KR20220019903A - Composition for improving skin condition or angiostenosis comprising oat extract or avenanthramide - Google Patents

Abstract

The present invention relates to a composition for improving skin condition comprising an oat extract or avenanthramide, which is an indicator component of oats, as an active ingredient. The composition of the present invention can be used as a functional cosmetic material for skin condition improvement such as skin whitening, skin regeneration, skin elasticity enhancement, and skin wrinkle improvement, and is expected to be utilized as a therapeutic agent for vascular stenosis and pigmentation diseases.

Description

Composition for improving skin condition or vascular stenosis comprising oat extract or avenanthramide {Composition for improving skin condition or angiostenosis comprising oat extract or avenanthramide}

The present invention relates to a composition for improving skin condition comprising an oat extract or avenanthramide, which is an indicator component of oats, as an active ingredient.

When skin aging occurs, skin elasticity decreases and skin wrinkles increase. The decrease in skin elasticity and the formation of skin wrinkles are caused by decreased collagen synthesis and accelerated expression of MMP (Matrix metalloproteinase), an enzyme that decomposes collagen. it is known

In addition, the production of inflammation-inducing factors increases in skin cells due to aging or UV rays, and the biosynthesis of MMP increases due to an inflammatory reaction, resulting in collagen degradation, a decrease in skin elasticity, and the formation of skin wrinkles. Therefore, in order to reduce skin wrinkles and maintain elasticity, it is necessary to protect the skin by suppressing the inflammatory reaction and promoting skin regeneration from wounds.

On the other hand, a person's skin color is genetically determined according to the concentration and distribution of melanin in the skin, but is also affected by environmental or physiological conditions such as solar ultraviolet rays, fatigue, and stress. Melanin is produced through a non-enzymatic oxidation reaction after the enzyme tyrosinase acts on tyrosine, a kind of amino acid, to convert it into DOPA and dopaquinone.

Commonly known whitening ingredients include substances that inhibit tyrosinase enzyme activity, such as kojic acid and arbutin, hydroquinone, vitamin C (L-Ascorbic acid) and derivatives thereof and various There are plant extracts. By inhibiting the synthesis of melanin pigment, they can brighten the skin tone and realize skin whitening, and can also be used to improve skin hyperpigmentation such as spots or freckles caused by ultraviolet rays, hormones, or heredity. However, there is a problem in that the amount of use is limited due to safety issues such as irritation and redness when applied to the skin, or the actual effect cannot be expected because the effect is insignificant.

Therefore, there is a demand for the development of ingredients that are safe for the living body, stable active ingredients, and above all, have excellent effects on skin regeneration, wrinkles, skin elasticity and whitening.

On the other hand, oats are known to be rich in soluble fiber and protein, as well as vitamins and minerals. Avenanthramide, phenolic acid, and flavonoid are known as the most important phenolic compounds in oats, and the biological activities of antioxidant, anti-inflammatory, anti-allergic and anti-carcinogenic properties have been reported for phenolic compounds. . The antioxidant and anti-inflammatory activities of oats and its index components, avenanthramides A, B and C, have been previously reported, but as a skin protectant through tyrosinase inhibition such as whitening, pigmentation inhibition, and blemish pigment production inhibition. The role is unknown.

Republic of Korea Patent Registration No. 10-1761080

The present inventors found that oat extract and its index component, avenanthramide, inhibit the expression and enzyme activity of MMP-9, a factor related to skin elasticity, skin regeneration, and skin aging, and the activity of tyrosinase, a factor related to skin whitening. By confirming that and inhibiting melanin production, the present invention was completed. In addition, the present inventors have completed the present invention by confirming that oat extract and avenanthramide can improve blood vessel stenosis.

Accordingly, it is an object of the present invention to provide a cosmetic composition for improving skin condition comprising, as an active ingredient, an oat extract, or avenanthramide, which is an indicator component of oats, or a cosmetically acceptable salt thereof.

Another object of the present invention is to provide a food composition for improving skin condition comprising an oat extract, or avenanthramide, which is an indicator component of oats, or a pharmaceutically acceptable salt thereof as an active ingredient.

Another object of the present invention is to provide a pharmaceutical composition for preventing or treating skin pigmentation diseases, comprising an oat extract, or avenanthramide, which is an indicator component of oats, or a pharmaceutically acceptable salt thereof, as an active ingredient. .

Another object of the present invention is to provide a food composition for the prevention or improvement of vascular stenosis comprising an oat extract or avenanthramide, which is an indicator component of oats, or a pharmaceutically acceptable salt thereof as an active ingredient.

Another object of the present invention is to provide a pharmaceutical composition for preventing or treating vascular stenosis comprising an oat extract, or avenanthramide, which is an indicator component of oats, or a pharmaceutically acceptable salt thereof, as an active ingredient.

However, the technical problem to be achieved by the present invention is not limited to the above-mentioned tasks, and other tasks not mentioned can be clearly understood by those of ordinary skill in the art to which the present invention belongs from the following description. will be.

In order to achieve the object of the present invention, the present invention provides a cosmetic composition for improving skin condition comprising an oat extract or avenanthramide, which is an indicator component of oats, or a cosmetically acceptable salt thereof, as an active ingredient.

In addition, the present invention provides a food composition for improving skin condition comprising an oat extract, or avenanthramide, which is an indicator component of oats, or a pharmaceutically acceptable salt thereof as an active ingredient.

Furthermore, the present invention provides a cosmetic method for improving the skin condition of an individual using the composition.

In addition, the present invention provides the use of the composition for improving skin condition.

Furthermore, the present invention provides the use of the composition for the preparation of a preparation for improving skin conditions.

In addition, the present invention provides a pharmaceutical composition for preventing or treating skin pigmentation diseases, comprising an oat extract, or avenanthramide, which is an indicator component of oats, or a pharmaceutically acceptable salt thereof, as an active ingredient.

Furthermore, the present invention provides a method for preventing or treating skin pigmentation disease, comprising administering the pharmaceutical composition to an individual in need thereof.

In addition, the present invention provides a use for preventing, improving or treating skin pigmentation diseases of the pharmaceutical composition.

Furthermore, the present invention provides the use of the pharmaceutical composition for the preparation of a preparation for the prevention, improvement or treatment of skin pigmentation diseases.

