Melanin production inhibitor

Abstract

To provide a new melanogenesis depressant.SOLUTION: The present invention discloses (1) a melanogenesis depressant comprising glucosylceramide as an active ingredient, wherein the glucosylceramide comprises a 18-20C fatty acid sphingadienine (4E,8Z or 8E) or sphingenine (4E) as substituents and (2) the melanogenesis depressant set forth in (1), wherein the glucosylceramide is at least one selected from G GlcCer (d18:2(4E,8Z) / C18:0), GlcCer (d18:2(4E,8Z) / C20:0), GlcCer (d18:2(4E,8E) / C20:0) and GlcCer (d18:1(4E) / C20:0).SELECTED DRAWING: None

Description

[Technical Field]

[0001] The present invention relates to a melanin production inhibitor containing glucosylceramide, which can be widely used in foods, pharmaceuticals, quasi-drugs, cosmetics, topical skin preparations, etc. [Background technology]

[0002] Sphingolipids are a general term for compounds in which a fatty acid and a long-chain base, sphingosine, are bonded via an acid amide bond, and a representative example is ceramide. Generally, ceramides include ceramide aglycones, which are composed of a fatty acid and a long-chain base, glucosylceramides, which are ceramide aglycones bound to glucose, and sphingomyelins, which are ceramide aglycones bound to phosphorylcholine. Glucosylceramide is a compound found in various plants, including rice, corn, and konjac. To date, various types of glucosylceramide have been isolated and identified, and it is known that the composition of glucosylceramide varies from plant to plant. Glucosylceramide is known to have functions such as moisturizing the skin, regulating moisture, maintaining elasticity, surface protection (barrier effect), collagen protection, and antioxidant properties (stabilization of vitamins C and E), and is widely used in the field of cosmetics. Recently, it has been revealed that glucosylceramide acts to block the entry of antigens from the outside world into the body and suppress atopic dermatitis, and its development as a pharmaceutical product is also underway (Patent Document 1).

[0003] [Patent Document 1] Patent Publication No. 2001-097983 DISCLOSURE OF THE INVENTION [Problem to be solved by the invention]

[0004] Under these circumstances, the present inventors have discovered that glucosylceramides in which a specific fatty acid and sphingosine are bound have an inhibitory effect on melanin production, and have completed the present invention. That is, an object of the present invention is to provide a novel melanin production inhibitor having excellent effects. [Means for solving the problem]

[0005] The features of the present invention for solving the above problems are as follows. 1. A melanin production inhibitor containing glucosylceramide as an active ingredient, The glucosylceramide is a melanin production inhibitor characterized in that it is a sphingosine substituted with any one of C18 to C20 fatty acids and (4E, 8Z or 8E) sphingadienine or (4E) sphingenine. 2. The melanin production inhibitor according to claim 1, wherein the glucosylceramide is at least one selected from GlcCer (d18:2(4E,8Z) / 18:0), GlcCer (d18:2(4E,8Z) / 20:0), GlcCer (d18:2(4E,8E) / 20:0), and GlcCer (d18:1(4E) / 20:0). [Brief explanation of the drawings]

[0006] [Figure 1] This is a chart of a preparative chromatogram of glucosylceramide (Oryza Ceramide). [Figure 2] 1 shows the relationship between compounds and HPLC fractions. [Figure 3] FIG. 1 shows the structure-activity relationship in the melanin production inhibitory effect of glucosylceramide. DETAILED DESCRIPTION OF THE INVENTION

[0007] The present invention will be described in detail below. The present invention is characterized by using glucosylceramide as an active ingredient. Glucosylceramide is a compound represented by the following chemical formula: [ka]

[0008] In the formula, R1 represents a straight-chain or branched-chain saturated or unsaturated hydrocarbon (hereinafter referred to as "sphingosine"), R2 represents a fatty acid residue, and G represents glucose.

[0009] The glucosylceramide is characterized in that R1 is substituted with sphingadienine (4E, 8Z or 8E).

[0010] Furthermore, the glucosylceramide is characterized in that R2 is substituted with any one of C17 to C19 fatty acids.

[0011] Examples of glucosylceramides that satisfy the above conditions include, but are not limited to, GlcCer (d18:2(4E,8Z) / C18:0), GlcCer (d18:2(4E,8Z) / C20:0), GlcCer (d18:2(4E,8E) / C20:0), and GlcCer (d18:1(4E) / C20:0). These may be used alone or in combination of two or more.

[0012] The above-mentioned GlcCer (d18:2(4E,8Z) / C18:0), GlcCer (d18:2(4E,8Z) / C20:0), GlcCer (d18:2(4E,8E) / C20:0), and GlcCer (d18:1(4E) / C20:0) are compounds represented by the following chemical formulas, respectively.

[0013] [ka] [ka] [ka] [ka]

[0014] The glucosylceramide can be obtained by isolating it from rice-derived glycosphingolipids, but is not limited to this method. As this rice-derived glycosphingolipid, for example, "Oryza Ceramide" manufactured by Oryza Oil & Fat Chemical Co., Ltd. can be used, but is not limited to this. Specifically, it can be obtained by, for example, the method described in the Examples of this specification, but is not limited to this method.

[0015] The melanin production inhibitor of the present invention can be used as an ingredient in various foods and beverages. Examples of foods and beverages include general foods such as confectioneries (gum, candy, caramel, chocolate, cookies, snacks, jelly, gummy candy, tablet candy, etc.), noodles (soba, udon, ramen, etc.), dairy products (milk, ice cream, yogurt, etc.), seasonings (miso, soy sauce, etc.), soups, beverages (juice, coffee, black tea, green tea, carbonated drinks, sports drinks, etc.), as well as health foods (tablets, capsules, etc.), and nutritional supplements (nutritional drinks, etc.). The agent of the present invention can be appropriately incorporated into these foods and beverages.

