JP7350236B2 - Hyaluronic acid production inhibitor and moisturizer, collagen production promoter and anti-wrinkle agent - Google Patents

Description

Article 30, Paragraph 2 of the Patent Act applies Website publication date: January 29, 2019 http://www. jwrs. org/wood2019/Prg_Oral. html Website publication date: January 29, 2019 http://www. jwrs. org/wood2019/Prg_Poster. html Website publication date: March 4, 2019 http://conference. wdc-jp. com/jwrs2019/2019/contents/oral. html#menu Website publication date: March 4, 2019 http://conference. wdc-jp. com/jwrs2019/2019/contents/common/pdf/M16-09-1030. pdf Website publication date: March 4, 2019 http://conference. wdc-jp. com/jwrs2019/2019/contents/poster. html#menu Website publication date: March 4, 2019 http://conference. wdc-jp. com/jwrs2019/2019/contents/common/pdf/M15-P-04. pdf

The present invention relates to a hyaluronic acid production promoter and a moisturizing agent. The present invention also relates to collagen production promoters and anti-wrinkle agents. The present invention also relates to a cell activator. The present invention also relates to melanin production inhibitors and whitening agents.

Plants contain components that have various functions and efficacy. Conventionally, efforts have been made to apply the functions of plant extracts and to elucidate new functionalities of plant extracts.
In recent years, abandoned bamboo forests have become a problem nationwide. Bamboo shoots were cultivated in various places through government bamboo forest promotion projects, and until the 1970s, bamboo shoots were an important source of income for rural villages, but prices collapsed due to the increase in imported foreign bamboo shoots. Due to changes in lifestyles, the introduction of substitutes, and a lack of successors, the number of abandoned bamboo forests is increasing. To solve this problem, new functionalities such as bamboo extracts are being developed and applied products using bamboo extracts are being developed (Patent Documents 1 and 2).

JP2018-095630A Japanese Patent Application Publication No. 11-269020

However, the functionality of bamboo extracts varies depending on the part of the bamboo from which it is sourced, and its functionality is not fully known.
An object of the present invention is to provide a new hyaluronic acid production promoter and collagen production promoter. Furthermore, an object of the present invention is to provide a new cell activator. Another object of the present invention is to provide a melanin production inhibitor.

As a result of extensive research in order to solve the above problems, the present inventors found that the following invention met the above objects, leading to the present invention. That is, the present invention relates to the following inventions.

<1> A hyaluronic acid production promoter containing bamboo stem epidermis extract as an active ingredient.
<2> A collagen production promoter containing bamboo stem epidermis extract as an active ingredient.
<3> A moisturizer containing the hyaluronic acid production promoter according to <1> above.
<4> An anti-wrinkle agent containing the collagen production promoter according to <2> above.
<5> β-sitosterol, beturinic acid, Friedelin, tricine, 4-hydroxybenzoic acid, glucopyranosyl dimethoxycinnamate, dihydroxypropiophenone glucoside, linolesinol glucoside, tachioside, 1,2-dilinolein, triacontanol, and linolene A cell activation agent containing as an active ingredient one or more selected from Group A consisting of acids.
<6> One or more selected from Group B consisting of beturinic acid, Friedelin, tricine, linolesinol glucoside, dihydroxypropiophenone glucoside, 1,2-dilinolein, triacontanol, linolenic acid, and tachioside as an active ingredient Contains melanin production inhibitor.
<7> A whitening agent containing the melanin production inhibitor according to <6> above.

Further, the present invention relates to the following inventions.
<X1> A moisturizer containing bamboo stem epidermis extract as an active ingredient.
<X2> An anti-wrinkle agent containing bamboo stem epidermis extract as an active ingredient.
<X3> One or more selected from Group B consisting of beturinic acid, Friedelin, tricine, linoresinol glucoside, dihydroxypropiophenone glucoside, 1,2-dilinolein, triacontanol, linolenic acid, and tachioside as an active ingredient Contains whitening agent.

According to the present invention, a new hyaluronic acid production promoter and collagen production promoter are provided. More specifically, a hyaluronic acid production promoter and a collagen production promoter containing a bamboo stem epidermis extract are provided. Further, according to the present invention, a new cell activating agent is provided. Further, according to the present invention, a melanin production inhibitor is provided.

MeOH extract of bamboo stem epidermis, hexane soluble part of MeOH extract of bamboo stem epidermis, ethyl acetate soluble part of MeOH extract of bamboo stem epidermis, and aqueous layer residue of MeOH extract of bamboo stem epidermis were added, respectively. It is a diagram plotting the hyaluronic acid production rate, cell survival rate, and hyaluronic acid production rate per cell for the system and control. MeOH extract of bamboo stem epidermis, hexane soluble part of MeOH extract of bamboo stem epidermis, ethyl acetate soluble part of MeOH extract of bamboo stem epidermis, and aqueous layer residue of MeOH extract of bamboo stem epidermis were added, respectively. It is a diagram plotting the collagen production rate, cell survival rate, and collagen production rate per cell of the system and control. FIG. 2 is a diagram plotting the hyaluronic acid production rate, cell survival rate, and hyaluronic acid production rate per cell in a system to which compounds 1 to 5 isolated from the methanol extract of bamboo stem epidermis were added, respectively, and a control system. FIG. 2 is a diagram plotting the hyaluronic acid production rate, cell survival rate, and hyaluronic acid production rate per cell in a system to which compounds 6 to 12 isolated from the methanol extract of bamboo stem epidermis were added, respectively, and a control system. FIG. 3 is a diagram plotting the collagen production rate, cell survival rate, and collagen production rate per cell in a system to which Compounds 1 to 5 isolated from the methanol extract of bamboo stem epidermis were added, respectively, and a control system. FIG. 2 is a diagram plotting the collagen production rate, cell survival rate, and collagen production rate per cell in a system to which compounds 6 to 12 isolated from the methanol extract of bamboo stem epidermis were added, respectively, and a control system. The melanin production rate per cell was plotted in the system to which beturinic acid, Friedelin, tricine, linolesinol glucoside, dihydroxypropiophenone glucoside, 1,2-dilinolein, triacontanol, linolenic acid, and tachyoside were added, and the control. It is a diagram. FIG. 2 is a diagram plotting the melanin production rate per cell in a system to which β-sitosterol, 4-hydroxybenzoic acid, and glucopyranosyl dimethoxycinnamate were added, respectively, and a control.