In addition, the present invention provides a food composition for preventing or improving vascular stenosis comprising an oat extract or avenanthramide, which is an indicator component of oats, or a pharmaceutically acceptable salt thereof as an active ingredient.

In addition, the present invention provides a pharmaceutical composition for preventing or treating vascular stenosis comprising an oat extract or avenanthramide, which is an indicator component of oats, or a pharmaceutically acceptable salt thereof, as an active ingredient.

Furthermore, the present invention provides a method for preventing, improving or treating vascular stenosis, comprising administering the composition to a subject in need thereof.

In addition, the present invention provides the use of the composition for preventing, improving or treating vascular stenosis.

Furthermore, the present invention provides the use of the composition for the preparation of a preparation for the prevention, improvement or treatment of vascular stenosis.

In one embodiment of the present invention, the improvement of the skin condition may be skin whitening, skin regeneration, skin elasticity enhancement, or skin wrinkle improvement, but is not limited thereto.

In another embodiment of the present invention, the skin whitening includes spots, freckles, black spots, age spots, spots, milk coffee spots, nevus Ota, nevus blue, spots with hyperpigmentation, hyperpigmentation after drug use, and gestational brown spots (gravidic) chloasma), and may be to relieve one or more selected from the group consisting of hyperpigmentation after inflammation due to wounds or dermatitis, but is not limited thereto.

In another embodiment of the present invention, the avenanthramide may be avenanthramide A, avenanthramide B or avenanthramide C, but is not limited thereto.

In another embodiment of the present invention, the oat extract is alcohol having 1 to 6 carbon atoms, water, acetone, ether, benzene, chloroform, ethyl acetate, methylene chloride, hexane (hexane), cyclohexane (cyclohexane) , petroleum ether (petroleum ether), subcritical fluid, supercritical fluid, and may be extracted with a solvent selected from the group consisting of a mixed solvent thereof, but is not limited thereto.

In another embodiment of the present invention, the active ingredient may inhibit the activity of tyrosinase, but is not limited thereto.

In another embodiment of the present invention, the active ingredient may inhibit melanin production, but is not limited thereto.

In another embodiment of the present invention, the active ingredient may inhibit the expression of matrix metallopeptidase 9 (MMP-9) or its enzymatic activity, but is not limited thereto.

In another embodiment of the present invention, the skin pigmentation disease is melasma, freckles, black spots, age spots, spots, milk coffee spots, nevus Ota, blue nevus, hyperpigmentation spots, hyperpigmentation after drug use, gestational brown It may be one or more selected from the group consisting of hyperpigmentation after inflammation and hyperpigmentation after inflammation due to wounds or dermatitis, but is not limited thereto.

According to the present invention, oat extract and avenanthramide, an indicator component of oats, inhibit MMP-9 expression and enzymatic activity, and inhibit tyrosinase activity and melanin production. Therefore, the composition comprising the oat extract and avenanthramide of the present invention can be used as a functional cosmetic material for skin condition improvement such as skin whitening, skin regeneration, skin elasticity enhancement and skin wrinkle improvement. It is also expected to be used as a treatment for deposit disease.

Figure 1a shows the chemical structures of avenanthramides A, B and C.
Figure 1b shows the results of the MMT experiment of avenanthramides performed using human smooth muscle cells.
2 shows the results of evaluation of the effect of avenanthramide on cell migration and wound healing performed using human smooth muscle cells activated with TNF-α.
3a shows the results of evaluating the effect of avenanthramide on MMP-9 protein expression in human smooth muscle cells.
3b shows the results of evaluating the effect of avenanthramide on MMP-9 mRNA expression in human smooth muscle cells.
Figure 3c shows the results of evaluating the effect of avenanthramide on the MMP-9 enzyme activity performed using human smooth muscle cells.
4 shows the results of the MMT experiment of avenanthramides performed using skin cells.
5a shows the results of evaluating the inhibition of tyrosinase activity of avenanthramide in vitro.
Figure 5b shows the results of evaluating the inhibition of tyrosinase activity of avenanthramide using α-MSH-treated skin cells.
6 shows the results of evaluating the melanogenesis inhibitory activity of avenanthramide in α-MSH-treated skin cells.
7 shows the prediction result of the binding site of avenanthramide and tyrosinase through molecular docking simulation analysis.
8 schematically shows a mechanism for regulating the activity and production of MMP-9 through the MAPKs signaling pathway of oat extract and avenanthramide, an indicator component of oats, and inhibiting melanogenesis.

The present invention provides a cosmetic composition for improving skin condition comprising an oat extract or avenanthramide, which is an indicator component of oats, or a cosmetically acceptable salt thereof, as an active ingredient.

In the present invention, oats can be used without damaging their original form or used after performing a pretreatment process in consideration of the process speed and process (manufacturing) efficiency intended by those skilled in the art, and the pretreatment process includes, for example, For example, it may be subjected to conventional screening, washing, cutting, powdering, drying, and the like steps.

In the present invention, the oat extract may be obtained from the whole or a part of oats, for example, it may be obtained from a portion comprising one or more selected from the group consisting of stems, roots, leaves, fruits and flowers.

The oat extract may be extracted by a known natural product extraction method. For example, it may be extracted with one or more solvents selected from the group consisting of water, an organic solvent having 1 to 6 carbon atoms, and a subcritical or supercritical fluid. The organic solvent having 1 to 6 carbon atoms is alcohol, acetone, ether, benzene, chloroform, ethyl acetate, methylene chloride, having 1 to 6 carbon atoms. chloride), hexane (hexane), cyclohexane (cyclohexane) and petroleum ether (petroleum ether) may be at least one selected from the group consisting of, but is not limited thereto.

In the present invention, the oat extract of the present invention may be extracted with ethanol. As the extraction method, conventional extraction methods such as cold extraction, warm soaking, and heating using the above solvent can be used. The concentration of the ethanol is not particularly limited, for example, 10 to 70%, 10 to 60%, 10 to 50%, 10 to 40%, 10 to 30%, 10 to 25%, 10 to 20%, 15 to 50%, 15 to 40%, 15 to 30%, 15 to 25% or 15 to 20% of the ethanol extract.

