JP7565717B2 - Composition - Google Patents

Description

The present invention relates to a composition, particularly to a composition for inhibiting melanin production.

Sunburn on the skin and the subsequent pigmentation, age spots, melasma, and senile lentigo are conditions in which melanin production is significantly increased due to activation of melanocytes (pigment cells) in the skin. The occurrence and worsening of these skin pigmentation-related problems hinders the beauty of the skin, so there has been active research into ingredients that have the effect of preventing or improving them, and active ingredients based on various mechanisms of action have been created.

Patent Document 1 discloses that there are factors that control multiple melanocyte activation factors, and further discloses that by using the activity of the control factors as an indicator, it is possible to more efficiently screen for materials that may have a stronger effect in inhibiting melanocyte activation.

JP 2019-004746 A

Although conventional pigmentation inhibitors have a certain degree of inhibitory effect, it is difficult to say that they have achieved a fully satisfactory effect. In addition, in preventing sunburn and pigmentation, other than the approach of using sunscreens containing ultraviolet protection agents, a new approach to creating skin that is less susceptible to sunburn and pigmentation that has excellent effects has not yet been found.
In view of the above circumstances, an objective of the present invention is to provide a composition that can prevent sunburn and pigmentation by inhibiting melanin production and making the skin less susceptible to these disorders.

As a result of intensive research conducted by the inventors to solve the above problems, they discovered that by combining specific plant extracts, they exert an excellent melanin production inhibitory effect that is not observed when the extracts are used alone, and thus completed the present invention.

That is, the present invention is as follows.
[1] A composition containing Chorei extract, Calendula officinalis extract and/or hop extract.
[2] The composition described in [1], which is for inhibiting melanin production.
[3] The composition described in [1] or [2], which is used to prevent sunburn and/or pigmentation.
[4] The composition according to any one of [1] to [3], wherein the composition is a food or beverage or an external preparation for skin.

The present invention provides a composition suitable for use in inhibiting melanin production. The composition can be in the form of an external skin preparation or a food or drink, and can be easily and continuously incorporated into daily life to create skin that is less prone to sunburn and pigmentation.

Graph showing the relative expression level of EHF gene mRNA in keratinocytes to which the test extract was added (Dunnett's test, **: p<0.01). Graph showing the relative expression level of EHF gene mRNA in keratinocytes to which the test extract was added (Dunnett's test, *: p<0.05). A graph showing the relative expression level of EHF gene mRNA in keratinocytes to which the test extract was added. A graph showing the relative expression level of EHF gene mRNA in keratinocytes to which the test extract was added.

The composition of the present invention contains Chorei extract, Calendula officinalis extract and/or hop extract.
Polyporus extract is an extract of Polyporus umbellata, the sclerotium of Polyporus umbellata Fries, a fungus in the Polyporaceae family of the Aphyllostearyl orders.
Calendula Officinalis Flower Extract is an extract of Calendula Officinalis Flower, which belongs to the Asteraceae family of the Asterales order.
Hop extract is an extract of the genus Humulus (Hops), which belongs to the family Cannabaceae, order Rosales.

The above extract refers collectively to not only the extract itself, but also fractions of the extract, purified fractions, extracts or fractions, and products from which the solvent has been removed from the purified product.

The extract may be any extract that is normally used in skin external preparations such as cosmetics and pharmaceuticals, or in compositions for oral ingestion, and may be extracted from a plant body by a conventional method.
For example, the whole plant may be used, or parts of the plant, such as the plant body, above-ground parts, rhizomes, trunks, leaves, stems, flowers, flower buds, and fruits, may be used for the extraction. In the case of Trichoderma edodes, the sclerotia are used for the extraction. Usually, these are crushed or cut into small pieces in advance to improve the extraction efficiency.
As the extraction solvent, one or more selected from polar solvents such as water, alcohols such as ethanol, isopropyl alcohol, butanol, polyhydric alcohols such as 1,3-butanediol, polypropylene glycol, ketones such as acetone, methyl ethyl ketone, ethers such as diethyl ether, tetrahydrofuran, etc. are preferably used.
A specific example of the extraction method is a method in which 1 to 30 parts by mass of a solvent is added to 1 part by mass of a part of a plant body or the like used for extraction or a dried product thereof, followed by immersion for several days at room temperature or several hours at a temperature near the boiling point, and then cooling to room temperature, after which insoluble matter and/or the solvent are removed as desired, and fractionation and purification are performed by column chromatography or the like.

