Hair Anti-Aging Agent

Abstract

To provide means for preventing or ameliorating gray hair; to provide means for preventing or ameliorating hair loss caused by aging; and to provide an agent for inhibiting or ameliorating DNA damage of hair follicle constituent cells, or an agent for inhibiting reduction of pigment stem cells.MEANS FOR SOLVING THE PROBLEM: It is found that berberbine, derivatives thereof, and salts thereof have an excellent preventive or ameliorating effect on gray hair and senile hair loss, and the effect is due to an effect of inhibiting or ameliorating DNA damage of hair follicle constituent cells and an effect of inhibiting reduction of pigment stem cells. That is, the present invention is a preventive or ameliorating agent for gray hair or senile hair loss, an inhibiting or ameliorating agent for DNA damage of hair follicle constituent cells, and an inhibiting agent for reduction of pigment stem cells, which contains at least one ingredient, as an active ingredient, selected from the group consisting of berberbine, derivatives thereof, and salts thereof.SELECTED DRAWING: Figure 1

Description

[Technical field]

[0001] The present invention relates to an agent for preventing or ameliorating gray hair or senile alopecia, an agent for suppressing or ameliorating DNA damage in hair follicle constituent cells, and an agent for suppressing the decrease in melanocyte stem cells. [Background technology]

[0002] The pigment (melanin) of hair is produced by pigment cells in the hair bulb, and epithelial cells that are supplied with this pigment form hair, resulting in the development of colored hair. When hair falls out, the hair bulb disappears, and the pigment cells in the hair bulb disappear as well. Hair follicle stem cells and melanocyte stem cells exist in a part of the hair follicle called the bulge region, and when a new hair cycle begins and new hair is produced, epithelial cells are supplied from the hair follicle stem cells, and melanocytes are supplied from the melanocyte stem cells to the hair bulb, and the above cycle is repeated again.

[0003] There are several causes of gray hair, including aging, stress, and vitiligo, and it occurs when an abnormality occurs in some of the above-mentioned processes. In the past, it was believed that gray hair could be prevented or improved by maintaining or improving the pigment production ability of the pigment cells in the hair bulb, and many substances have been found that activate enzymes such as tyrosinase, which is involved in pigment synthesis in pigment cells (Patent Documents 1 to 5).

[0004] In recent years, the mechanism of gray hair development due to aging has been elucidated. Melanocyte stem cells that have accumulated DNA damage to an extent that cannot be repaired differentiate while still in the bulge region (this differentiation is called ectopic differentiation), lose their self-maintenance function, and disappear (Non-Patent Document 1). As a result, pigment cells cannot be supplied to the hair bulb, and epithelial cells that do not have pigment produce hair, resulting in gray hair. In other words, in the majority of gray hair caused by aging, pigment cells do not exist in the hair bulb in the first place, and activation of enzymes involved in pigment synthesis cannot improve the condition. Gray hair with pigment cells remaining in the hair bulb is thought to be able to be improved by conventional approaches, but survey results show that only about 20% of gray hair (not necessarily caused by aging) has pigment cells in the hair bulb (Non-Patent Document 2).

[0005] Furthermore, it has been elucidated that melanocyte stem cells are maintained by the function of hair follicle stem cells (Non-Patent Document 3). Hair follicle stem cells produce factors including TGF-β and WNT ligands, thereby forming an environment (melanocyte stem cell niche) in which melanocyte stem cells can exist while maintaining their stemness, and controlling the maintenance and activation of melanocyte stem cells (Non-Patent Documents 3, 4). In other words, in order to prevent or improve gray hair, it is important to maintain hair follicle stem cells and further maintain the melanocyte stem cell niche function to prevent the disappearance of melanocyte stem cells.