In the present invention, the avenanthramide may be avenanthramide A, avenanthramide B or avenanthramide C, but is not limited thereto. Avenanthramide is known as an indicator component of oats, and the chemical structures of avenanthramides A, B and C are shown in FIG. 1A.

The cosmetic composition of the present invention may include a cosmetically acceptable salt of avenanthramide. As used herein, the term “cosmetically acceptable salt” includes salts derived from cosmetically acceptable inorganic acids, organic acids or bases. In addition, the term "pharmaceutically acceptable salt" includes salts derived from pharmaceutically acceptable inorganic acids, organic acids or bases, and the term "food pharmaceutically acceptable salts" refers to pharmaceutically acceptable organic acids, inorganic acids or bases. salts derived from

Examples of suitable acids include hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, perchloric acid, fumaric acid, maleic acid, phosphoric acid, glycolic acid, lactic acid, salicylic acid, succinic acid, toluene-p-sulfonic acid, tartaric acid, acetic acid, citric acid, methanesulfonic acid, formic acid , benzoic acid, malonic acid, gluconic acid, naphthalene-2-sulfonic acid, benzenesulfonic acid, and the like. Acid addition salts can be prepared by conventional methods, for example, by dissolving the compound in an aqueous solution of an excess of acid, and precipitating the salt using a water-miscible organic solvent such as methanol, ethanol, acetone or acetonitrile. It can also be prepared by heating an equimolar amount of the compound and an acid or alcohol in water and then evaporating the mixture to dryness, or by suction filtration of the precipitated salt.

Salts derived from suitable bases may include, but are not limited to, alkali metals such as sodium and potassium, alkaline earth metals such as magnesium, and ammonium. The alkali metal or alkaline earth metal salt can be obtained, for example, by dissolving the compound in an excess alkali metal hydroxide or alkaline earth metal hydroxide solution, filtering the undissolved compound salt, and then evaporating and drying the filtrate. In this case, it is pharmaceutically suitable to prepare a sodium, potassium or calcium salt as the metal salt, and the corresponding silver salt can be obtained by reacting an alkali metal or alkaline earth metal salt with a suitable silver salt (eg, silver nitrate).

As used herein, the term “improvement” refers to any action of reducing a parameter related to a desired skin condition, for example, the degree of a symptom by administration of the composition according to the present invention.

In the present invention, the skin condition improvement may be skin whitening, skin regeneration, skin elasticity enhancement, or skin wrinkle improvement, but is not limited thereto.

In the present invention, the skin whitening is, for example, spots, freckles, black spots, age spots, spots, milk coffee spots, nevus Ota, nevus blue, hyperpigmentation spots, hyperpigmentation after drug use, gestational brown spots (gravidic) chloasma), or to relieve (ameliorate) hyperpigmentation after inflammation due to wounds (or dermatitis).

In the present invention, the active ingredient may inhibit the activity of tyrosinase, but is not limited thereto (see Example 6).

In addition, the active ingredient may inhibit melanin production, but is not limited thereto (see Example 7).

In addition, the active ingredient may inhibit the expression of MMP-9 or its enzymatic activity, but is not limited thereto (see Example 4).

The dosage form of the cosmetic composition according to the present invention includes skin lotion, skin softener, skin toner, astringent, lotion, milk lotion, moisture lotion, nourishing lotion, massage cream, nourishing cream, mist, moisture cream, hand cream, hand lotion, foundation, It may be in the form of essence, nutritional essence, pack, soap, cleansing foam, cleansing lotion, cleansing cream, cleansing oil, cleansing balm, body lotion or body cleanser.

The cosmetic composition of the present invention may further include a composition selected from the group consisting of water-soluble vitamins, oil-soluble vitamins, high molecular weight peptides, high molecular weight polysaccharides, and sphingolipids.

As water-soluble vitamins, any type of vitamin can be used as long as it can be formulated in cosmetics. Examples include vitamin B1, vitamin B2, vitamin B6, pyridoxine, pyridoxine hydrochloride, vitamin B12, pantothenic acid, nicotinic acid, nicotinic acid amide, folic acid, vitamin C, and vitamin H. and their salts (thiamine hydrochloride, sodium ascorbate salt, etc.) and derivatives (ascorbic acid-2-phosphate sodium salt, ascorbic acid-2-phosphate magnesium salt, etc.) are also water-soluble vitamins that can be used in the present invention. Included. Water-soluble vitamins can be obtained by conventional methods, such as a microbial transformation method, a purification method from a microbial culture, an enzymatic method, or a chemical synthesis method.

The oil-soluble vitamin may be any type of vitamin that can be formulated in cosmetics. Examples thereof include vitamin A, carotene, vitamin D2, vitamin D3, and vitamin E (d1-alpha tocopherol, d-alpha tocopherol, d-alpha tocopherol). , their derivatives (ascorbine palmitate, ascorbine stearate, ascorbine dipalmitate, dl-alpha tocopherol acetate, dl-alpha tocopherol nicotinic acid, vitamin E, DL-pantothenyl alcohol, D-pantothenyl alcohol, pantothenyl ethyl ethers, etc.) are also included in the oil-soluble vitamins used in the present invention. Oil-soluble vitamins can be obtained by conventional methods such as a microbial transformation method, a purification method from a microbial culture, and an enzyme or chemical synthesis method.

The macromolecular peptide may be any compound as long as it can be formulated in cosmetics, and examples thereof include collagen, hydrolyzed collagen, gelatin, elastin, hydrolyzed elastin, and keratin. Polymeric peptides can be purified and obtained by conventional methods such as purification from a microbial culture solution, enzyme method or chemical synthesis method, or can be purified and used from natural products such as dermis of pigs or cattle, silk fiber of silkworms, etc.

The high molecular weight polysaccharide may be any compound as long as it can be formulated in cosmetics, and examples thereof include hydroxyethyl cellulose, xanthan gum, sodium hyaluronate, chondroitin sulfate or a salt thereof (sodium salt, etc.). For example, chondroitin sulfate or a salt thereof can be used after purification from mammals or fish.

The sphingolipid may be any as long as it can be formulated in cosmetics, and examples thereof include ceramide, phytosphingosine, and sphingoglycolipid. Sphingo lipids can be purified by conventional methods from mammals, fish, shellfish, yeast or plants, or can be obtained by chemical synthesis.