The content of the Chorei extract in the composition of the present invention is preferably 0.0001 to 0.09 mass %, more preferably 0.0005 to 0.03 mass %, and even more preferably 0.001 to 0.003 mass %, calculated as solid matter based on the total composition.
The content of the calendula extract in the composition of the present invention is preferably 0.0001 to 0.1 mass %, more preferably 0.0006 to 0.04 mass %, and even more preferably 0.001 to 0.004 mass %, calculated as solid matter based on the total mass of the composition.
The content of the hop extract in the composition of the present invention is preferably 0.0001 to 0.05 mass %, more preferably 0.0002 to 0.02 mass %, and even more preferably 0.0005 to 0.0015 mass %, calculated as solid matter based on the total mass of the composition.

The mass ratio (in terms of solid matter) of the contents of the Calendula officinalis extract and the Calendula officinalis extract in the composition of the present invention is preferably 5:17 to 15:2, more preferably 10:17 to 5:1, and
A ratio of 5:17 to 5:2 is even more preferred.
The mass ratio (in terms of solid matter) of the contents of the corn syrup extract and the hop extract in the composition of the present invention is preferably from 2:9 to 18:1, more preferably from 1:3 to 12:1, and even more preferably from 2:3 to 6:1.

By setting the content or the mass ratio within the above range, the desired effects can be easily obtained and the freedom in formulation design can be ensured.
The above content can be appropriately adjusted according to the administration route, such as transdermal administration or oral administration, as described below.

As shown in the Examples below, the combination of the Chorei extract and the Calendula officinalis extract, and the combination of the Chorei extract and the hop extract enhance the expression of the EHF gene (Gene ID: 26298) in keratinocytes.
The expression of the EHF gene is not enhanced.
The EHF gene is a factor that suppresses multiple melanocyte activating factors. Enhanced expression of EHF in keratinocytes suppresses melanocyte activating factors such as endothelin 1, IL-1α, GRO, NRG1, SCF, TNF, ADM, IL-6, and GM-CSF. It is known that factors that activate melanocytes activate melanocytes by enhancing their expression, and promote melanin production by tyrosinase.
Therefore, the composition of the present invention, which enhances the expression of the EHF gene, has the effect of suppressing melanin production through the suppression of melanocyte activating factor.

The composition of the present invention can prevent sunburn and pigmentation by inhibiting melanin production. Sunburn is a condition in which the skin becomes red and inflamed after exposure to ultraviolet rays.
and the subsequent sun tanning, which is the deposition of melanin pigment in the epidermis. The pigmentation that can be prevented by the composition of the present invention is not limited to that caused by exposure to ultraviolet rays, but may be pigmentation caused by excessive production of melanin due to various causes, such as skin inflammation such as acne, aging, melasma, drugs, and various diseases.
Furthermore, the composition of the present invention can prevent pigmentation, thereby resulting in a whitening effect on the skin.

The route of administration of the composition of the present invention is not particularly limited and may be transdermal, oral, nasal, intravenous, etc., but it is preferably taken transdermally or orally.
The dosage is not particularly limited, but taking into consideration the desired effect and safety, it is preferable to take 0.3 to 300 μg/day in solid terms as the total amount of the Chorei extract, Calendula officinalis extract, and/or hop extract in one or several divided doses. In addition to taking it once, it is also preferable to take it continuously or intermittently for several weeks to several months.

When the composition of the present invention is administered transdermally, it is preferably used as a composition for external application to the skin. The form of the composition for external application to the skin is not particularly limited as long as it is applied externally to the skin, but preferred examples include cosmetics (including quasi-drugs) and medicines, and more preferred are cosmetics.
The areas to which the topical skin preparation is applied are not particularly limited, but are usually the face, limbs, neck, and décolleté.

Examples of dosage forms of topical skin compositions include, but are not limited to, lotion dosage forms, emulsion dosage forms such as lotions and creams, oil dosage forms, gel dosage forms, packs, cleansers, etc.

When the composition of the present invention is in the form of a composition for external use on the skin, ingredients that are usually used in the formulation of cosmetics, quasi-drugs, pharmaceuticals and the like can be optionally blended in the preparation thereof.
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 sulfates; 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 and gelling agents; antioxidants; ultraviolet absorbers; coloring agents; preservatives; and powders.

