JP6740548B2 - Hair growth agent containing urolithins, and 5α reductase activity inhibitor containing urolithins - Google Patents

Description

TECHNICAL FIELD The present invention relates to a hair growth agent containing urolithins and a 5α reductase activity inhibitor containing urolithins.

The mechanism of hair loss is that the male hormone testosterone is converted to dihydrotestosterone (DHT) (or sometimes referred to as 5α-dihydrotestosterone, (5α-DHT)) by the action of 5α reductase. It starts by DHT accumulates in the sebaceous glands and stimulates them to secrete excess sebum. This excessively secreted sebum blocks pores and inhibits hair growth. On the other hand, DHT causes hair mother cells to die. Therefore, in order to suppress the production of DHT, 5α reductase activity inhibitors have been developed (Patent Documents 1 to 3). As 5α-reductase activity inhibitors, in addition to progesterone (non-patent document 1), which is a female hormone, unsaturated fatty acids (non-patent document 2) and the like are being studied, but safety problems and sufficient effects are expected. Since it does not play, it does not meet the user's requirements.

Patent Document 4 describes a composition for external application to hair containing a pomegranate extract as an active ingredient. The pomegranate extract is effective for preventing hair loss and hair growth, and has a remarkable hair regeneration effect and firmness on hair. It is described that it gives a firmness.
Further, Patent Literature 5 describes a hair cosmetic composition containing estradiol obtained from a pomegranate seed extract as an active ingredient. Estradiol is a type of female hormone that is secreted synthetically in the body, and other female hormones such as estrogen, estriol, and estrone are also included.
Furthermore, Patent Document 6 describes a hair suppressor, a hair papilla cell growth inhibitor, a testosterone 5α-reductase activity inhibitor, and an antiandrogen that contain an extract from pomegranate as an active ingredient.

On the other hand, it is known that a substance called urolithin exists. Urolitin represented by urolithin A is known as a metabolite of ellagic acid derived from ellagitannin contained in pomegranate, raspberry, blackberry, cloudberry, strawberry, walnut and the like. Ellagitannin is classified as a hydrolyzable tannin, and it is known that when ingested, it is hydrolyzed in the body and converted into ellagic acid. Although such ellagitannin and ellagic acid have very low intestinal absorbability in the body, it is known that when they are ingested, they are further metabolized by the human colonic microbiota to be converted into urolithins. The urolithins thus produced are one of the most important compounds in the living body. In recent years, the intestinal bacterium has been identified as Gordonibacterium urolithinfaciens (Non-Patent Document 3).

It has been reported in rats and humans that urolithin A is the main metabolite after ingestion of ellagitannin and ellagic acid, and it has been reported that urolithin A has anti-inflammatory action, anticancer action and the like. Patent Document 7 describes urolithins such as urolithin A and urolithin B, and an effective amount of urolithins for the treatment or prevention of symptoms selected from obesity, decreased metabolic rate, metabolic syndrome, etc. Foods, etc. containing it are listed.

Thus, in addition to the mechanism of hair loss, there are many findings concerning hair growth, hair cosmetics, uses of urolithins, biosynthesis of urolithins, etc., but urolithins have an effect on hair growth, And, it is not known to have an effect of inhibiting 5α reductase activity.

JP, 2014-159421, A Japanese Patent Publication No. 2007-516209 Japanese Patent Publication No. 2003-520229 JP, 2002-020242, A JP, 2011-162513, A JP, 2010-013442, A Japanese Patent Publication No. 2014-501764

Rabe T et al., “Inhibition of skin 5 alpha-reductase by oral contraceptive progestins in vitro.” Gynecol Endocrinol. 2000 Aug;14(4):223-30. Liang T et al., “Inhibition of steroid 5 alpha-reductase by specific fatty unsaturated fatty acids.” Biochem J. 1992 Jul 15;285 (Pt 2):557-62. Int. J. Syst. Eval. Microbiol., 64, 2346-2352 (2014)

The present invention has been made under the above circumstances, and an object of the present invention is to provide a novel hair growth agent containing urolithins, and a novel 5α reductase activity inhibitor containing urolithins.