In the cosmetic composition of the present invention, in addition to the above essential ingredients, other ingredients usually blended in cosmetics may be blended as needed.

Other ingredients that may be added include oils and fats, moisturizers, emollients, surfactants, organic and inorganic pigments, organic powders, UV absorbers, preservatives, bactericides, antioxidants, plant extracts, pH adjusters, alcohols, colorants, fragrances, A blood circulation promoter, a cooling agent, a restrictive agent, purified water, etc. are mentioned.

Examples of the fats and oils include ester fats and oils, hydrocarbon oils, silicone oils, fluorine oils, animal fats, and vegetable oils.

As ester-based fats and oils, tri2-ethylhexanoate glyceryl, 2-ethylhexanoate cetyl, isopropyl myristate, butyl myristate, isopropyl palmitate, ethyl stearate, octyl palmitate, isocetyl isostearate, stearic acid Butyl, ethyl linoleate, isopropyl linoleate, ethyl oleate, isocetyl myristate, isostearyl myristate, isostearyl palmitate, octyldodecyl myristate, isocetyl isostearate, diethyl sebacate, adipine Diisopropyl acid, isoalkyl neopentanoate, tri(caprylic acid, capric acid) glyceryl, trimethylol propane tri-2-ethylhexanoate, trimethylol propane triisostearate, pentaelysitol tetra2-ethylhexanoate , cetyl caprylate, decyl laurate, hexyl laurate, decyl myristate, myristyl myristate, cetyl myristate, stearyl stearate, decyl oleate, cetyl ricinoleate, isostea laurate Lil, isotridecyl myristate, isocetyl palmitate, octyl stearate, isocetyl stearate, isodecyl oleate, octyldodecyl oleate, octyldodecyl linoleate, isostearic acid isopropyl, 2-ethylhexanoate cetoster Aryl, 2-ethyl stearyl hexanoate, hexyl isostearate, ethylene glycol dioctanoate, ethylene glycol dioleate, propylene glycol dicapric acid, di(caprylic acid, capric acid) propylene glycol, dicaprylic acid propylene glycol, dicapric acid Neopentyl glycol phosphate, neopentyl glycol dioctanoate, glyceryl tricaprylate, glyceryl triundecyl, glyceryl triisopalmitate, glyceryl triisostearate, octyldodecyl neopentanoate, isostearyl octanoate , isononanoate octyl, neodecanoate hexyldecyl, neodecanoate octyldodecyl, isostearate isocetyl, isostearyl isostearate, isostearic acid octyldecyl, polyglycerol oleate, polyglycerol isostearate, Triisocetyl citrate, triisoalkyl citrate, triisooctyl citrate, lauryl lactate, myristyl lactate, cetyl lactate, octyldecyl lactate, triethyl citrate, acetyl triethyl citrate, acetyl tributyl citrate, trioctyl citrate, dimalate Isostearyl, 2-ethylhexyl hydroxystearate, di2-ethylhexyl succinate, diisobutyl adipate, diisopropyl sebacate, dioctyl sebacate, Cholesteryl stearate, cholesteryl isostearate, cholesteryl hydroxystearate, cholesteryl oleate, dihydrocholesteryl oleate, phytsteryl isostearate, phytsteryl oleate, 12-stealoyl Ester type, such as hydroxy stearate isocetyl, 12-stealoyl hydroxy stearyl stearate, and 12-stealoyl hydroxy stearate isostearyl, etc. are mentioned.

Examples of hydrocarbon-based fats and oils include hydrocarbon-based fats and oils such as squalene, liquid paraffin, alpha-olefin oligomer, isoparaffin, ceresin, paraffin, liquid isoparaffin, polybudene, microcrystalline wax, petrolatum.

Examples of silicone-based oils and fats include polymethyl silicone, methylphenyl silicone, methylcyclopolysiloxane, octamethylpolysiloxane, decamethylpolysiloxane, dodecamethylcyclosiloxane, dimethylsiloxane/methylcetyloxysiloxane copolymer, dimethylsiloxane/methylstealloxysiloxane copolymer, and alkyl Modified silicone oil, amino modified silicone oil, etc. are mentioned.

Perfluoropolyether etc. are mentioned as fluorine-type fats and oils.

As animal or vegetable oils, avocado oil, almond oil, olive oil, sesame oil, rice bran oil, saffron oil, soybean oil, corn oil, rapeseed oil, passerine oil, palm kernel oil, palm oil, castor oil, sunflower oil, grape seed oil , Cottonseed Oil, Palm Oil, Kukui Nut Oil, Wheat Germ Oil, Rice Germ Oil, Shea Butter, Wormwood Colostrum, Marcus Damien Nut Oil, Meadow Oil, Egg Yolk Oil, Tallow Oil, Horse Oil, Mink Oil, Orange Raffy Oil, Jojoba Oil , animal or plant oils and fats such as candelabra wax, carnauba wax, liquid lanolin, and hydrogenated castor oil.

Examples of the moisturizing agent include a water-soluble low-molecular moisturizer, a fat-soluble molecular moisturizer, a water-soluble polymer, and a fat-soluble polymer.

As water-soluble low molecular weight moisturizers, serine, glutamine, sorbitol, mannitol, pyrrolidone-sodium carboxylate, glycerin, propylene glycol, 1,3-butylene glycol, ethylene glycol, polyethylene glycol B (polymerization degree n = 2 or more), polypropylene Glycol (polymerization degree n=2 or more), polyglycerol B (polymerization degree n=2 or more), lactic acid, lactic acid salt, etc. are mentioned.

Cholesterol, cholesterol ester, etc. are mentioned as a fat-soluble low molecular weight humectant.

Examples of the water-soluble polymer include carboxyvinyl polymer, polyaspartate, tragacanth, xanthan gum, methyl cellulose, hydroxymethyl cellulose, hydroxyethyl cellulose, hydroxypropyl cellulose, carboxymethyl cellulose, water-soluble chitin, chitosan, and dextrin. can

Examples of the fat-soluble polymer include polyvinylpyrrolidone/eicocene copolymer, polyvinylpyrrolidone/hexadecene copolymer, nitrocellulose, dextrin fatty acid ester, and high molecular weight silicone.