When the composition of the present invention is in the form of a food or drink, ingredients that are usually used in the production of food or drink can be optionally blended therein.
Examples of such optional ingredients include proteins, carbohydrates, fats, nutrients, seasonings, and flavorings. Examples of carbohydrates include monosaccharides, such as glucose, fructose, and the like; disaccharides, such as maltose, sucrose, and oligosaccharides; and polysaccharides, such as dextrin, cyclodextrin, and other common sugars, and sugar alcohols, such as xylitol, sorbitol, and erythritol. Examples of flavorings include natural flavorings (thaumatin, stevia extract, and the like) and synthetic flavorings (saccharin, aspartame, and the like). In addition, additives used in the above-mentioned pharmaceutical compositions that are also added to normal foods may be used in the same manner.

The form of the food or drink may be liquid, paste, solid, powder, granules, etc. Tablets, liquid food, feed, etc. are also included in the form of food or drink.

In addition, it may be contained in other general foods and beverages, for example, wheat flour products such as bread, macaroni, spaghetti, noodles, cake mix, fried chicken flour, and breadcrumbs; instant foods such as instant noodles, cup noodles, retort/prepared foods, pre-cooked foods, microwave foods, instant soups/stews, instant miso soup/clean soups, canned soups, and freeze-dried foods; agricultural processed products such as canned agricultural products, canned fruit, jams/marmalades, pickles, boiled beans, dried agricultural products, and cereals (processed grain products); marine processed products such as canned seafood, fish ham/sausages, marine paste products, marine delicacies, and tsukudani (food stew); livestock processed products such as canned livestock products/pastes, and livestock ham/sausages; milk and dairy products such as processed milk, milk drinks, yogurt, lactic acid bacteria drinks, cheese, ice cream, modified milk powder, cream, and other dairy products; butter, Fats and oils such as margarines and vegetable oils; basic seasonings such as soy sauce, miso, sauces, processed tomato seasonings, mirin, and vinegars; complex seasonings and foods such as cooking mixes, curry bases, sauces, dressings, noodle soups, spices, and other complex seasonings; frozen foods such as frozen ingredients, semi-cooked frozen foods, and cooked frozen foods; confectioneries such as caramel, candy, chewing gum, chocolate, cookies, biscuits, cakes, pies, snacks, crackers, Japanese sweets, rice confectioneries, bean confectioneries, and dessert confectioneries; beverages such as carbonated drinks, natural fruit juices, fruit juice drinks, soft drinks containing fruit juice, fruit pulp drinks, fruit drinks containing fruit grains, vegetable drinks, soy milk, soy milk drinks, coffee drinks, tea drinks, powdered drinks, concentrated drinks, sports drinks, nutritional drinks, and alcoholic drinks; and foods other than these, the composition of the present invention may be added.

Examples of food compositions containing the composition of the present invention include ordinary foods, beverages, functional foods such as functional foods and foods for specified health uses, and supplements, with functional foods being particularly preferred.
When the food composition according to the present invention is used for preventing sunburn or pigmentation, the usefulness or functionality of the composition may be indicated when the product is manufactured.
Such "display" acts include all acts for informing consumers of the above-mentioned uses, and any expression that can recall or infer uses such as "for those who wish to suppress melanin production,""for skin that is less susceptible to sunburn," or "for preventing pigmentation" falls under the "display" acts of the present invention, regardless of the purpose of the display, the content of the display, the object or medium on which it is displayed, etc.
In addition, it is preferable that the "labeling" is done in an expression that allows consumers to directly recognize the above-mentioned use. Specifically, examples include the act of transferring, delivering, displaying for transfer or delivery, or importing a food or drink product or a product package, container, etc. on which the above-mentioned use is written, and the act of displaying or distributing the above-mentioned use in advertising materials at the sales site such as advertisements, price lists, catalogs, pamphlets, POP, etc., or in transaction documents, or the act of providing the above-mentioned use in information containing the above-mentioned use by electromagnetic (Internet, etc.) methods. Note that, when the food composition of the present invention is approved under various systems set by the government such as health functional foods and is implemented under such approval, it is preferable to label it in a manner based on the approval.

The present invention will be described in more detail below with reference to examples, but the present invention is not limited to the following examples as long as the gist of the invention is not exceeded.

