A method for preventing or improving blemishes and screening for the same.

Abstract

To provide a new agent for preventing or improving blotches, and a screening method for skin-whitening materials for use in the agent.SOLUTION: An agent for preventing or improving blotches comprises at least one extract selected from Ocimum tenuiflorum L. extract, Tamarindus indica L. extract, Geranium robertianum extract, Ginkgo biloba extract, Equisetum arvense extract, Rosa roxburghii extract, Phellodendron amurense extract, and Paeonia suffruticosa extract as an active ingredient. There is also provided a screening method for skin-whitening materials using RGMB as an index.SELECTED DRAWING: None

Description

Technical Field

[0001] The present invention relates to a freckle prevention or improvement agent and a screening method thereof, etc.

Background Art

[0002] Skin pigmentation symptoms such as freckles greatly affect the appearance of the face, so there is high interest in their prevention and improvement. Conventionally, whitening agents with various mechanisms of action have been developed, but there is a high demand for more effective whitening agents, and new whitening materials are required. In order to develop new whitening materials, screening of whitening materials has been attempted (Patent Document 1), but further development of whitening materials is required.

[0003] On the other hand, Repulsive guidance molecule BMP co-receptor (RGM), which is a nerve cell secretion factor, is a protein known as a nerve repulsive factor and is also known to be involved in cell proliferation and differentiation. Protein expression in humans has also been confirmed, and mRNA expression in the brain and skin has been confirmed. The RGM family includes three subtypes called RGMA, RGMB, and RGMC. However, the relationship between the RGM family and freckles is not known.

Prior Art Documents

Patent Documents

[0004]

Patent Document 1

Summary of the Invention

Problems to be Solved by the Invention

[0006] The inventors of this invention conducted research in search of new blemish prevention or improvement agents and whitening materials to be used in such agents. They identified RGMB as a new causative factor of blemishes and developed a screening method for whitening materials using RGMB as an indicator. Furthermore, they discovered that the whitening materials obtained through this screening method can be used as blemish prevention or improvement agents. Based on these findings, the inventors completed the present invention. In other words, the gist of this invention is as follows:

[0007] [1] An RGMB secretion inhibitor containing one or more extracts selected from holy basil extract, Indian date extract, Geranium robertianum extract, Ginkgo biloba extract, Equisetum arvense extract, Rosa rugosa extract, Phellodendron amurense extract, and Paeonia suffruticosa extract as active ingredients. [2] A melanin production inhibitor containing the RGMB secretion inhibitor described in [1]. [3] A blemish prevention or improvement agent containing the RGMB secretion inhibitor described in [1]. A whitening composition comprising the RGMB secretion inhibitor described in [1], the melanin production inhibitor described in [2], or the spot prevention or improvement agent described in [3]. [5] The whitening composition described in [4], which is for whitening by suppressing melanin production. [6] A whitening composition according to [3] or [4], for the prevention or improvement of blemishes. [7] A whitening composition according to any one of [4] to [6], which is a cosmetic. [8] A whitening composition according to any of [4] to [6], which is a food or beverage. [9] The amount of RGMB in cells to which the test substance was added was greater than in cells to which the test substance was not added. A step of selecting a test substance that reduces the amount of RGMB compared to the amount of RGMB, A screening method for whitening materials, including [specific material].

[0008]

[10] The step of performing the screening method described in [9], and A method for designing a whitening composition, comprising the step of incorporating a substance selected by the above step. A method for producing a whitening composition, comprising the step of preparing a whitening composition containing a whitening material obtained by the screening method described in

[11] [9]. A whitening method (excluding medical procedures) comprising the step of administering a whitening composition prepared by the method for producing a whitening composition described in

[11] to a subject of interest. [Effects of the Invention]

[0009] According to the present invention, it is possible to provide a novel RGMB secretion inhibitor, a melanin production inhibitor, and a spot prevention or improvement agent, a whitening composition containing the same, and a screening method for whitening materials used in the same. [Brief explanation of the drawing]

[0010] [Figure 1] Figure 1 shows the melanin production-promoting effect of RGMB. [Figure 2] Figure 2 shows the RGMB secretion inhibitory effects of horsetail extract, geranium extract, ginkgo extract, rose extract, and Phellodendron amurense extract. [Figure 3] Figure 3 shows the effect of peony extract on suppressing RGMB secretion. [Figure 4] Figure 4 shows the effect of Indian dates and holy basil extracts on suppressing RGMB secretion. [Modes for carrying out the invention]

[0011] The present invention will be described below. <Screening Method> One aspect of the present invention relates to a screening method for skin-whitening materials (hereinafter sometimes referred to as "the screening method of the present invention"), which includes the step of selecting a test substance to which the amount of RGMB in cells to which the test substance is added is reduced compared to the amount of RGMB in cells to which the test substance is not added.

[0012] As described above, the inventors of the present invention conducted studies to find a new whitening material and identified RGMB, a nerve cell secretion factor, as a melanocyte pigment formation promoting factor. Based on such findings, a screening method for a whitening material using RGMB as an index was found. In addition, it was found that the whitening material obtained by the screening method can be used as an RGMB secretion inhibitor, a melanin production inhibitor, and a freckle prevention or improvement agent. Thus, the present invention was completed.

[0013] ≪RGMB≫ RGMB is a nerve cell secretion factor, known as a nerve repulsive factor, and is a protein known to be involved in cell growth and differentiation. However, the relationship between RGM and melanocyte pigment formation promotion and freckles has not been known conventionally and has been newly found by the inventors of the present invention. According to the present invention based on such findings, RGMB can be used as an index for a screening method of a whitening material.