The present inventors have searched for a novel hair growth agent and a novel inhibitor that inhibits 5α reductase activity, and found that urolithins inhibit the conversion of testosterone to dihydrotestosterone, and completed the present invention. Let That is, the present invention is as follows.
<1>
A hair restorer containing urolithins.
<2>
A 5α reductase activity inhibitor containing urolithins.
<3>
The hair restorer according to <1>, wherein the urolithins are urolithin A, urolithin B, urolithin C, urolithin D or urolithin E.
<4>
The 5α reductase activity inhibitor according to <2>, wherein the urolithins are urolithin A, urolithin B, urolithin C, urolithin D or urolithin E.

ADVANTAGE OF THE INVENTION According to this invention, as a novel hair growth agent and a new inhibitor which inhibits 5(alpha) reductase activity, the hair growth agent containing urolithins and the 5(alpha) reductase activity inhibitor containing urolithins are provided. it can.

FIG. 3 is a graph showing the relationship between the urolithin A concentration and the 5α reductase activity inhibition rate in one embodiment of the present invention. FIG. 3 is a graph showing the relationship between urolithin B concentration and 5α reductase activity inhibition rate in one embodiment of the present invention. FIG. 4 is a graph showing the relationship between the urolithin C concentration and the 5α reductase activity inhibition rate in one embodiment of the present invention. FIG. 3 is a graph showing the relationship between urolithin D concentration and 5α reductase activity inhibition rate in one embodiment of the present invention. FIG. 4 is a graph showing the relationship between the urolithin E concentration and the 5α reductase activity inhibition rate in one embodiment of the present invention.

The present invention includes the following first invention and second invention.
The first invention is a hair restorer containing urolithins.
The second invention is a 5α reductase activity inhibitor containing urolithins.

<1. Hair growth agent containing urolithins>
The first invention of the present invention is a hair restorer containing urolithins.

(Urolithins)
The urolithins in the first invention of the present invention are not particularly limited, but their structures are substances represented by the above general formula (1). Further, as shown in Table 1, urolithins are represented by R1 to R6 in the chemical formula, and urolithin A, urolithin B, urolithin C, urolithin D, urolithin E, urolithin M3, urolithin M4, urolithin M5, urolithin M6, urolithin M7, and Isourolithin A and the like can be mentioned.

Of these, urolithin A, urolithin B, urolithin C, urolithin D, and urolithin E are preferable, and urolithin A is more preferable, since they have a high effect of inhibiting 5α reductase activity.

The method for obtaining urolithins is not particularly limited, and commercially available products may be used, or may be synthesized by chemical synthesis.

Examples of commercially available urolithins include urolithin A, urolithin B, urolithin C, urolithin D, and urolithin E (manufactured by Dalton Pharma).

In addition, as a synthetic method by chemical synthesis, a conventional method can be used. For example, as described in Examples of the present specification, 2-bromo-5-methoxybenzoic acid and aluminum chloride are used as raw materials for synthesis. There is a method.

Alternatively, punicalagin, which is a type of ellagitannin, may be extracted from the plant and hydrolyzed to ellagic acid, or ellagic acid may be extracted and converted into urolithins using a microorganism.

The type of plant is not particularly limited, and examples thereof include pomegranate, raspberry, blackberry, cloudberry, boysenberry, strawberry, walnut, ginger ginger and the like. Of these, pomegranate, boysenberry, and ginger ginger are preferable, and pomegranate is more preferable, since they contain a high content of ellagitannin and/or ellagic acid.
Any one of these plants may be used, or two or more thereof may be used in combination. Further, the extraction method and the extraction conditions from the plant are not particularly limited, and may be a conventional method. For example, known extraction methods such as water extraction, hot water extraction, warm water extraction, alcohol extraction, supercritical extraction and the like can be used.

When solvent extraction is performed, examples of the solvent include water; alcohols such as lower alcohols such as methanol and ethanol, and polyhydric alcohols such as propylene glycol and 1,3-butylene glycol (whether anhydrous or hydrous. ); Ketones such as acetone, diethyl ether, dioxane, acetonitrile, esters such as ethyl acetate, xylene and the like can be mentioned, and water and ethanol are preferable. These solvents may be used alone or in combination of two or more.