Examples of the emollient include long-chain acylglutamic acid cholesteryl ester, hydroxystearic acid cholesteryl, 12-hydroxystearic acid, stearic acid, rosin acid, and lanolin fatty acid cholesteryl ester.

Nonionic surfactants, anionic surfactants, cationic surfactants, amphoteric surfactants, etc. are mentioned as surfactant.

Nonionic surfactants include self-emulsifying glycerin monostearate, propylene glycol fatty acid ester, glycerin fatty acid ester, polyglycerol fatty acid ester, sorbitan fatty acid ester, POE (polyoxyethylene) sorbitan fatty acid ester, POE sorbitan fatty acid ester, POE Glycerin fatty acid ester, POE alkyl ether, POE fatty acid ester, POE hydrogenated castor oil, POE castor oil, POE/POP (polyoxyethylene/polyoxypropylene) copolymer, POE/POP alkyl ether, polyether-modified silicone, lauric acid alkanolamides, alkylamine oxides, hydrogenated soybean phospholipids, and the like.

Examples of the anionic surfactant include fatty acid soap, alpha-acyl sulfonate, alkyl sulfonate, alkyl allyl sulfonate, alkyl naphthalene sulfonate, alkyl sulfate, POE alkyl ether sulfate, alkyl amide sulfate, alkyl phosphate, POE alkyl phosphate, and alkyl amide phosphate. , alkyloylalkyltaurine salts, N-acylamino acid salts, POE alkylethercarboxylate salts, alkylsulfosuccinate salts, alkylsulfoacetate sodium, acylated hydrolyzed collagen peptide salts, perfluoroalkylphosphate esters, and the like. .

Examples of the cationic surfactant include alkyltrimethylammonium chloride, stearyltrimethylammonium chloride, stearyltrimethylammonium bromide, cetostearyltrimethylammonium chloride, distearyldimethylammonium chloride, stearyldimethylbenzylammonium chloride, behenyltrimethylammonium bromide, chloride Benzalkonium, diethylaminoethylamide stearate, dimethylaminopropylamide stearate, quaternary ammonium salt of lanolin derivative, etc. are mentioned.

Examples of the amphoteric surfactant include carboxybetaine type, amidebetaine type, sulfobetaine type, hydroxysulfobetaine type, amidesulfobetaine type, phosphobetaine type, aminocarboxylate type, imidazoline derivative type, amidamine type, etc. Amphoteric surfactant etc. are mentioned.

Examples of organic and inorganic pigments include silicic acid, silicic anhydride, magnesium silicate, talc, sericite, mica, kaolin, bengala, clay, bentonite, titanium-coated mica, bismuth oxychloride, zirconium oxide, magnesium oxide, zinc oxide, titanium oxide, aluminum oxide. inorganic pigments such as calcium sulfate, barium sulfate, magnesium sulfate, calcium carbonate, magnesium carbonate, iron oxide, ultramarine blue, chromium oxide, chromium hydroxide, calamine, and complexes thereof; Polyamide, polyester, polypropylene, polystyrene, polyurethane, vinyl resin, urea resin, phenol resin, fluororesin, silicon resin, acrylic resin, melamine resin, epoxy resin, polycarbonate resin, divinylbenzene/styrene copolymer, and organic pigments such as silk powder, cellulose, CI pigment yellow and CI pigment orange, and composite pigments of these inorganic pigments and organic pigments.

Examples of the organic powder include metal soaps such as calcium stearate; alkyl phosphate metal salts such as sodium zinc cetylate, zinc laurylate, and calcium laurylate; acylamino acid polyvalent metal salts such as calcium N-lauroyl-beta-alanine, zinc N-lauroyl-beta-alanine, and calcium N-lauroyl glycine; amidesulfonic acid polyvalent metal salts such as calcium N-lauroyl-taurine and calcium N-palmitoyl-taurine; N such as N-epsilon-lauroyl-L-lysine, N-epsilon-palmitoylizine, N-alpha-paritoylolnithine, N-alpha-lauroylarginine, N-alpha-hydrogenated beef fatty acid acylarginine, etc. -acyl basic amino acids; N-acyl polypeptides such as N-lauroylglycylglycine; alpha-amino fatty acids such as alpha-aminocaprylic acid and alpha-aminolauric acid; Polyethylene, polypropylene, nylon, polymethyl methacrylate, polystyrene, a divinylbenzene-styrene copolymer, ethylene tetrafluoride, etc. are mentioned.

As UV absorbers, para-aminobenzoic acid, para-aminobenzoate ethyl, para-aminobenzoate amyl, para-aminobenzoate octyl, ethylene glycol salicylate, phenyl salicylate, octyl salicylate, benzyl salicylate, butyl salicylate, homomentyl salicylate, benzyl cinnamate , para-methoxycinnamic acid-2-ethoxyethyl, para-methoxycinnamic acid octyl, dipara-methoxycinnamic acid mono-2-ethylhexaneglyceryl, para-methoxycinnamic acid isopropyl, diisopropyl diisopropyl cinnamic acid ester mixture, uro Cannic acid, ethyl urokanate, hydroxymethoxybenzophenone, hydroxymethoxybenzophenonesulfonic acid and salts thereof, dihydroxymethoxybenzophenone, sodium dihydroxymethoxybenzophenonedisulfonate, dihydroxybenzophenone , tetrahydroxybenzophenone, 4-tert-butyl-4'-methoxydibenzoylmethane, 2,4,6-trianilino-p-(carbo-2'-ethylhexyl-1'-oxy)-1 ,3,5-triazine, 2-(2-hydroxy-5-methylphenyl)benzotriazole, etc. are mentioned.

Examples of disinfectants include hinokitiol, triclosan, trichlorohydroxydiphenyl ether, chlorhexidine gluconate, phenoxyethanol, resorcin, isopropylmethylphenol, azulene, salicylic acid, zinc phyllithione, benzalkonium chloride, photosensitizer. So, No. 301, mononitroguaial sodium, undecyrenic acid, etc. are mentioned.

Examples of the antioxidant include butylhydroxyanisole, propyl gallic acid, and ellisorbic acid.