[0016] These foods and beverages can contain various ingredients depending on their type, such as glucose, fructose, sucrose, maltose, sorbitol, stevioside, corn syrup, lactose, citric acid, tartaric acid, malic acid, succinic acid, lactic acid, L-ascorbic acid, dl-α-tocopherol, sodium erythorbate, glycerin, propylene glycol, glycerin fatty acid esters, polyglycerin fatty acid esters, sucrose fatty acid esters, sorbitan fatty acid esters, propylene glycol fatty acid esters, gum arabic, carrageenan, casein, gelatin, pectin, agar, B vitamins, nicotinamide, calcium pantothenate, amino acids, calcium salts, colorants, flavorings, and preservatives.

[0017] Specifically, the agent of the present invention can be spray-dried or freeze-dried together with powdered cellulose, and then formed into a powder, granules, tablets, or solution, which can be easily incorporated into foods and beverages (e.g., instant foods). The agent of the present invention can also be dissolved in, for example, fats and oils, ethanol, glycerin, or a mixture thereof to form a liquid, which can then be added to beverages or solid foods. If necessary, the agent can also be mixed with a binder such as gum arabic or dextrin to form a powder or granules, which can then be added to beverages or solid foods.

[0018] When the agent of the present invention is applied to food and drink, the amount of the active ingredient added is preferably 1 to 20 wt % in total relative to the food and drink.

[0019] The melanin production inhibitor of the present invention may be used as a material for pharmaceuticals (including pharmaceuticals and quasi-drugs). Pharmaceutical preparations can be produced by appropriately blending the agent of the present invention with raw materials for pharmaceutical preparations. Examples of pharmaceutical raw materials that can be incorporated into the agent of the present invention include excipients (glucose, lactose, sucrose, sodium chloride, starch, calcium carbonate, kaolin, crystalline cellulose, cocoa butter, hardened vegetable oil, kaolin, talc, etc.), binders (distilled water, saline, ethanol water, simple syrup, glucose solution, starch solution, gelatin solution, carboxymethylcellulose, potassium phosphate, polyvinylpyrrolidone, etc.), disintegrants (sodium alginate, agar, sodium bicarbonate, calcium carbonate, sodium lauryl sulfate, stearate monoglyceride, starch, lactose, gum arabic powder, gelatin, ethanol, etc.), disintegration inhibitors (sucrose, stearin, cocoa butter, hydrogenated oil, etc.), absorption enhancers (quaternary ammonium base, sodium lauryl sulfate, etc.), adsorbents (glycerin, starch, lactose, kaolin, bentonite, silicic acid, etc.), lubricants (purified talc, stearates, polyethylene glycol, etc.).

[0020] The melanin production inhibitor of the present invention can generally be administered orally in the form of tablets, pills, soft or hard capsules, fine granules, powders, granules, liquids, etc., but may also be administered parenterally. When administered parenterally, it can be administered in the form of a solution or with the addition of a dispersing agent, suspending agent, stabilizer, etc., by local tissue administration, intradermal, subcutaneous, intramuscular, or intravenous injection, etc. It may also be in the form of a suppository, etc.

[0021] The dosage may vary depending on the administration method, the condition, the age of the patient, etc., but typically, adults can be given 0.5 to 5000 mg of the active ingredient per day, and children can be given 0.5 to 3000 mg. The compounding ratio of the melanin production inhibitor can be changed as appropriate depending on the dosage form, but is usually about 0.3 to 15.0 wt% when administered orally or via mucosal absorption, and about 0.01 to 10 wt% when administered parenterally. Note that the dosage varies depending on various conditions, so in some cases a smaller dosage than the above is sufficient, and in other cases it may be necessary to administer more than the range.

[0022] The melanin production inhibitor of the present invention can be expected to have a melanin production inhibitory effect even when used as an external skin preparation (including cosmetics, pharmaceuticals, and quasi-drugs). Examples of the forms of external skin preparations that can be blended with the melanin production inhibitor of the present invention include emulsions, soaps, facial cleansers, bath additives, creams, emulsions, lotions, colognes, shaving creams, shaving lotions, cosmetic oils, suntan / sunscreen lotions, face powders, foundations, perfumes, packs, nail creams, enamel, enamel removers, eyebrow pencils, blushers, eye creams, eye shadows, mascara, eyeliners, lipsticks, lip balms, shampoos, rinses, hair dyes, dispersions, cleansers, etc. Examples of the forms of pharmaceuticals or quasi-drugs that can be blended with the melanin production inhibitor of the present invention include ointments, creams, external liquids, etc.

[0023] In addition to the melanin production inhibitor of the present invention, the topical skin preparations in the above forms can contain ingredients that are incorporated into topical skin preparations such as cosmetics and quasi-drugs, as long as the melanin production inhibitory effect is not impaired, such as oils, higher alcohols, fatty acids, ultraviolet absorbers, powders, pigments, surfactants, polyhydric alcohols, sugars, polymers, physiologically active ingredients, solvents, antioxidants, fragrances, preservatives, etc. Examples are listed below, but the present invention is not limited to these examples.

