MSR gene expression enhancer

Abstract

To provide a technique for reducing sulfoxidized methionine in proteins in epidermis.SOLUTION: One or more selected from the group consisting of loofah extract, cornflower flower extract, ginkgo biloba leaf extract, raspberry juice, Salicornia herbacea extract, and Fragaria chiloensis juice is used as an active ingredient that enhances the expression of the methionine sulfoxide reductase (MSR) gene. The extract can promote, in epidermis, the reduction of methionine sulfoxidized in keratinocyte growth factor (KGF), which is a factor for promoting epidermal turnover, and in keratin of keratinocytes, and therefore can be suitably blended into a topical skin composition for improving epidermal turnover and / or skin flexibility.SELECTED DRAWING: Figure 1

Description

Technical Field

[0001] The present invention relates to a methionine sulfoxide reductase (hereinafter referred to as MSR) gene expression enhancer, and further to a topical skin composition for improving epidermal turnover and / or skin flexibility containing the same.

Background Art

[0002] Skin is exposed to oxidative stress caused by various factors such as ultraviolet rays, air pollutants, and stress in daily life. Oxidation phenomena in the skin include various ones such as glycation, carbonylation, nitration, and sulfoxidation, which lead to deterioration of various skin conditions such as spots, wrinkles, dullness, and lack of transparency, which is not preferable. Among these, focusing on the sulfoxidation of methionine, when methionine of keratinocyte growth factor (KGF), which is one of the factors promoting epidermal turnover in the skin, is sulfoxidized, the turnover is disrupted, and skin dryness, dullness, reduced transparency, and roughness occur. It is also known that when methionine of keratin in epidermal cells is sulfoxidized, skin flexibility decreases. Therefore, means for counteracting the sulfoxidation of methionine in epidermal cells are desired.

[0003] It is known that sulfoxidized methionine is reduced by the reductase methionine sulfoxide reductase (MSR). However, MSR decreases or its activity decreases with ultraviolet rays. Due to such background, it is considered that enhancing the expression of MSR or enhancing its activity is useful for improving skin conditions. So far, it has been disclosed that lipochroman-6 is used as an active ingredient to restore the expression of the gene encoding MSR and the functionality of the protein (Patent Document 1).

Prior Art Documents

Patent Documents

[0004]

Patent Document 1

Summary of the Invention

Problems to be Solved by the Invention

[0005] An object of the present invention is to provide a technique for reducing sulfoxidized methionine of a protein in the epidermis. Another object is to improve the skin condition by reducing sulfoxidized methionine.

Means for Solving the Problems

[0006] As a result of intensive studies to solve the above problems, the present inventors have found that six types of plant extracts have an action of enhancing the expression of the MSR gene and can solve the above problems, and have completed the present invention.

[0007] That is, the present invention is as follows. [1] An MSR gene expression enhancer comprising one or more selected from the group consisting of loofah extract, calendula flower extract, ginkgo leaf extract, strawberry juice, acanthopanax extract, and Fragaria × ananassa juice. [4] The MSR gene expression enhancer according to [1], which enhances the expression of the MSR gene in keratinocytes. [3] A topical skin composition for improving epidermal turnover and / or skin flexibility, containing the MSR gene expression enhancer according to [1] or [2].

Effects of the Invention

[0008] The present invention provides a component that enhances the expression of the MSR gene in stratum corneum cells. Since MSR can promote the reduction of sulfoxidized methionine of keratin in KGF, which is a factor promoting epidermal turnover, in the epidermis, a composition containing such a component can be used as a composition for improving epidermal turnover and / or skin flexibility.