[0014] The amount of RGMB in the screening method of the present invention may be, for example, the expression or secretion amount of RGMB protein, the expression amount of the gene encoding RGMB protein, or the like.

[0015] An example of the procedure in the screening method of the present invention is given below. However, it is not limited to the following content as long as it does not depart from the gist of the present invention, and can be implemented with appropriate modifications. can be

[0016] The test substance is added to nerve cells and incubated for 12 to 72 hours. Then, mRNA is extracted from the cells, and the expression level of the RGMB gene encoding the indicator RGMB is quantitatively measured by RT-PCR using primers that specifically detect the gene. The expression level of the gene is also measured in cells that were not treated with the test substance as a control. If the expression level of the gene in cells treated with the test substance is lower than the expression level in cells that were not treated with the test substance (control), the test substance is determined to be a skin-whitening material and to have a skin-whitening effect. Alternatively, the intracellular abundance or secretion amount of RGMB protein encoded by the RGMB gene in the cell supernatant can be quantitatively measured using conventional methods and determined as the amount of RGMB.

[0017] Alternatively, RGMB and the test substance are added to melanocytes and incubated for 12 to 72 hours. Afterward, the amount of melanin produced in the melanocytes is quantitatively measured. Melanin production is also measured in cells that did not receive the test substance as a control. If the amount of melanin produced in cells with the test substance is lower than the amount of melanin produced in cells without the test substance (control), the test substance is determined to be a skin-whitening material and to have a skin-whitening effect.

[0018] Here, the degree of reduction in RGMB amount or melanin production is sufficient if the amount in cells to which the test substance was added is lower than the amount in cells to which the test substance was not added. For example, 12 to 72 hours after the addition of the test substance, the amount in cells to which the test substance was added may be reduced to 90% or less, 80% or less, 70% or less, 60% or less, 50% or less, 40% or less, 30% or less, 20% or less, 10% or less, or 0% compared to the amount in cells to which the test substance was not added.

[0019] ≪Cell≫ The cells used in the screening method of the present invention are nerve cells, melanocytes (melanin cells), etc. Their origin is not particularly limited, and they may be derived from humans, non-human mammals (such as mice, humans, pigs, dogs, rabbits, etc.), and can be induced cells from somatic stem cells such as iPS cells, embryonic stem cells (ES cells), mesenchymal stem cells, hematopoietic stem cells, intestinal stem cells, established cell lines, etc. The cell culture method can be applied without particular limitation as long as the culture conditions do not interfere with the execution of the screening method of the present invention, and a conventionally known cell culture method can be used.

[0020] The test substance targeted by the screening method of the present invention is not limited in its form, origin, etc., and may be a pure substance, an extract derived from animals or plants, or a mixture thereof, etc. Extracts derived from animals or plants include not only the extracts themselves derived from animals or plants, but also fractions of the extracts, purified fractions, solvent-removed products of the extracts, fractions, or purified products. Plant-derived extracts include extracts using plants growing wild or cultivated, extracts using those sold as crude Chinese herbal medicine raw materials, commercially available extracts, etc.

[0021] <RGMB secretion inhibitor, melanin production inhibitor, and freckle prevention or improvement agent> Another aspect of the present invention relates to an RGMB secretion inhibitor (hereinafter sometimes referred to as "the RGMB secretion inhibitor of the present invention") containing as an active ingredient one or more extracts selected from holy basil extract, Indian date extract, Japanese butterbur extract, ginkgo extract, Japanese horsetail extract, Japanese rose extract, peony extract, and button extract. Note that one or a combination of two or more of these extracts can be used as an active ingredient.

[0022] Also, another aspect of the present invention relates to a melanin production inhibitor (hereinafter sometimes referred to as "the melanin production inhibitor of the present invention") containing the RGMB secretion inhibitor of the present invention. ​A further aspect of the present invention relates to a melanin production inhibitor containing one or more extracts selected from holy basil extract, Indian date extract, Geranium robertianum extract, Ginkgo biloba extract, Equisetum arvense extract, Rosa rugosa extract, Phellodendron amurense extract, and Paeonia suffruticosa extract as active ingredients. One or more of these extracts can be used in combination as active ingredients.

[0023] Another aspect of the present invention relates to a spot-preventing or improving agent (hereinafter sometimes referred to as "the spot-preventing or improving agent of the present invention") that contains the RGMB secretion inhibitor of the present invention. A further aspect of the present invention relates to a spot-preventing or improving agent containing one or more extracts selected from holy basil extract, Indian date extract, Geranium robertianum extract, Ginkgo biloba extract, Equisetum arvense extract, Rosa rugosa extract, Phellodendron amurense extract, and Paeonia suffruticosa extract as active ingredients. One or more of these extracts can be used in combination as active ingredients.

[0024] The present inventors have discovered, through the screening method of the present invention, that holy basil extract, Indian date extract, Geranium robertianum extract, Ginkgo biloba extract, Equisetum arvense extract, Rosa rugosa extract, Phellodendron amurense extract, and Paeonia suffruticosa extract have RGMB secretion inhibitory effects, and therefore have melanin production inhibitory effects, as well as effects of preventing or improving blemishes. Thus, the RGMB secretion inhibitor, melanin production inhibitor, and blemish prevention or improvement agent of the present invention have been discovered.