[0006] It is known that by inflicting DNA damage in black mice using X-rays, it is possible to induce white hair that reflects the above-mentioned mechanism of aging, and this is widely used as an age-related mouse model of gray hair (Patent Document 6). In addition, in this model, X-rays damage both hair follicle stem cells and melanocyte stem cells, and it has been elucidated that the main target is hair follicle stem cells (impairment of melanocyte stem cell niche function) (Non-Patent Document 5). [Prior art documents] [Patent documents]

[0007] [Patent Document 1] JP 2001-288052 A [Patent Document 2] JP 2001-131032 A [Patent Document 3] JP 2007-8888 A [Patent Document 4] JP 2011-157317 A [Patent Document 5] JP 2002-47130 A [Patent Document 6] JP 2015-193550 A [Non-patent literature]

[0008] [Non-Patent Document 1] Inomata K, Aoto T, Binh NT, Okamoto N, Tanimura S, Wakayama T, Iseki S, Hara E, Masunaga T, Shimizu H, Nishimura EK, Genotoxic stress abrogates renewal of melanocyte stem cells by triggering their differentiation, Cell.2009;137(6):1088-99. [Non-Patent Document 2] "Anti-aging Series 1: The Actual Situation of Gray Hair, Hair Loss, and Hair Growth" by Tatsuro Ideta et al., NTS, July 4, 2005, p.49-61 [Non-Patent Document 3] Tanimura S, Tadokoro Y, Inomata K, Binh NT, Nishie W, Yamazaki S, Nakauchi H, Tanaka Y, McMillan JR, Sawamura D, Yancey K, Shimizu H, Nishimura EK, Hair follicle stem cells provide a functional niche for melanocyte stem cells, Cell Stem Cell. 2011;8(2):177-87. [Non-Patent Document 4] Rabbani P, Takeo M Chou W, Myung P, Bosenberg M, Chin L, Taketo MM, Ito M, Coordinated activation of Wnt in epithelial and melanocyte stem cells initiates pigmented hair regeneration, Cell. 2011 Jun 10;145(6):941-955. [Non-Patent Document 5] Aoki H, Hara A, Motohashi T, Kunisada T, Keratinocyte stem cells but not melanocyte stem cells are the primary target for radiation-induced hair graying, J Invest Dermatol. 2013;133(9):2143-51. [Non-Patent Document 6] Matsumura H, Mohri Y, Binh NT, Morinaga H, Fukuda M, Ito M, Kurata S, Hoeijmakers J, Nishimura EK, Hair follicle aging is driven by transepidermal elimination of stem cells via COL17A1 proteolysis, Science. 2016;351(6273):aad4395. Summary of the Invention [Problem to be solved by the invention]

[0009] An object of the present invention is to provide a means for preventing or ameliorating gray hair.An object of the present invention is to provide a means for preventing or ameliorating hair loss caused by aging.An object of the present invention is to provide an agent for suppressing or ameliorating DNA damage in hair follicle constituent cells.An object of the present invention is to provide an agent for suppressing the decrease in melanocyte stem cells. [Means for solving the problem]

[0010] Therefore, the inventors conducted extensive research and discovered that berberrubine and its derivatives, as well as salts thereof, have excellent preventive or ameliorative effects on gray hair or age-related alopecia, and that this effect is due to an effect of inhibiting or ameliorating DNA damage in hair follicle constituent cells, and an effect of inhibiting the decrease in melanocyte stem cells, thereby completing the present invention.

[0011] That is, the present invention provides (1) An agent for preventing or improving gray hair, comprising as an active ingredient at least one selected from the group consisting of berberrubine, its derivatives (excluding berberine), and salts thereof; (2) An agent for preventing or improving senile alopecia, comprising as an active ingredient at least one selected from the group consisting of berberrubine, its derivatives, and salts thereof; (3) An agent for suppressing or improving DNA damage in hair follicle constituent cells, comprising at least one active ingredient selected from the group consisting of berberrubine, its derivatives, and salts thereof; (4) The agent for suppressing or improving DNA damage according to (3), wherein the hair follicle-constituting cells are hair follicle stem cells or melanocyte stem cells. (5) An agent for suppressing the decrease in melanocyte stem cells, comprising at least one active ingredient selected from the group consisting of berberrubine, its derivatives, and salts thereof; It is. Effect of the Invention

[0012] The agent for preventing or ameliorating gray hair and hair loss of the present invention can prevent or ameliorate the occurrence of gray hair and age-related hair loss through an action of inhibiting or ameliorating DNA damage in hair follicle constituent cells or an action of inhibiting the decrease in melanocyte stem cells. [Brief description of the drawings]