The method for hydrolyzing the extracted ellagitannin such as punicalagin to ellagic acid is not particularly limited, but includes a method of hydrolyzing with an acid, an enzyme, or a microorganism.

The method for converting ellagic acid to urolithins using a microorganism is not particularly limited, but, for example, a known method described in Food Funct., 5, 8, 1779-1784 (2014) can be used. ..

The obtained urolithins can be used as they are, but may be dried and used in powder form. If necessary, the obtained urolithins may be subjected to purification, concentration treatment and the like. As the purification treatment, filtration or adsorption or decolorization treatment using an ion exchange resin, an activated carbon column or the like can be performed. Further, as the concentration treatment, a conventional method such as an evaporator can be used.

Further, a method of subjecting the obtained urolithins (or a purified product or a concentrate) to freeze-drying to be powdered, or an excipient such as dextrin, corn starch, or gum arabic is added to be powdered by spray-drying. You may pulverize according to a well-known method, such as a method. After that, it may be dissolved in pure water, ethanol or the like, if necessary.

(Hair growth agent)
The first invention of the present invention is a hair-growing agent containing the above-mentioned urolithins. The hair restorer according to the present invention may contain one kind or a plurality of kinds of the above-mentioned urolithins.

As used herein, hair growth includes prevention, treatment, and delay of progression of male pattern baldness (AGA). Therefore, the hair-growth effect in this specification may refer to the hair loss prevention effect, and the hair growth agent may refer to the hair loss prevention agent. In androgenetic alopecia (AGA), the male hormone testosterone plays an important role, and it is considered that testosterone progresses due to dihydrotestosterone (DHT) converted by 5α reductase.

The first invention of the present invention is a hair restorer containing urolithins, wherein urolithins inhibit the activity of 5α reductase and testosterone, which is a male hormone, produces dihydrotestosterone (DHT) which is converted by the activity of 5α reductase. By suppressing, a hair growth effect is exhibited.
From this, the hair restorer according to the present invention is used for cosmetics, pharmaceuticals, foods, and supplements, and is particularly preferably used as cosmetics.

The hair restorer according to the present invention includes known excipients other than urolithins, fragrances, coloring agents, emulsifiers, stabilizers, thickeners, enzymes, preservatives, lubricants, surfactants, disintegrating agents, and disintegrating agents. Inhibitors, binders, absorption promoters, adsorbents, moisturizers, solubilizers, preservatives, flavoring agents, sweeteners, etc. are blended, if necessary, within a range that does not impair the effects of the first invention of the present invention. can do.

The content of urolithins relative to the total amount of the hair restorer is not particularly limited as long as the desired effect of the first invention of the present invention is exhibited, but the total amount of urolithins is usually 0.0001 to 10% by mass, and preferably Is 0.001 to 1% by mass, and more preferably 0.01 to 0.1% by mass.

<Cosmetic>
When the hair restorer according to the first aspect of the present invention is used as a material for cosmetics, the hair restorer is a liquid such as an aqueous solution, lotion, spray solution, suspension and emulsion; semisolid such as cream and paste; It can be prepared into cosmetics having various desired dosage forms such as gel. Such cosmetics are useful as various cosmetics such as hair cosmetics and body cosmetics.
The cosmetic containing the hair restorer according to the first aspect of the present invention can be produced by a conventional method. Further, the amount of the hair-growth agent to be added to the cosmetic, the mixing method, and the mixing time can be appropriately selected. Further, if necessary, it can be enclosed in an appropriate container such as a bottle, a bag, a can, a spray can, a spray container, a box and a pack.

When the hair restorer according to the first aspect of the present invention is used as a material for cosmetics, the content of the hair restorer with respect to the total amount of the cosmetic is particularly limited as long as the desired effect according to the first aspect of the present invention is achieved. However, the total amount of urolithins is usually 0.0001 to 10% by mass, preferably 0.001 to 1% by mass, and more preferably 0.01 to 0.1% by mass.