[0024] (1) Examples of oil Ester-based oil phase ingredients: glyceryl tri-2-ethylhexanoate, cetyl 2-ethylhexanoate, isopropyl myristate, butyl myristate, isopropyl palmitate, ethyl stearate, octyl palmitate, isocetyl isostearate, butyl stearate, butyl myristate, ethyl linoleate, isopropyl linoleate, ethyl oleate, isocetyl myristate, isostearyl myristate, isostearyl palmitate, octyldodecyl myristate, isocetyl isostearate, diethyl sebacate, diisopropyl adipate Isopropyl, Isoarachyl Neopentanoate, Caprylic / Capric Triglyceride, Trimethylolpropane Tri-2-Ethylhexanoate, Trimethylolpropane Triisostearate, Pentaerythritol Tetra-2-Ethylhexanoate, Cetyl Caprylate, Decyl Laurate, Hexyl Laurate, Decyl Myristate, Myristyl Myristate, Cetyl Myristate, Stearyl Stearate, Decyl Oleate, Cetyl Ricinoleate, Isostearyl Laurate, Isotridecyl Myristate, Isocetyl Myristate, Isostearate Myristate Tearyl, Isocetyl Palmitate, Isostearyl Palmitate, Octyl Stearate, Isocetyl Stearate, Isodecyl Oleate, Octyldodecyl Oleate, Octyldodecyl Linoleate, Isopropyl Isostearate, Cetostearyl 2-Ethylhexanoate, Stearyl 2-Ethylhexanoate, Hexyl Isostearate, Ethylene Glycol Dioctanoate, Ethylene Glycol Dioleate, Propylene Glycol Dicaprate, Propylene Glycol Dicaprylate, Propylene Glycol Dicaprylate, Dicaprin Neopentyl glycol acid, neopentyl glycol dioctanoate, glyceryl tricaprylate, glyceryl triundecylate, glyceryl triisopalmitate, glyceryl triisostearate, octyldodecyl neopentanoate, isostearyl octanoate, octyl isononanoate, hexyldecyl neodecanoate, octyldodecyl neodecanoate, isocetyl isostearate, isostearyl isostearate, octyldecyl isostearate, polyglycerin oleate, polyglycerin isostearate, dipropyl carbonate,Dialkyl carbonate (C12-18), triisocetyl citrate, triisoarachidyl citrate, triisooctyl citrate, lauryl lactate, myristyl lactate, cetyl lactate, octyldecyl lactate, triethyl citrate, acetyltriethyl citrate, acetyltributyl citrate, trioctyl citrate, diisostearyl malate, 2-ethylhexyl hydroxystearate, di-2-ethylhexyl succinate, diisobutyl adipate, diisopropyl sebacate propyl, dioctyl sebacate, cholesteryl stearate, cholesteryl isostearate, cholesteryl hydroxystearate, cholesteryl oleate, dihydrocholesteryl oleate, phytosteryl isostearate, phytosteryl oleate, isocetyl 12-stearoylhydroxystearate, stearyl 12-stearoylhydroxystearate, isostearyl 12-stearoylhydroxystearate, etc. Hydrocarbon-based oil phase components: squalane, liquid paraffin, α-olefin oligomer, isoparaffin, ceresin, paraffin, liquid isoparaffin, polybutene, microcrystalline wax, petrolatum, etc. Animal and vegetable oils and their hydrogenated oils, and naturally derived waxes: beef tallow, hydrogenated beef tallow, lard, hydrogenated lard, horse oil, hydrogenated horse oil, mink oil, orange roughy oil, fish oil, hydrogenated fish oil, egg yolk oil, and other animal oils and their hydrogenated oils, avocado oil, almond oil, olive oil, cocoa butter, kiwi seed oil, apricot kernel oil, kukui nut oil, sesame oil, wheat germ oil, rice germ oil, rice bran oil, safflower oil, shea butter, soybean oil, evening primrose oil, perilla oil, tea seed oil, camellia oil, corn oil, rapeseed oil, Examples of suitable oils include vegetable oils and their hydrogenated oils such as hydrogenated rapeseed oil, palm kernel oil, hydrogenated palm kernel oil, palm oil, hydrogenated palm oil, peanut oil, hydrogenated peanut oil, castor oil, hydrogenated castor oil, sunflower oil, grape seed oil, jojoba oil, hydrogenated jojoba oil, macadamia nut oil, medlar oil, cottonseed oil, hydrogenated cottonseed oil, coconut oil, and hydrogenated coconut oil; and waxes such as beeswax, high acid value beeswax, lanolin, reduced lanolin, hydrogenated lanolin, liquid lanolin, carnauba wax, and montan wax. Silicone-based oil phase components: dimethylpolysiloxane, methylphenylpolysiloxane, methylcyclopolysiloxane, octamethylpolysiloxane, decamethylpolysiloxane, dodecamethylcyclosiloxane, methylhydrogenpolysiloxane, polyether-modified organopolysiloxane, dimethylsiloxane-methylcetyloxysiloxane copolymer, dimethylsiloxane-methylstearoxysiloxane copolymer, alkyl-modified organopolysiloxane, terminal-modified organopolysiloxane, amino-modified silicone oil, amino-modified organopolysiloxane, dimethiconol, silicone gel, acrylic silicone, trimethylsiloxysilicate, silicone RTV rubber, etc. Fluorine-based oil phase components: perfluoropolyether, fluorine-modified organopolysiloxane, fluorinated pitch, fluorocarbon, fluoroalcohol, fluoroalkyl-polyoxyalkylene co-modified organopolysiloxane, etc.

[0025] (2) Examples of higher alcohols Examples include lauryl alcohol, myristyl alcohol, cetyl alcohol, stearyl alcohol, isostearyl alcohol, oleyl alcohol, behenyl alcohol, 2-ethylhexanol, hexadecyl alcohol, and octyldodecanol.

[0026] (3) Examples of fatty acids Examples of the acid include caprylic acid, capric acid, undecylenic acid, lauric acid, myristic acid, palmitic acid, palmitoleic acid, stearic acid, isostearic acid, oleic acid, linoleic acid, linolenic acid, arachidic acid, arachidonic acid, behenic acid, erucic acid, and 2-ethylhexanoic acid.