The embodiments of the present invention will be described in detail below, but the explanation of the constituent elements described below is an example (representative example) of the embodiments of the present invention, and the present invention will be described below unless the gist thereof is changed. is not limited to the content. Note that when the expression "~" is used in this specification, it is used as an expression including the numerical values before and after it.

[Hyaluronic acid production promoter]
The hyaluronic acid production promoter of the present invention contains bamboo stem epidermis extract as an active ingredient.
Hyaluronic acid is a main component of the extracellular matrix and is involved in various cell-cell interactions. Hyaluronic acid is an essential element for maintaining moisture in the skin, and it is thought that by producing and accumulating hyaluronic acid, the skin prevents the loss of water in the body and protects against physical stimuli. . By promoting hyaluronic acid production, you can keep your skin moisturized and healthy. The present inventors have discovered that a bamboo stem epidermis extract has the effect of promoting hyaluronic acid production. Since the bamboo stem epidermis extract is a naturally derived component, the hyaluronic acid production promoter of the present invention can also be highly safe.

[Collagen production promoter]
The collagen production promoter of the present invention contains bamboo stem epidermis extract as an active ingredient.
Collagen is a structural protein that forms connective tissues such as animal skin. Collagen is important for maintaining skin strength and keeping it wrinkle-free and youthful. By promoting the production of collagen, it gives firmness to the skin, maintains skin elasticity, and keeps the skin youthful and healthy. The present inventors have discovered that a bamboo stem epidermis extract has the effect of promoting collagen production. Furthermore, since the bamboo stem epidermis extract is a naturally derived component, the collagen production promoter of the present invention can also be highly safe.

(Bamboo stem epidermis extract)
The bamboo stem epidermis extract used in the hyaluronic acid production promoter of the present invention and the collagen production promoter of the present invention is extracted from the bamboo stem epidermis using an extraction solvent.
The bamboo trunk epidermis refers to the green skin on the surface of the bamboo trunk, and is approximately 1 mm thick. As mentioned above, bamboo stem epidermal extract is effective in promoting hyaluronic acid production and collagen production.

Bamboo that is the source of the bamboo trunk skin includes Madake, Mosouchiku, Hachiku, Hoteichiku, Kikkouchiku, Horaichiku, Narihiradake, Chishimazasa, Tochiku, Shihouchiku, Kanchiku, Yadake, and Medake. The stem epidermis of Prunus japonicus is preferred because it accumulates in large quantities and is easily available.

To prepare a bamboo stem epidermis extract, the bamboo stem epidermis is crushed or powdered to an appropriate size, dried as appropriate, and then immersed in an extraction solvent to bring the bamboo stem epidermis into contact with the extraction solvent. It can be done with

As the extraction solvent, water, an organic solvent, etc. can be used. As the water, water, physiological saline, a buffer solution, or the like can be used. Examples of organic solvents include methanol, ethanol, n-propyl alcohol, i-propyl alcohol, butyl alcohol, pentanol, ethylene glycol, propylene glycol, 1,3-butylene glycol, glycerin, acetone, methyl ethyl ketone, ethyl acetate, isopropyl acetate, Hexane, chloroform, diethyl ether, etc. can be used.

Further, the extraction solvent may be a mixed solvent of water and an organic solvent. In the case of a mixed solvent of water and an organic solvent, the content of the organic solvent can be 1% by volume or more, 10% by volume or more, 20% by volume or more, or 30% by volume or more based on the total volume. Further, the content of the organic solvent can be 99.8% by volume or less, 95% by volume or less, 90% by volume or less, or 80% by volume or less based on the total volume.

The extraction temperature and time may be adjusted as appropriate depending on the shape of the bamboo trunk epidermis, the type of extraction solvent used, the extraction method, etc.
The extraction temperature can be 0°C or higher, 10°C or higher, 20°C or higher, or the like. Further, the upper limit of the extraction temperature can be 100°C or lower, 90°C or lower, or 80°C or lower.
The extraction time can be 10 minutes or more, 30 minutes or more, 1 hour or more, 5 hours or more, 10 hours or more, etc. The upper limit can be 350 hours or less, 250 hours or less, 120 hours or less, 100 hours or less, 75 hours or less, 50 hours or less, 40 hours or less, 30 hours or less, 20 hours or less, etc. By performing extraction while stirring or shaking, the active ingredients can be efficiently extracted from the bamboo stem epidermis, and the extraction time can be shortened.

After the extraction solvent and the bamboo stem epidermis are brought into contact to obtain a solution in which the active ingredients are extracted, the extract obtained by separating the solution from the bamboo stem epidermis may be used as a bamboo stem epidermis extract as it is. Further, the obtained extract may be used as a bamboo stem epidermis extract by adjusting the pH, etc., or may be used as a concentrated liquid by concentrating the extract in order to increase the concentration of active ingredients. Alternatively, a dried product obtained by drying the obtained extract may be used as a bamboo stem epidermis extract.

In addition, the bamboo stem epidermis extract and its dried product are subjected to further treatments such as activated carbon treatment, liquid-liquid distribution, column chromatography, liquid chromatography, size exclusion chromatography, thin layer chromatography, and gel permeation. The highly active fraction may be collected and used as a bamboo stem epidermis extract.

For example, an example of a method for obtaining a dried bamboo stem epidermis extract includes the step (i) of contacting bamboo stem epidermis with an extraction solvent to obtain a bamboo stem epidermis extract; and removing the solvent from the bamboo stem epidermis extract. A manufacturing method including a step (ii) of removing.

In step (i), the contact between the bamboo trunk epidermis and the extraction solvent can be carried out, for example, by immersing the bamboo trunk epidermis in the extraction solvent. The amount of solvent can be set as appropriate depending on the shape and amount of the bamboo trunk skin, but generally it is 1 to 1000 times, 1 to 500 times, or 5 to 100 times that of the bamboo trunk skin in terms of mass ratio. , 10 to 50 times, 10 to 25 times, 5 to 20 times, etc. After the bamboo stem epidermis is immersed in the extraction solvent, it is stirred or shaken for a predetermined extraction time, thereby obtaining an extract in which the active ingredients are extracted from the bamboo stem epidermis into the extraction solvent. In order to enhance the effect of promoting hyaluronic acid production and collagen, the extraction solvent should be a lower alcohol with 1 to 5 carbon atoms such as methanol, ethanol, n-propyl alcohol, i-propyl alcohol, butyl alcohol, pentanol, or a carbon It is preferably a mixed solvent of a lower alcohol of numbers 1 to 5 and water, and preferably one selected from the group consisting of methanol, ethanol, a mixed solvent of methanol and water, and a mixed solvent of ethanol and water. More preferred.