<Pharmaceutical>
When the hair-growth agent according to the first aspect of the present invention is used as a material for a drug, it can be used as a drug for the prevention and treatment of male pattern baldness (AGA), for example. The dosage form can be selected according to the disease to be prevented or treated, the usage form of the drug, the administration route, and the like. Examples thereof include tablets, coated tablets, pills, capsules, granules, powders, solutions, suspensions, emulsions, syrups, injections, suppositories, dips, decoctions and tinctures. These various preparations include fillers, fillers, excipients, binders, moisturizers, disintegrants, surfactants, lubricants, colorants, and flavoring agents as necessary for the main drug according to the conventional method. , A solubilizing agent, a suspending agent, a coating agent, and the like, which can be used for formulation. In addition, a coloring agent, a preservative, a flavoring agent, a flavoring agent, a sweetening agent and the like and other pharmaceuticals may be contained in the pharmaceutical preparation.

When the hair restorer according to the first aspect of the present invention is used as a raw material for a drug, the content of the hair restorer with respect to the total amount of the drug is not particularly limited as long as the desired effect according to the first aspect of the present invention is exhibited. The total amount of urolithins is usually 0.0001 to 10% by mass, preferably 0.001 to 1% by mass, and more preferably 0.01 to 0.1% by mass.

<Food>
When the hair restorer according to the first aspect of the present invention is used as a material for foods, it can be used as general foods, foods for specified health use, dietary supplements, functional foods, foods for patients, food additives, etc. .. As the form of food, a soft drink containing the hair restorer according to the first aspect of the present invention, milk, pudding, jelly, candy, gum, gummy, yogurt, chocolate, soup, cookies, snacks, ice cream, popsicle, Examples thereof include bread, cake, cream puff, ham, meat sauce, curry, stew, cheese, butter and dressing.

The hair restorer according to the first aspect of the present invention can use water, proteins, sugars, lipids, vitamins, minerals, organic acids, organic bases, fruit juices, flavors and the like as main components. Examples of the proteins include whole milk powder, skim milk powder, partially skimmed milk powder, casein, soybean protein, egg protein, meat protein and other animal and vegetable proteins, and hydrolysates and butters thereof. Examples of the sugar include saccharides, modified starch (in addition to dextrin, soluble starch, British starch, oxidized starch, starch ester, starch ether, etc.) and dietary fiber. Examples of lipids include vegetable oils such as lard, safflower oil, corn oil, rapeseed oil, coconut oil, fractionated oils thereof, hydrogenated oils, transesterified oils and the like. Examples of vitamins include vitamin A, carotene, vitamin B group, vitamin C, vitamin D group, vitamin E, vitamin K group, vitamin P, vitamin Q, niacin, nicotinic acid, pantothenic acid, biotin, inositol, choline. , Folic acid and the like, and examples of the minerals include calcium, potassium, magnesium, sodium, copper, iron, manganese, zinc, selenium and whey minerals. Examples of the organic acid include malic acid, citric acid, lactic acid, tartaric acid and the like. These components may be used in combination of two or more kinds, and synthetic products and/or foods containing a large amount thereof may be used.

The food containing the hair restorer according to the first aspect of the present invention can be manufactured by a conventional method. Further, the amount of the hair-growth agent to be added to the food, the mixing method, and the mixing time can be appropriately selected. Furthermore, if necessary, it can be enclosed in an appropriate container such as a bottle, bag, can, box, or pack.
When the hair restorer according to the first aspect of the present invention is used as a material for food, the content of the hair restorer with respect to the total amount of the food is not particularly limited as long as the desired effect according to the first aspect of the present invention is exhibited. The total amount of urolithins is usually 0.0001 to 10% by mass, preferably 0.001 to 1% by mass, and more preferably 0.01 to 0.1% by mass.

<Supplement>
A supplement is one of the food categories consisting of dietary supplements, and in the present specification, it refers to a functional auxiliary substance capable of providing a hair growth effect and the like.
When the hair-growth agent according to the first aspect of the present invention is used as a material for supplements, it can be in the form of a solid, a gel, or a liquid. The form of the supplement can be, for example, various processed foods and drinks, powders, tablets, pills, capsules, jellies, granules and the like.