[0025] The extract used as the expression promoter of the present invention is not limited as long as it has RGMB secretion inhibitory activity, melanin production inhibitory activity, and blemish prevention or improvement activity. The term "extract" refers not only to the animal or plant-derived extract itself, but also to fractions of the extract, purified fractions, extracts, fractions, or solvent-removed products of the purified product, and plant-derived extracts include extracts using wild or cultivated plants, extracts using plants sold as raw materials for herbal medicines, and commercially available extracts.

[0026] The extraction process can use the whole plant, or parts such as the plant body, above-ground parts, rhizomes, trunks, leaves, stems, flower spikes, and flower buds. However, it is preferable to crush or finely chop these parts beforehand to improve extraction efficiency. Suitable extraction solvents include one or more selected from alcohols such as water, ethanol, isopropyl alcohol, and butanol; polyhydric alcohols such as 1,3-butanediol and polypropylene glycol; ketones such as acetone and methyl ethyl ketone; ethers such as diethyl ether and tetrahydrofuran; and polar solvents such as dimethyl sulfoxide (DMSO). A specific extraction method may involve adding 1 to 30 parts by mass of the solvent to 1 mass of the plant body or its dried form, immersing it for several days at room temperature or for several hours at a temperature near its boiling point, cooling it to room temperature, removing insoluble matter and / or solvent as desired, and then fractionating and purifying it by column chromatography or the like.

[0027] More specifically, for example, horsetail extract can be an extract of the whole plant, leaves, roots, fruits, seeds, or flowers of the horsetail plant (Equisetum arvense), which belongs to the genus Equisetum in the family Equisetaceae. More specifically, for example, Geranium robertianum extract refers to extracts from the whole plant, or leaves, roots, fruits, seeds, and flowers of Geranium robertianum, a plant belonging to the genus Geranium in the family Geraniaceae. More specifically, ginkgo extracts include extracts from the whole plant, leaves, roots, fruits, seeds, and flowers (preferably leaves) of the Ginkgo biloba plant, which belongs to the genus Ginkgo in the family Ginkgoaceae. More specifically, for example, Izayoi rose extract is derived from the Izayoi rose, a plant belonging to the genus Rosa in the family Rosaceae. This includes extracts from the whole plant or leaves, roots, fruits, seeds, and flowers of the rose (Rosa roxburghii). More specifically, for example, Phellodendron amurense extract can be an extract of the whole plant, or leaves, roots, fruits, seeds, or flowers of the Phellodendron amurense plant, which belongs to the genus Phellodendron in the Rutaceae family. More specifically, for example, peony extract can be an extract of the whole plant or leaves, roots, fruits, seeds, or flowers of the peony plant (Paeonia suffruticosa) belonging to the genus Paeoniaceae. More specifically, for example, Indian date extract (sometimes simply called "Indian dates" as an ingredient name, but this is also included in the present invention) includes extracts of the whole plant or leaves, roots, fruits, seeds, and flowers of the plant Tamarindus indica L., a member of the genus Tamarindus in the legume family. More specifically, for example, holy basil extract (sometimes called "holy basil extract" as an ingredient name, but this is also included in the present invention) includes extracts of the whole plant or leaves, roots, fruits, seeds, and flowers of the plant Ocimum tenuiflorum L., a member of the Lamiaceae family.

[0028] The RGMB secretion inhibitory effect, melanin production inhibitory effect, and blemish prevention or improvement effect can be confirmed, for example, based on methods for measuring the amount of RGMB or melanin production, a blemish model, and evaluations by panelists.

[0029] <Whitening composition> The whitening materials screened by the screening method of the present invention have the effect of reducing or inhibiting the expression, secretion, or expression of RGMB, a pigment formation promoting factor in melanocytes, and exhibit melanin production suppression effects and blemish prevention or prevention effects. As described later, they can be incorporated into cosmetics, foods and beverages, pharmaceuticals, etc., including quasi-drugs, to exhibit whitening effects. In other words, they can be used as whitening compositions.

[0030] In other words, one aspect of the present invention relates to a whitening composition (hereinafter sometimes referred to as "the whitening composition of the present invention") containing the RGMB secretion inhibitor of the present invention, the melanin production inhibitor of the present invention, or the blemish prevention or improvement agent of the present invention. One or more of these agents can be used in combination as active ingredients. Furthermore, one embodiment of the present invention relates to a whitening composition that whitens the skin by suppressing melanin production. Furthermore, a further embodiment of the whitening composition of the present invention relates to a whitening composition for preventing or improving blemishes. Another aspect of the present invention relates to a method for designing a whitening composition of the present invention (hereinafter sometimes referred to as "the design method of the present invention"), which includes the steps of performing the screening method of the present invention and incorporating the substance selected by the said step.

[0031] The whitening composition of the present invention comprises the RGMB secretion inhibitor of the present invention, the melanin production inhibitor of the present invention, or the spot prevention or improvement agent of the present invention, and may further contain ingredients used in cosmetics, food and beverages, pharmaceuticals, etc., including quasi-drugs, as long as they do not interfere with the effects thereof. The whitening composition can be manufactured by formulating the above-mentioned agents and any ingredients used in cosmetics, food and beverages, pharmaceuticals, etc., including quasi-drugs, according to conventional methods.

[0032] The whitening composition of the present invention is applied as a cosmetic, food or beverage, or pharmaceutical, including quasi-drugs. In the case of cosmetics and pharmaceuticals, the whitening composition is ingested by applying it to the skin, while in the case of food or beverages, it is ingested orally.