[0027] (4) Examples of UV absorbers Para-aminobenzoic acid, amyl para-aminobenzoate, ethyl dihydroxypropyl para-aminobenzoate, glyceryl para-aminobenzoate, ethyl para-aminobenzoate, octyl para-aminobenzoate, octyldimethyl para-aminobenzoate, ethylene glycol salicylate, octyl salicylate, triethanolamine salicylate, phenyl salicylate, butylphenyl salicylate, benzyl salicylate, homomenthyl salicylate, benzyl cinnamate, octyl para-methoxycinnamate, 2-ethylhexyl para-methoxycinnamate, glyceryl di-para-methoxycinnamate mono-2-ethylhexanoate, isopropyl para-methoxycinnamate, diethanolamine para-methoxyhydrocinnamate, diisopropyl and diisopropyl cinnamate mixture, urocanic acid, ethyl urocanate, hydroxymethoxybenzophenone, hydroxymethoxybenzophenone sulfonic acid and its salts, di Examples of the hydroxymethoxybenzophenone include sodium dihydroxymethoxybenzophenone disulfonate, dihydroxybenzophenone, dihydroxydimethoxybenzophenone, hydroxyoctoxybenzophenone, tetrahydroxybenzophenone, butylmethoxydibenzoylmethane, 2,4,6-trianilino-p-(carbo-2-ethylhexyl-1-oxy)-1,3,5-triazine, 2-(2-hydroxy-5-methylphenyl)benzotriazole, methyl-O-aminobenzoate, 2-ethylhexyl-2-cyano-3,3-diphenylacrylate, phenylbenzimidazole sulfate, 3-(4-methylbenzylidene)camphor, isopropyldibenzoylmethane, 2-ethylhexyl 4-(3,4-dimethoxyphenylmethylene)-2,5-dioxo-1-imidazolidinepropionate, and polymer derivatives and silane derivatives thereof.

[0028] (5) Examples of powders and pigments Dyes such as Red No. 104, Red No. 201, Yellow No. 4, Blue No. 1, Black No. 401, lake dyes such as Yellow No. 4 AL Lake and Yellow No. 203 BA Lake, nylon powder, silk powder, urethane powder, Teflon (registered trademark) powder, silicone powder, polymethyl methacrylate powder, cellulose powder, starch, silicone elastomer spherical powder, polymers such as polyethylene powder, yellow iron oxide, red iron oxide, black iron oxide, chromium oxide, carbon black, colored pigments such as ultramarine and iron blue, zinc oxide, titanium oxide, acid Examples of suitable powders include white pigments such as cerium chloride, extender pigments such as talc, mica, sericite, kaolin, and plate-like barium sulfate, pearl pigments such as titanium dioxide, metal salts such as barium sulfate, calcium carbonate, magnesium carbonate, aluminum silicate, and magnesium silicate, inorganic powders such as silica and alumina, metal soaps such as aluminum stearate, magnesium stearate, zinc palmitate, zinc myristate, magnesium myristate, zinc laurate, and zinc undecylenate, bentonite, smectite, and boron nitride. There are no particular limitations on the shape (spherical, rod-like, needle-like, plate-like, irregular, flaky, spindle-like, etc.) and particle size of these powders. These powders may or may not have been previously surface-treated by conventionally known surface treatments, such as fluorine compound treatment, silicone treatment, silicone resin treatment, pendant treatment, silane coupling agent treatment, titanium coupling agent treatment, oil treatment, N-acylated lysine treatment, polyacrylic acid treatment, metal soap treatment, amino acid treatment, lecithin treatment, inorganic compound treatment, plasma treatment, mechanochemical treatment, etc.

[0029] (6) Examples of surfactants Anionic surfactants: fatty acid soaps, α-acylsulfonates, alkylsulfonates, alkylarylsulfonates, alkylnaphthalenesulfonates, alkyl sulfates, POE alkyl ether sulfates, alkylamide sulfates, alkyl phosphates, POE alkyl phosphates, alkylamide phosphates, alkyloylalkyl taurine salts, N-acylamino acid salts, POE alkyl ether carboxylates, alkyl sulfosuccinates, sodium alkylsulfoacetates, acylated hydrolyzed collagen peptide salts, perfluoroalkyl phosphate esters, etc. Cationic surfactants: alkyltrimethylammonium chloride, stearyltrimethylammonium chloride, stearyltrimethylammonium bromide, cetostearyltrimethylammonium chloride, distearyldimethylammonium chloride, stearyldimethylbenzylammonium chloride, behenyltrimethylammonium bromide, benzalkonium chloride, behenic acid amidopropyldimethylhydroxypropylammonium chloride, stearic acid diethylaminoethylamide, stearic acid dimethylaminopropylamide, lanolin derivative quaternary ammonium salts, etc. Amphoteric surfactants: Examples thereof include carboxybetaine type, amidobetaine type, sulfobetaine type, hydroxysulfobetaine type, amidosulfobetaine type, phosphobetaine type, aminocarboxylate type, imidazoline derivative type, and amidoamine type. Nonionic surfactants: Propylene glycol fatty acid esters, glycerin fatty acid esters, polyglycerin fatty acid esters, sorbitan fatty acid esters, POE sorbitan fatty acid esters, POE sorbitan fatty acid esters, POE glycerin fatty acid esters, POE alkyl ethers, POE fatty acid esters, POE hydrogenated castor oil, POE castor oil, POE-POP copolymers, POE-POP alkyl ethers, polyether-modified silicone lauric acid alkanolamide, alkylamine oxide, hydrogenated soybean phospholipids, etc. Natural surfactants: Examples include lecithin, saponin, sugar-based surfactants, etc.

