Medicinal application of nardosinone in prevention or treatment of alopecia and promotion of hair growth

Abstract

The invention relates to a drug application of nardosinone in preventing or treating alopecia and promoting hair growth. The medicine for preventing or treating alopecia comprises nardosinone or pharmaceutically usable salts, esters or glycosides thereof. The nardosinone is used as a unique active component, 75% ethanol is adopted to prepare a solution with the concentration range of 1.0-3.0 mg / mL, the solution is externally applied to scalp, and the administration dosage range of the solution for preventing or treating alopecia of adults is 1-10 mL per day. The existing medicine for hair loss and alopecia is poor in treatment effect and has side effects. Research finds that nardosinone serving as an active component can achieve the application effect of preventing or treating alopecia and white hair by regulating and controlling the level of TGF beta family genes closely related to hair development and growth. The invention verifies that the active monomer nardosinone derived from rhizoma nardostachyos has the potential of preventing or treating the alopecia for the first time, the alopecia can be prevented or treated through skin administration of a certain dose of nardosinone, and a new method is provided for preventing or treating the alopecia.

Description

Technical Field

[0001] The invention belongs to the technical field of biomedicine, and in particular relates to the application of naringenin in the preparation of a drug for preventing or treating hair loss. Background Art

[0002] With the increase of multiple stress factors such as society, work, and life, the number of people suffering from hair loss is increasing, and the trend is getting younger. In developed countries in Europe and America, the hair loss rate of adult men is 32%, and that of women is 28%; in Asian countries, the hair loss rate of adult men is 25%, and that of women is 22%. Although hair loss is not a serious disease that threatens human life, it brings great mental pain to patients. Some patients, especially young patients, develop a serious inferiority complex, which affects their personal self-confidence and social interaction.

[0003] At present, there are two main types of drug treatments for hair loss: one is finasteride and steroid hormone drugs. However, finasteride has a series of side effects, including sexual dysfunction, moderate to severe depression, mood disorders, sleep and eating disorders, and increases the risk of breast cancer in men. Another commonly used drug is vasodilators such as minoxidil. However, the use of minoxidil can cause allergic reactions in some patients, accompanied by itchy scalp, increased dandruff, etc. A small number of patients develop hirsutism, and severe cases can cause chest tightness and dizziness. In summary, due to the toxic and side effects of chemical drugs, the demand for safer and gentler plant alternative treatments has increased significantly. Therefore, it is of great social significance to find a natural product that is effective in treating or preventing hair loss and is high-quality, low-cost, and easy to use.

[0004] As a sesquiterpenoid compound derived from the traditional Chinese medicinal material Nardostachys grandiflora, Nardostachys grandiflora has a wide range of and precise pharmacological activities, such as sedation, anti-epileptic, anti-depressant, promotion of nerve growth, and lowering of blood pressure.

[0005] So far, there is no report on the research and development of the application of gadolinium to treat hair loss. Summary of the invention

[0006] The present invention aims at the problem that existing hair loss and alopecia drugs have poor therapeutic effects and side effects. Through research, it is found that galanthin, as an active ingredient, can achieve the effect of preventing or treating hair loss and gray hair by regulating the level of TGFβ family genes that are closely related to hair development and growth. Therefore, the present invention proposes the use of galanthin in products or drugs for preventing or treating hair loss and promoting hair growth.

[0007] The technical solution adopted by the present invention is:

[0008] As an active ingredient, galanthin (theoretically, including its pharmaceutically usable salts, esters or glycosides) is used for preventing or treating hair loss and promoting hair growth.

[0009] In the application, nardostrobin is the only active ingredient, and 75% ethanol is used to prepare a nardostrobin solution with a concentration range of 1.0-3.0 mg / mL.

[0010] The naringasol solution is applied externally to the scalp, and the dosage for preventing or treating hair loss in adults (60 kg) is 1-10 mL per day.

[0011] Application of naringenone or its pharmaceutically usable salts, esters or glycosides in the preparation of products or medicines for preventing or treating hair loss and promoting hair growth.

[0012] In the drug for preventing or treating hair loss, the content of narostane or its pharmaceutically usable salts, esters or glycosides as an active ingredient ranges from 1 to 100 mg.

[0013] The application is to prepare a naringenone solution with 75% ethanol, which is applied externally to the scalp. The dosage range for preventing or treating hair loss in adults (60 kg) should be 1-30 mg per day.

[0014] The medicine is in the form of an external dosage form and can be a microemulsion, emulsion, cream, spray, gel, tincture or liniment.

[0015] Beneficial effects of the invention:

[0016] 1. The present invention verifies for the first time that the active monomer naringenin derived from naringenin has the potential to prevent or treat hair loss. A certain dose of naringenin can be administered through the skin to prevent or treat hair loss, providing a new method for preventing or treating hair loss. Naphthoxin is the main active ingredient in naringenin, which can be extracted from natural plants, and the extraction method is mature. In addition, naringenin can be administered through the skin or orally, which is easy to be accepted and conducive to clinical promotion and application.