In step (ii), the solvent can be removed from the bamboo trunk epidermis extract obtained in step (i) by drying under reduced pressure, freeze drying, spray drying, or the like.

Further, as an example of a method for obtaining a dried product, a manufacturing process including a step (iii) of performing liquid-liquid distribution using a mixed solvent after step (ii) can be mentioned. Liquid-liquid distribution may be performed multiple times. As a mixed solvent for liquid-liquid distribution, two or more mixed solvents having different polarities and being incompatible with each other can be used. Hexane or the like can be used as a low polar solvent, ethyl acetate or the like can be used as a medium polar solvent, and water can be used as a highly polar solvent. The mixed solvent may contain three or more kinds of solvents, and for example, a mixed solvent of hexane, ethyl acetate, and water can be used.
By further subjecting the dried product obtained in step (ii) to liquid-liquid distribution, it is possible to separate the components in the dried product obtained in step (ii) and obtain a highly active fraction.

Further, as an example of a method for obtaining a dried product, a manufacturing process including a step (iv) of performing chromatography after step (iii) may be mentioned. As the chromatography, liquid chromatography, thin layer chromatography, size exclusion chromatography, etc. can be used, and multiple chromatography methods may be used in combination. By processing by chromatography, highly active components can be further concentrated.

Because it has an excellent effect on promoting hyaluronic acid production and collagen, bamboo stem epidermal extract is extracted with a lower alcohol with 1 to 5 carbon atoms or a mixed solvent of lower alcohol with 1 to 5 carbon atoms and water. It is preferably an extract extracted with one selected from the group consisting of methanol, ethanol, a mixed solvent of methanol and water, and a mixed solvent of ethanol and water.

Specific examples of bamboo stem epidermis extracts include the following extracts (a) to (d).
(a) Methanol extract obtained by extracting bamboo stem epidermis with methanol or a mixed solvent of methanol-water (b) Methanol obtained by extracting bamboo stem epidermis with methanol or a mixed solvent of methanol-water The extract was further subjected to liquid-liquid distribution with a mixed solvent of hexane and water, and the hexane layer was collected to obtain a hexane-soluble product (c) bamboo trunk epidermis in methanol or a mixed solvent of methanol and water. The methanol extract obtained by the extraction was subjected to liquid-liquid partitioning with a mixed solvent of hexane and water, and then the obtained aqueous layer was subjected to liquid-liquid partitioning with a mixed solvent of ethyl acetate and water, and the ethyl acetate layer was separated. Ethyl acetate soluble material obtained by recovering (d) The methanol extract obtained by extracting the bamboo trunk epidermis with methanol or a mixed solvent of methanol-water, is dissolved in a liquid-liquid manner with a mixed solvent of hexane-water. Then, the resulting aqueous layer is subjected to liquid-liquid distribution with a mixed solvent of ethyl acetate and water, and the aqueous layer is collected.

The main components constituting the hexane soluble material (b) above are linolenic acid, β-sitosterol, 1,2-dilinolein, beturinic acid, triacontanol, and tricine.
Further, the main components constituting the ethyl acetate soluble material (c) above are Friedelin and 4-hydroxybenzoic acid.
Further, the main components constituting the water-soluble material (d) above are glucopyranosyl dimethoxycinnamate, linoresinol glucoside, dihydroxypropiophenone glucoside, and tachyoside.

The hyaluronic acid production promoter of the present invention promotes the production of hyaluronic acid, which is an essential component for maintaining moisture in the skin, and therefore can be suitably used as a moisturizer. In other words, a moisturizer containing bamboo stem epidermis extract may be used.

The uses of the hyaluronic acid production promoter and humectant of the present invention are not particularly limited, and can be used in cosmetics, pharmaceuticals, food and beverages, and the like.

The hyaluronic acid production promoter of the present invention and the humectant of the present invention may contain other components such as known additives in addition to the bamboo stem epidermis extract. In the hyaluronic acid production promoter of the present invention and the moisturizer of the present invention, the content of the bamboo stem epidermis extract is not particularly limited as long as the purpose of the present invention can be achieved, but for example, 0.01 to 100 μg/ mg (0.01 to 100 μg of bamboo stem epidermis extract in dry weight per 1 mg of the hyaluronic acid production promoter of the present invention or the moisturizer of the present invention). Further, the content of the bamboo stem epidermis extract is preferably 0.05 to 50 μg/mg, more preferably 0.05 to 10 μg/mg. Further, the content of the bamboo stem epidermis extract may be 0.05 to 1 μg/mg.

In addition, the collagen production promoter of the present invention has the effect of maintaining skin elasticity by promoting collagen production, and can be suitably used as an anti-wrinkle agent for the purpose of preventing and improving wrinkles. can. In other words, it may be used as an anti-wrinkle agent containing bamboo stem epidermis extract.

The uses of the collagen production promoter of the present invention and the anti-wrinkle agent of the present invention are not particularly limited, and can be used in cosmetics, pharmaceuticals, food and drink products, and the like.

The collagen production promoter of the present invention and the anti-wrinkle agent of the present invention may contain other components such as known additives in addition to the bamboo stem epidermis extract. In the collagen production promoter of the present invention and the anti-wrinkle agent of the present invention, the content of bamboo stem epidermis extract is not particularly limited as long as the purpose of the present invention can be achieved, but for example, 0.01 to 100 μg/ mg (0.01 to 100 μg dry weight of the bamboo stem epidermis extract per 1 mg of the collagen production promoter of the present invention or the anti-wrinkle agent of the present invention). Further, the content of the bamboo stem epidermis extract is preferably 0.05 to 50 μg/mg, more preferably 0.05 to 10 μg/mg. Further, the content of the bamboo stem epidermis extract may be 0.05 to 1 μg/mg.

[Cell activation agent]
The cell activator of the present invention includes β-sitosterol, beturinic acid, Friedelin, tricine, 4-hydroxybenzoic acid, glucopyranosyl dimethoxycinnamate, dihydroxypropiophenone glucoside, linolesinol glucoside, tachyoside, 1,2-dilinolein, Contains one or more selected from Group A consisting of triacontanol and linolenic acid as an active ingredient. The present inventors have investigated the effects of β-sitosterol, beturinic acid, Friedelin, tricine, 4-hydroxybenzoic acid, glucopyranosyl dimethoxycinnamate, dihydroxypropiophenone glucoside, linolesinol glucoside, tachyoside, 1,2-dilinolein, triacontanol. discovered that linolenic acid has a cell activation function.