Brief Description of the Drawings

[0009]

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Mode for Carrying Out the Invention

[0010] <1> MSR gene expression enhancer The agent of the present invention contains, as an active ingredient, one or more selected from the group consisting of loofah extract, safflower flower extract, ginkgo leaf extract, strawberry juice, acanthopanax extract, and Fragaria × ananassa juice. Loofah extract is an extract of Luffa cylindrica / Luffa aegyptiaca of the genus Luffa in the Cucurbitaceae family. Cornflower flower extract is an extract of the flower part of Centaurea Cyanus of the genus Centaurea in the Asteraceae family. Ginkgo leaf extract is an extract of the leaf part of Ginkgo biloba of the genus Ginkgo in the Ginkgoaceae family. product. Strawberry juice is an extract of the fruit and / or juice of Rubus Idaeus of the genus Rubus in the Rosaceae family. Salicornia extract is an extract of Salicornia herbacea / Salicornia europaea of the genus Salicornia in the Amaranthaceae family. Fragaria chiloensis juice is an extract of the fruit and / or juice of Fragaria chiloensis of the genus Fragaria in the Rosaceae family.

[0011] The above-mentioned extract means not only the extract itself, but also the general term for fractions of the extract, purified fractions, extracts or fractions, and solvent-removed products of purified products.

[0012] Also, as the extract, any extract that is usually used in skin external preparations or oral intake compositions such as cosmetics and pharmaceuticals may be used, and an extract obtained from a plant body by a conventional method can be used. As the extraction source, for example, in addition to using the whole plant, parts such as the plant body, above-ground part, rhizome part, trunk part, leaf part, stem part, flower, flower bud, fruit, and fruit juice are used for the extraction operation. However, for cornflower flower extract, ginkgo leaf extract, strawberry juice, and Fragaria chiloensis juice, the extraction operation is carried out using a predetermined part.

[0013] As the extraction solvent, one or more selected from polar solvents such as water, alcohols such as ethanol, isopropyl alcohol, and butanol, polyhydric alcohols such as 1,3 - butanediol and polypropylene glycol, ketones such as acetone and methyl ethyl ketone, and ethers such as diethyl ether and tetrahydrofuran are preferably used.

[0014] As a specific extraction method, for example, 1 to 30 parts by mass of a solvent is added to 1 part by mass of a part used for extraction of a plant body or the like or its dried product, immersed at room temperature for several days or at a temperature near the boiling point for several hours, cooled to room temperature, and then, if desired, insoluble matters and / or the solvent are removed, and fractionation purification is carried out by column chromatography or the like.

[0015] The total content of one or more plant extracts selected from the group consisting of Momordica charantia extract, Hieracium umbellatum flower extract, Ginkgo biloba leaf extract, strawberry juice, Actinidia polygama extract, and Fragaria × ananassa juice in the agent of the present invention is preferably 0.00001 to 0.09% by mass in terms of solid content based on the whole composition, more preferably 0.00005 to 0.03% by mass, and still more preferably 0.0001 to 0.003% by mass. By setting the content within the above range, a desired effect can be easily obtained, and the degree of freedom in formulation design can be ensured. It should be noted that the above content can be appropriately adjusted according to the administration route described later and the form of the composition to be contained.

[0016] Momordica charantia extract, Hieracium umbellatum flower extract, Ginkgo biloba leaf extract, strawberry juice, Actinidia polygama extract, and Fragaria × ananassa juice all have the effect of enhancing the expression of the MSR gene alone. That is, each of these extracts is an active ingredient of an MSR gene expression enhancer. The MSR gene in this specification includes MsrA and MsrB.

[0017] In addition, the MSR gene expression enhancing effect of any substance can be confirmed by the fact that the MSR gene expression level in cells to which the target substance is added is higher than that in cells to which the substance is not added, usually 110% or more, preferably 120% or more, more preferably 130% or more. The expression level of the MSR gene can be determined, for example, by performing PCR using a DNA fragment having a sequence that specifically binds to the sequence of the MSR gene as a primer and measuring the amount of mRNA. Further, for example, the intracellular abundance of the protein encoded by the MSR gene may be quantitatively measured by a conventional method and used as the expression level of the MSR gene.