[0017] The androgenic alopecia model of mice was established by subcutaneously injecting testosterone propionate 5 mg / kg / d into the hair loss area of ​​mice for 18 consecutive days. An equal volume of the corresponding drug was given by applying the prepared naringenin solution to the skin once a day for 18 consecutive days to observe the effect of naringenin on hair growth in mice with androgenic alopecia model. It was found that naringenin can effectively alleviate the inhibitory effect of testosterone propionate on hair growth of C57BL / 6 mice.

[0018] 2. Nardone has a significant therapeutic effect on the hair loss of mice caused by testosterone propionate, and it can significantly promote the hair growth of mice and increase the number of hair follicles. The research results of the present invention show that in the mouse hair loss model caused by testosterone propionate, after 18 days of skin administration of ardone to mice, the patchy hair loss on the back of mice caused by testosterone propionate can be significantly reduced, and the hair of mice can be made shiny, soft and denser; the results of pathological sections show that the administration of ardone can increase the number of skin hair follicles in testosterone propionate model mice and promote hair follicle growth; qPCR test results show that ardone can inhibit the expression of TGFβ1 and TGFβ2mRNA caused by testosterone propionate. The results of the effect of ardone on the expression of TGFβ1 and TGFβ2mRNA in the skin tissue of mice with androgenic alopecia model show that ardone can promote hair growth by inhibiting the expression of TGFβ1 and TGF-β2mRNA. By experimentally observing the effect of naringenin on the recovery of hair follicle tissue in mice with androgenic alopecia, it can be confirmed that naringenin can improve the reduction in the number of hair follicles caused by testosterone propionate and maintain the normal morphology of hair follicles. The above results show that naringenin, a natural compound derived from naringenin, has the potential to prevent or treat hair loss.

[0019] 3. The present invention is applied to products for preventing or treating hair loss. As an active ingredient, naringenin can achieve the effect of preventing or treating hair loss and gray hair by regulating the level of TGFβ family genes that are closely related to hair development and growth. The raw material cost of naringenin is low and easy to obtain, and it has a good prospect for use in drugs for preventing or treating hair loss. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1 This is a comparison of the differences in skin appearance of C57BL / 6 mice in each group in the efficacy test;

[0021] Figure 2 It is a comparison of the skin tissue pathological sections (HE staining) of C57BL / 6 mice in each group in the effect test (10×);

[0022] In the figure, A: HE staining of the cross-section of the skin of mice in each group; B: HE staining of the longitudinal section of the skin of mice in each group;

[0023] Figure 1 , Figure 2 In the table, Control: blank control group; Model: model group; L-Nar: low-dose narostane group; H-Nar: high-dose narostane group; Minoxidil: minoxidil group. DETAILED DESCRIPTION

[0024] In order to make the technical concept and advantages of the present invention to achieve its invention purpose more clearly, the technical solution of the present invention is further described in detail below with reference to the accompanying drawings. It should be understood that the following embodiments are only used to explain and illustrate the preferred implementation of the present invention, and should not be regarded as and do not constitute a limitation on the scope of patent protection claimed by the present invention. In addition, the technical features involved in each implementation described below can be combined with each other as long as they do not conflict with each other.

[0025] Embodiment and application: Experimental study on the treatment of androgenic alopecia model mice with naringenone

[0026] 1. Materials:

[0027] 1. Animals: 60 C57BL / 6 male mice, 6 weeks old, weighing 18-22 g, provided by Guangzhou Yancheng Biotechnology Co., Ltd., and kept for use after inspection and quarantine. Animal production license number SCXK (Guangdong) 2017-0174.

[0028] 2. Drugs and reagents: Preparation of naringenin solution: naringenin solution containing 1.0 and 3.0 mg / mL naringenin was prepared with 75% ethanol; 5% minoxidil tincture (Xiamen Meishang Pharmaceutical Co., Ltd.).

[0029] The chemical structure of naringenone is as follows:

[0030]

[0031] 3. Instruments: Thermo Fisher Evos M5000 microscope, Huawei mobile phone.

[0032] 2. Test methods:

[0033] 1. Preparation of mouse hair loss model: After healthy SPF-grade C57BL / 6 mice were fed for one week, they were anesthetized by intraperitoneal injection of 0.3% sodium pentobarbital solution. A 2×4 cm area was selected at 0.5 cm from the proximal end of the tail. 2 After the long side of the mouse is parallel to the depilatory area of ​​the spine, use a pet shaver to shave all the hair in the depilatory area on the back short, until the back skin is vaguely visible. Then apply the depilatory cream evenly to the shaved area. After further soaking the cotton ball with saline, wipe off the depilatory cream and residual hair with a wet cotton ball, and confirm that the hair follicles on the back skin of the mouse are in the resting phase, that is, the back skin of the mouse can be observed to be pink with the naked eye. Testosterone propionate 5 mg / kg / d was subcutaneously injected into the depilatory area of ​​the mouse for 18 consecutive days to establish the mouse androgenic alopecia model.

