Use of nardosinone in drug for preventing or treating alopecia and promoting hair growth

Abstract

The present invention relates to the use of nardosinone in a product or drug for preventing or treating alopecia and promoting hair growth. The drug for preventing or treating alopecia comprises nardosinone or a pharmaceutically acceptable salt, ester or glycoside thereof. As the only active ingredient, nardosinone is prepared with 75% ethanol into a solution with a concentration of 1.0-3.0 mg / mL. The solution is applied topically to the scalp, and the daily dosage for preventing or treating alopecia in adults is 1-10 mL. Existing drugs for treating hair loss and alopecia show poor therapeutic effects and have side effects. Studies find that nardosinone as an active ingredient can achieve the application effect of preventing or treating alopecia and canities by means of regulating the level of TGFβ family genes which are closely related to hair development and growth. The present invention verifies for the first time that nardosinone, an active monomer derived from Nardostachys jatamansi, has the potential to prevent or treat alopecia. The administration of a certain dose of nardosinone to the skin can prevent or treat alopecia, thus providing a new method for preventing or treating alopecia.

Description

Nardostachysone in the prevention or treatment of hair loss and in the promotion of hair growth. Technical Field

[0001] This invention belongs to the field of biomedical technology, and in particular relates to the application of naringinone in the preparation of drugs for the prevention or treatment of hair loss. Background Technology

[0002] With increasing pressure from social, work, and life factors, the number of people suffering from hair loss is rising and showing a trend towards affecting younger people. In developed countries in Europe and America, the hair loss rate for adult men is 32%, and for women it is 28%; in Asian countries, the rate for adult men is 25%, and for women it is 22%. Although hair loss is not a life-threatening disease, it causes immense psychological distress to sufferers. Some patients, especially young patients, develop severe feelings of inferiority, which affects their self-confidence and social interactions.

[0003] Currently, drug treatments for hair loss mainly fall into two categories: 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 abnormalities, and an increased risk of breast cancer in men. The other commonly used drug is vasodilators such as minoxidil. However, minoxidil can cause allergic reactions in some patients, accompanied by itching, increased dandruff, and in a few cases, hirsutism; severe cases can cause chest tightness and dizziness. In summary, due to the toxic side effects of chemical drugs, the demand for safer and gentler plant-based alternative treatments has increased significantly. Therefore, finding a high-quality, affordable, and easy-to-use natural product that effectively treats or prevents hair loss is of significant social importance.

[0004] Nardostachysone, a sesquiterpene compound derived from the traditional Chinese medicine Nardostachys jatamansi, has a wide range of definite pharmacological activities, such as sedation, anti-epileptic, antidepressant, nerve growth promotion, and blood pressure reduction.

[0005] To date, there are no reports of research and development on the use of naringin in the treatment of hair loss. Technical issues

[0006] This invention addresses the problem that existing hair loss and alopecia treatments are ineffective and have side effects. Through research, it has been found that naringinone, as an active ingredient, can prevent or treat hair loss and gray hair by regulating the level of TGFβ family genes that are closely related to hair development and growth. Therefore, this invention proposes the application of naringinone in products or drugs for the prevention or treatment of hair loss and the promotion of hair growth. Technical solutions

[0007] The technical solution adopted in this invention is as follows:

[0008] Nardostachysone (theoretically, including its pharmaceutically usable salts, esters, or glycosides) is used as an active ingredient to prevent or treat hair loss and promote hair growth.

[0009] In the aforementioned application, nardostachys is the sole active ingredient, and a nardostachys solution with a concentration range of 1.0-3.0 mg / mL is prepared using 75% ethanol.

[0010] The naringinone solution is applied topically to the scalp. For adults (60 kg) to prevent or treat hair loss, the dosage is 1-10 mL per day.

[0011] The use of naringinone or its pharmaceutically usable salts, esters or glycosides in the preparation of products or medicines for the prevention or treatment of hair loss or the promotion of hair growth.

[0012] The drug for preventing or treating hair loss, wherein the content of the active ingredient, naringinone or its pharmaceutically usable salt, ester or glycoside, ranges from 1 to 100 mg.

[0013] The application described herein involves preparing a 75% ethanol solution of naringinone and applying it topically to the scalp. The dosage range for adults (60 kg) for the prevention or treatment of hair loss should be 1-30 mg per day.

[0014] The drug is a topical dosage form and may be a microemulsion, emulsion, cream, spray, gel, tincture, or liniment. Beneficial effects

[0015] 1. This invention is the first to verify the potential of spikenone, an active monomer derived from Nardostachys jatamansi, to prevent or treat hair loss. Administering a specific dose of spikenone through the skin can prevent or treat hair loss, providing a new method for its prevention and treatment. Spironolone is the main active ingredient in Nardostachys jatamansi, which can be extracted from natural plants using well-established extraction methods. Furthermore, spikenone can be administered through the skin or orally, making it easily acceptable and conducive to clinical promotion and application.