[0018] When methionine of KGF, which is one of the factors promoting epidermal turnover in the skin, is sulfoxidized, the function of KGF is impaired, resulting in disrupted turnover, dryness, dullness, reduced transparency, and a rough feeling of the skin. By enhancing the expression level of MSR, the sulfoxidized methionine of KGF can be reduced, KGF can be normally repaired, and epidermal turnover can be improved. In addition, it is known that when methionine of keratin is sulfoxidized in epidermal cells, the flexibility of the skin decreases. By enhancing the expression level of MSR, the sulf oxidized methionine of keratin can be reduced, and the flexibility of the skin can be improved. Conventionally, when a topical skin preparation such as a cosmetic imparts flexibility to the skin, it is by imparting water retention ability or moisturizing the epidermis. The skin flexibility in the present invention is distinguished from the conventional ones in that it is obtained by changing the properties of the protein itself of the skin (epidermis).

[0019] The administration route of the agent of the present invention is not particularly limited, such as transdermal, oral, nasal, intravenous injection, etc., but transdermal administration is preferred. Here, "administration" may be replaced with "ingestion". Although the dosage of the agent and composition of the present invention is not particularly limited, considering the desired effects and safety, as the total amount of one or more selected from loofah extract, safflower flower extract, ginkgo leaf extract, strawberry fruit juice, houttuynia cordata extract, and Fragaria × ananassa juice, it is preferably ingested in one or several divided doses at 0.3 to 300 μg / day in terms of solid content. In addition to single-dose ingestion, it is preferably ingested continuously or intermittently for several weeks to several months.

[0020] The form of the topical skin composition of the present invention is not particularly limited as long as it is topically applicable to the skin, but preferably includes cosmetics (including quasi-drugs), pharmaceuticals, etc., and more preferably cosmetics. The site for applying the topical skin composition of the present invention is not particularly limited, but usually includes the face, limbs, neck, and décolletage.

[0021] Examples of the dosage form of the topical skin composition include, but are not particularly limited to, lotion dosage form, emulsifier dosage forms such as emulsions and creams, oil dosage form, gel dosage form, pack, cleansing agent, etc. Also, as the form of the topical skin composition, either a leave-on type or a leave-off type may be used. Since MSR decreases or its activity decreases with ultraviolet rays, the form of the topical skin composition applied to the skin during the day, such as sunscreen cosmetics, is particularly preferred.

[0022] In the production of the topical skin composition of the present invention, components usually used in the formulation of cosmetics, quasi-drugs, pharmaceuticals, etc. can be arbitrarily blended. Examples of such optional components include hydrocarbons such as squalane, petrolatum, and microcrystalline wax; esters such as jojoba oil, carnauba wax, and octyldodecyl oleate; triglycerides such as olive oil, beef tallow, and coconut oil; fatty acids such as stearic acid, oleic acid, and retinoic acid; higher alcohols such as oleyl alcohol, stearyl alcohol, and octyldodecanol; anionic surfactants such as sulfosuccinate esters and sodium polyoxyethylene alkyl sulfate; amphoteric surfactants such as alkyl betaine salts; cationic surfactants such as dialkyl ammonium salts; nonionic surfactants such as sorbitan fatty acid esters, fatty acid monoglycerides, polyoxyethylene adducts thereof, polyoxyethylene alkyl ethers, and polyoxyethylene fatty acid esters; polyhydric alcohols such as polyethylene glycol, glycerin, and 1,3 - butanediol; thickening / gelling agents, antioxidants, ultraviolet absorbers, colorants, preservatives, powders, etc. can be arbitrarily blended.

Examples

[0023] Hereinafter, the present invention will be described in more detail with reference to examples. However, the present invention is not limited to the following examples as long as the gist thereof is not exceeded.