[0034] 2. Grouping and administration: 60 mice were randomly divided into a blank group (Control), a model group (Model), a low-dose naringenone group (L-Nar, 200 μL, equivalent to naringenone 0.2 mg), a high-dose naringenone group (H-Nar, 200 μL, equivalent to naringenone 0.6 mg) and a positive drug group (Minoxidil, 200 μL, equivalent to minoxidil 10.0 mg), with 12 mice in each group. The blank group was given multiple subcutaneous injections of normal saline in the depilated area after hair removal, and the model group and the drug group were given multiple subcutaneous injections of testosterone propionate 5 mg / kg / d in the depilated area. In addition, the naringenone drug group and the positive drug group were given equal volumes of corresponding drugs by skin application, once a day, for 18 consecutive days.

[0035] 3. Observation of hair growth: 18 days after administration, the hair growth of the depilatory area of ​​each group of mice was recorded by taking photos with a Huawei mobile phone and stored for later use.

[0036] 4. Determination of hair quality and length: The mice were killed by dislocating the neck, and a 2×4 cm hair removal area on the back of each mouse was shaved with a pet shaver. 2 The hair of each area was weighed with an analytical balance to obtain the average value. At the same time, 10 new hairs were randomly selected from each mouse in each group and the hair length was measured with a vernier caliper to calculate the average value and compare the hair growth of each group of mice.

[0037] 5. Histological observation: 18 days after administration, 3 mice were randomly selected from each group and killed by dislocation of the neck. The skin tissue of the depilated area was fixed with 4% paraformaldehyde solution, dehydrated step by step, transparent, embedded, and sliced ​​for routine HE staining. Five high-power fields (×10) were randomly selected for hair follicle counting under a Thermo Fisher Evos M5000 microscope, and the average value was taken.

[0038] 6. qPCR detection of related genes: Three mice were randomly selected from each group, and the back skin was taken to extract total RNA. The mRNA expression of TGF-β1 and TGF-β2 was detected by qPCR. The results were expressed as multiples of gene expression relative to the normal control group, with GAPDH as the internal reference.

[0039] 7. Statistical analysis: The results of this experiment are shown in the figure, and the results are expressed as mean ± SD. SPSS17.0 statistical software was used for data analysis, and one-way analysis of variance was used for comparison between groups.

[0040] 3. Test results:

[0041] 1. Effect of naringenone on hair growth in androgenic alopecia model mice

[0042] Figure 1: is a comparison chart of the differences in skin appearance of each group of C57BL / 6 mice in the effect test; Table 1 is the statistics of hair growth of each group of C57BL / 6 mice in the effect test.

[0043] Table 1: Statistics of hair growth in each group of mice

[0044]

[0045] Note: Compared with the Control group: * p<0.05, ** p<0.01, compared with Model group: # p<0.05, n=12. Control: blank control group; Model: model group; L-Nar: low-dose naringenin group; H-Nar: low- and high-dose naringenin groups; Minoxidil: minoxidil group.

[0046] like Figure 1 As shown in Table 1, compared with the Control group, the hair length and hair quality of the Model group mice were significantly reduced (p < 0.01). Compared with the Model group, the high-dose galanthin group and the minoxidil group could significantly increase the hair length and hair quality (p < 0.05), and there was no significant difference between the high-dose galanthin group and the minoxidil group (p > 0.05); while the low-dose group galanthin slightly increased, but without statistical significance (p > 0.05).

[0047] The above results indicate that galanthin can alleviate the inhibitory effect of testosterone propionate on hair growth in C57BL / 6 mice.

[0048] 2. Effect of naringapine on the recovery of hair follicle tissue in mice with androgenic alopecia

[0049] Figure 2 : is a comparison of skin tissue pathological sections (HE staining) of C57BL / 6 mice in each group in the effect test (10×); Table 2 is the statistics of the number of hair follicles per unit field of view of C57BL / 6 mice in each group in the effect test.

[0050] Table 2: Statistics of hair follicle numbers per unit field of view in each group of mice

[0051]

[0052] Note: Compared with the Control group: * p<0.05; compared with Model group: # p<0.05, n=3.

[0053] Control: blank control group; Model: model group; L-Nar: low-dose narostane group; H-Nar: high-dose narostane group; Minoxidil: minoxidil group.