When the hair restorer is used as a material for supplements, excipients such as dextrin, preservatives such as vitamin C, flavoring agents such as vanillin, pigments such as safflower pigments, monosaccharides, oligosaccharides and polysaccharides (eg, It may contain additives such as glucose, fructose, sucrose, sucrose, and sugars containing these), acidulants, flavors, oils and fats, emulsifiers, milk powder, or agar. These components may be used in combination of two or more kinds, and may be a synthetic product and/or a large amount thereof.

When the hair restorer is used as a material for supplements, it can be manufactured according to a conventional method. Further, the amount to be added to the supplement, the mixing method, and the mixing time can be appropriately selected. Furthermore, if necessary, it can be enclosed in an appropriate container such as a bottle, bag, can, box, or pack.
When the hair restorer is used as a material for a supplement, the content of the hair restorer with respect to the total amount of the supplement is not particularly limited as long as the above effects are exhibited, but as a urolithin, the total amount is usually 0.0001 to 10% by weight. And preferably 0.0005 to 1% by weight, and more preferably 0.001 to 0.1% by weight.

Cosmetics, pharmaceuticals, foods, supplements, etc. containing the hair-growth agent according to the first aspect of the present invention are cosmetics, pharmaceuticals, foods, supplements, etc. with a label indicating that they are used for hair-growth. Can be sold.

The "display" means all actions to inform the consumer of the above-mentioned use, and if it is a display that can recall or analogize the above-mentioned use, the purpose of the display, the content of the display, the display Regardless of the object, medium, etc. to be processed, they all correspond to the "display" of the first invention of the present invention. However, it is preferable to display the expression so that the consumer can directly recognize the use. Specifically, the act of stating the above-mentioned use in a product or packaging of a product related to cosmetics, pharmaceuticals, foods, supplements, etc., and the transfer, delivery, transfer or delivery of a product or packaging of a product stating the above-mentioned use. For the purpose of exhibiting, importing, advertising regarding products, listing the above-mentioned uses in price lists or transaction documents, or displaying or distributing them, or by describing the above-mentioned uses in the information containing these electromagnetic ( An example of the act, etc. provided by a method such as the Internet).

On the other hand, the display is preferably a display approved by the government (for example, a display approved in accordance with various systems established by the government and displayed in a form based on such approval), and particularly packaging and containers. It is preferable to display it on advertising materials such as catalogs, pamphlets, POP, etc. at sales sites, and other documents.

Further, for example, if it is a food containing the hair restorer according to the first aspect of the present invention, the display as a health food, a functional food, a special purpose food, a nutritionally functional food, a quasi-drug etc. is exemplified. In addition, labeling approved by the Ministry of Health, Labor and Welfare, for example, labeling approved by foods for specified health uses or similar systems can be exemplified. Examples of the latter include labeling as a food for specified health use, labeling as a food for conditional health care, labeling that affects the structure and function of the body, labeling for reducing disease risk, etc. Speaking of which, the indication as a food for specified health use (in particular, the indication of the purpose of health use) stipulated in the Health Promotion Law Enforcement Regulations (April 30, 2003, Ministry of Health, Labor and Welfare Ordinance No. 86 of Japan), and the like The display can be listed as a typical example.

<2. 5α-reductase activity inhibitor containing urolithins>
The second invention of the present invention is a 5α reductase activity inhibitor containing urolithins.

5α-reductase is also called steroid 5α-reductase or testosterone-5α-reductase, and among the androgens having androgen action, the double bond between 4-position and 5-position of testosterone, which is a typical one, becomes the α-type at 5 position. Is an enzyme that produces dihydrotestosterone, which is the active male hormone. The 5α reductase activity inhibitor suppresses the production of dihydrotestosterone by inhibiting the reducing activity of 5α reductase.

As described above, hair loss is initiated by the conversion of the male hormone testosterone into dihydrotestosterone (DHT) by the action of 5α reductase. Therefore, for example, from the viewpoint of hair growth, urolithins inhibit the 5α reductase activity, and testosterone, which is a male hormone, suppresses the production of dihydrotestosterone (DHT) that is converted by the 5α reductase activity, thereby improving the hair growth effect. Play.