[0033] The content (amount) of the RGMB secretion inhibitor, the melanin production inhibitor, or the spot prevention or improvement agent of the present invention in the whitening composition is not limited, but is usually 0.00001% by mass or more, preferably 0.0001% by mass or more, more preferably 0.001% by mass or more, and is usually 10 The amount is less than or equal to % by mass, preferably 5% by mass or less, and more preferably 1% by mass or less. By setting it within the above range, a whitening composition that preferably exhibits a whitening effect can be obtained. Furthermore, the type of RGMB secretion inhibitor, melanin production inhibitor, or blemish prevention or improvement agent of the present invention included in the whitening composition may be not limited to one type, but may be two or more types.

[0034] Whitening compositions are particularly preferred for use in cosmetics or food and beverages.

[0035] Cosmetics The following describes the ingredients that may be included in cosmetics when applied to cosmetics. When applied to cosmetics, it is possible to include a wide range of ingredients commonly used in cosmetics, and there are no limitations on their dosage form or use.

[0036] In addition to the RGMB secretion inhibitor, melanin production inhibitor, or blemish prevention or improvement agent of the present invention described above, cosmetics may also contain other active ingredients having the same or different functions. Examples of active ingredients include whitening ingredients, wrinkle-improving ingredients, anti-inflammatory ingredients, the RGMB secretion inhibitor of the present invention, the melanin production inhibitor of the present invention, or plant and animal extracts other than the spot-preventing or improving agent of the present invention.

[0037] As for whitening ingredients, there are no particular limitations as long as they are commonly used in cosmetics. Examples include 4-n-butylresorcinol, ascorbic acid glucoside, 3-O-ethyl ascorbic acid, tranexamic acid, arbutin, 1-triphenylmethylpiperidine, 1-triphenylmethylpyrrolidine, 2-(triphenylmethyloxy)ethanol, 2-(triphenylmethylamino)ethanol, 2-(triphenylmethyloxy)ethylamine, triphenylmethylamine, triphenylmethanol, triphenylmethane and aminodiphenylmethane, N-(p-toluyl)cysteic acid, and N-(p-methoxybenzoyl)cysteic acid. Other whitening ingredients include N-benzoyl-serine, N-(p-methylbenzoyl)serine, N-(p-ethylbenzoyl)serine, N-(p-methoxybenzoyl)serine, N-(p-fluorobenzoyl)serine, N-(p-trifluoromethylbenzoyl)serine, N-(2-naphthoyl)serine, N-(4-phenylbenzoyl)serine, N-(p-methylbenzoyl)serine methyl ester, N-(p-methylbenzoyl)serine ethyl ester, N-(2-naphthoyl)serine methyl ester, N-benzoyl-O-methylserine, N-(p-methylbenzoyl)-O-methylserine, N-(p-methylbenzoyl)-O-acetylserine, and N-(2-naphthoyl)-O-methylserine.

[0038] These whitening ingredients are already commercially available, or can be obtained through synthesis. For example, 3-O-ethyl ascorbic acid can be synthesized by a known method described in Japanese Patent Publication No. 8-134055. There are also commercially available products ("VC Ethyl" manufactured by Nippon Seika Co., Ltd.), so it is possible to obtain and use these. 1-triphenylmethylpiperidine, 1-triphenylmethylpyrrolidine, 2-(triphenylmethyloxy)ethanol, 2-(triphenylmethylamino)ethanol, 2-(triphenylmethyloxy)ethylamine, triphenylmethylamine, triphenylmethanol, triphenylmethane, and aminodiphenylmethane are described in Patent Document WO / 2010 / 074052, and N-(o-toluyl)cysteic acid, N-(m-toluyl)cysteic acid, N-(p-toluyl)cysteic acid, N-(p-methoxyb N-(4-phenylbenzoyl)cysteic acid, N-(p-toluyl)homocysteic acid are listed in WO / 2011 / 058730 as N-benzoyl-serine, N-(p-methylbenzoyl)serine, N-(p-ethylbenzoyl)serine, N-(p-methoxybenzoyl)serine, N-(p-fluorobenzoyl)serine, N-(p-trifluoromethylbenzoyl)serine, N-(2-naphthoyl)serine, N-(4-phenylbenzoyl)serine, N-(p-methylbenzoyl)serine. Methyl esters, N-(p-methylbenzoyl)serine ethyl esters, N-(2-naphthoyl)serine methyl esters, N-benzoyl-O-methylserine, N-(p-methylbenzoyl)-O-methylserine, N-(p-methylbenzoyl)-O-acetylserine, N-(2-naphthoyl)-O-methylserine, etc., have their respective synthesis methods published in WO / 2011 / 074643, and can be synthesized according to said disclosure. The amount of whitening ingredients in cosmetics is usually 0.01 to 30% by mass, preferably 0.1 to 10% by mass, and more preferably 1 to 5% by mass.

[0039] There are no particular limitations on the wrinkle-improving ingredients, as long as they are commonly used in cosmetics. For example, vitamin A or its derivatives include retinol, retinal, retinoic acid, tretinoin, isotretinoin, tocopherol retinoic acid, retinyl palmitate, retinyl acetate, benzyl ursolate, ursolic acid phosphate, benzyl betulinate, benzyl acid phosphate, glycolic acid, alpha-glycolic acid such as lactic acid, etc. The amount of wrinkle-improving ingredients in cosmetics is usually 0.01 to 30% by mass, preferably 0.1 to 10% by mass, and more preferably 1 to 5% by mass.

