd.; D-mannose was purchased from MCE; dimethyl sulfoxide (DMSO) was purchased from Sigma-Aldrich; and melanin staining solution (ferrous sulfate method) was purchased from Zhuhai Beso Biotechnology Co., Ltd.

[0024] Example 1 Establishment and drug administration of AGA mouse model (1) Experimental preparation Before the experiment, male C57BL / 6 mice were acclimated for 7 days and then randomly divided into 4 experimental groups, with 5 mice in each group.

[0025] The four experimental groups are: Control group (CK): hair removal was performed without any other treatment.

[0026] DHT group: DHT was applied to the hair removal area on the back to induce the AGA model.

[0027] Saline / DHT group: DHT was applied to the hair removal area on the back and saline treatment was given (saline was injected intraperitoneally).

[0028] D-mannose / DHT group: DHT was applied to the hair removal area on the back, and mannose treatment was supplemented (D-mannose was injected intraperitoneally).

[0029] (2) Model establishment and drug administration One day before the experiment (day 0): shave the backs of all mice and apply depilatory cream to remove 2 cm × 2 cm of hair.

[0030] Day 1 to Day 21 of the experiment: Starting from the first day of the experiment, for 21 consecutive days, mice in the DHT group, saline / DHT group, and D-mannose / DHT group were topically smeared with 200 µL of DHT solution on their backs every day. At the same time, mice in the D-mannose / DHT group received an additional intraperitoneal injection of 200 µL of D-mannose solution every day. The daily dose of D-mannose injected into mice was 2 g of D-mannose per kg of mice; mice in the saline / DHT group received an additional intraperitoneal injection of saline equal to the dose of D-mannose solution every day.

[0031] (3) Observation and recording Photographing records: On the 21st day of the experiment, the treated area on the back of the mice was photographed to record the growth of hair. Figure 1 shown.

[0032] Day 21: At the end of the experiment, dorsal skin samples of all mice were collected for strong histological detection in Examples 2 and 3.

[0033] Example 2 Hematoxylin-eosin (H&E) staining of mouse skin tissue Skin samples of 1 cm2 (1 cm×1 cm) were obtained from the back of mice and fixed in 4% paraformaldehyde solution, followed by dehydration and paraffin embedding. The samples were cut into 5 µm sections and dried in an oven at 60°C. The sections were treated with xylene for 15 minutes to remove paraffin and immersed a second time to ensure thorough dewaxing. Then, the sections were stained in a hematoxylin aqueous solution for 5 minutes and then rinsed with running water for 15 minutes. Next, the sections were dehydrated in 75% and 90% ethanol, each for 10 minutes. After that, the sections were immersed in 0.5% ethanol-eosin staining solution for 2 minutes for staining and rinsed with running water for 1 minute. After staining, the sections were dehydrated again in anhydrous ethanol, then transparentized in xylene for 3 minutes, and this step was repeated. Finally, the sections were mounted with neutral resin and observed and photographed under a microscope.

[0034] On the 21st day after the start of the experiment, the H&E staining results of the back skin of mice in each group were as follows: Figure 2 As shown, including (a) H&E staining observation results, (b) hair follicle number statistics and (c) skin thickness statistics.

[0035] Example 3 Melanin staining of mouse skin In this example, melanin staining observation and analysis of the experimental mouse skin was performed according to the following steps: (1) Slice preparation: Place the slices of Example 2 in a 55°C incubator for preheating for 30 minutes to complete dewaxing and antigen retrieval.

[0036] (2) Cleaning: Clean the sections once with distilled water.

[0037] (3) Melanin staining: Treat the sections with ferrous sulfate solution for 1 hour. Washing: Wash again with distilled water 3 times.

[0038] (4) Reaction: Add potassium ferrocyanide acetic acid solution dropwise and treat for 30 minutes.

[0039] (5) Cleaning: Clean with glacial acetic acid solution for 3 seconds and shake dry.

[0040] (6) Van Gieson staining: stain for 1 minute.

[0041] (7) Dehydration and transparency: first treat with 95% ethanol for 30 seconds, then dehydrate with anhydrous ethanol, and finally transparentize with xylene.