Examples of the pH adjuster include citric acid, sodium citrate, malic acid, sodium malate, fmalic acid, sodium fumarate, succinic acid, sodium succinate, sodium hydroxide, sodium monohydrogen phosphate, and the like.

As alcohol, higher alcohols, such as cetyl alcohol, are mentioned.

In addition, blending components that may be added other than this are not limited thereto, and any of the above components can be blended within a range that does not impair the object and effect of the present invention, but 0.01-5% by weight or 0.01-3 based on the total weight % by weight.

When the formulation of the present invention is a lotion, paste, cream or gel, animal fiber, vegetable fiber, wax, paraffin, starch, tracanth, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide, etc. can be used

When the formulation of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used as a carrier component. In particular, in the case of a spray, additional chlorofluorohydrocarbon, propane /may contain propellants such as butane or dimethyl ether.

When the formulation of the present invention is a solution or emulsion, a solvent, solvating agent or emulsifying agent is used as a carrier component, for example, water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1 ,3-butylglycol oil, glycerol fatty esters, fatty acid esters of polyethylene glycol or sorbitan.

When the formulation of the present invention is a suspension, as a carrier component, water, a liquid diluent such as ethanol or propylene glycol, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, microcrystalline Cellulose, aluminum metahydroxide, bentonite, agar or tracanth may be used.

When the formulation of the present invention is a surfactant-containing cleansing agent, aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyl taurate, sarcosinate, fatty acid amide as carrier components Ether sulfate, alkylamidobetaine, aliphatic alcohol, fatty acid glyceride, fatty acid diethanolamide, vegetable oil, linolin derivative or ethoxylated glycerol fatty acid ester and the like may be used.

As another aspect of the present invention, the present invention provides a food composition for improving skin condition comprising an oat extract, or avenanthramide, which is an indicator component of oats, or a pharmaceutically acceptable salt thereof, as an active ingredient.

In addition, as another aspect of the present invention, the present invention provides a food composition for preventing or improving vascular stenosis comprising an oat extract or avenanthramide, which is an indicator component of oats, or a pharmaceutically acceptable salt thereof as an active ingredient. to provide.

In the present invention, the food composition of the present invention may be a health functional food composition. The health functional food may have the advantage of having a more excellent effect by ingesting it in the form of an inner beauty food. The inner beauty (inner beauty) refers to food that is called 'eating cosmetics or beauty food', and refers to food that absorbs various ingredients good for the skin into the body and changes the skin constitution to make it healthy. You can choose and consume inner beauty food that is suitable for each individual by considering your skin condition and lifestyle. For example, when cosmetics and inner beauty food are mixed, the effect is significantly increased compared to using only cosmetics or pharmaceuticals, so that it can have the advantage of being able to see a more effective skin condition improvement effect.

When the active ingredient is used as a food additive, the active ingredient may be added as it is or used together with other foods or food ingredients, and may be appropriately used according to a conventional method. The mixed amount of the active ingredient may be appropriately determined according to the purpose of use (prevention, health or therapeutic treatment).

There is no particular limitation on the type of the food. Examples of foods to which the above substances can be added include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gums, dairy products including ice cream, various soups, beverages, tea, drinks, There are alcoholic beverages and vitamin complexes, and includes all health functional foods in the ordinary sense.

The health beverage composition according to the present invention may contain various flavoring agents or natural carbohydrates as additional ingredients, as in a conventional beverage. The above-mentioned natural carbohydrates are monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol and erythritol. As the sweetener, natural sweeteners such as taumartin and stevia extract, synthetic sweeteners such as saccharin and aspartame, and the like can be used. The proportion of the natural carbohydrate is generally about 0.01-0.20 g or about 0.04-0.10 g per 100 mL of the composition of the present invention.

In addition to the above, the composition of the present invention includes various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, Carbonating agents used in carbonated beverages, etc. may be contained. In addition, the composition of the present invention may contain pulp for the production of natural fruit juice, fruit juice beverage, and vegetable beverage. These components may be used independently or in combination. The proportion of these additives is not critical, but is generally selected in the range of 0.01-0.20 parts by weight per 100 parts by weight of the composition of the present invention.

As another aspect of the present invention, the present invention is for preventing or treating skin pigmentation disease or vascular stenosis comprising an oat extract or avenanthramide, which is an indicator component of oats, or a pharmaceutically acceptable salt thereof as an active ingredient A pharmaceutical composition is provided.

Further, as another aspect of the present invention, the present invention provides a method for preventing or treating a skin pigmentation disease or vascular stenosis comprising administering the pharmaceutical composition to an individual.

As used herein, the term "subject" refers to a subject in need of treatment for a disease, and more specifically, a human or non-human primate, mouse, rat, dog, cat, horse. , and mammals such as cattle.

As used herein, the term “administration” refers to providing a given composition of the present invention to a subject by any suitable method.

As used herein, the term “prevention” refers to any action that inhibits or delays the onset of a desired disease, and “treatment” refers to a desired disease and metabolic abnormalities resulting therefrom by administration of the pharmaceutical composition according to the present invention. It refers to any action that improves or changes to a favorable condition.

As used herein, the term "skin pigmentation disease" refers to any disease or disease of the skin in which discoloration of the skin or proliferation of abnormally pigmented cells occurs, and skin pigmentation may include excessive or unwanted pigmentation. can Factors such as aging, environmental stress and UV exposure can be potential causes of skin pigmentation. The skin pigmentation of the present invention may be due to an increase in one or more various types of melanin biosynthesized in the skin and/or hair follicles and deposited on the hair or skin compared to the patient's basic pigmentation, such as melasma, freckles, black spots, age spots , moles, milk coffee spots, nevus of Ota, nevus blue, spots of hyperpigmentation, hyperpigmentation after drug use, gravidic chloasma, and hyperpigmentation after inflammation due to wound or dermatitis It may be one or more, but is not limited thereto.

The content of the active ingredient in the composition of the present invention can be appropriately adjusted depending on the symptoms of the disease, the degree of progression of the symptoms, the condition of the patient, etc., for example, 0.0001 to 99.9% by weight, or 0.001 to 50% by weight based on the total weight of the composition. However, the present invention is not limited thereto. The content ratio is a value based on the dry amount from which the solvent is removed.