[0040] Other than the RGMB secretion inhibitor, melanin production inhibitor, or blemish prevention or improvement agent of the present invention, the plant and animal-derived extracts are not particularly limited as long as they are commonly used in pharmaceuticals, cosmetics, food and beverages, etc. For example, Akebia extract, Thujopsis dolabrata extract, Asparagus extract, Avocado extract, Hydrangea macrophylla extract, Almond extract, Arnica extract, Aloe extract, Aronia extract, Apricot extract, Indian kino extract, Fennel extract, Aralia cordata extract, Rosehip extract, Eleutherococcus senticosus extract, Enmeisou extract, Scutellaria baicalensis extract, Coptis japonica extract, Panax ginseng extract, Hypericum perforatum extract, Lamium album extract, Orange extract, Citrus aurantium extract, Pueraria lobata extract, Chamomile extract, Carrot extract, Artemisia princeps extract, licorice extract, kiwi extract, cucumber extract, guava extract, Sophora flavescens extract, gardenia extract, Sasa veitchii extract, Sophora flavescens extract, walnut extract, grapefruit extract, black rice extract, chlorella extract, mulberry extract, Alpinia speciosa extract, Alpinia zerumbet extract, gentian extract, Geranium thunbergii extract, black tea extract, burdock extract, rice extract, rice fermentation extract, rice bran fermentation extract, rice germ oil, lingonberry extract, sage extract, soapwort extract, bamboo grass extract Hawthorn extract, sunflower extract, Japanese pepper extract, shiitake mushroom extract, rehmannia extract, lithospermum extract, perilla extract, linden extract, meadowsweet extract, peony extract, ginger extract, calamus root extract, birch extract, stevia extract, stevia ferment, ivy extract, hawthorn extract, elderberry extract, yarrow extract, peppermint extract, sage extract, mallow extract, chuanthera extract, swertia japonica extract, mulberry extract Kiss extract, rhubarb extract, soybean extract, jujube extract, thyme extract, dandelion extract, tea extract, clove extract, citrus peel extract, sweet tea extract, chili pepper extract, angelica extract, calendula extract, peach kernel extract, spruce extract, Houttuynia cordata extract, tomato extract, natto extract, carrot extract, garlic extract, wild rose extract, hibiscus extract, Ophiopogon extract, lotus extract, parsley extract, birch extract, witch hazel extract, Isodon japonicus extract, cypress extract, loquat extract,Preferred extracts include coltsfoot extract, butterbur extract, poria extract, butcher's broom extract, grape extract, grape seed extract, loofah extract, safflower extract, peppermint extract, linden extract, hop extract, pine extract, horse chestnut extract, skunk cabbage extract, soapberry extract, lemon balm extract, seaweed extract, peach extract, cornflower extract, eucalyptus extract, saxifrage extract, yuzu extract, lily extract, coix seed extract, mugwort extract, lavender extract, green tea extract, apple extract, rooibos tea extract, reishi mushroom extract, lettuce extract, lemon extract, forsythia extract, astragalus extract, rose extract, rosemary extract, Roman chamomile extract, royal jelly extract, and Sanguisorba officinalis extract. The content of plant and animal extracts other than the RGMB secretion inhibitor, melanin production inhibitor, or spot prevention or improvement agent of the present invention in the cosmetic composition is usually 0.01 to 30% by mass, preferably 0.1 to 10% by mass, and more preferably 1 to 5% by mass.

[0041] Examples of anti-inflammatory components include clarinon, glabridin, glycyrrhizic acid, glycyrrhetinic acid, and pantothenyl alcohol, with glycyrrhizic acid and its salts being preferred, alkyl glycyrrhetinate and its salts, and glycyrrhetinic acid and its salts. The content of anti-inflammatory ingredients in cosmetics is usually 0.01 to 30% by mass, preferably 0.1 to 10% by mass, and more preferably 1 to 5% by mass.

[0042] Other ingredients that may be included in cosmetics are listed below as examples. Examples of oily components include polar oils and volatile hydrocarbon oils. As polar oils, synthetic ester oils include isopropyl myristate, cetyl octanoate, octyldodecyl myristate, isopropyl palmitate, butyl stearate, hexyl laurate, myristyl myristate, decyl oleate, hexyldecyl dimethyloctanoate, cetyl lactate, myristyl lactate, lanolin acetate, isocetyl stearate, isocetyl isostearate, cholesteryl 12-hydroxystearylate, and ethylene glycol di-2-ethylhexylate. Examples include dipentaerythritol fatty acid ester, N-alkyl glycol monoisostearate, neopentyl glycol dicaprate, diisostearyl malate, glyceryl di-2-heptylundecanoate, trimethylolpropane tri-2-ethylhexylate, trimethylolpropane triisostearate, pentaneerythritol tetra-2-ethylhexylate, glyceryl tri-2-ethylhexylate, and trimethylolpropane triisostearate. Furthermore, other examples include cetyl 2-ethylhexanoate, 2-ethylhexyl palmitate, glyceryl trimyristate, glyceride tri-2-heptylundecanoate, methyl castor oil fatty acid ester, oleic acid oil, cetostearyl alcohol, acetoglyceride, 2-heptylundecyl palmitate, diisobutyl adipate, 2-octyldodecyl N-lauroyl-L-glutamate, di-2-heptylundecyl adipate, ethyl laurate, di-2-ethylhexyl sebatate, 2-hexyldecyl myristate, 2-hexyldecyl palmitate, 2-hexyldecyl adipate, diisopropyl sebatate, 2-ethylhexyl succinate, ethyl acetate, butyl acetate, amyl acetate, triethyl citrate, octyl methoxycinnamate, and the like. Other examples of natural oils include avocado oil, camellia oil, turtle oil, macadamia nut oil, corn oil, mink oil, olive oil, rapeseed oil, egg yolk oil, sesame oil, peach kernel oil, wheat germ oil, sasanqua oil, castor oil, linseed oil, safflower oil, cottonseed oil, elm oil, soybean oil, peanut oil, tea seed oil, kaya oil, rice bran oil, cinnamon oil, Japanese tung oil, jojoba oil, wheat germ oil, triglycerin, glyceryl trioctanoate, and glyceryl triisopalmitate.