In particular, the cell activation agent of the present invention can be used as an activation agent for fibroblasts in the skin dermis. Fibroblasts in the skin dermis are cells that produce important components of the skin, such as ``hyaluronic acid'' and ``collagen.'' Decline in the function of dermal fibroblasts is a factor that causes skin aging symptoms such as wrinkles and sagging. By promoting the proliferation and activation of fibroblasts, it is expected to prevent and improve skin aging.

Compounds of group A (β-sitosterol, beturinic acid, Friedelin, tricine, 4-hydroxybenzoic acid, glucopyranosyl dimethoxycinnamate, dihydroxypropiophenone glucoside, linolesinol glucoside, tachyoside, 1,2-dilinolein, triacontanol, Linolenic acid) can be extracted and isolated from plant materials, for example, from bamboo stem epidermis extract.

The use of the cell activator of the present invention is not particularly limited, and it can be used in cosmetics, pharmaceuticals, food and drink products, and the like.

The cell activation agent of the present invention can contain other components such as known additives in addition to the compound of group A. In the cell activation agent of the present invention, the content of the group A compound is not particularly limited as long as it can achieve the object of the present invention.

When the cell activator of the present invention contains as an active ingredient any one of the group consisting of β-sitosterol, beturinic acid, Friedelin, tricine, Friedelin, 1,2-dilinolein, triacontanol, and linolenic acid, The content is preferably, for example, 0.01 to 100 μg/mg (0.01 to 100 μg of the active ingredient in dry weight per 1 mg of the cell activator of the present invention). Further, the content of the active ingredient is preferably 0.05 to 50 μg/mg, more preferably 0.05 to 10 μg/mg. Further, the content of the active ingredient may be 0.05 to 1 μg/mg.

When the cell activation agent of the present invention contains as an active ingredient any one selected from the group consisting of 4-hydroxybenzoic acid, glucopyranosyl dimethoxycinnamate, dihydroxypropiophenone glucoside, linolesinol glucoside, and tachioside; The content thereof is preferably, for example, 0.1 to 1000 μg/mg (0.1 to 1000 μg dry weight of the active ingredient per 1 mg of the cell activator of the present invention). Furthermore, the content of the active ingredient is preferably 0.1 to 500 μg/mg, more preferably 0.5 to 100 μg/mg. Further, the content of the active ingredient may be 0.5 to 10 μg/mg.

[Melanin production inhibitor]
The melanin production inhibitor of the present invention is selected from Group B consisting of beturinic acid, Friedelin, tricine, linoresinol glucoside, dihydroxypropiophenone glucoside, 1,2-dilinolein, triacontanol, linolenic acid, and tachyoside. Contains one or more as active ingredients.
Melanin is a general term for substances that are pigmented by polymerizing phenolic substances. The skin is divided into the epidermis and dermis, and melanin cells (melanocytes) located in the basal layer of the epidermis synthesize melanin. Melanin is produced using tyrosine as a raw material when stimulated by ultraviolet rays, and is excreted into the epidermal cells due to skin turnover, but remaining in the epidermal cells causes age spots. By suppressing the production of melanin, which causes age spots, it is expected to have whitening effects such as preventing and improving age spots. Awareness about skin whitening has been increasing in recent years, and there has been a demand for new agents and whitening agents that have a melanin production suppressing effect.
The present inventors have discovered that beturinic acid, Friedelin, tricine, linoresinol glucoside, dihydroxypropiophenone glucoside, 1,2-dilinolein, triacontanol, linolenic acid, and tachioside have a melanin production suppressing function.

Group B compounds (beturinic acid, Friedelin, tricine, linolesinol glucoside, dihydroxypropiophenone glucoside, 1,2-dilinolein, triacontanol, linolenic acid, tachyoside) must be extracted and isolated from plant materials, respectively. For example, it can be extracted and isolated from bamboo stem epidermis extract.

Since the melanin production inhibitor of the present invention is effective in preventing and improving age spots by inhibiting melanin production, the melanin production inhibitor of the present invention can be suitably used as a whitening agent. That is, it contains as an active ingredient one or more selected from Group B consisting of beturinic acid, Friedelin, tricine, linoresinol glucoside, dihydroxypropiophenone glucoside, 1,2-dilinolein, triacontanol, linolenic acid, and tachyoside. It can also be used as a whitening agent.

The use of the melanin production inhibitor of the present invention and the whitening agent of the present invention is not particularly limited, and they can be used in cosmetics, pharmaceuticals, food and drink products, and the like.

The melanin production inhibitor of the present invention and the skin whitening agent of the present invention may contain other components such as known additives in addition to the compound of group B. In the melanin production inhibitor of the present invention and the skin whitening agent of the present invention, the content of the compound of group B is not particularly limited as long as it can achieve the object of the present invention.

When the melanin production inhibitor of the present invention or the whitening agent of the present invention contains beturinic acid as an active ingredient, the content is, for example, 0.01 to 1000 μg/mg (per 1 mg of the melanin production inhibitor of the present invention). The dry weight of beturinic acid is preferably 0.01 to 1000 μg). Further, the content of beturinic acid is preferably 0.01 to 500 μg/mg, more preferably 0.05 to 100 μg/mg.

When the melanin production inhibitor of the present invention or the whitening agent of the present invention contains any one selected from the group consisting of tricin, friedelin, and triacontanol as an active ingredient, the content thereof is, for example, 0.01 to The amount is preferably 100 μg/mg (0.01 to 100 μg of the active ingredient in dry weight per 1 mg of the melanin production inhibitor of the present invention). Further, the content of the active ingredient is preferably 0.01 to 50 μg/mg, more preferably 0.05 to 10 μg/mg.

When the melanin production inhibitor of the present invention or the whitening agent of the present invention contains linolesinol glucoside as an active ingredient, the content is, for example, 0.1 to 1000 μg/mg (1 mg of the melanin production inhibitor of the present invention). It is preferable that the dry weight of linolesinol glucoside is 0.1 to 1000 μg). Further, the content of linolesinol glucoside is preferably 0.1 to 500 μg/mg, more preferably 0.5 to 200 μg/mg. Further, the content of linoresinol glucoside may be 0.5 to 100 μg/mg or 0.5 to 10 μg/mg.

When the melanin production inhibitor of the present invention or the whitening agent of the present invention contains dihydroxypropiophenone glucoside as an active ingredient, the content is, for example, 0.1 to 100 μg/mg (the melanin production inhibitor of the present invention It is preferable that the dry weight of dihydroxypropiophenone glucoside is 0.1 to 100 μg per 1 mg of dihydroxypropiophenone glucoside. Further, the content of dihydroxypropiophenone glucoside is preferably 0.5 to 50 μg/mg, more preferably 0.5 to 10 μg/mg.