[0013] [Figure 1] FIG. 1 is a graph showing the lightness of mouse back hair when berberrubine was evaluated in Test Example 1. [Diagram 2] FIG. 2 is a graph showing the lightness of mouse back hair when berberine was evaluated in Test Example 1. [Diagram 3] FIG. 3 is a graph showing the hair growth score of mice 15 days after hair removal when berberine was evaluated in Test Example 2. [Figure 4] FIG. 4 is a graph showing the hair growth score of mice 17 days after hair removal when berberine was evaluated in Test Example 2. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0014] Berberrubin in the present invention is a compound represented by 5,6-dihydro-9-hydroxy-10-methoxybenzo[g]-1,3-benzodioxolo[5,6-a]quinolizinium. The derivative in the present invention refers to a compound obtained by changing a small part in the molecule of the original compound. An example of a derivative is, but is not limited to, berberine (2,3-methylenedioxy-9,10-dimethoxy-5,6-dihydrodibenzo[a,g]quinolizinium), in which one of the hydroxyl groups of berberrubine is changed to a methoxy group. The salt in the present invention includes, but is not limited to, for example, chlorides, salts with hydrochloric acid, sulfuric acid, and tannic acid.

[0015] Berberubine, its derivatives, and salts thereof used in the present invention can be produced by known synthesis methods, or commercially available products such as berberubine chloride (Nagara Science Co., Ltd., Kishida Chemical Co., Ltd.), berberine chloride (Sigma-Aldrich), berberine hydrochloride (Nacalai Tesque), berberine chloride hydrate (Alps Pharmaceutical Co., Ltd.), and berberine tannate (Alps Pharmaceutical Co., Ltd.) can be used.

[0016] In the present invention, hair follicle-constituting cells refer to any cells that constitute hair follicle tissue, such as epithelial cells, hair follicle stem cells, outer root sheath cells, inner root sheath cells, and hair matrix cells, pigment cells, melanocyte stem cells, chromatoblasts, and pigment cells, and mesenchymal cells, hair papilla cells, and dermal root sheath cells.

[0017] In the present invention, hair follicle stem cells refer to stem cells of epithelial cells that are present in the bulge region of the hair follicle and in the secondary hair germ.

[0018] In the present invention, the melanocyte stem cells refer to stem cells of pigment cells that are present in the bulge region of the hair follicle and in the secondary hair germ.

[0019] The preventive or ameliorative effects of the present invention on gray hair and senile alopecia are believed to be due to the effect of maintaining or improving the niche function of epithelial cells on pigment cells, the effect of inhibiting ectopic differentiation of melanocyte stem cells, and the effect of inhibiting the decrease in hair follicle stem cells and melanocyte stem cells. Ectopic differentiation refers to the phenomenon in which melanocyte stem cells differentiate into melanocytes while remaining in the hair follicle bulge region where they should originally exist as stem cells. Niche function refers to the function of a certain type of cell to construct and control the environment in which other cells exist.

[0020] The agent for preventing or improving gray hair of the present invention can be provided as a quasi-drug, a medicine, or a food or drink. It can also be used as a reagent (positive control, etc.). The form of administration is not particularly limited, but includes external use and internal administration, and is preferably external use applied to the skin including the scalp.

[0021] When the present invention is applied externally, examples of the dosage form include lotions, liquids, creams, ointments, gels, sprays, shampoos, conditioners, soaps, etc., and when the present invention is applied internally, examples of the dosage form include tablets, powders, granules, liquids, capsules, dry syrups, jellies, liquid foods, semi-solid foods, solid foods, etc. These can be produced by known methods. During production, various additives that can be contained in cosmetics, quasi-drugs, medicines, foods and beverages, or reagents can be blended within the range that does not impair the effects of the present invention.

[0022] Furthermore, the agent for preventing or improving gray hair of the present invention can be used in combination with hair growth substances such as Swertia japonica extract, carrot extract, capsicum tincture, ginger tincture, loquat leaf extract, glycyrrhetinic acid, dipotassium glycyrrhizinate, monoammonium glycyrrhizinate, pantothenic acid, panthenol, vitamin E and its derivatives, hinokitiol, salicylic acid, piroctone olamine, minoxidil, adenosine, t-flavanone, cytopurine, pentadecanoic acid glyceride, allantoin, and nicotinic acid amide.By combining, hair production is accelerated, and the effect of preventing or improving gray hair can be obtained at an early stage.