[0043] Examples of volatile hydrocarbon oils include isododecane and isohexadecane.

[0044] The surfactants include fatty acid soaps (sodium laurate, sodium palmitate, etc.), anionic surfactants such as potassium lauryl sulfate and alkyl sulfate triethanolamine ether, cationic surfactants such as stearyltrimethylammonium chloride, benzalkonium chloride and laurylamine oxide, betaine-based surfactants (alkyl betaine, amide betaine, sulfobetaine, etc.), imidazoline-based amphoteric surfactants (2-cocoyl-2-imidazolinium hydroxide-1-carboxyethyloxy disodium salt, etc.), amphoteric surfactants such as acylmethyl taurine, and sorbitan fatty acid esters (sorbitan mo Nostearate, sorbitan sesquioleate, etc.), glycerin fatty acids (glyceryl monostearate, etc.), propylene glycol fatty acid esters (propylene glycol monostearate, etc.), hydrogenated castor oil derivatives, glycerin alkyl ethers, POE sorbitan fatty acid esters (POE sorbitan monooleate, polyoxyethylene sorbitan monostearate, etc.), POE sorbitol fatty acid esters (POE-sorbitol monolaurate, etc.), POE glycerin fatty acid esters (POE-glycerin monoisostearate, etc.), POE Examples include fatty acid esters (polyethylene glycol monooleate, POE distearate, etc.), POE alkyl ethers (POE 2-octyldodecyl ether, etc.), POE alkylphenyl ethers (POE nonylphenyl ether, etc.), Pluronic® type, POE·POP alkyl ethers (POE·POP 2-decyltetradecyl ether, etc.), Tetronic, POE castor oil / hydrogenated castor oil derivatives (POE castor oil, POE hydrogenated castor oil, etc.), sucrose fatty acid esters, alkyl glucosides and other nonionic surfactants.

[0045] Examples of polyhydric alcohols include polyethylene glycol, glycerin, 1,3-butylene glycol, erythritol, sorbitol, xylitol, maltitol, propylene glycol, dipropylene glycol, diglycerin, isoprene glycol, 1,2-pentanediol, 2,4-hexylene glycol, 1,2-hexanediol, and 1,2-octanediol.

[0046] Examples of thickening agents include guar gum, quince seed, carrageenan, galactan, gum arabic, pectin, mannan, starch, xanthan gum, curdlan, methylcellulose, hydroxyethylcellulose, carboxymethylcellulose, methylhydroxypropylcellulose, chondroitin sulfate, dermatan sulfate, glycogen, heparan sulfate, hyaluronic acid, sodium hyaluronate, tragacanth gum, keratan sulfate, chondroitin, mucoitin sulfate, hydroxyethyl guar gum, carboxymethyl guar gum, dextran, kerato sulfate, locust bean gum, succinoglycan, carotenoid acid, chitin, chitosan, carboxymethyl chitin, agar, polyvinyl alcohol, polyvinylpyrrolidone, carboxyvinyl polymer, alkyl-modified carboxyvinyl polymer, sodium polyacrylate, polyethylene glycol, and bentonite.

[0047] Powders include powders such as mica, talc, kaolin, synthetic mica, calcium carbonate, magnesium carbonate, anhydrous silicic acid (silica), aluminum oxide, and barium sulfate, which may have surface treatments; inorganic pigments such as red iron oxide, yellow iron oxide, black iron oxide, cobalt oxide, ultramarine, Prussian blue, titanium dioxide, and zinc oxide, which may have surface treatments; pearlescent agents such as titanium mica, fish scale foil, and bismuth oxychloride, which may have surface treatments; and Examples include organic dyes such as Red 202, Red 228, Red 226, Yellow 4, Blue 404, Yellow 5, Red 505, Red 230, Red 223, Orange 201, Red 213, Yellow 204, Yellow 203, Blue 1, Green 201, Violet 201, and Red 204, which may be carboxylated; organic powders such as polyethylene powder, polymethyl methacrylate, nylon powder, and organopolysiloxane elastomers.

[0048] Examples of UV absorbers include para-aminobenzoic acid-based UV absorbers, anthranilic acid-based UV absorbers, salicylic acid-based UV absorbers, cinnamic acid-based UV absorbers, benzophenone-based UV absorbers, sugar-based UV absorbers, 2-(2'-hydroxy-5'-t-octylphenyl)benzotriazole, 4-methoxy-4'-t-butyldibenzoylmethane, and other UV absorbers.