[0030] (7) Examples of polyhydric alcohols and sugars Examples of suitable sugars include ethylene glycol, diethylene glycol, polyethylene glycol, propylene glycol, dipropylene glycol, polypropylene glycol, glycerin, diglycerin, polyglycerin, 3-methyl-1,3-butanediol, 1,3-butylene glycol, sorbitol, mannitol, raffinose, erythritol, glucose, sucrose, fructose, xylitol, lactose, maltose, maltitol, trehalose, alkylated trehalose, mixed isomerized sugars, sulfated trehalose, pullulan, etc. Chemically modified versions of these sugars may also be used.

[0031] (8) Examples of polymers Acrylate / methacrylate copolymer (Plussize, manufactured by GOO Chemical Co., Ltd.), vinyl acetate / crotonic acid copolymer (Resin 28-1310, manufactured by NSC), vinyl acetate / crotonic acid / vinyl neodecanoate copolymer (28-2930, manufactured by NSC), methyl vinyl ether maleic acid half ester (Gantrez ES, manufactured by ISP), t-butyl acrylate / ethyl acrylate / methacrylic acid copolymer (Ruvimer, manufactured by BASF), vinylpyrrolidone / vinyl acetate / vinyl propionate copolymer (Ruviscol VAP, manufactured by BASF), vinyl acetate Vinyl acetate / crotonic acid copolymer (Rubyset CA, BASF), vinyl acetate / crotonic acid / vinyl pyrrolidone copolymer (Rubyset CAP, BASF), vinyl pyrrolidone / acrylate copolymer (Rubyflex, BASF), acrylate / acrylamide copolymer (Ultrahold, BASF), vinyl acetate / butyl maleate / isobornyl acrylate copolymer (Advantage, ISP), carboxyvinyl polymer (Carbopol, BFGoodrich), acrylic acid / alkyl methacrylate copolymer (Pemulen, BF Examples of such polymers include anionic polymers such as acetic acid amphoteric dialkylaminoethyl methacrylate polymer (Yukaformer, manufactured by Mitsubishi Chemical Corporation) and amphoteric polymers such as octylacrylamide acrylate / hydroxypropyl acrylate / butylaminoethyl methacrylate copolymer (AMPHOMER, manufactured by NSC Corporation); cationic polymers such as quaternized vinylpyrrolidone / dimethylaminoethyl methacrylate (GAFQUAT, manufactured by ISP Corporation) and methylvinylimidazolium chloride / vinylpyrrolidone copolymer (Rubycoat, manufactured by BASF Corporation); and nonionic polymers such as polyvinylpyrrolidone (Ruviscol K, manufactured by BASF Corporation), vinylpyrrolidone / vinyl acetate copolymer (Ruviscol VA, manufactured by BASF Corporation), vinylpyrrolidone / dimethylaminoethyl methacrylate copolymer (Copolymer 937, manufactured by ISP Corporation), and vinylcaprolactam / vinylpyrrolidone / dimethylaminoethyl methacrylate copolymer (Copolymer VC713, manufactured by ISP Corporation).Also suitable for use are naturally occurring polymeric compounds such as cellulose or derivatives thereof, keratin and collagen or derivatives thereof, calcium alginate, pullulan, agar, gelatin, tamarind seed polysaccharides, xanthan gum, carrageenan, high methoxyl pectin, low methoxyl pectin, guar gum, gum arabic, crystalline cellulose, arabinogalactan, karaya gum, tragacanth gum, alginic acid, albumin, casein, curdlan, gellan gum, and dextran.