[0016] A mouse androgenic alopecia model was established by subcutaneously injecting 5 mg / kg / d of testosterone propionate into the alopecia area for 18 consecutive days. An equal volume of the corresponding drug was then administered via skin application of a prepared naringinone solution once daily for 18 consecutive days. The effect of naringinone on hair growth in the androgenic alopecia model mice was observed, and it was found that it could effectively alleviate the inhibitory effect of testosterone propionate on hair growth in C57BL / 6 mice.

[0017] 2. Nardostachysone has a significant therapeutic effect on testosterone propionate-induced alopecia in mice, significantly promoting hair growth and increasing the number of hair follicles. The results of this invention show that in a testosterone propionate-induced alopecia mouse model, 18 days of transdermal administration of nardostachysone significantly reduced patchy hair loss on the back of mice induced by testosterone propionate, and made the mouse hair shiny, soft, and denser. Pathological section results showed that nardostachysone administration increased the number of hair follicles in the skin of testosterone propionate-induced model mice and promoted hair follicle growth. qPCR results showed that nardostachysone could inhibit the expression of TGFβ1 and TGFβ2 mRNA induced by testosterone propionate. The effect of nardostachysone on the expression of TGFβ1 and TGFβ2 mRNA in the skin tissue of androgenetic alopecia model mice indicates that nardostachysone can promote hair growth by inhibiting the expression of TGFβ1 and TGFβ2 mRNA. Experimental observation of the effect of nardostachysone on hair follicle tissue recovery in a mouse model of androgenetic alopecia confirmed that nardostachysone can increase the reduction in hair follicle count induced by testosterone propionate and maintain normal hair follicle morphology. These results indicate that nardostachysone, a natural compound derived from spikenard, has the potential to prevent or treat hair loss.

[0018] 3. This invention is applied to products for the prevention or treatment of hair loss. The active ingredient, naringinone, regulates the levels of TGFβ family genes closely related to hair development and growth, thus achieving the effect of preventing or treating hair loss and gray hair. Narsalisinone is a low-cost and readily available raw material, showing promising potential in the prevention and treatment of hair loss. Attached Figure Description

[0019] Figure 1 is a comparison of the differences in skin appearance among the C57BL / 6 mice in each group during the efficacy test;

[0020] Figure 2 is a comparison of skin tissue pathological sections (HE staining) of C57BL / 6 mice in each group during the efficacy test (10×).

[0021] In the figure, A: HE staining of transverse sections of mouse skin in each group; B: HE staining of longitudinal sections of mouse skin in each group;

[0022] In Figures 1 and 2, Control: blank control group; Model: model group; L-Nar: low-dose naringin group; H-Nar: high-dose naringin group; Minoxidil: minoxidil group. Embodiments of the present invention

[0023] To make the technical concept and advantages of this invention clearer for achieving its invention purpose, the technical solution of this invention will be further described in detail below with reference to the accompanying drawings. It should be understood that the following embodiments are only for explaining and illustrating the preferred embodiments of this invention, and should not be regarded as and do not constitute a limitation on the scope of patent protection required by this invention. In addition, the technical features involved in the following described embodiments can be combined with each other as long as they do not conflict with each other.

[0024] Examples and Applications: Therapeutic Experiment of Nardosinone on Androgenetic Alopecia Model Mice

[0025] I. Materials:

[0026] 1. Animals: 60 male C57BL / 6 mice, 6 weeks old, weighing 18 - 22 g, provided by Guangzhou Yancheng Biotechnology Co., Ltd., and reserved after inspection and quarantine. Animal production license number: SCXK (Guangdong) 2017 - 0174.

[0027] 2. Drugs and Reagents: Preparation of Nardosinone Solution: Nardosinone was prepared into nardosinone solutions containing 1.0 and 3.0 mg / mL of nardosinone with 75% ethanol; 5% minoxidil tincture (Xiamen Meishang Pharmaceutical Co., Ltd.).

[0028] The chemical structural formula of nardosinone is as follows:

[0029] .

[0030] 3. Instruments: Thermo Fisher Evos M5000 microscope, Huawei mobile phone.