[0054] Results from cross-sectional and longitudinal sections of the skin ( Figure 2 ) showed that compared with the Control group, the inner hair root sheath disappeared in the depilatory area of ​​the Model group mice, the outer hair root sheath gradually keratinized, the shape was short and sparse, the hair papilla became thinner, the inner hair root sheath partially disappeared, and the hair had a rod-shaped end. Compared with the Model group, the high-dose galanthin group and minoxidil group had more hair follicles in the depilatory area, the hair follicles were long and large, the melanin formation was obvious, and the sebaceous glands were slightly reduced; while the low-dose group had no obvious improvement.

[0055] Furthermore, the number of hair follicles in 5 different areas was observed under a 10x optical microscope. As shown in Table 2, the number of hair follicles in the Model group was significantly reduced compared with the Control group (p < 0.05). The number of hair follicles in the high-dose galanthin group and the minoxidil group was significantly increased compared with the Model group (p < 0.05).

[0056] The above research results show that galanthin can improve the reduction in hair follicle number caused by testosterone propionate and maintain the normal morphology of hair follicles.

[0057] 3. Effect of naringapolone on the expression of TGFβ1 and TGFβ2 mRNA in the skin tissue of androgenic alopecia model mice Table 3 is the statistical expression of TGF-β1 and TGF-β2 mRNA in the skin tissue of C57BL / 6 mice in each group in the effect test.

[0058] Table 3: Statistics of TGFβ1 and TGFβ2 mRNA expression in skin tissue of mice in each group

[0059]

[0060] Note: Compared with the Control group: * p<0.05, ** p<0.01; compared with Model group: # p<0.05, n=3.

[0061] Control: blank control group; Model: model group; L-Nar: low-dose narostane group; H-Nar: high-dose narostane group; Minoxidil: minoxidil group.

[0062] As shown in Table 3, compared with the Control group, the expression of TGFβ1 and TGFβ2 mRNA in the skin tissue of the Model group was significantly increased (p < 0.01). Compared with the Model group, the expression of TGFβ1 and TGFβ2 mRNA in the skin tissue of mice in the high-dose and low-dose galanthin groups and the positive drug group was significantly decreased (p < 0.05). This result shows that galanthin can promote hair growth by inhibiting the expression of TGFβ1 and TGF-β2 mRNA.

[0063] Based on the above experiments, narostane or its pharmaceutically acceptable salts, esters or glycosides can be used to prevent or treat hair loss and to prepare products or drugs that promote hair growth. The content of narostane or its pharmaceutically acceptable salts, esters or glycosides as an active ingredient ranges from 1 to 100 mg.

[0064] The only active ingredient is galanthin, which is used to prepare a drug for preventing or treating hair loss. 75% ethanol is used to prepare a 1.0-3.0 mg / mL galanthin solution. It is applied to the scalp for external use. The dosage range for preventing or treating hair loss in adults (60 kg) should be 1-30 mg per day.

[0065] Of course, the products or drugs for preventing or treating hair loss and promoting hair growth may also include pharmaceutically acceptable excipients; the dosage form of the drug for preventing or treating hair loss is an oral dosage form, an injection dosage form or an external dosage form; the oral dosage form is a tablet, a granule, a capsule, a powder, an emulsion, a suspension or a solution; the external dosage form is a solution, an ointment, a cream, a lotion, a spray or a gel. The drug for preventing or treating hair loss or promoting hair growth is an immediate release dosage form or a slow release dosage form. The product for preventing or treating hair loss is a drug, a health product or a nutritious food for preventing or treating hair loss.

Claims

1. Nardostachysin or its pharmaceutically usable salts, esters or glycosides are used for preventing or treating hair loss and promoting hair growth.

2. The use according to claim 1, characterized in that: Nardostachysinone is the only active ingredient, and 75% ethanol is used to prepare a nardostachysinone solution with a concentration range of 1.0 to 3.0 mg / mL.

3. The use according to claim 2, characterized in that: The solution of chlorpheniramine is applied topically to the scalp, and the dosage for adults (60 kg) to prevent or treat hair loss is 1-10 mL per day.

4. The use according to claim 1, characterized in that: The solution of galanthin is prepared with 75% ethanol and applied externally to the scalp. The dosage for preventing or treating hair loss in adults (60 kg) is 1-30 mg per day.

5. Use of naringenone or its pharmaceutically usable salts, esters or glycosides in the preparation of drugs for preventing or treating hair loss and promoting hair growth.

6. The use according to claim 5, characterized in that: The drug for preventing or treating hair loss and promoting hair growth, wherein the content of narostane or its pharmaceutically usable salts, esters or glycosides as an active ingredient ranges from 1 to 100 mg.

7. The use according to claim 5 or 6, characterized in that: The drug for preventing or treating hair loss and promoting hair growth is in the form of an external dosage form and can be a microemulsion, emulsion, cream, spray, gel, tincture or liniment.

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