[0042] (8) Sealing: Use neutral gum for sealing.

[0043] (9) Observe and photograph the melanin staining results of the skin of each group of mice, and perform quantitative analysis of melanin staining. The analysis method is as follows: Use the IHC Toolbox tool in the Image J software to select the positive area, and the software will automatically outline the melanin-stained positive area. Next, adjust the appropriate color threshold, convert the image to 8-bit format, and select the positive area and the entire stained area by adjusting the threshold to obtain the area data. Finally, calculate the percentage of the melanin-stained positive area to the total stained area to obtain the final result.

[0044] Melanin staining results of mouse skin in each group are shown in Figure 3 .

[0045] Results Analysis Figure 1 The back hair growth of mice in each group on days 0 and 21 is shown. By day 21, the back hair of mice in the CK group was almost completely regenerated, covering most of the area; and the mice in the D-mannose / DHT group also showed significant hair regeneration, and their hair coverage was significantly higher than that of the DHT group and the saline / DHT group. Overall, the mice in the D-mannose / DHT group showed a clear advantage in hair regeneration, close to the CK group, suggesting that D-mannose may have the effect of promoting hair regeneration. The results showed that D-mannose can promote hair regeneration in DHT-induced AGA model mice.

[0046] Figure 2 H&E staining results of each treatment group are shown. Figure 2 The staining results of a and 2b showed that the length of the growing hair in the D-mannose / DHT group was significantly longer than that in the DHT group and the saline / DHT group, and the difference was statistically significant (P<0.005). At the same time, the degree of hair regeneration in the D-mannose / DHT group was similar to that in the CK group, showing the potential of D-mannose in promoting hair regeneration. Comparison of the dermal thickness results of each treatment group ( Figure 2 a and 2c) showed that the skin thickness of mice in the CK group and D-mannose / DHT group was significantly higher than that in the DHT group and saline / DHT group, and the difference was statistically significant (P<0.05). However, there was no statistically significant difference in skin thickness between the CK group and the D-mannose / DHT group. The results showed that D-mannose could intervene in the inhibition of hair growth and thinning of skin thickness caused by DHT, and had the effect of promoting hair regeneration.

[0047] Figure 3 Results of the ferrous sulfate method for assessing skin melanin levels in different mouse groups are presented. Figure 3It can be seen that there is very little melanin staining in the skin of mice in the DHT group and the saline / DHT group, while the melanin levels in the CK group and the D-mannose / DHT group are significantly higher than those in the DHT group, and the difference is statistically significant (P<0.05). In addition, the melanin distribution in the D-mannose / DHT group is also significantly higher than that in the CK group, and the difference is also statistically significant (P<0.001). This shows that D-mannose can promote the production of melanin.

[0048] Based on the above results, D-mannose was shown to promote hair regeneration, increase hair length, and promote melanin production in hair follicles. It can be seen that D-mannose can effectively resist hair loss caused by DHT, showing the potential value of D-mannose in the treatment of AGA and providing a new solution for the treatment of AGA.

[0049] The raw materials and equipment used in the present invention, unless otherwise specified, are all commonly used raw materials and equipment in the art; the methods used in the present invention, unless otherwise specified, are all conventional methods in the art.

[0050] 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 falls within the protection scope of the technical solution of the present invention.

Claims

1. Use of D-mannose in preparing products for preventing and / or treating androgenic alopecia.

2. The use according to claim 1, characterized in that: The product is medicine, daily chemical product, food, or food additive.

3. The use according to claim 2, characterized in that: The product is a medicine, daily chemical product, food, or food additive for resisting the inhibition of hair follicle growth caused by dihydrotestosterone.

4. A product for treating androgenic alopecia, characterized in that: The active ingredient of the preparation includes D-mannose.

5. A product for treating androgenic alopecia according to claim 4, characterized in that: The active ingredient of the product is D-mannose.

6. A product for treating androgenic alopecia according to claim 4, characterized in that: The product is medicine, daily chemical product, food, or food additive.

7. A product for treating androgenic alopecia according to claim 6, characterized in that: The product is a medicine, daily chemical product, food, or food additive for resisting the inhibition of hair follicle growth caused by dihydrotestosterone.

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