[0031] II. Test Methods:

[0032] 1. Preparation of Mouse Alopecia Model: After acclimating healthy SPF - level C57BL / 6 mice for 1 week, the mice were anesthetized by intraperitoneal injection of 0.3% sodium pentobarbital solution. Select a depilation area about 2×4 cm with the long side parallel to the spine at a distance of 0.5 cm from the proximal end of the tail tip, and then use a pet hair clipper to cut all the hair in the dorsal depilation area short until the dorsal skin is faintly visible. Then evenly apply depilatory cream to the area where the hair has been removed. Further, after wetting a cotton ball with physiological saline, wipe off the depilatory cream and residual hair with the wet cotton ball, and confirm that the hair follicles on the dorsal skin of the mouse are all in the resting phase, that is, the dorsal skin of the mouse can be observed to be pink with the naked eye. Subcutaneously inject testosterone propionate at 5 mg / kg / d for 18 consecutive days in the depilated area of the mouse to establish an androgenetic alopecia model of the mouse.

[0033] ​​2. Grouping and Drug Administration: Sixty mice were randomly divided into four groups: a control group, a model group, a low-dose naringin group (L-Nar, 200 μL, equivalent to 0.2 mg of naringin), a high-dose naringin group (H-Nar, 200 μL, equivalent to 0.6 mg of naringin), and a positive control group (Minoxidil, 200 μL, equivalent to 10.0 mg of minoxidil), with 12 mice in each group. After hair removal, the control group received multiple subcutaneous injections of saline in the hair removal area. The model group and the drug-treated groups received multiple subcutaneous injections of testosterone propionate (5 mg / kg / day) in the hair removal area. Additionally, the naringin-treated group and the positive control group received an equal volume of the corresponding drug applied topically once daily for 18 consecutive days.

[0034] 3. Observation of hair growth: 18 days after administration, the hair growth of mice in the bald areas of each group was recorded by taking photos with a Huawei mobile phone and saved for later use.

[0035] 4. Measurement of hair quality and length: Mice were euthanized by dislocation of the neck, and 2×4 cm of hair was shaved from the back of each mouse using a pet hair clipper. 2 The area of ​​hair was measured using an analytical balance, and the average value was calculated. At the same time, 10 newly grown hairs were randomly selected from each mouse in each group, and the length of the hairs was measured using calipers. The average value was calculated, and the hair growth of mice in each group was compared.

[0036] 5. Histological observation: Eighteen days after drug administration, three mice were randomly selected from each group and euthanized by cervical dislocation. Skin tissue from the hair-removed area was collected, fixed with 4% paraformaldehyde solution, dehydrated stepwise, cleared, embedded, sectioned, and stained with hematoxylin and eosin (HE). Hair follicles were observed under a Thermo Fisher Evos M5000 microscope, and five high-power fields (×10) were randomly selected for counting, and the average value was taken.

[0037] 6. qPCR detection of related genes: Three mice were randomly selected from each group, and total RNA was extracted from their dorsal skin. The mRNA expression levels of TGF-β1 and TGF-β2 were detected by qPCR. Results were expressed as fold changes in gene expression relative to the normal control group, using GAPDH as an internal control.

[0038] 7. Statistical Analysis: The results of this experiment are shown in the figure. The results are expressed as mean ± SD. Data analysis was performed using SPSS 17.0 statistical software, and one-way ANOVA was used for comparisons between groups.

[0039] III. Experimental Results:

[0040] 1. Effects of naringin on hair growth in androgenetic alopecia model mice

[0041] Figure 1 shows a comparison of the differences in skin appearance among the C57BL / 6 mice in each group during the efficacy test; Table 1 shows the statistics of hair growth among the C57BL / 6 mice in each group during the efficacy test.

[0042] Table 1: Statistics on hair growth in mice of each group

[0043]

[0044] Note: Compared with the Control group: * p < 0.05 ** p < 0.01, compared with the Model group: # p < 0.05, n = 12. Control: blank control group; Model: model group; L-Nar: low-dose naringin group; H-Nar: low- and high-dose naringin group; Minoxidil: minoxidil group.

[0045] As shown in Figure 1 and Table 1, compared with the Control group, the hair length and hair quality of mice in the Model group were significantly reduced (p < 0.01). Compared with the Model group, the high-dose naringin group and the minoxidil group significantly increased hair length and hair quality (p < 0.05), and there was no significant difference between the high-dose naringin group and the minoxidil group (p > 0.05); while the low-dose naringin group showed a slight increase, it was not statistically significant (p > 0.05).

[0046] The above results indicate that naringinone can alleviate the inhibitory effect of testosterone propionate on hair growth in C57BL / 6 mice.

[0047] 2. Effects of naringin on hair follicle tissue recovery in androgenetic alopecia model mice

[0048] Figure 2 is a comparison of skin tissue pathological sections (HE staining) of C57BL / 6 mice in each group during the efficacy test (10×); Table 2 is a statistical table of the number of hair follicles per unit field of view of C57BL / 6 mice in each group during the efficacy test.