[0032] (9) Examples of physiologically active ingredients Examples of physiologically active ingredients include substances that impart some kind of physiological activity to the skin when applied to the skin. Examples include whitening ingredients, immunostimulants, anti-aging agents, UV protection agents, slimming agents, tightening agents, antioxidants, hair growth agents, hair restoration agents, moisturizers, blood circulation promoters, antibacterial agents, disinfectants, drying agents, cooling agents, warming agents, vitamins, amino acids, wound healing promoters, irritation relievers, analgesics, cell activators, enzyme components, etc. Examples of suitable ingredients include angelica extract, avocado extract, hydrangea extract, althea extract, arnica extract, aloe extract, apricot extract, apricot kernel extract, ginkgo extract, fennel extract, turmeric extract, oolong tea extract, angelica tree extract, echinacea leaf extract, Scutellaria root extract, Phellodendron bark extract, Coptis japonica extract, barley extract, St. John's wort extract, white nettle extract, watercress extract, orange extract, Dried seawater, seaweed extract, hydrolyzed elastin, hydrolyzed wheat powder, hydrolyzed silk, chamomile extract, carrot extract, artemisia capillaris extract, licorice extract, kalk extract, kakyoku extract, cinchona extract, cucumber extract, guanosine, gardenia extract, kumazasa extract, sophora flavescens extract, walnut extract, grapefruit extract, clematis extract, chlorella extract, mulberry extract, gentian extract, black tea extract, yeast extract, burdock extract, Fermented rice bran extract, rice germ oil, comfrey extract, collagen, bilberry extract, Chinese radish extract, Bupleurum extract, umbilical cord extract, salvia extract, soapwort extract, bamboo extract, hawthorn extract, Japanese pepper extract, shiitake mushroom extract, rehmannia root extract, Lithospermum root extract, perilla extract, linden extract, meadowsweet extract, peony extract, calamus root extract, white birch extract, horsetail extract, ivy extract, hawthorn extract, se Sambucus extract, yarrow extract, peppermint extract, sage extract, mallow extract, cnidium extract, Swertia japonica extract, soybean extract, Chinese laurel extract, thyme extract, tea extract, clove extract, Imperata cylindrica extract, tangerine 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,Examples of such extracts include burdock root extract, parsley extract, honey, witch hazel extract, parietaria extract, burdock root extract, bisabolol, loquat extract, coltsfoot extract, butterbur stalk extract, poria cocos extract, butcher's broom extract, grape extract, propolis, loofah extract, safflower extract, peppermint extract, linden extract, peony extract, hop extract, pine extract, horse chestnut extract, skunk cabbage extract, soapberry extract, melissa extract, peach extract, cornflower extract, eucalyptus extract, saxifrage extract, coix seed extract, mugwort extract, lavender extract, apple extract, lettuce extract, lemon extract, astragalus extract, rose extract, rosemary extract, Roman chamomile extract, and royal jelly extract. In addition, biopolymers such as deoxyribonucleic acid, mucopolysaccharides, sodium hyaluronate, sodium chondroitin sulfate, collagen, elastin, chitin, chitosan, and hydrolyzed eggshell membrane; moisturizing ingredients such as amino acids, hydrolyzed peptides, sodium lactate, urea, sodium pyrrolidonecarboxylate, betaine, whey, and trimethylglycine; oily ingredients such as sphingolipids, ceramides, phytosphingosine, cholesterol, cholesterol derivatives, and phospholipids; immunostimulants such as ε-aminocaproic acid, glycyrrhizic acid, β-glycyrrhetinic acid, lysozyme chloride, guaiazulene, and hydrocortisone; vitamins such as vitamin A, vitamin B2, vitamin B6, vitamin C, vitamin D, vitamin E, calcium pantothenate, biotin, nicotinamide, and vitamin C ester; allantoin and diisopropyl alcohol. Active ingredients such as mindichloroacetate and 4-aminomethylcyclohexanecarboxylic acid, antioxidants such as tocopherol, carotenoids, flavonoids, tannins, lignans, and saponins, cell activators such as alpha-hydroxy acids and beta-hydroxy acids, circulation promoters such as gamma-oryzanol and vitamin E derivatives, wound healing agents such as retinol and retinol derivatives, skin whitening agents such as arbutin, kojic acid, placenta extract, sulfur, ellagic acid, linoleic acid, tranexamic acid, and glutathione, cepharanthine, licorice extract, capsicum tincture, hinokitiol, iodized garlic extract, pyridoxine hydrochloride, DL-alpha-tocopherol, DL-alpha-tocopherol acetate, nicotinic acid, nicotinic acid derivatives, calcium pantothenate, and D-pantothenyl Examples of hair growth agents include alcohol, acetyl pantothenyl ethyl ether, biotin, allantoin, isopropyl methylphenol, estradiol, ethinyl estradiol, capronium chloride, benzalkonium chloride, diphenhydramine hydrochloride, tacanal, camphor, salicylic acid, nonylic acid vanillylamide, nonanoic acid vanillylamide, piroctone olamine, glyceryl pentadecanoate, L-menthol, mononitroguaiacol, resorcinol, γ-aminobutyric acid, benzethonium chloride, mexiletine hydrochloride, auxin, female hormones, cantharides tincture, cyclosporine, zinc pyrithione, hydrocholorthisone, minoxidil, polyoxyethylene sorbitan monostearate, peppermint oil, and Sasanishiki extract.

[0033] (10) Examples of antioxidants Examples of antioxidants include sodium bisulfite, sodium sulfite, erythorbic acid, sodium erythorbate, dilauryl thiodipropionate, tocopherol, tolylbiguanide, nordihydroguaiaretic acid, parahydroxyanisole, butylhydroxyanisole, dibutylhydroxytoluene, ascorbyl stearate, ascorbyl palmitate, octyl gallate, propyl gallate, carotenoids, flavonoids, tannins, lignans, saponin, and plant extracts known to have antioxidant effects, such as apple extract and clove extract.

[0034] (11) Examples of solvents Examples include purified water, ethanol, lower alcohols, ethers, LPG, fluorocarbons, N-methylpyrrolidone, fluoroalcohols, volatile linear silicones, and next-generation chlorofluorocarbons. [Example]

[0035] Examples of the present invention will be described below. Note that the examples shown below are provided to confirm the various actions and effects of the agent of the present invention obtained by the present invention, and the scope of the present invention is not limited to these products and manufacturing methods.

[0036] Example: Extract preparation Oryza Ceramide (registered trademark) manufactured by Oryza Oil & Fat Chemical Co., Ltd. was used as the starting material. The concentrate (10 g) was adsorbed onto silica gel (22 g) to produce Mabushi. This was fractionated by medium-pressure flash chromatography (Yamazen Corporation). Specifically, Mabushi was applied to a medium-pressure silica gel flash column (Universal Column Silica Gel, 3 L) and fractionated sequentially with the following solvents (1. hexane:ethyl acetate, 2. chloroform:methanol). An ELSD detector was used for detection. The separation conditions are shown in Table 1. [Table 1]

[0037] Fractions 26–33 eluted with chloroform:methanol (80:20) (10–20 min) were concentrated to obtain crude glucosylceramide. The resulting fractions were separated and purified by HPLC, and the peaks HPLC-2 to HPLC-11 shown in Figure 1 were collected (HPLC-12 appears after HPLC-11). An ODS column (PrepODS 20 × 250 mm, Inertsil) was used, and a differential refractometer was used for detection. The mobile phase was methanol, and the flow rate was 9 mL / min. The recorder settings were ATN 8 and chart speed 5 mm / min. Each peak was concentrated and dried to give HPLC-2 (23.6 mg), HPLC-3 (16 mg), HPLC-4 (117.6 mg), HPLC-5 (56.5 mg), HPLC-6 (32.2 mg), HPLC-6.5 (6.0 mg), HPLC-7 (29.1 mg), HPLC-7.5 (4.5 mg), HPLC-8 (5.8 mg), HPLC-9 (28.7 mg), HPLC-10 (8.9 mg), HPLC-11 (8.8 mg), and HPLC-12 (3.9 mg). The structure of glucosylceramide was determined by comparing the proton and carbon-13 NMR spectra and mass spectra of each component with literature values ​​(References 1 to 11 below) (Figure 2). Note that for HPLC-7, the structure was determined from data on similar components because literature was unavailable.