[0049] Furthermore, when applied as a cosmetic, the dosage form can be any of the commonly known forms, such as lotion, emulsion, essence, cream, or powder-containing dosage form. The form of the cosmetic product is not limited to, but examples include lotions, emulsions, serums, creams, gels, sun care products, etc.

[0050] ≪Food and beverages≫ The following describes the ingredients that may be included in food and beverages when applicable. When applicable to food and beverages, it is possible to include a wide range of ingredients commonly used in food and beverages, and there are no limitations on their form or other aspects. Furthermore, in the production of the food and beverages of the present invention, any ingredients commonly used in food production can be arbitrarily incorporated. Examples of such optional components include proteins, carbohydrates, fats, nutrients, seasonings, and flavorings. Examples of carbohydrates include monosaccharides such as glucose and fructose; disaccharides such as maltose, sucrose, and oligosaccharides; and polysaccharides such as dextrin and cyclodextrin, as well as sugar alcohols such as xylitol, sorbitol, and erythritol. Examples of flavorings include natural flavorings (thaumatin, stevia extract, etc.) and synthetic flavorings (saccharin, aspartame, etc.). In addition, additives used in pharmaceutical compositions that are commonly added to foods may also be used.

[0051] The form of food and beverages can be liquid, paste, solid, powder, granules, etc. Furthermore, tablets, liquid foods, and animal feed are also included in the definition of food and beverages.

[0052] The types of food and beverages are not limited to, but include, for example, wheat flour products such as bread, macaroni, spaghetti, noodles, cake mix, fried chicken batter, and breadcrumbs; instant noodles, cup noodles, retort / prepared foods, canned prepared foods, microwaveable foods, instant soups / stews, instant miso soup / clear soup, canned soups, freeze-dried foods, and other instant foods; canned agricultural products, canned fruits, jams / marmalades, pickles, boiled beans, and dried agricultural products. Processed agricultural products such as cereals (grain products); processed marine products such as canned seafood, fish ham and sausages, processed seafood products, seafood delicacies, and tsukudani (simmered seafood); processed livestock products such as canned and paste meats, and meat ham and sausages; processed milk, milk beverages, yogurts, lactic acid bacteria beverages, cheese, ice cream, powdered milk, cream, and other dairy products; fats and oils such as butter, margarine, and vegetable oils; soy sauce and miso. Basic seasonings such as sauces, tomato-based seasonings, mirin, and vinegars; compound seasonings and foods such as cooking mixes, curry bases, sauces, dressings, noodle soup bases, spices, and other compound seasonings; frozen foods such as raw frozen foods, semi-cooked frozen foods, and cooked frozen foods; confectionery such as caramel, candy, chewing gum, chocolate, cookies, biscuits, cakes, pies, snacks, crackers, Japanese sweets, rice sweets, bean sweets, and dessert sweets; beverages such as carbonated drinks, natural fruit juices, fruit juice drinks, fruit juice-containing soft drinks, fruit pulp drinks, fruit juice drinks with fruit pulp, vegetable drinks, soy milk, soy milk drinks, coffee drinks, tea drinks, powdered drinks, concentrated drinks, sports drinks, nutritional drinks, and alcoholic beverages; the RGMB secretion inhibitor of the present invention, the melanin production inhibitor of the present invention, or the blemish prevention or improvement agent of the present invention may be added to foods other than those listed above.

[0053] Embodiments of foods and beverages containing the RGMB secretion inhibitor of the present invention, the melanin production inhibitor of the present invention, or the blemish prevention or improvement agent of the present invention include ordinary foods, beverages, foods with functional claims, health functional foods such as foods for specified health uses, and supplements, with foods with functional claims being particularly preferred. When the food or beverage of the present invention is intended for skin whitening purposes, a label indicating its usefulness and functionality may be attached to the product during manufacturing. Such "display" acts include all acts that inform consumers of the aforementioned uses, and any expression that can evoke or infer uses such as "for skin whitening," "for inhibiting melanin production," or "for preventing or improving blemishes" falls under the "display" acts of the present invention, regardless of the purpose of the display, the content of the display, or the object or medium on which it is displayed. Furthermore, it is preferable that the "display" be made in a way that allows consumers to directly recognize the above-mentioned use. Specifically, the above-mentioned use should be indicated on food and beverage products or their packaging, containers, etc. Examples of actions that include transferring, delivering, displaying for transfer or delivery, or importing the described items; displaying or distributing advertisements, price lists, catalogs, brochures, POP displays, or other promotional materials at sales sites, or transaction documents that include the above-mentioned uses; or providing information containing such materials by electromagnetic means (such as the Internet) that includes the above-mentioned uses. Furthermore, if the food and beverages of the present invention are approved under various systems established by the government, such as health functional foods, and are implemented under such approval, it is preferable to display them in a manner based on said approval.

[0054] The preferred intake of the food and beverage of the present invention, when consumed by an adult, is preferably 0.001 to 1000 mg / kg per day in solid matter terms of the RGMB secretion inhibitor, melanin production inhibitor, or blemish prevention or improvement agent of the present invention, from the viewpoint of achieving sufficient efficacy. More preferably, it is 0.01 to 100 mg / kg, and particularly preferably 0.1 to 10 mg / kg. This daily amount can be taken all at once or divided into several doses. In addition to single doses, it is also preferable to take it continuously or intermittently for several weeks to several months. [Examples]

[0055] The present invention will be specifically described below with reference to examples, but these are merely illustrative examples of the present invention, and the scope of the present invention is not limited thereto.