The pharmaceutical composition according to the present invention may further include suitable carriers, excipients and diluents commonly used in the preparation of pharmaceutical compositions. The excipient may be, for example, at least one selected from the group consisting of a diluent, a binder, a disintegrant, a lubricant, an adsorbent, a humectant, a film-coating material, and a controlled-release additive.

The pharmaceutical composition according to the present invention can be prepared according to a conventional method, respectively, in powders, granules, sustained-release granules, enteric granules, liquids, eye drops, elsilic, emulsions, suspensions, alcohols, troches, fragrances, and limonaade. , tablets, sustained release tablets, enteric tablets, sublingual tablets, hard capsules, soft capsules, sustained release capsules, enteric capsules, pills, tinctures, soft extracts, dry extracts, fluid extracts, injections, capsules, perfusates, It can be formulated and used in the form of external preparations such as warning agents, lotions, pasta agents, sprays, inhalants, patches, sterile injection solutions, or aerosols, and the external preparations are creams, gels, patches, sprays, ointments, warning agents. , lotion, liniment, pasta, or cataplasma.

Carriers, excipients and diluents that may be included in the pharmaceutical composition according to the present invention include lactose, dextrose, sucrose, oligosaccharide, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum acacia, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil.

In the case of formulation, it is prepared using commonly used diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrants, and surfactants.

Corn starch, potato starch, wheat starch, lactose, sucrose, glucose, fructose, di-mannitol, precipitated calcium carbonate, synthetic aluminum silicate, phosphoric acid as additives for tablets, powders, granules, capsules, pills, and troches according to the present invention Calcium monohydrogen, calcium sulfate, sodium chloride, sodium hydrogen carbonate, purified lanolin, microcrystalline cellulose, dextrin, sodium alginate, methyl cellulose, sodium carboxymethyl cellulose, kaolin, urea, colloidal silica gel, hydroxypropyl starch, hydroxypropyl methyl excipients such as cellulose, 1928, 2208, 2906, 2910, propylene glycol, casein, calcium lactate, and Primogel; Gelatin, gum arabic, ethanol, agar powder, cellulose acetate phthalate, carboxymethylcellulose, calcium carboxymethylcellulose, glucose, purified water, sodium caseinate, glycerin, stearic acid, sodium carboxymethylcellulose, sodium methylcellulose, methylcellulose, microcrystalline cellulose, dextrin , hydroxycellulose, hydroxypropyl starch, hydroxymethylcellulose, purified shellac, starch powder, hydroxypropylcellulose, hydroxypropylmethylcellulose, polyvinyl alcohol, polyvinylpyrrolidone, etc. Hydroxypropyl methylcellulose, corn starch, agar powder, methylcellulose, bentonite, hydroxypropyl starch, sodium carboxymethylcellulose, sodium alginate, calcium carboxymethylcellulose, calcium citrate, sodium lauryl sulfate, silicic anhydride, 1-hydroxy Propyl cellulose, dextran, ion exchange resin, polyvinyl acetate, formaldehyde treated casein and gelatin, alginic acid, amylose, guar gum, sodium bicarbonate, polyvinylpyrrolidone, calcium phosphate, gelled starch, gum arabic, Disintegrants such as amylopectin, pectin, sodium polyphosphate, ethyl cellulose, sucrose, magnesium aluminum silicate, di-sorbitol solution, light anhydrous silicic acid; Calcium stearate, magnesium stearate, stearic acid, hydrogenated vegetable oil, talc, lycopodite, kaolin, petrolatum, sodium stearate, cacao butter, sodium salicylate, magnesium salicylate, polyethylene glycol 4000,6000, liquid paraffin, hydrogenated soybean oil (Lubri) wax), aluminum stearate, zinc stearate, sodium lauryl sulfate, magnesium oxide, macrogol, synthetic aluminum silicate, silicic anhydride, higher fatty acid, higher alcohol, silicone oil, paraffin oil, polyethylene glycol fatty acid ether, starch, sodium chloride, Lubricating agents such as sodium acetate, sodium oleate, dl-leucine, light anhydrous silicic acid may be used.

As additives for the liquid formulation according to the present invention, water, diluted hydrochloric acid, diluted sulfuric acid, sodium citrate, monostearate sucrose, polyoxyethylene sorbitol fatty acid esters (Twinester), polyoxyethylene monoalkyl ethers, lanolin ethers, Lanolin esters, acetic acid, hydrochloric acid, aqueous ammonia, ammonium carbonate, potassium hydroxide, sodium hydroxide, prolamine, polyvinylpyrrolidone, ethyl cellulose, sodium carboxymethyl cellulose, etc. can be used.

In the syrup according to the present invention, a sucrose solution, other sugars or sweeteners may be used, and if necessary, a fragrance, colorant, preservative, stabilizer, suspending agent, emulsifying agent, thickening agent, etc. may be used.

Purified water may be used in the emulsion according to the present invention, and if necessary, an emulsifier, preservative, stabilizer, fragrance, etc. may be used.

Suspending agents such as acacia, tragacantha, methylcellulose, carboxymethylcellulose, sodium carboxymethylcellulose, microcrystalline cellulose, sodium alginate, hydroxypropylmethylcellulose, 1828, 2906, and 2910 may be used in the suspending agent according to the present invention, If necessary, surfactants, preservatives, stabilizers, colorants, and fragrances may be used.

The injection according to the present invention includes distilled water for injection, 0.9% sodium chloride injection, Ringel injection, dextrose injection, dextrose + sodium chloride injection, PEG (PEG), lactated Ringel injection, ethanol, propylene glycol, non-volatile oil-sesame oil , solvents such as cottonseed oil, peanut oil, soybean oil, corn oil, ethyl oleate, isopropyl myristate, and benzene benzoate; Solubilizing aids such as sodium benzoate, sodium salicylate, sodium acetate, urea, urethane, monoethylacetamide, butazolidine, propylene glycol, tweens, nijeongtinamide, hexamine, and dimethylacetamide; Weak acids and their salts (acetic acid and sodium acetate), weak bases and their salts (ammonia and ammonium acetate), organic compounds, proteins, buffers such as albumin, peptone, gum; isotonic agents such as sodium chloride; Stabilizers such as sodium bisulfite (NaHSO ₃ ) carbon dioxide gas, sodium metabisulfite (Na ₂ S ₂ O ₃ ), sodium sulfite (Na ₂ SO ₃ ), nitrogen gas (N ₂ ), ethylenediaminetetraacetic acid; sulphating agents such as sodium bisulfide 0.1%, sodium formaldehyde sulfoxylate, thiourea, disodium ethylenediaminetetraacetate, acetone sodium bisulfite; analgesic agents such as benzyl alcohol, chlorobutanol, procaine hydrochloride, glucose, and calcium gluconate; suspending agents such as SiMC sodium, sodium alginate, Tween 80, or aluminum monostearate.