Similarly, it is known that prostatic hypertrophy and acne vulgaris (acne) are initiated by conversion of the male hormone testosterone into dihydrotestosterone (DHT) by the action of 5α reductase. Therefore, for example, from the viewpoint of treatment or prevention of diseases or symptoms of benign prostatic hyperplasia and acne vulgaris (acne), urolithins inhibit the 5α reductase activity, and the male hormone testosterone is converted by the 5α reductase activity. By suppressing the production of dihydrotestosterone (DHT), it is possible to treat or prevent diseases or symptoms of benign prostatic hyperplasia and acne vulgaris (acne).

From this, the 5α reductase activity inhibitor according to the second invention of the present invention is preferably used for hair growth, as well as for treating or preventing diseases or symptoms of prostatic hypertrophy and acne vulgaris (acne). .. The 5α-reductase activity inhibitor according to the second aspect of the present invention is used in cosmetics, pharmaceuticals, foods and supplements, and is particularly preferably used as cosmetics.

The content of urolithins relative to the total amount of the 5α reductase activity inhibitor according to the second invention of the present invention is not particularly limited as long as the desired effect according to the second invention of the present invention is exhibited, but as the total amount of urolithins, The amount is usually 0.0001 to 10% by mass, preferably 0.001 to 1% by mass, and more preferably 0.01 to 0.1% by mass.

When the 5α reductase activity inhibitor according to the second invention of the present invention is used as a material for cosmetics, the content of the 5α reductase activity inhibitor with respect to the total amount of the cosmetic is such that the desired effect according to the second invention of the present invention is obtained. The total amount of urolithins is usually 0.0001 to 10% by mass, preferably 0.001 to 1% by mass, and more preferably 0.01 to 0.1% by mass, as long as it is played. Is.

When the 5α reductase activity inhibitor according to the second invention of the present invention is used as a raw material of a drug, the content of the 5α reductase activity inhibitor with respect to the total amount of the drug is such that the desired effect according to the second invention of the present invention is achieved. The total amount of urolithins is usually 0.0001 to 10% by mass, preferably 0.001 to 1% by mass, and more preferably 0.01 to 0.1% by mass, though not particularly limited. ..

When the 5α reductase activity inhibitor according to the second invention of the present invention is used as a food material, the content of the 5α reductase activity inhibitor with respect to the total amount of the food is such that the desired effect according to the second invention of the present invention is achieved. The total amount of urolithins is usually 0.0001 to 10% by mass, preferably 0.001 to 1% by mass, and more preferably 0.01 to 0.1% by mass, though not particularly limited. ..

When the 5α reductase activity inhibitor according to the second invention of the present invention is used as a material for supplements, the content of the 5α reductase activity inhibitor with respect to the total amount of the supplement is such that the desired effect according to the second invention of the present invention is achieved. The total amount of urolithins is usually 0.0001 to 10% by mass, preferably 0.001 to 1% by mass, and more preferably 0.01 to 0.1% by mass, though not particularly limited. ..

Regarding the other aspects of the second invention of the present invention, the above-mentioned description of the first invention of the present invention is applied.

Hereinafter, the present invention will be described in more detail with reference to specific examples, but the present invention is not limited to these examples.

(Method for analyzing urolithin)
A case where urolithin A is used as an example of urolithins will be described. The analysis of urolithin A was performed using HPLC. That is, a solution prepared by dissolving urolithin A (manufactured by Dalton Farma) in a suitable solvent was analyzed under the following HPLC conditions, and the purity (%) (A) and the peak area value (B) in HPLC were used. The factor of urolithin A and the urolithin A concentration of the sample were calculated by the following calculation formulas (1) and (2).

