Application of mitochondrial quinone in the intervention and treatment of androgenetic alopecia

Abstract

This invention relates to the field of pharmaceutical technology and discloses the application of mitochondrial quinone in the intervention and treatment of androgenetic alopecia. This invention explores the application and related molecular mechanisms of mitochondrial quinone in alleviating hair loss in mice stimulated by DHT, clarifying the effectiveness of mitochondrial quinone in inhibiting abnormal hair follicle growth and promoting hair regeneration in mice induced by dihydrotestosterone. This provides experimental evidence for the clinical application of mitochondrial quinone in the intervention and treatment of hair loss diseases, especially androgenetic alopecia, and offers another safe and effective substance for the effective prevention and treatment of androgenetic alopecia.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of medicine, in particular to the application of mitochondrial quinone in intervention and treatment of androgenetic alopecia. BACKGROUND

[0002] Androgenetic alopecia (AGA) is also known as seborrheic alopecia or alopecia precox, which is a hair loss disease occurring in adolescence or after adolescence, and is also one of the most common causes of progressive hair loss. In men, AGA mainly manifests as backward movement of the frontal hairline, progressive reduction of the hair on the top of the head, and thinning of the hair, and the area of hair loss gradually increases; in women, AGA is less likely to accumulate in the frontal hairline and temporal region, but the hair on the top of the head is diffusely reduced, thinned, and the hairline is widened. The main pathological manifestations of AGA are shortening of the anagen phase of the hair follicle, which leads to miniaturization of the hair follicle, and the terminal hair rich in pigment and medulla is replaced by vellus hair which is thin, soft, and has little pigment. Studies have shown that the pathogenesis of AGA involves many factors, including genetic susceptibility, abnormal androgen metabolism, gene changes, abnormal molecular signal transduction pathways, and inflammatory reactions in the microenvironment of the hair follicle region.

[0003] The therapeutic drugs and methods for AGA each have advantages and limitations, and the treatment scheme of oral drugs combined with external drugs is generally used in clinical practice. At present, the drugs approved by the Food and Drug Administration (FDA) include finasteride and minoxidil, which are respectively used for male AGA and female AGA. Oral drugs for men also include dutasteride, which, like finasteride, mainly inhibits type II 5α-reductase to inhibit the reduction of testosterone to dihydrotestosterone (DHT), improve the microcirculation of hair follicles, and promote hair growth; women can mainly use external minoxidil to treat, which can open the potassium channels of vascular smooth muscle and hair follicles, induce the relaxation of micro- and small blood vessels, and stimulate the microcirculation around the hair follicle; they can also choose oral spironolactone and sex hormones (ethinyl estradiol and large doses of cyproterone acetate) to counteract androgens. In recent years, injection of platelet-rich plasma, microneedle, low-intensity laser therapy, hair transplantation, and fat-derived stem cell component extract as emerging treatment methods can be used as an adjunct to mainstream drugs.

[0004] The level of androgens in AGA patients is usually within the normal range, because in addition to the hormone synthesis pathway in the gonads and other organs, sex hormones can be independently synthesized and interact in the hair follicle to affect the transcription and expression of genes. AGA is accompanied by aging of androgen-sensitive DPCs located at the base of the hair follicle. High levels of DHT can cause DPCs to age or degenerate.

[0005] Healthy and beautiful hair can give people a good image, and make people more confident in interpersonal communication. People pay more and more attention to the prevention and treatment of hair loss. In this field, it is still necessary to find a safe and efficient drug to effectively prevent and treat androgenetic alopecia. SUMMARY

[0006] In order to solve the above technical problems of preventing and treating androgenetic alopecia, the application provides the application of mitochondrial quinone in intervention and treatment of androgenetic alopecia. The application explores the application of mitochondrial quinone in relieving hair loss of mice under the stimulation of DHT and the related molecular mechanism, and determines the effectiveness of mitochondrial quinone in inhibiting abnormal growth of mouse hair follicles caused by dihydrotestosterone and promoting hair regeneration, thereby providing experimental basis for intervention or treatment of alopecia, especially androgenetic alopecia, and giving the application of mitochondrial quinone in intervention and treatment of androgenetic alopecia.

[0007] The specific technical scheme of the application is as follows:

[0008] In one aspect, the application provides the application of mitochondrial quinone in the preparation of products for intervention and / or treatment of androgenetic alopecia.