[0049] Table 2: Number of hair follicles per unit field of view in each group of mice

[0050]

[0051] Note: Compared with the Control group: * p < 0.05; compared with the Model group: # p < 0.05, n = 3.

[0052] Control: Blank control group; Model: Model group; L-Nar: Low-dose naringin group; H-Nar: High-dose naringin group; Minoxidil: Minoxidil group.

[0053] As shown in the cross-sectional and longitudinal sections of the skin (Figure 2), compared with the Control group, the Model group mice showed that the hair root sheath disappeared in the hair-removed area, the outer hair root sheath gradually keratinized, became shorter and sparser, the dermal papilla became thinner, the inner hair root sheath partially disappeared, and the hairs had rod-shaped ends. Compared with the Model group, the high-dose naringin group and the minoxidil group showed a greater number of hair follicles in each hair-removed area, the hair follicles were longer and larger, melanin formation was obvious, and the sebaceous glands were slightly reduced; while the low-dose group showed no significant improvement.

[0054] Furthermore, the number of hair follicles in five different regions was observed under a 10x optical microscope. As shown in Table 2, the number of hair follicles in the Model group was significantly lower than that in the Control group (p < 0.05). Compared with the Model group, the number of hair follicles in the high-dose naringin group and the minoxidil group was significantly increased (p < 0.05).

[0055] The above research results show that naringinone can increase the reduction in the number of hair follicles caused by testosterone propionate and maintain the normal morphology of hair follicles.

[0056] 3. Effects of naringin on the expression of TGFβ1 and TGFβ2 mRNA in skin tissue of mice with androgenetic alopecia

[0057] Table 3 shows the statistical analysis of TGF-β1 and TGF-β2 mRNA expression levels in the skin tissue of C57BL / 6 mice in each group during the efficacy test.

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

[0059]

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

[0061] Control: Blank control group; Model: Model group; L-Nar: Low-dose naringin group; H-Nar: High-dose naringin 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- and low-dose naringin groups and the positive control group was significantly decreased (p < 0.05). These results indicate that naringin can promote hair growth by inhibiting the expression of TGFβ1 and TGFβ2 mRNA.

[0063] Based on the above experiments, naringinone or its pharmaceutically usable salts, esters, or glycosides can be used in the prevention or treatment of hair loss and in the preparation of products or drugs that promote hair growth. The content of naringinone or its pharmaceutically usable salts, esters, or glycosides as the active ingredient ranges from 1 to 100 mg.

[0064] Nardostachysone is the sole active ingredient used in the preparation of drugs for the prevention or treatment of hair loss. It is prepared as a 1.0–3.0 mg / mL solution using 75% ethanol. For external application to the scalp, the dosage range for adults (60 kg) for the prevention or treatment of hair loss should be 1–30 mg daily.

[0065] Of course, products or drugs used to prevent or treat hair loss and promote hair growth may also include pharmaceutically acceptable excipients; the dosage form of the hair loss prevention or treatment drug may be an oral dosage form, an injectable dosage form, or a topical dosage form; the oral dosage form may be a tablet, granule, capsule, powder, emulsion, suspension, or solution; the topical dosage form may be a solution, ointment, cream, lotion, spray, or gel. The hair loss prevention or hair growth promotion drug may be an immediate-release dosage form or a slow-release dosage form. The hair loss prevention or treatment product may be a drug, health product, or nutritional food for preventing or treating hair loss.

Claims

1. Nardostachysone or its pharmaceutically usable salts, esters or glycosides are used to prevent or treat hair loss and promote hair growth.

2. Use according to claim 1, characterized in that: Nardostachys is the only active ingredient, and it is prepared into a solution with a concentration range of 1.0 to 3.0 mg / mL using 75% ethanol.

3. Use according to claim 2, characterized in that: Nardostachysone solution is applied topically to the scalp. For adults (60 kg) to prevent or treat hair loss, the dosage is 1-10 mL per day.

4. Use according to claim 1, characterized in that: The solution of naringinone is prepared using 75% ethanol and applied topically to the scalp. The dosage for adults (60 kg) to prevent or treat hair loss is 1-30 mg per day.

5. The use of naringinone or its pharmaceutically usable salts, esters or glycosides in the preparation of medicines for the prevention or treatment of hair loss and for promoting hair growth.

6. Use according to claim 5, characterized in that: The drug for preventing or treating hair loss and promoting hair growth contains, in which the content of the active ingredient, naringinone or its pharmaceutically usable salt, ester or glycoside, ranges from 1 to 100 mg.

7. Use according to claim 5 or 6, characterized in that: The medications used to prevent or treat hair loss and promote hair growth are topical dosage forms, which may be microemulsions, emulsions, creams, sprays, gels, tinctures, or liniments.

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