(Urolithin A factor calculation formula)
Factor of urolithin A=(B)/(concentration of standard solution of urolithin A (mg/L)×(A)/100) (1)
(Sample urolithin A concentration calculation formula)
Urolithin A concentration of sample (mg/L)=peak area value of urolithin A in sample/factor of urolithin A (2)

(Analysis conditions)
Analytical column: Inertsil ODS-3 (250 x 4.6 mm) (manufactured by GL Science)
Detection wavelength: 305nm
Mobile phase: water/acetonitrile/acetic acid=74/25/1
Column temperature: 40°C
Flow rate: 1.0 mL/min
Under the above conditions, urolithin A has a retention time of 16.5 minutes.

<Preparation of urolithin A>
5-Bromo-5-methoxybenzoic acid (5 g, manufactured by Wako Pure Chemical Industries, Ltd.) and aluminum chloride (15 g) were refluxed in 150 mL of chlorobenzene for 2.5 hours. After cooling, the reaction solution was transferred to ice water and extracted three times with 250 mL of diethyl ether. The obtained extract was concentrated under reduced pressure and diethyl ether was distilled off to obtain 4.2 g of 2-bromo-5-hydroxybenzoic acid. The obtained 3.9 g of 2-bromo-5-hydroxybenzoic acid and 3.9 g of resorcinol (manufactured by Tokyo Chemical Industry Co., Ltd.) were heated in 9 mL of 4M NaOH aqueous solution at 60° C. for 30 minutes. After adding 1.8 mL of a 10% copper sulfate aqueous solution to this reaction liquid, heating was further performed at 80° C. for 10 minutes. The generated precipitate was collected by filtration to obtain a white powder of urolithin A.

<Measurement of 5α reductase activity inhibitory activity>
[Example 1]
10 mM Tris-HCl buffer (pH 7.2), 0.185 mg/mL testosterone (manufactured by Sigma), 20 μg/mL rat liver S-9 enzyme solution (manufactured by Oriental Yeast Co., Ltd.), 0.14 mg/mL NADPH. , And a final concentration of 1 mg/L, 10 mg/L, 100 mg/L, or 1000 mg/L of urolithin A as a test sample, a total of 1 mL of reaction solution was prepared, and enzyme reaction was performed at 37° C. for 30 minutes.
After the enzymatic reaction, 2 mL of dichloromethane was added to stop the reaction, and 0.2 mL of 0.1 mg/mL p-hydroxybenzoic acid-n-hexyl ester (manufactured by Tokyo Kasei Kogyo Co., Ltd.) was added as an internal standard substance to give 10. After shaking for 1 minute, the mixture was centrifuged (3000 rpm, 10 minutes). After removing the supernatant, dichloromethane was distilled off from the dichloromethane layer by nitrogen concentration, and methanol was added to prepare a measurement sample. The 5α reductase activity inhibition rate was calculated by measuring the amount of testosterone in the measurement sample by HPLC.

[Comparative Example 1]
The same one as in Example 1 except that urolithin A was not used was used as a control (Comparative Example 1).

A blank similar to that of Example 1 was used except that the enzyme 5α reductase was not added.

(Testosterone analysis conditions)
Analytical column: Inertsil ODS-4 (150 x 4.6 mm) (manufactured by GL Science)
Detection wavelength: 240nm
Mobile phase: water/methanol = 60/40
Column temperature: 35°C
Flow rate: 1.0 mL/min

The 5α reductase activity inhibition rate was calculated by the following calculation formula (3).
Inhibition rate of 5α reductase activity (%)={1-(AB)/(AC)}×100
・(3)
A: residual amount of testosterone in blank (no enzyme added, test sample added) B: residual amount of testosterone in test reaction solution (added enzyme, added test sample) C: residual testosterone amount in control (added enzyme, no test sample added)

A graph showing the relationship between the urolithin A concentration and the 5α reductase activity inhibition rate is shown in FIG. 1. As shown in FIG. 1, the 5α reductase activity inhibition rate was found to increase in a concentration-dependent manner of urolithin A, and it was confirmed that the protein acts as a 5α reductase inhibitor.

[Example 2]
Example except that urolithin B (manufactured by Dalton Pharma) was used as a test sample and the reaction solution was prepared so that the final concentration was 15.6 mg/L, 62.5 mg/L, 250 mg/L, or 1000 mg/L. The same method as 1 was used.