[0009] Mitochondrial quinone is a fat-soluble substance existing in mitochondria, and is an endogenous substance in the human body. In the prior art, mitochondrial quinone is applied as a mitochondrial targeted antioxidant. The application takes the hair follicles and cells in the hair follicle site of mouse skin as the research object, and determines the role of mitochondrial quinone in resisting mitochondrial dysfunction of dermal papilla cells and abnormal hair growth cycle of AGA model mice. Based on this, the application of mitochondrial quinone in intervention and treatment of androgenetic alopecia is given, and another safe and efficient substance for effectively preventing and treating androgenetic alopecia is provided.

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

[0011] As a preferred embodiment of the above technical scheme of the 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 drug, a daily chemical product, a food or a food additive that resists the inhibition of hair follicle growth caused by DHT.

[0012] The application explores the application of mitochondrial quinone in relieving hair loss of mice under the stimulation of DHT and the related molecular mechanism, determines the effectiveness of mitochondrial quinone in inhibiting abnormal growth of mouse hair follicles caused by dihydrotestosterone, and proves the feasibility of mitochondrial quinone in promoting hair regeneration, thereby providing a basis for intervention or treatment of androgenetic alopecia, and giving the feasibility of mitochondrial quinone in intervening and / or treating androgenetic alopecia by resisting the inhibition of hair follicle growth caused by DHT.

[0013] In one aspect, the present application provides a preparation for treating androgenetic alopecia, wherein the active ingredient of the preparation comprises a mitochondrial quinone.

[0014] The present application takes the hair follicle and the cells in the hair follicle of the mouse skin as the research object, and determines the role of the mitochondrial quinone in resisting the mitochondrial function abnormality of the dermal papilla cells and the abnormality of the hair growth cycle of the AGA model mouse. Based on this, the application of the mitochondrial quinone in the intervention and treatment of androgenetic alopecia is given.

[0015] As a preferred embodiment of the above technical scheme of the present application, the active ingredient of the preparation is a mitochondrial quinone.

[0016] The present application determines the effectiveness of the mitochondrial quinone in inhibiting the abnormal growth of the mouse hair follicle caused by dihydrotestosterone, and proves the feasibility of the mitochondrial quinone in promoting hair regeneration, thereby providing a basis for the intervention or treatment of androgenetic alopecia, and giving the feasibility of the mitochondrial quinone in the intervention and / or treatment of androgenetic alopecia by resisting the inhibition of the hair follicle growth caused by DHT.

[0017] As a preferred embodiment of the above technical scheme of the present application, the preparation is a medicine, a daily chemical product, a food, or a food additive.

[0018] As a preferred embodiment of the above technical scheme of the present application, the preparation is a medicine, a daily chemical product, a food, or a food additive.

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

[0020] The present application determines the effectiveness of the mitochondrial quinone in inhibiting the abnormal growth of the mouse hair follicle caused by dihydrotestosterone, and proves the feasibility of the mitochondrial quinone in promoting hair regeneration, thereby providing a basis for the intervention or treatment of androgenetic alopecia, and giving the feasibility of the mitochondrial quinone in the intervention and / or treatment of androgenetic alopecia by resisting the inhibition of the hair follicle growth caused by DHT. BRIEF DESCRIPTION OF DRAWINGS

[0021] Figure 1 The figure is the hair growth condition of the mouse in different periods in Example 1 of the present application;

[0022] Figure 2 The figure is the H&E staining condition of the back skin of the mouse in each group after being treated for 19 days in Example 3 of the present application;

[0023] Figure 3Figure of Ki67 immunofluorescence staining results of each group of mice in Example 4 of the present application;

[0024] Figure 4 Figure of the observation results of melanin staining of the skin of each group of mice in Example 5 of the present application. DETAILED DESCRIPTION

[0025] The present application will be further described below in conjunction with examples and drawings. In the examples of the present application, the mitochondria quinone is abbreviated as MitoQ.

[0026] The sources and processing of the materials related to the examples of the present application are as follows:

[0027] ① Experimental materials Experimental animals: clean grade 6-week-old male C57BL / 6 mice (purchased from Zhejiang Academy of Medical Sciences Animal Experimental Center). The mice were randomly divided into the following groups: untreated group (control), DHT group, DHT+MitoQ group, 4 in each group, and were raised in separate cages.

[0028] ② Raising

[0029] The Zhejiang Academy of Medical Sciences Animal Experimental Center was responsible for the daily raising of the mice.