[0023] The amount of the berberrubine derivatives and salts thereof of the present invention to be incorporated when provided as cosmetics, quasi-drugs, pharmaceuticals, foods, beverages, or reagents is 0.000001 to 20 mass%, preferably 0.0001 to 10 mass%, and more preferably 0.01 to 1 mass%, based on the total composition.

[0024] In addition, the agent for preventing or improving gray hair, the agent for preventing or improving senile alopecia, the agent for inhibiting or improving DNA damage to hair follicle constituent cells, and the agent for inhibiting the decrease in melanocyte stem cells of the present invention can be labeled on a product (medicine, quasi-drug, food, beverage, or reagent, etc.) or its instruction manual containing the agent for preventing or improving gray hair, preventing or improving senile alopecia, inhibiting or improving DNA damage to hair follicle constituent cells, inhibiting or improving DNA damage to hair follicle stem cells, inhibiting or improving DNA damage to melanocyte stem cells, or inhibiting the decrease in melanocyte stem cells. Here, "labeled on the product or its instruction manual" means that the product body, container, packaging, etc. are labeled, or that the instruction manual, package insert, promotional material, application materials, other printed matter, or advertisements disclosing information about the product are labeled. In addition, these labels can include information about the use for preventing or improving gray hair, preventing or improving senile alopecia, or information about the use for preventing or treating diseases or symptoms caused by DNA damage to hair follicle constituent cells, DNA damage to hair follicle stem cells, DNA damage to melanocyte stem cells, or inhibiting the decrease in melanocyte stem cells. EXAMPLES

[0025] The present invention will be described in more detail below with reference to examples and test examples, but the present invention is not limited to these examples.

[0026] (Test Example 1) Evaluation of anti-hair graying effect using X-ray mouse gray hair model <Test Method> Seven-week-old C57BL / 6 mice were anesthetized and the hair on their backs was shaved off with clippers, leaving a few millimeters. After shaving, mice with good back condition were selected and randomized (using EXSUS Professional Version 7.7.1 (CAC Exicare)) to ensure that the mean and variance of body weight in each group were approximately equal, and the mice were divided into groups. The mice were treated with 200 μL of a dosing solution prepared by dissolving berberrubine chloride or berberine hydrochloride in ethanol to a concentration of 0.1% (w / v), or with ethanol as a solvent, once a day for one week, starting the day after hair shaving. Six days after hair shaving, hair removal was performed on the back under anesthesia. Hair removal was performed by applying a hair remover (Epilat (Kracie)) to the shaved area. The day after hair removal, the mice were irradiated with a total dose of 5 Gy of X-rays using an X-ray irradiator (RX-650 (Faxitron X-ray)). After raising the mice until all the body hair had grown in, the probe of a color difference meter (CR-400, manufactured by Konica Minolta) was placed on the body hair to measure the lightness (L value), which was used as an index of the rate of white hair.

[0027] <Test Results> Figures 1 and 2 show the results of evaluating berberrubin and berberine, respectively. When DNA damage was caused to hair follicle stem cells and melanocyte stem cells by X-ray irradiation, the number of melanocyte stem cells decreased, leading to an increase in hair brightness and induction of hair whitening (Non-Patent Document 1). However, application of berberrubin or berberine inhibited the increase in hair brightness and inhibited hair whitening. In other words, the effects of berberrubin and berberine in preventing and improving gray hair were confirmed. It is presumed that both compounds suppress and improve DNA damage in hair follicle stem cells and melanocyte stem cells, thereby maintaining and improving the melanocyte stem cell niche function of hair follicle stem cells, and as a result, ectopic differentiation of melanocyte stem cells is suppressed and melanocyte stem cells are maintained (Non-Patent Document 1). In other words, both compounds are presumed to have the effects of inhibiting and improving DNA damage, maintaining and improving the niche function of epithelial cells for pigmentary cells, and inhibiting and maintaining ectopic differentiation of melanocyte stem cells. It has been confirmed that berberrubine and berberine do not directly absorb or interfere with X-rays.