When the melanin production inhibitor of the present invention or the whitening agent of the present invention contains 1,2-dilinolein as an active ingredient, the content is, for example, 0.01 to 10 μg/mg (the melanin production inhibitor of the present invention It is preferable that the dry weight of 1,2-dilinolein is 0.01 to 10 μg per 1 mg. Further, the content of 1,2-dilinolein is preferably 0.05 to 5 μg/mg, more preferably 0.05 to 1 μg/mg.

When the melanin production inhibitor of the present invention or the whitening agent of the present invention contains linolenic acid as an active ingredient, the content is, for example, 0.01 to 5 μg/mg (per 1 mg of the melanin production inhibitor of the present invention). The dry weight of linolenic acid is preferably 0.01 to 5 μg). Further, the content of linolenic acid is preferably 0.05 to 1 μg/mg, more preferably 0.05 to 0.5 μg/mg.

When the melanin production inhibitor of the present invention or the whitening agent of the present invention contains tachyoside as an active ingredient, the content is, for example, 1 to 1000 μg/mg (1 mg of the melanin production inhibitor of the present invention, tachyoside is preferably 1 to 1,000 μg/mg in terms of dry weight, more preferably 1 to 500 μg/mg. Furthermore, the content of tachyoside is preferably 5 to 400 μg/mg, more preferably 10 to 400 μg/mg, and even more preferably 50 to 400 μg/mg.

[Melanin production promoter]
The present invention relates to a melanin production promoter containing as an active ingredient one or more selected from Group C consisting of β-sitosterol, 4-hydroxybenzoic acid, and glucopyranosyl dimethoxycinnamate. Increased melanin production causes skin to turn brown and hair to turn black. Ingredients that enhance melanin production are expected to have effects such as preventing and improving skin browning, vitiligo, and gray hair. The present inventors have discovered that β-sitosterol, 4-hydroxybenzoic acid, and glucopyranosyl dimethoxycinnamate have a melanin production promoting function.
Group C compounds (β-sitosterol, 4-hydroxybenzoic acid, glucopyranosyl dimethoxycinnamate) can be extracted and isolated from plant materials, for example, extracted from bamboo stem epidermis extract. I can do it.

Since the melanin production promoter of the present invention enhances melanin production, it can be suitably used as a skin browning agent or a gray hair prevention and improvement agent. That is, it may be used as a skin browning agent or gray hair prevention and improvement agent containing as an active ingredient one or more selected from Group C consisting of β-sitosterol, 4-hydroxybenzoic acid, and glucopyranosyl dimethoxycinnamate.
These uses are not particularly limited, and can be used in cosmetics, medicines, food and drinks, etc.

The melanin production promoter of the present invention can contain other components such as known additives in addition to the compound of Group C. In the melanin production promoter of the present invention, the content of the group C compound is not particularly limited as long as the object of the present invention can be achieved.

When the melanin production promoter of the present invention contains β-sitosterol as an active ingredient, the content is, for example, 0.01 to 100 μg/mg (β-sitosterol per 1 mg of the cell activator of the present invention). is preferably 0.01 to 100 μg (dry weight). Further, the content of β-sitosterol is more preferably 0.05 to 50 μg/mg, and even more preferably 0.05 to 10 μg/mg. Further, the content of β-sitosterol may be 0.05 to 1 μg/mg, or 0.05 to 0.1 μg/mg.

When the melanin production promoter of the present invention contains 4-hydroxybenzoic acid as an active ingredient, the content is, for example, 0.1 to 1000 μg/mg (4-hydroxybenzoic acid per 1 mg of the cell activator of the present invention). The dry weight of -hydroxybenzoic acid is preferably 0.1 to 1000 μg, more preferably 0.5 to 500 μg/mg, and even more preferably 0.5 to 100 μg/mg. Further, the content of 4-hydroxybenzoic acid is preferably 5 to 100 μg/mg, more preferably 50 to 100 μg/mg.

When the melanin production promoter of the present invention contains glucopyranosyl dimethoxycinnamate as an active ingredient, the content is, for example, 0.1 to 1000 μg/mg (per 1 mg of the cell activator of the present invention, glucopyranosyl dimethoxycinnamate). The dry weight of glucopyranosyl acid is preferably 0.1 to 1000 μg). Further, the content of glucopyranosyl dimethoxycinnamate is preferably 0.5 to 500 μg/mg, more preferably 0.5 to 100 μg/mg.

[Cosmetics]
Various agents of the present invention (hyaluronic acid production promoter of the present invention, moisturizing agent of the present invention, collagen production promoter of the present invention, anti-wrinkle agent of the present invention, cell activation agent of the present invention, melanin production inhibitor of the present invention) agent, the whitening agent of the present invention, the melanin production promoter of the present invention, the skin browning agent of the present invention, or the gray hair prevention and improvement agent of the present invention) can be used in cosmetics. Among these, the various agents of the present invention are useful as external preparations for skin and external preparations for scalp use.

The form of the cosmetic is not particularly limited, and may be in the form of powder, solid, liquid, emulsion, or the like. Specifically, for example, skin care cosmetics such as soaps, lotions, serums, milky lotions, lotions, essences, creams, gels, and packs, and makeup products such as powders, powder foundations, cream foundations, blushers, and lipsticks. It can be used as a cosmetic. It can also be used as hair cosmetics such as shampoos, conditioners, treatments, and conditioners.

Cosmetics containing the various agents of the present invention can contain, in addition to the various agents of the present invention, one or more components that can be used in ordinary cosmetics as long as the object of the present invention is not impaired. For example, liquid oils, solid oils, various surfactants, metal soaps, whitening agents, humectants, gelling agents, water-soluble oils, which are usually used as base ingredients or additive ingredients in cosmetics or for the purpose of imparting functionality. Polymers, lower alcohols, polyhydric alcohols, sugars, ultraviolet absorbers, film-forming agents, resins, clathrate compounds, deodorants, amino acids, vitamins, drugs, extracts derived from plants, animals, and microorganisms, organic Examples include acids, organic amines, sequestering agents, antioxidants, antibacterial agents, preservatives, pH adjusters, refreshing agents, fragrances, colorants, antibacterial agents, and thickeners.