[Comparative Example 2]
The same one as in Example 2 except that urolithin B was not added was used as a control (Comparative Example 2).
A blank similar to that in Example 2 was used except that the enzyme 5α reductase was not added.

A graph showing the relationship between the urolithin B concentration and the 5α reductase activity inhibition rate is shown in FIG. 2. As shown in FIG. 2, the urolithin B concentration-dependent increase in the 5α reductase activity inhibition rate was observed, and it was confirmed that the urolithin B acts as a 5α reductase inhibitor.

[Example 3]
The same method as in Example 1 was performed except that urolithin C (manufactured by Dalton Pharma) was used as a test sample.

[Comparative Example 3]
A control (Comparative Example 3) was prepared in the same manner as in Example 3 except that urolithin C was not added.
A blank similar to that of Example 3 was used except that the enzyme 5α reductase was not added.

A graph showing the relationship between the concentration of urolithin C and the inhibition rate of 5α reductase activity is shown in FIG. As shown in FIG. 3, the urolithin C concentration-dependent increase in the 5α-reductase activity inhibition rate was observed, and it was confirmed that the urolithin C acts as a 5α-reductase inhibitor.

[Example 4]
The same method as in Example 1 was performed except that urolithin D (manufactured by Dalton Pharma) was used as a test sample.

[Comparative Example 4]
The same one as in Example 4 except that urolithin D was not used was used as a control (Comparative Example 4).
Also, a blank similar to that of Example 4 was used except that the enzyme 5α reductase was not added.

A graph showing the relationship between the urolithin D concentration and the 5α reductase activity inhibition rate is shown in FIG. As shown in FIG. 4, an increase in the 5α-reductase activity inhibition rate was observed depending on the concentration of urolithin D, and it was confirmed that the protein acts as a 5α-reductase inhibitor.

[Example 5]
The same method as in Example 1 was performed except that urolithin E (manufactured by Dalton Pharma) was used as a test sample.

[Comparative Example 5]
The same one as in Example 5 except that urolithin E was not used was used as a control (Comparative Example 5).
A blank similar to that in Example 5 was used except that the enzyme 5α reductase was not added.

A graph showing the relationship between the concentration of urolithin E and the inhibition rate of 5α reductase activity is shown in FIG. As shown in FIG. 5, the urolithin E concentration-dependent increase in the 5α reductase activity inhibition rate was observed, and it was confirmed that the urolithin E acts as a 5α reductase inhibitor.

[Production Example 1 Hair tonic for hair growth]
Based on the formulations (Formulation Example 1) shown in Table 2, hair tonics for hair growth of Test Product A and Control Product A were manufactured. All were manufactured by dissolving components 1 to 3 in component 4.

[Example 6, Comparative Example 6 (hair growth effect test)]
A sensory test (hair growth effect test) was carried out using the hair tonics for hair growth of Test Product A (Example 6) and Control Product A (Comparative Example 6) manufactured in Production Example 1.
The sensory test was carried out by 10 men. As a control period for the first two months, a hair tonic containing no urolithin A (control product A, Comparative Example 6) was applied to the scalp twice a day, about 3 ml each time. The next two months was a transition period, and a hair tonic containing urolithin A (test product A) was similarly applied. A hair tonic containing urolithin A (test product A, Example 6) was applied in the same manner for the next 2 months as a test period.

In the final week of the control period and the final week of the test period, the hair was washed once a day for a total of 4 days, and the number of hairs that had fallen during hair washing was collected, and the number of hairs was measured to calculate the number of hairs lost per hair wash. ..
As a result, it was confirmed that in 7 out of 10 subjects, the number of hair loss at the time of washing the hair was decreased by 40 or more per wash, which was effective for hair growth.

INDUSTRIAL APPLICABILITY The present invention can be applied to formulation technology for cosmetics, medicines, foods, supplements and the like.

Claims (2)

A hair restorer containing urolithin A, urolithin B, urolithin C, urolithin D or urolithin E. A 5α reductase activity inhibitor containing urolithin A, urolithin B, urolithin C, urolithin D or urolithin E.