[0030] ③ Reagents

[0031] The hematoxylin-eosin staining kit was purchased from Shanghai Biyun Tian Biotechnology Co., Ltd.; DHT (androstanolone) was purchased from Shanghai Maikelin Biotechnology Co., Ltd.; MitoQ was purchased from Xi'an Ruixi Biological Technology Co., Ltd.; dimethyl sulfoxide (DMSO) was purchased from Sigma-Aldrich Company; Anti-Ki67 antibody was purchased from Abeam (Shanghai) Trading Co., Ltd.; fluorescent secondary antibody was purchased from Nanjing Aisiyi Biological Technology Co., Ltd.; melanin staining solution (ferrous sulfate method) was purchased from Zhuhai Baisuo Biological Technology Co., Ltd.

[0032] Example 1 Establishment of mouse model

[0033] After the purchased mice were adaptively fed for 7 days, they were randomly divided into groups: control group (shaved only), DHT group (DHT applied on the nape), DHT+MitoQ group (DHT applied on the nape and MitoQ injected intraperitoneally), 4 in each group, a total of 12, raised in separate cages. On the experimental day (recorded as Day 0), the mice were shaved on the back and applied depilatory cream to remove hair, with an area of about 2cm×2cm. The next day (recorded as Day 1), 200μl of DHT solution was applied to the depilated area, and the treatment group was injected with 200μl of MitoQ solution intraperitoneally, once a day, for a total of 19 days. During the 19 days, the corresponding treatment was implemented at a fixed time every day according to different groups. On the 19th day, the back skin of the mice was collected for subsequent index detection.

[0034] Photographs were taken on the 0th, 10th, and 19th days, and the results are as followsFigure 1 Results are shown in FIG. 1.

[0035] Example 2 Preparation of wax blocks and sectioning

[0036] Fresh dorsal skin tissue (1 cm x 1 cm) of mice was fixed with 4% paraformaldehyde, dehydrated and then embedded in paraffin. The wax blocks were cut into sections with a thickness of 4 pm and then placed in an oven at 60°C for baking, for use in subsequent experiments.

[0037] Example 3 Hematoxylin and eosin (H&E) staining

[0038] The sections obtained in Example 2 were soaked in xylene for 15 min, repeated once, and then dehydrated in a graded ethanol series and placed in a hematoxylin aqueous solution for 5 min. The sections were then rinsed with running water for 15 min, dehydrated in 75% and 90% ethanol for 10 min each, and then taken out and placed in a 0.5% ethanol- eosin staining solution for 2 min. The sections were then rinsed with running water for 1 min, and then dehydrated in anhydrous ethanol.

[0039] The sections were then transparentized in xylene for 3 min, repeated once, and then mounted with neutral resin. The sections were observed under a microscope and the results are shown in FIG. 3, where a is a graph of the observation results of H&E staining, b is a graph of the number of hair follicles, and c is a graph of the thickness of the skin. (** represents P < 0.01, *** represents P < 0.001, and ns represents no statistical difference) Figure 2

[0040] Example 4 Immunofluorescence observation

[0041] The sections obtained in Example 2 were placed in a thermostat at 55°C for 30 min. After deparaffinization and completion of antigen repair, the sections were incubated with 5% goat serum, and then placed in a wet box at 37°C for 30 min in the dark. Then, 1:200 diluted Ki67 rabbit anti-antibody (Abeam, Cambridge, UK) was added, and the sections were incubated at 4°C overnight. The next day, after rewarming for 15 min, the primary antibody was removed and the sections were washed with PBS 3 times, each for 5 min. Then, 1:5000 diluted orange fluorescent mouse anti-antibody (ACE, Nanjing, China) was added, and the sections were incubated at room temperature in the dark for 1 h. Then, the secondary antibody was removed and the sections were washed with PBS 3 times, each for 5 min. Then, DAPI working solution was added, and the sections were incubated at room temperature in the dark for 10 min. Then, the sections were washed with PBS 3 times, each for 5 min. After mounting with anti-fluorescence quencher, the sections were observed under a fluorescence microscope (IX70-141 inverted microscope, Olympus, Japan), and the results are shown in FIG. 4. Figure 3

[0042] Example 5 Melanin staining

[0043] Melanin staining was performed according to the melanin staining kit (ferrous sulfate method), and the specific steps were as follows:

[0044] ​​The section obtained in Example 2 was placed in a thermostat at 55°C for 30 min, dewaxed and antigen repaired, washed once with distilled water, treated with ferrous sulfate solution for 1 h, washed three times with distilled water, and then treated with potassium ferricyanide-acetic acid solution for 30 min. After washing with acetic acid solution for 3 s, the section was dried, dyed with Van Gieson solution for 1 min, removed from the dye, placed in 95% ethanol for 30 s, dehydrated with anhydrous ethanol, cleared with xylene, and mounted.

[0045] The results of observing the melanin staining of the skin of the mice in each group are shown in Figure 4 .