[0056] <Example Test 1> Identification of factors that promote melanin production in melanocytes The inventors confirmed that human neuronal secretory factors promote melanocyte pigment formation (melanin production). Therefore, they conducted a comprehensive analysis of secretory proteins from human sensory nerve cells and found that RGMB, a human neuronal secretory factor, significantly promotes melanin production in melanocytes. Furthermore, the melanin production-promoting effect of RGMB addition was confirmed as follows.

[0057] (method) (1) Human melanocytes 4.5 × 10 4 Seed cells per well were seeded into a 24-well plate and incubated overnight at 37°C in a 5% CO2 environment. (2) Remove the supernatant by aspirating, and add 0.5 μCi [ 14 Medium containing 10, 50, and 100 ng / mL of [C]-2-Thiouracil and RGMB was added. α-MSH was added at 1 μM as a positive control for melanin production enhancement. (3) After culturing for 72 hours at 37°C in a 5% CO2 environment, the "cell count" is measured using WST-8. (4) Remove the supernatant by aspirating, wash with PBS (-), and add 110 μL of 100% TCA solution. (5) Add ice-cold water to make a 10-15% TCA solution and collect in a tube. (6) Leave undisturbed in ice for 15 minutes. (7) After removing the supernatant by centrifugation, add 1 mL of ice-cold 10% TCA. (8) After removing the supernatant by centrifugation, add 1 mL of INSTA-GEL PLUS and mix. (9) Using a scintillation counter 14 Measuring C activity and measuring "melanin production" (10) Calculate the amount of melanin produced per cell from (3) and (9) (melanin production / number of cells). Evaluate the melanin production of each additive group, with the control mean set to 100%.

[0058] (result) As shown in Figure 1, melanin production was significantly enhanced in the groups treated with RGMB at 10 ng / mL and 50 ng / mL.

[0059] <Example 1> Screening of whitening materials using RGMB as an indicator We screened skin-whitening materials using RGMB as an indicator, as follows:

[0060] (material) Cells used: Human iPS cell-derived sensory nerve precursor cells (AXOL Bioscience) Culture media used: Seedling medium, maintenance medium (AXOL Bioscience) RGMB quantification method: ELISA (using polyclonal rabbit anti-human RGMB antibody (LifeSpan Biosciences)) The extracts to be tested are shown in the table below.

[0061] [Table 1]

[0062] (method) (1) Human iPS cell-derived sensory nerve precursor cells 1 x 10 5 Seed cells per well into a 48-well plate, cultured until Day 24 according to the manufacturer's protocol, and then guided to develop sensory nerve cells. (2) Day 25: Collect half of the culture supernatant, add half of the extract-containing medium (extract-added group) or solvent-containing medium (solvent control group), and incubate for 3 days. (3) Day 28: Half of the culture supernatant is collected and visually confirmed to be free of cytotoxicity. (4) The amount of RGMB in the culture supernatant on Day 25 and Day 28 was quantified using an ELISA kit. (5) Analyze the amount of RGMB secreted by sensory nerve cells over a 3-day period to evaluate the effects of the extract.

[0063] (result) As shown in Figures 2 and 3, horsetail extract, geranium extract, ginkgo extract, rose extract, Phellodendron bark extract, and peony extract were selected as substances that suppress the secretion of RGMB from sensory nerve cells. Furthermore, as shown in Figure 4, holy basil extract and Indian dates were selected as substances that suppress the secretion of RGMB from sensory nerve cells.

[0064] The amount of RGMB secreted in the extract-treated group was calculated as follows. RGMB secretion in the extract-treated group (%) = RGMB secretion in the extract-treated group (RGMB secretion on Day 28 - RGMB secretion on Day 25) / RGMB secretion in the solvent control group (RGMB secretion on Day 28 - RGMB secretion on Day 25) x 100

[0065] As described above, RGMB is a melanin production promoting factor in melanocytes. Therefore, the screening method of the present invention, which uses the RGMB secretion inhibitory effect as an indicator, can be used to screen for whitening materials, such as those for treating blemishes, by inhibiting melanin production in melanocytes. [Industrial applicability]

[0066] This invention can be applied to cosmetics, pharmaceuticals, food and beverages, and the like, including quasi-drugs.

Claims

1. An RGMB secretion inhibitor containing one or more extracts selected from holy basil extract, Indian date extract, Geranium robertianum extract, Ginkgo biloba extract, Equisetum arvense extract, Rosa rugosa extract, Phellodendron amurense extract, and Paeonia suffruticosa extract as active ingredients.

2. A melanin production inhibitor containing the RGMB secretion inhibitor described in claim 1, and used for melanin production inhibition applications based on its RGMB secretion inhibitory effect.

3. A blemish prevention or improvement agent containing the RGMB secretion inhibitor described in claim 1, used for the prevention or improvement of blemishes based on its RGMB secretion inhibitory effect.

4. A whitening composition comprising the RGMB secretion inhibitor described in claim 1, the melanin production inhibitor described in claim 2, or the spot prevention or improvement agent described in claim 3, and used for whitening purposes based on its RGMB secretion inhibitory effect.

5. The whitening composition according to claim 4, which is for whitening by suppressing melanin production based on its RGMB secretion inhibitory effect.

6. The whitening composition according to claim 4, which is for preventing or improving blemishes based on its RGMB secretion inhibitory effect.

7. A cosmetic composition for skin whitening according to any one of claims 4 to 6.

8. A whitening composition according to any one of claims 4 to 6, which is a food or beverage.

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