[0038] HPLC-2: GlcCer (d18:2(4E,8Z) / C18:0) Reference 1, HPLC-3: GlcCer (t18:1(8Z) / C20:0) Reference 5, HPLC-4: GlcCer (d18:2(4E,8Z) / C20:0) Reference 2, HPLC-5: GlcCer (d18:2(4E,8E) / C20:0) Reference 2, HPLC-6: GlcCer (t18:1(8Z) / C22:0) Reference 6, HPLC-6.5: GlcCer (d18:1(4E) / C20:0) Reference 8, HPLC-7: GlcCer (d18:2(4E,8Z) / C22:0), HPLC-7.5: GlcCer (d18:2(4E,8E) / C22:0) Reference 9, HPLC-8: GlcCer (t18:1(8Z) / C24:0) Reference 7, HPLC-9: GlcCer (d18:2(4E,8Z) / C24:0) Reference 3, HPLC-10: GlcCer (d18:2(4E,8E) / C24:0) Reference 4, HPLC-11 GlcCer (t18:1(8Z) / C26:0) Reference 11, HPLC-12: GlcCer (d18:2(4E,8Z) / C26:0) Reference 10 Identification literature) 1) Inagaki. et al., Chem. Pharm. Bull., 52(11), 1307-1311 (2004) 2) Jung JH et al., J. Nat. Prod., 59, 319-322 (1996) 3) Liu H. et al., Phytochemistry., 49(8), 2403-2408 (1998) 4) Ryu J. et al., Arch. Pharm. Res., 26(2), 138-142 (2003) 5) Tuntiwachwuttikul P. et al., Chem. Pharm. Bull., 52(1), 27-32 (2004) 6) Kang SS et al., Chem. Pharm. Bull., 49, 321-323 (2001) 7) Luo Y. et al., Lipids, 39(9), 907-913 (2004) 8) CAS No. 943147-66-8 9) Chen X. et al., Tetrahedron Lett., 43, 3529-3532 (2002). 10) Chinese patent CN103417555 A 11) Zhang WK ​​et al., Chem. Physiol. Lipids, 148, 77-83 (2007).

[0039] Test example: Evaluation of the melanin production inhibitory effect of glucosylceramide using B16 melanoma cells 1. Test Method (1) Cultivation method of mouse skin-derived melanoma cells (B16 cells) B16 (JCRB 0202) cells derived from mouse melanoma purchased from the JCRB Cell Bank were cultured and used for the experiment. The medium used was Dulbecco's Modified Eagles Medium (DMEM containing 1000 mg / L glucose) supplemented with 10 (v / v)% FBS (fetal bovine serum), 100 units / mL penicillin G, and 100 μg / mL streptomycin. Cells were cultured in 75 cm 2 The subculture was carried out in a culture flask at 37°C in the presence of 5% CO2. The cultured cells were washed twice with PBS (-) and then detached from the flask with 0.25 w / v% Trypsin-1 mM EDTA·4Na solution containing phenol red for use in the experiment. (2) Evaluation method for theophylline-induced melanin production inhibition test using B16 cells B16 cell suspension (5.0 × 10 4 Cells (100 μL / well) were seeded into 96-well plates (100 μL / well) and 48-well plates (200 μL / well) and cultured for one day. After culture, the medium was replaced with medium containing theophylline (2 mM) and the test substance. The test substance was dissolved in DMSO, and the final DMSO concentration in the medium was adjusted to 0.5% (v / v). After 3 days of culture, an MTT assay (96-well plate, cytotoxicity) and a melanin quantification assay (48-well plate) were performed. (3) MTT test To evaluate cytotoxicity, an MTT test was performed as follows. Specifically, 10 μL / well of MTT solution (5 mg / mL in PBS(-)) was added to the 96-well plate after incubation. After 4 hours of incubation, the formation of formazan crystals was confirmed and the medium was removed by aspiration. DMSO solution (100 μL / well) was then added. After stirring for approximately 30 minutes using a plate mixer, the OD value was measured using a microplate reader (measurement wavelength: 570 nm, reference wavelength: 660 nm). (4) Melanin quantification To evaluate the inhibitory effect on melanin production, melanin was quantified as follows. Specifically, after culturing, the medium in the 48-well plate was aspirated and distilled water (100 μL / well) was added. Then, ultrasonic treatment was performed to disrupt the melanin in the cells, and 6N sodium hydroxide (20 μL / well) was added. After the bubbles had subsided, the solution (100 μL / well) was transferred to a new 96-well plate, and the OD value was measured using a microplate reader (measurement wavelength: 405 nm, reference wavelength: 655 nm). The melanin production inhibition rate was calculated using the following formula.

number

[0040] Tables 2 and 3 show the evaluation results of kojic acid (positive control), which is known as a skin-whitening ingredient. Table 1 shows the results of melanin production inhibitory activity, and Table 2 shows the results of cytotoxicity (MTT test). [Table 2]

[0041] [Table 3]

[0042] GlcCer (d18:2 (4E, 8Z) / C18:0), GlcCer (t18:1 (8Z) / C20:0), GlcCer (d18:2 (4E, 8Z) / C20:0), GlcCer (d18:2 (4E, 8E) / C20:0), GlcCer (d18:1 (4E) / C20:0), GlcCer (t18:1(8Z) / C22:0), GlcCer (d18:2 (4E, 8Z) / C22:0), GlcCer (d18:2 (4E, 8E) / C22:0), GlcCer (t18:1 (8Z) / C24:0), GlcCer (d18:2 (4E, The evaluation results for GlcCer (d18:2 (4E, 8E) / C24:0), GlcCer (d18:2 (4E, 8Z) / C16:0), and GlcCer (d18:2 (4E, 8E) / C16:0) are shown in Tables 4 and 5 below. Table 4 shows the melanin production inhibitory effect, and Table 5 shows the results of the cytotoxicity test (MTT test).