[0024] <Reference Example 1> Various extracts were prepared according to the following procedure. Loofah extract: The plant body of Luffa of the genus Luffa in the Cucurbitaceae family was pulverized, 10 times the amount of a 30% 1,3 - butylene glycol aqueous solution was added, and the reflux - extracted liquid was powdered by freeze - drying. Sunflower flower extract: The flower part of Helianthus of the genus Helianthus in the Asteraceae family was pulverized, 10 times the amount of a 45% 1,3 - butylene glycol aqueous solution was added, and the reflux - extracted liquid was freeze - dried to be powdered. Ginkgo leaf extract: The leaf part of Ginkgo of the genus Ginkgo in the Ginkgoaceae family was pulverized, 10 times the amount of a 45% 1,3 - butylene glycol aqueous solution was added, and the reflux - extracted liquid was powdered by freeze - drying. to be powdered. Strawberry juice: The fruits of Fragaria × ananassa Duch. (Rosaceae, Fragaria) were pressed, 10 times the amount of a 60% 1,3-butylene glycol aqueous solution was added, and the solution extracted by reflux was powdered by freeze-drying. Aeschynomene indica extract: The plant body of Aeschynomene indica L. (Euphorbiaceae, Aeschynomene) was pulverized, 10 times the amount of a 40% 1,3-butylene glycol aqueous solution was added, and the solution extracted by reflux was powdered by freeze-drying. Fragaria × ananassa Duch. cv. Toyonoka juice: The fruits of Fragaria × ananassa Duch. cv. Toyonoka (Rosaceae, Fragaria) were pressed, 10 times the amount of a 40% 1,3-butylene glycol aqueous solution was added, and the solution extracted by reflux was powdered by freeze-drying.

[0025] <Test Example 1> Examination of the effect of the test extract on the MSR gene expression level in epidermal keratinocytes Normal human epidermal keratinocytes (NHEK: Normal Human Epidermal Keratinocytes, neonatal, male, Caucasian, KURABO, #06445) stored at -150 °C were revived and cultured for 5 days in an environment of 37 °C and 5% CO2. The NHEK were collected and diluted using NHEK medium HUMedia-KG2 (KURABO Industries Ltd.) to a concentration of 4.0 × 10 4 cells / mL. Each medium was seeded at 1 mL / well in a 24-well plate (4.0 × 10 4The cells / wells were cultured overnight in an environment of 37°C and 5% CO2. After culturing, the medium was removed, and the medium for NHEK with each test extract added to reach a predetermined solid content concentration, or the medium for NHEK containing 1,3-BG instead of the test extract was used for replacement, and the cells were cultured for an additional 24 hours or 48 hours. After culturing, the cells were washed with PBS(-) and recovered, and RNA was extracted using QIAshredder and RNeasy Mini Kit (manufactured by QIAGEN) with QIAcube (manufactured by QIAGEN). The extracted RNA was reverse-transcribed into cDNA using SuperScript VILO cDNA Synthesis Kit (manufactured by Invitrigen), and RT-qPCR was performed using SYBR (manufactured by QIAGEN) with QuantStudio 7 (manufactured by Applied Biosystems) to analyze the expression levels of each MSR gene. The β-Actin gene was used as the endogenous control. As the test extracts, those prepared by the method of Reference Example 1 were all used.

[0026] In FIGS. 1 to 6, the relative values of the mRNA expression levels of each MSR gene in epidermal keratinocytes to which each extract was added, with the mRNA expression level of each gene in epidermal keratinocytes of the solvent control (control) set as 1, are shown. In epidermal keratinocytes to which any of the extracts was added, it was confirmed that the mRNA expression levels of each MSR gene significantly increased compared to the control. From this result, it was shown that each of these extracts has an action of enhancing MSR gene expression.

Industrial Applicability

[0027] According to the present invention, a component that enhances the expression of the MSR gene in epidermal keratinocytes is provided. Further, by incorporating such a component into a composition, it can be made into a topical skin composition for improving epidermal turnover and / or skin flexibility. These inventions contribute to the development in the beauty industry and meet the needs of consumers, and are very useful industrially.

Claims

Claim 1 A methionine sulfoxide reductase (MSR) gene expression enhancer comprising one or more selected from the group consisting of loofah extract, calendula flower extract, ginkgo leaf extract, strawberry juice, acanthopanax senticosus extract, and fragaria chiloensis juice. Claim 2 The MSR gene expression enhancer according to claim 1, which enhances the expression of the MSR gene in keratinocytes. Claim 3 A topical skin composition for improving epidermal turnover and / or skin flexibility, containing the MSR gene expression enhancer according to claim 1 or 2.

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