[0046] Data analysis

[0047] (1) Analysis of the data results of Example 1

[0048] Figure 1 The results of observing the back hair growth of the mice in each group at 0 days, 10 days and 19 days of drug treatment are shown in

[0049] As can be seen from Figure 1 , at 10 days, hair regeneration was observed on the backs of the mice in the control group and the DHT+MitoQ group, while the skin of the mice in the DHT group was pink and no hair regeneration was observed. At 19 days, the hair on the backs of the mice in the control group had almost completely regenerated, the hair of the mice in the DHT+MitoQ group had regenerated significantly and the skin had turned grayish black, and the skin of the mice in the DHT group had not changed significantly. This shows that MitoQ has the effect of promoting hair regeneration.

[0050] (2) Analysis of the data results of Example 3

[0051] Figure 2 The results of observing the skin of the mice in each group on day 19 are shown in

[0052] As shown in Figure 2 a, the number of guard hairs in the DHT group was significantly increased compared with the control group and the DHT+MitoQ group, while the number of terminal hairs was reduced compared with the control group and the DHT+MitoQ group, and it was observed that the DHT+MitoQ group could restore the abnormal morphology of the skin tissue caused by DHT. As shown in Figure 2 a and 2b, a large number of hair follicles were observed in the control group and the DHT+MitoQ group, and the number of hair follicles in the DHT group was significantly reduced compared with the control group and the DHT+MitoQ group. As shown in Figure 2 a and 2c, the thickness of the dermis in the DHT group was significantly thinner than that in the control group and the DHT+MitoQ group, thus showing that the hair growth period of the mice in the DHT group was delayed, and MitoQ can promote hair to enter the growth period.

[0053] Thus, it can be seen that MitoQ has the effect of increasing the number of hair follicles and terminal hairs.

[0054] (3) Analysis of the data results of Example 4

[0055] Figure 3 The images show the results of Ki67 immunofluorescence staining in mice of each group.

[0056] like Figure 3 As shown, significant cell proliferation was observed in the hair follicle area in the control group and the DHT+MitoQ group, while no significant proliferation was observed in the DHT group. The hair on the back of the mice in the DHT group had not yet entered the growth phase. Therefore, it can be concluded that the intervention of MitoQ can promote the proliferation of hair follicle cells.

[0057] (4) Analysis of the data results of Example 5

[0058] Figure 4 The image shows the results of melanin staining observation on the skin of mice in each group.

[0059] like Figure 4 As shown, obvious melanin distribution was observed in the hair follicles of both the control group and the DHT+MitoQ group, while melanin staining was rare in the hair follicles of the DHT group, indicating that MitoQ promotes melanin production in hair follicles.

[0060] In summary, mitochondrial quinone promotes hair regeneration, increases the number of hair follicles and terminal hairs, promotes hair follicle cell proliferation, and promotes melanin production in hair follicles. Furthermore, it demonstrates that mitochondrial quinone can inhibit dihydrotestosterone-induced abnormal hair follicle growth in mice, effectively promote hair regeneration, and proves the feasibility of mitochondrial quinone in intervening in androgenetic alopecia by resisting DHT-induced hair follicle growth inhibition.

[0061] The above embodiments further explore the application and related molecular mechanisms of MitoQ in alleviating DHT-induced hair loss in mice at the animal experimental level. By administering MitoQ via intraperitoneal injection combined with application of DHT solution to the hair-removed area on the back, the effectiveness of MitoQ in inhibiting abnormal hair follicle growth induced by dihydrotestosterone (DHT) and promoting hair regeneration in mice was clarified. This provides experimental evidence for the clinical application of MitoQ in the intervention or treatment of hair loss diseases, especially androgenetic alopecia, and provides a theoretical basis for its use as a drug, daily chemical product, food, food additive, or its active ingredient in the intervention and / or treatment of androgenetic alopecia.

[0062] Unless otherwise specified, the raw materials and equipment used in this invention are all commonly used in the field; unless otherwise specified, the methods used in this invention are all conventional methods in the field.

[0063] The above description is merely a preferred embodiment of the present invention and is not intended to limit the present invention in any way. Any simple modifications, alterations, and equivalent transformations made to the above embodiments based on the technical essence of the present invention shall still fall within the protection scope of the present invention.

Claims

1. Use of a mitochondrial quinone for the preparation of a medicinal product for the treatment of androgenetic alopecia, characterized in that: The mitochondrial quinone is MitoQ.

2. Use according to claim 1, characterized in that: The pharmaceutical product is against DHT caused hair follicle growth inhibition.

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