[0043] [Table 4]

[0044] [Table 5]

[0045] 2. Measurement results and effects of the embodiment As can be seen from Table 2, kojic acid was found to have a concentration-dependent inhibitory effect on melanin production. Therefore, it was determined that this experiment was valid as an evaluation test for melanin production inhibitory activity. As shown in Table 4, GlcCer (d18:2(4E,8Z) / C18:0), GlcCer (d18:2(4E,8Z) / C20:0), GlcCer (d18:2(4E,8E) / C20:0), and GlcCer (d18:1(4E) / C20:0), which have substituted sphingadienine (4E, 8Z, or 8E) or sphingenine (4E), exhibited melanin production inhibitory effects (Table 4). Furthermore, these glucosylceramides did not exhibit cytotoxicity (Table 5). From the above, it was confirmed that glucosylceramides in which sphingadienine (4E, 8Z, or 8E) or sphingenine (4E) was substituted are useful as melanin production inhibitors (FIG. 3).

[0046] Formulation examples of the melanin production inhibitor (glucosylceramide) of the present invention are shown below. Note that the following formulation examples do not limit the present invention. Formulation example 1: Chewing gum Sugar 53.0wt% Gum base 20.0 Glucose 10.0 Starch syrup 16.0 Fragrance 0.5 Melanin production inhibitor 0.5 100.0wt%

[0047] Mixing example 2: Gummies Reduced starch syrup 40.0wt% Granulated sugar 20.0 Glucose 20.0 Gelatin 4.7 Wednesday 9.68 Yuzu juice 4.0 Yuzu flavor 0.6 Dye 0.02 Melanin production inhibitor 1.0 100.0wt%

[0048] Mixing example 3: Candy Sugar 50.0wt% Starch syrup 33.0 Wednesday 14.4 organic acid 2.0 Fragrance 0.2 Melanin production inhibitor 0.4 100.0wt%

[0049] Blending example 4: Yogurt (hard / soft) Milk 41.5wt% Skim milk powder 5.8 Sugar 8.0 Agar 0.15 Gelatin 0.1 Lactic acid bacteria 0.005 Melanin production inhibitor 0.4 fragrance trace amount water residue 100.0wt%

[0050] Formulation example 5: Soft drink High fructose corn syrup 30.0wt% Emulsifier 0.5 Melanin production inhibitor 0.3 Fragrance (appropriate amount) Purified water remainder 100.0wt%

[0051] Formulation example 6: Tablet confectionery Sugar 76.4wt% Glucose 19.0 Sucrose fatty acid ester 0.2 Melanin production inhibitor 0.5 Purified water 3.9 100.0wt%

[0052] Formulation example 7: Soft capsule Brown rice germ oil 47.0wt% Yuzu seed oil 40.0 Emulsifier 12.0 Melanin production inhibitor 1.0 100.0wt%

[0053] Formulation example 8: Tablets Lactose 54.0wt% Microcrystalline cellulose 30.0 Starch decomposition product 10.0 Glycerin fatty acid ester 5.0 Melanin production inhibitor 1.0 100.0wt%

[0054] Formulation example 9: Cosmetic cream Squalane 20.0wt% Beeswax 5.0 Refined Jojoba Oil 5.0 Glycerin 5.0 Glycerin Monostearate 2.0 Polyoxyethylene (20) sorbitan Monostearate 2.0 Melanin Production Inhibitor 2.0 Preservatives (appropriate amount) Fragrance (appropriate amount) Purified water remainder 100.0wt%

[0055] Formulation example 10: Lotion Ethanol 5.0wt% Glycerin 2.0 1,3-butylene glycol 2.0 Polyethylene oleyl ether 0.5 Sodium citrate 0.1 Citric acid 0.1 Melanin production inhibitor 0.1 Purified water remainder 100.0wt%

[0056] Formulation example 11: Body gel Macadamia nut oil 2.0wt% Octyldodecyl myristate 10.0 Methylphenylpolysiloxane 5.0 Behenyl Alcohol 3.0 Stearic Acid 3.0 Batyl alcohol 1.0 Glyceryl Monostearate 1.0 Polyoxyethylene sorbitol tetraoleate 2.0 Hydrogenated soybean phospholipid 1.0 Ceramide 0.1 Retinol Palmitate 0.1 Preservatives (appropriate amount) Centella asiatica extract 1.0 Melanin production inhibitor 1.0 1,3-butylene glycol 5.0 Purified water remainder 100.0wt%

[0057] Formulation example 12: Emulsion Squalane 4.0wt% Vaseline 2.5 Cetyl alcohol 2.0 Glycerin 2.0 Lipophilic Glyceryl Monostearate 1.0 Stearic Acid 1.0 L-Arginine 1.0 Melanin production inhibitor 0.5 Potassium hydroxide 0.1 fragrance trace amount Purified water remainder 100.0wt%

[0058] Formulation example 13: Bath additive (liquid) Propylene glycol 50.0wt% Ethanol 20.0 Sodium sulfate 5.0 Melanin production inhibitor 0.5 Lanolin 0.5 Avocado oil 0.5 Dye 1.5 Fragrance 22.0 100.0wt% [Industrial Applicability]

[0059] As described above, the present invention can provide a novel melanin production inhibitor.

Claims

[Claim 1] A melanin production inhibitor characterized by being GlcCer (d18:1(4E) / C20:0).

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