<Reference Example>
The various extracts were prepared according to the following procedure.
Chorei extract: 500 g of dried sclerotium of Choreimaitake mushroom of the Polyporaceae family of the Aphyllostearylles orders was added to 5 L of 50% aqueous ethanol solution, and the extract was left at room temperature for 1 week, from which insoluble matter was removed by filtration.
Calendula officinalis extract: 100 g of dried flower heads of Calendula officinalis (Asteraceae family) was added to 1 L of water, and the mixture was extracted at 90°C for 2 hours with gentle stirring, after which insoluble matter was removed by filtration. The filtrate was then dried under reduced pressure, and the resulting powder was redissolved in a 50% aqueous solution of 1,3-butylene glycol.
Hop extract: 100 g of ground dried female inflorescences of hops of the genus Lupinus in the family Cannabaceae, order Rosales, was added to 1 L of water, and extraction was carried out for 2 hours at 80°C using a reflux extractor. The extract obtained after filtration was dried under reduced pressure and redissolved in a 50% aqueous solution of 1,3-butylene glycol.
Scutellaria root extract: After removing the periderm from the root of Scutellaria baicalensis (Lamiaceae family) and drying, 0.5 L of warm water was added to 100 g of the crushed powder and heated for 3 hours, and 1 L of ethanol was added and extracted at room temperature overnight, after which the precipitate was removed by filtration. The filtrate was then dried under reduced pressure, and the dried residue was redissolved in a 50% aqueous solution of 1,3-butylene glycol.
Althaea extract: 100 g of ground dried roots of Marshmallow (Mallow family, Malvaceae) was added to 1 L of water and extracted at room temperature for 3 days with gentle stirring, after which the precipitate was removed by filtration. The filtrate was then dried under reduced pressure, and the dry residue was redissolved in 50% 1,3-butylene glycol aqueous solution.

Test Example: Examination of the effect of various extracts on the amount of mRNA expression of the EHF gene The amount of mRNA expression of the EHF gene in keratinocytes to which a test extract had been added was measured according to the following procedure.
Normal human keratinocytes (NHEK, Kurabo Industries, Lot. 2568) were seeded in a 24-well plate (7.5×10 ⁴ cells/well) and cultured in HuMedia KG2 (Kurabo Industries, Ltd.) for 24 hours at 37° C. under 5% CO ₂ conditions.
After the incubation, the medium was replaced with an evaluation medium (HuMedia KG2 from which HC, hEGF, BPE, antibiotics, and phenol red had been removed) (900 μL/well).
Furthermore, the test extract adjusted to a solid content concentration of 1.0% by mass was added to each well to the concentration (volume %) shown in Table 1, and the cells were cultured for 6 hours at 37°C under 5% _CO2 conditions.
After the culture, the cells were collected using QIAshredder (QIAGEN) and RNeasy Mini Kit (QIAGEN), and RNA was extracted using QIAcube (QIAGEN).
SuperScript VILO cDNA Synthesis Kit (Ther
The expression level of the EHF gene was analyzed by real-time PCR using cDNA synthesized using a PCR kit (manufactured by Mo Fisher SCIENTIFIC).

Figures 1 to 4 show the mRNA expression level of the EHF gene in keratinocytes as a relative value when the mRNA expression level of the EHF gene in keratinocytes of the solvent control (#1) is set to 1.

No significant changes in EHF gene expression were observed when Chorei extract, Calendula officinalis extract, or hop extract were used alone, but a significant increase in EHF gene expression was observed when Chorei extract was combined with Calendula officinalis extract, and when Chorei extract was combined with hop extract.
No enhanced expression of the EHF gene was observed in the combination of Chorei extract and Scutellaria root extract, or in the combination of Chorei extract and Althea extract.

The present invention provides a composition suitable for use in inhibiting melanin production. The composition can be in the form of an external skin preparation or a food or drink, and can be easily and continuously incorporated into daily life to create skin that is less prone to sunburn and pigmentation, making it extremely useful industrially.

Claims (4)

A composition for enhancing EHF gene expression , comprising an extract of Polygala officinalis, an extract of Calendula officinalis and/or an extract of hops,
the mass ratio of the contents of the calendula extract and the calendula extract is 5:17 to 15:2, and the mass ratio of the contents of the calendula extract and the hop extract is 2:9 to 18:1;
The composition is a food or beverage, or an external skin preparation . The composition according to claim 1, which is for inhibiting melanin production. The composition according to claim 1 or 2, which is used to prevent sunburn and/or pigmentation. The content of the choy rei extract is 0.0001 to 0.09% by mass in terms of solid matter relative to the total mass of the composition;
The composition according to any one of claims 1 to 3, wherein the content of calendula extract, when contained, is 0.0001 to 0.1 mass % in terms of solid matter relative to the entire composition, and the content of hop extract, when contained, is 0.0001 to 0.05 mass % in terms of solid matter relative to the entire composition .

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