The amount of the various agents of the present invention varies depending on the aspect of the cosmetic, but as a dry weight, it may be 0.00001% by mass or more, 0.001% by mass or more, or 0.01% by mass based on the total mass. % or more, and 0.1% by mass or more. Further, the dry weight of the various agents of the present invention can be 20% by mass or less, 10% by mass or less, or 5% by mass or less based on the total mass.

The amount of cosmetics containing the various agents of the present invention to be used is appropriately selected depending on the gender, weight, age, purpose of use, etc. of the subject. For adult humans, it can be 0.01 to 1000 mg/kg (body weight)/day or more, 0.1 to 500 mg/kg (body weight)/day or more, 1 to 100 mg/kg (body weight)/day or more, It can be used once a day or divided into multiple doses.

[Pharmaceutical composition]
The various agents of the present invention can be combined with a pharmaceutically acceptable carrier to form a pharmaceutical composition. In addition, in this application, a pharmaceutical composition means a drug useful for at least one of prevention, treatment, and improvement of symptoms of a target disease, and includes not only pharmaceuticals but also quasi-drugs.

The dosage form of the pharmaceutical of the present invention is not particularly limited, and may be a solid preparation or a liquid preparation. Specific examples include powders, granules, tablets, capsules, troches, suspensions, emulsions, syrups, elixirs, injections, drops, and ointments. Pharmaceutically acceptable carriers include, for example, binders, stabilizers, excipients, diluents, pH buffers, thickeners, colorants, dispersants, emulsifiers, suspending agents, preservatives, Examples include disintegrants, solubilizers, solubilizing agents, and the like.

The amount of the various agents of the present invention in a pharmaceutical product is determined as appropriate depending on the dosage form and administration method of the preparation, but the amount may be 0.01 mass or more based on the total mass as a dry weight, or 0. .1% by mass or more, and 1% by mass or more. Moreover, the dry weight of the various agents of the present invention can be 95% by mass or less, 80% by mass or less, or 60% by mass or less based on the total mass. Sufficient effects can be achieved by blending within this range.

The dosage of pharmaceuticals containing the various agents of the present invention is appropriately selected depending on the gender, weight, age, purpose of use, etc. of the subject to be administered, but for adult humans, it is 0.01 to 1000 mg/kg. (body weight)/day or more, 0.1 to 500 mg/kg (body weight)/day or more, or 1 to 100 mg/kg (body weight)/day or more.

In addition, the administration method of the pharmaceutical composition is appropriately selected depending on the gender, weight, age, purpose of use, etc. of the subject, and may be administered orally or parenterally. good. Further, the pharmaceutical composition may be administered once a day or in multiple doses, or may be administered continuously over several days, weeks, or months.

[Food and drink composition]
The various agents of the present invention can be blended into food and drink products to form food and drink compositions. By preparing the food and drink compositions, the various agents of the present invention can be easily ingested on a daily basis. Specifically, it can be general food, functional food, food for specified health use, nutritional functional food, health food, nutritional supplement, food with functional claims, special food, food additive, supplement, pet food, etc. can.

The form of the food and beverage composition containing the various agents of the present invention is not particularly limited, and may include jelly, candy, mousse, cake, cookie, biscuit, candy, chewing gum, frozen dessert, yogurt, ice cream, cheese, cream, bread, and sausage. , ham, pasta, various teas, soft drinks, carbonated drinks, powdered drinks, coffee drinks, alcoholic beverages, energy drinks, and other known forms that can be taken orally. When used as a supplement, it may be in any form such as tablets, powders, granules, pills, capsules, sugar-coated tablets, films, troches, chewables, solutions, emulsions, and suspensions.

When the various agents of the present invention are blended into a food/beverage composition, the amount of the various agents of the present invention varies depending on the aspect of the food/beverage product, etc., but as a dry weight, 0.001% of the total mass The content can be at least 0.01% by mass, or at least 0.1% by mass. Moreover, the dry weight of the various agents of the present invention can be 50% by mass or less, 30% by mass or less, 10% by mass or less, or 5% by mass or less based on the total mass.

The intake amount of the food and drink composition containing the various agents of the present invention is appropriately selected depending on the gender, weight, age, purpose of use, etc. of the subject, but for adult humans, it is 0.01 to 0.01. The amount can be 1000 mg/kg (body weight)/day or more, 0.1 to 500 mg/kg (body weight)/day or more, or 1 to 100 mg/kg (body weight)/day or more. In addition, the food and drink composition can be taken once a day or in multiple doses, or can be taken continuously over several days, weeks, or months.

EXAMPLES Hereinafter, the present invention will be explained in more detail with reference to Examples, but the present invention is not limited to the following Examples unless the gist thereof is changed.

Example 1. Evaluation of bamboo stem epidermis extract
Bamboo stem epidermal extract was used to evaluate its ability to promote hyaluronic acid production and collagen production.

1-1. Production of bamboo stem epidermis extract [methanol extract]
15 L of 99.5% methanol was added to 2.7 kg of powdered bamboo trunk epidermis (manufactured by Kazei Bussan Co., Ltd.), and extraction was performed three times (24 hours each) while shaking at room temperature. The extract was filtered, and the solvent was distilled off from the filtrate by vacuum concentration to obtain a methanol extract (223 g).

[Hexane soluble matter, ethyl acetate soluble matter, aqueous layer residue]
Liquid-liquid partition extraction was performed on 220 g of methanol extract with 300 mL of water and hexane (2 L x 3 times) to obtain 60 g of hexane-soluble portion, and further liquid-liquid partition extraction was performed with ethyl acetate (2 L x 3 times). 40 g of ethyl acetate soluble portion and 45 g of aqueous layer residue were obtained.

1-2. Evaluation The collagen production promoting function of the methanol extract (MeOH extract), hexane soluble material (hexane soluble part), ethyl acetate soluble material (ethyl acetate soluble part), and aqueous layer residue obtained in 1-1 The function of promoting hyaluronic acid production was evaluated using the following procedures (A1) to (A5).

(A1) Preparation of Samples Dimethyl sulfoxide (DMSO) or chloroform was added to each of the methanol extract (MeOH extract), hexane soluble material, ethyl acetate soluble material, and aqueous layer residue. Thereafter, it was stirred and treated with ultrasonic waves to completely dissolve it. Each of the obtained samples was diluted as necessary and subjected to respective functional evaluations.