[0028] (Test Example 2) Evaluation of anti-alopecia effect using X-ray mouse senile alopecia model It has been elucidated that X-rays can induce alopecia through a mechanism similar to that of age-related alopecia (senile alopecia) in addition to age-related gray hair (Non-Patent Document 6). Senile alopecia differs from male pattern baldness caused by male hormones in that hair follicle stem cells are gradually lost due to the accumulation of DNA damage in hair follicle stem cells associated with aging, causing hair follicle miniaturization. X-ray irradiated mice can also be used as a model for this senile alopecia. <Test Method> Seven-week-old C57BL / 6 mice were anesthetized and the hair on their backs was shaved off with clippers, leaving a few millimeters. After shaving, mice with good back condition were selected and randomized (using EXSUS Professional Version 7.7.1 (CAC Exicare)) to ensure that the mean and variance of body weight in each group were approximately equal, and the mice were divided into groups. The administration solution, prepared by dissolving berberine hydrochloride in ethanol to a concentration of 0.1% (w / v), or the solvent ethanol was applied to the mice once a day at 200 μL for one week from the day after hair shaving. Six days after hair shaving, hair removal was performed on the back under anesthesia. Hair removal was performed by applying a hair remover (Epilat (Kracie)) to the shaved area. The day after hair removal, the mice were irradiated with a total dose of 5 Gy of X-rays using an X-ray irradiator (RX-650 (Faxitron X-ray)). On the 15th and 17th days after hair removal, when differences in the degree of hair growth were observed, a hair growth score was given according to the following scoring criteria to evaluate the degree of hair growth.

[0029] Score 1: Pink skin Score 2: Gray discoloration of skin (less than 30% of shaved area) Score 3: Gray discoloration of the skin (30% to less than 60% of the shaved area) or hair growth (less than 30% of the shaved area) Score 4: Gray discoloration of the skin (more than 60% of the shaved area) or hair growth (more than 30% but less than 60% of the shaved area) Score 5: Hair growth is observed (more than 60% of the shaved area)

[0030] <Test Results> Figures 3 and 4 show the results of evaluating berberine. When hair follicle stem cells were exposed to X-rays and DNA damage was caused, the generation of new hair was delayed. However, when berberine was applied, the delay in the generation of new hair was no longer observed. In other words, berberine was confirmed to have the effect of preventing and improving hair loss associated with aging, including senile alopecia. This is because berberine suppresses and improves DNA damage in hair follicle stem cells, thereby maintaining hair follicle stem cells. Based on the results of Test Example 1, berberrubin, which has the effect of suppressing and improving DNA damage in hair follicle stem cells, also has these actions. [Industrial Applicability]

[0031] The agent for preventing or improving gray hair and hair loss of the present invention can be used in the fields of quasi-drugs, medicines, or foods and beverages for preventing or improving gray hair and hair loss caused by aging or DNA damage. In addition, the agent for suppressing or improving DNA damage in hair follicle constituent cells and the agent for suppressing the decrease in melanocyte stem cells can also be used in the fields of quasi-drugs, medicines, or foods and beverages where these effects are required.

Claims

1. An agent for preventing or improving gray hair, comprising at least one active ingredient selected from the group consisting of berberrubine and its salts (however, excluding agents for preventing or improving gray hair containing a Coptis japonica extract and agents for preventing or improving gray hair containing Phellodendron Bark or Phellodendron Bark extract).

2. An agent for preventing or improving senile alopecia, comprising at least one active ingredient selected from the group consisting of berberrubine, berberine, and salts thereof (excluding, however, an agent for preventing or improving senile alopecia containing a Coptis japonica extract, and an agent for preventing or improving senile alopecia containing Phellodendron Bark or an extract of Phellodendron Bark).

3. An agent for suppressing or improving DNA damage in hair follicle constituent cells, comprising at least one active ingredient selected from the group consisting of berberrubine, berberine, and salts thereof (provided that the agent contains a coptis japonica extract) (Excluding agents for suppressing or improving DNA damage to hair follicle constituent cells, and agents for suppressing or improving DNA damage to hair follicle constituent cells that contain Phellodendron Bark or Phellodendron Bark extract).

4. 4. The method according to claim 3, wherein the hair follicle constituent cells are hair follicle stem cells or melanocyte stem cells. Improver.

5. An inhibitor of the decrease in melanocyte stem cells, comprising at least one active ingredient selected from the group consisting of berberrubine, berberine, and salts thereof (excluding inhibitors of the decrease in melanocyte stem cells containing a coptis japonica extract and inhibitors of the decrease in melanocyte stem cells containing Phellodendron Bark or an extract of Phellodendron Bark).

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