(A2) Cell culture Adult human dermal fibroblasts (NHDF-Ad) were used as model cells. NHDF-Ad cells were grown in a φ10 cm dish using Dulbecco's Modified Eagle Medium (D-MEM) (high glucose) containing 1% penicillin-streptomycin and 10% fetal bovine serum (FBS) until confluent. Precultured. Thereafter, the cells were washed with phosphate buffered saline (PBS), resuspended in the medium, and then seeded in a 96-well plate at a concentration of 0.5 x 10 ⁴ cells/well in a CO ₂ incubator (37°C, 5% CO ₂ ) was cultured overnight.

(A3) Sample addition After overnight culture, the medium was replaced with a serum-free medium (containing 1% penicillin-streptomycin) containing the sample prepared in (A1), and cultured in a CO ₂ incubator for 72 hours. The MeOH extract, hexane soluble material, and ethyl acetate soluble material were added at final concentrations of 0.1 μg/mL, 1 μg/mL, and 10 μg/mL. The aqueous layer residue was added at a final concentration of 0.1 μg/mL and 1 μg/mL. DMSO or chloroform was used as a control.

(A4) Hyaluronic acid production test/collagen production test Collect the culture supernatant 72 hours after sample addition, and measure the amount of hyaluronic acid or collagen produced using a commercially available ELISA kit (Human Collagen Type I ELISA kit (Acel Co., Ltd.) , QnE Hyaluronic Acid (HA) ELISA Assay Kit (Biotech Trading Partners, LLC)) to calculate the hyaluronic acid production rate or collagen production rate.
The hyaluronic acid production rate or collagen production rate of the control was set as 100%, and the hyaluronic acid production rate or collagen production rate upon addition of each sample was calculated as a relative ratio of the control.

(A5) Cell viability measurement (MTT assay)
After collecting the culture supernatant 72 hours after sample addition, 100 μL of serum-free medium (containing 1% penicillin-streptomycin) containing no sample was added to each well of the 96-well plate. Next, 20 μL of MTT staining solution (5 mg/mL in PBS) was added and cultured for 4 hours in a CO ₂ incubator. The medium was removed, and 100 μL of hydrochloric acid-isopropanol solution was added to each well, and the formazan was completely dissolved by pipetting. Absorbance at 570 nm was measured using a microplate reader, and cell survival rate was calculated.
The cell survival rate of the control was set as 100%, and the cell survival rate upon addition of each sample was calculated based on the relative ratio of the control.

FIG. 1 shows the hyaluronic acid production rate, cell survival rate, and hyaluronic acid production rate per cell (hyaluronic acid production rate/cell survival rate) when the control and each sample were added. FIG. 2 shows the collagen production rate, cell survival rate, and collagen production rate per cell (collagen production rate/cell survival rate) when the control and each sample were added. The numerical values on the horizontal axis in FIGS. 1 and 2 are the final concentrations of each sample.
As shown in FIG. 1, regarding the hyaluronic acid production rate per cell, a significant increase was observed in MeOH extract, hexane soluble material, ethyl acetate soluble material, and aqueous layer residue compared to the control. As shown in FIG. 2, a significant increase in the collagen production rate per cell was observed in MeOH extract, hexane soluble material, ethyl acetate soluble material, and aqueous layer residue compared to the control.

Example 2. Evaluation of isolated components from bamboo stem epidermis extract 2-1. Production of isolated components In the same manner as in Example 1, 1-1, a methanol extract of bamboo stem epidermis was subjected to liquid-liquid distribution to obtain a hexane soluble material, an ethyl acetate soluble material, and an aqueous layer residue.

The obtained hexane-soluble matter was subjected to open column chromatography (normal phase column, eluent: ethyl acetate-hexane system), medium pressure preparative chromatography (MPLC), and reversed phase column (eluent: methanol-water system). ) and a normal phase column (eluent: ethyl acetate-hexane system), and the eluted components were confirmed by thin layer chromatography (TLC). Components with the same Rf value were grouped. After that, the components were isolated by reverse-phase preparative TLC (developing solvent: methanol-water system), fatty acids linolenic acid, 1,2-dilinolein, and triacontanol, plant steroids β-sitosterol and beturinic acid, and phenol. The compound tricine was obtained. Identification of these components was performed by nuclear magnetic resonance (NMR) and mass spectrometry (MS).

The obtained ethyl acetate soluble material was analyzed using open column chromatography (normal phase column, eluent: ethyl acetate-hexane system) and MPLC, and reversed phase column (eluent: methanol-water system) and normal phase column (eluent: ethyl acetate-hexane system). Fractionation was performed using a liquid (ethyl acetate-hexane system), and the eluted components were confirmed by TLC. Components with the same Rf value were grouped. Thereafter, the components were isolated by reverse phase preparative TLC (developing solvent: methanol-water system) to obtain Friedelin, a triterpene, and 4-hydroxybenzoic acid, a phenol compound. Identification of these components was performed by nuclear magnetic resonance (NMR) and mass spectrometry (MS).

The resulting aqueous layer residue was subjected to open column chromatography (normal phase column, eluent: ethyl acetate-hexane system) and MPLC using a reverse phase column (eluent: methanol-water system) and a normal phase column (eluent: The components were isolated by fine fractionation using ethyl acetate-hexane (ethyl acetate-hexane system) to obtain phenolic compounds glucopyranosyl 3,4-dimethoxycinnamate, lionyresinol glucoside, dihydroxypropiophenone glucoside, and tachyoside. Identification of these components was performed by nuclear magnetic resonance (NMR) and mass spectrometry (MS).

Tables 1 and 2 list the compound names and structures of the isolated components.

2-2. Evaluation Dimethyl sulfoxide (DMSO) or chloroform was added to each of the components obtained in 2-1. Thereafter, it was stirred and treated with ultrasonic waves to completely dissolve it. Each sample was prepared by diluting as necessary. Using the prepared samples, the functionality of each was evaluated in the same manner as in Example 1 1-2 (A2) to (A5).
The samples were added at final concentrations of 0.1 μg/mL, 1 μg/mL, and 10 μg/mL for Compound 1, Compound 2, Compound 5, Compound 6, and Compound 7. Compound 3, Compound 4, Compound 8, Compound 9, Compound 10, Compound 11, and Compound 12 were added at final concentrations of 1 μg/mL, 10 μg/mL, and 100 μg/mL. DMSO or chloroform was used as a control.

3 and 4 show the hyaluronic acid production rate, cell survival rate, and hyaluronic acid production rate per cell (hyaluronic acid production rate/cell survival rate) when the control and each sample were added. 5 and 6 show the collagen production rate, cell survival rate, and collagen production rate per cell (collagen production rate/cell survival rate) when the control and each sample were added. The numerical values on the horizontal axis in FIGS. 3 to 6 are the final concentrations of each sample.

As shown in FIGS. 3 to 6, it can be seen that many of the components contained in the bamboo stem epidermis extract have a higher cell survival rate than the control and have a cell activation function.

In addition, Compound 6 was found to increase the hyaluronic acid production rate per cell and the collagen production rate per sample. Compound 10 was found to have an increased collagen production rate per sample. From this, it is considered that a composition containing tricin is useful as a hyaluronic acid production promoter. Furthermore, a composition containing tricin and/or linoresinol glucoside is considered to be useful as a collagen production promoter.

Example 3. Evaluation of melanin production suppressing function The melanin production suppressing function of beturinic acid, Friedelin, Tricine, linolesinol glucoside, dihydroxypropiophenone glucoside, 1,2-dilinolein, triacontanol, linolenic acid, and tachioside was evaluated using the following (B1). It was evaluated according to the procedure of ~(B5). Each component used in the evaluation was isolated from a bamboo stem epidermis extract.

(B1) Sample preparation Add dimethyl sulfoxide (DMSO) or chloroform to beturinic acid, Friedelin, tricine, linoresinol glucoside, dihydroxypropiophenone glucoside, 1,2-dilinolein, triacontanol, linolenic acid, and tachyoside. added. Thereafter, it was stirred and treated with ultrasonic waves to completely dissolve it. Each sample was prepared by diluting as necessary.

(B2) Cell culture B16 melanoma cells were precultured in a 75 cm ² flask using D-MEM (low glucose) (containing 1% penicillin-streptomycin and 10% fetal bovine serum (FBS)) until confluent. Thereafter, 1 mL of the cells were seeded in a 24-well plate at a concentration of 0.5×10 ⁵ cells/ml, and cultured overnight in a CO ₂ incubator (37° C., 5% CO ₂ ).

(B3) Sample addition After overnight culture, the medium was aspirated and each sample prepared in (B1) was added to a 24-well plate in Dulbecco's Modified Eagle Medium (D-MEM) (low glucose), which is a serum-free medium. 500 μL of a diluted solution diluted 200 times with 1% penicillin-streptomycin) was added and cultured in a CO ₂ incubator for 24 hours.
Note that each sample was added at a final concentration shown on the horizontal axis in FIG. DMSO or chloroform was used as a control.

(B4) Cell viability measurement (Cell Counting kit-8)
Forty-eight hours after sample addition, the sample-containing medium was removed, and 500 μL of D-MEM (low glucose) (containing 1% penicillin-streptomycin and 10% fetal bovine serum (FBS)) was added to each well of the 24-well plate. Next, 30 μL of Cell Counting kit-8 solution was added to each well, and the cells were incubated for 1 hour in a CO ₂ incubator. 150 μL of the medium was taken and dispensed into a 96-well plate, and the absorbance at 450 nm was measured using a microplate reader to calculate the cell survival rate.
The cell survival rate upon addition of each sample was calculated as a relative ratio of the control, with the cell survival rate of the control being 100%.

(B5) Measurement of melanin production After measuring the cell survival rate, the medium in each 24-well plate was removed using an aspirator, the cells were detached with a 0.25% trypsin-EDTA solution, and the cells were collected from each dish. After centrifugation and washing, the cells were transferred to a 1.5 mL tube, centrifuged again, the supernatant was removed, and 200 μL of 1M NaOH solution was added. After vortexing, the reaction was carried out at 100° C. for 10 minutes. After confirming that the melanin was completely dissolved, the reaction solution was centrifuged and 150 μL of each supernatant was transferred to a 96-well plate to avoid removing precipitates, and the absorbance at 405 nm was measured using a plate reader.
The melanin production rate upon addition of each sample was calculated as a relative ratio of the control, with the melanin production rate of the control being 100%.

FIG. 7 shows a plot of the melanin production rate per cell (melanin production rate/cell survival rate) in each sample. As shown in Figure 7, the system containing beturinic acid, Friedelin, tricine, linoresinol glucoside, dihydroxypropiophenone glucoside, 1,2-dilinolein, triacontanol, linolenic acid, and tachyoside was more effective than the control. , the melanin production rate per cell was low.

(Formulation example for cosmetics)
Tables 3 and 4 show examples of formulations of cosmetics containing ethanol extracts obtained by extracting active ingredients from bamboo trunk epidermis using ethanol as an extraction solvent and drying them.

(Evaluation of melanin production promoting function)
The melanin production rate per cell of β-sitosterol, 4-hydroxybenzoic acid, and glucopyranosyl dimethoxycinnamate was evaluated. Samples of β-sitosterol, 4-hydroxybenzoic acid, and glucopyranosyl dimethoxycinnamate were prepared using dimethyl sulfoxide (DMSO) or chloroform and evaluated in the same manner as in steps (B2) to (B5) of Example 3. . The results are shown in FIG. Note that each sample was added at a final concentration shown on the horizontal axis in FIG.
As shown in FIG. 8, compared to the control, the melanin production rate per cell was higher in the system to which β-sitosterol, 4-hydroxybenzoic acid, and glucopyranosi dimethoxycinnamate were added.

The hyaluronic acid production promoter of the present invention, the collagen production promoter of the present invention, and the like can be used in various applications such as cosmetics, food and drink products, and are industrially useful.

Claims (4)

A hyaluronic acid production promoter containing bamboo stem epidermis extract as an active ingredient,
The bamboo stem epidermis extract is
The methanol extract obtained by extracting bamboo trunk epidermis with methanol or a mixed solvent of methanol and water is subjected to liquid-liquid partitioning with a mixed solvent of hexane and water, and then the resulting aqueous layer is mixed with ethyl acetate and water. A hyaluronic acid production promoter which is an ethyl acetate soluble material obtained by liquid-liquid distribution with a mixed solvent of and collecting the ethyl acetate layer . A collagen production promoter containing bamboo stem epidermis extract as an active ingredient,
The bamboo stem epidermis extract is
The methanol extract obtained by extracting bamboo trunk epidermis with methanol or a mixed solvent of methanol and water is subjected to liquid-liquid partitioning with a mixed solvent of hexane and water, and then the resulting aqueous layer is mixed with ethyl acetate and water. A collagen production promoter which is an ethyl acetate soluble material obtained by liquid-liquid distribution with a mixed solvent of and collecting the ethyl acetate layer . A moisturizing agent containing the hyaluronic acid production promoter according to claim 1 . An anti-wrinkle agent containing the collagen production promoter according to claim 2 .