The suppository according to the present invention includes cacao fat, lanolin, witepsol, polyethylene glycol, glycerogelatin, methyl cellulose, carboxymethyl cellulose, a mixture of stearic acid and oleic acid, Subanal, cottonseed oil, peanut oil, palm oil, cacao butter + Cholesterol, Lecithin, Lanet Wax, Glycerol Monostearate, Tween or Span, Imhausen, Monolene (Propylene Glycol Monostearate), Glycerin, Adeps Solidus, Butyrum Tego -G), Cebes Pharma 16, Hexalide Base 95, Cotomar, Hydroxote SP, S-70-XXA, S-70-XX75 (S-70-XX95), Hydro Hydrokote 25, Hydrokote 711, Idropostal, Massa estrarium, A, AS, B, C, D, E, I, T, Massa-MF, Masupol, Masupol-15, Neosupostal-N, Paramound-B, Suposiro (OSI, OSIX, A, B, C, D, H, L), Suppository IV type (AB, B, A, BC, BBG, E, BGF, C, D, 299), supostal (N, Es), Wecobi (W, R, S, M, Fs), tester triglyceride base (TG-95, MA, 57) and The same mechanism may be used.

Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc., and these solid preparations include at least one excipient in the extract, for example, starch, calcium carbonate, sucrose ( It is prepared by mixing sucrose), lactose, or gelatin. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used.

Liquid formulations for oral administration include suspensions, internal solutions, emulsions, syrups, etc. In addition to water and liquid paraffin, which are commonly used simple diluents, various excipients such as wetting agents, sweeteners, fragrances, and preservatives may be included. there is. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Non-aqueous solvents and suspending agents include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate.

The pharmaceutical composition according to the present invention is administered in a pharmaceutically effective amount. In the present invention, "pharmaceutically effective amount" means an amount sufficient to treat a disease at a reasonable benefit/risk ratio applicable to medical treatment, and the effective dose level is determined by the type, severity, drug activity, and type of the patient's disease; Sensitivity to the drug, administration time, administration route and excretion rate, treatment period, factors including concurrent drugs and other factors well known in the medical field may be determined.

The pharmaceutical composition according to the present invention may be administered as an individual therapeutic agent or may be administered in combination with other therapeutic agents, may be administered sequentially or simultaneously with conventional therapeutic agents, and may be administered singly or multiple times. In consideration of all of the above factors, it is important to administer an amount capable of obtaining the maximum effect with a minimum amount without side effects, which can be easily determined by a person skilled in the art to which the present invention pertains.

The pharmaceutical composition of the present invention may be administered to an individual by various routes. All modes of administration can be contemplated, for example, oral administration, subcutaneous injection, intraperitoneal administration, intravenous injection, intramuscular injection, paraspinal space (intrathecal) injection, sublingual administration, buccal administration, rectal insertion, vaginal It can be administered according to internal insertion, ocular administration, ear administration, nasal administration, inhalation, spraying through the mouth or nose, skin administration, transdermal administration, and the like.

Hereinafter, preferred examples are presented to help the understanding of the present invention. However, the following examples are only provided for easier understanding of the present invention, and the contents of the present invention are not limited by the following examples.

[Example]

Example 1. Experimental method

test material

The 20% alcohol oat extract preparation was prepared by adding 500 grams of oats to a final concentration of 20% (g/vol), crushing, mixing and extracting the supernatant by centrifugation, followed by freeze-drying. Avenanthramides (A, B, C) were isolated from 20% alcoholic oat extract by HPLC. Specifically, 20% ethanol oat extract was loaded onto a reverse phase HPLC (RPHPLC) column (CAPCELL-PAK C18, 4.6 mm Х 250 mm, Shiseido Co., Japan) to obtain a concentration equalization gradient (5-65%). acetonitrile/0.1% TFA) solvent for 60 minutes at a flow rate of 1 ml/min. The dried sample extract (100 μg) was dissolved in 5 μl of dimethylsulfoxide (DMSO), and 2 μl of each was injected. Avenanthramide (A, B, C) was purchased from Sigma-Aldrich (Saint-Louis, Mo, USA) in some cases. Corning trans-well Wright-Giemsa stain solution and DAPI (6-diamidino-2-phenylindole dihydrochloride) were purchased from Sigma-Aldrich (Saint-Louis, Mo, USA). An ELISA assay kit for checking cytokine secretion was purchased from Affymetrix eBioscience (USA). Human recombinant TNF-α protein was purchased from PeproTech (Rocky Hill, NJ, USA), antibody NFkB from Abcam (Cambridge, MA, USA), p-IkB, IkB from Santa Cruz Biotechnology (Paso Robles, CA, USA) purchased from Another antibody pERK, ERK, pp38, pJNK, and JNK were purchased from Cell signaling technology (Danvers, MA, USA), and MMP-9, MMP-2, p38, and β-actin were purchased from Santa Cruz Biotechnology (Paso Robles, CA, USA) was purchased.

cell culture

Human skin cancer cells (SK-MEL-2) and human smooth muscle cells (HASMC) were purchased from the American Type Culture Collection (ATCC; Rockville, MD, USA). Cells were cultured in DMEM (WelGENE Co., Daegu, Korea), and each medium was treated with 10% Fetal bovine serum (FBS), 100 U/ml penicillin, and 100 mg/ml streptomycin (P/S). Incubated in an incubator at 37° C. and 5% CO ₂ .

Reverse Transcription Polymerase Chain Reaction (RT-PCR)

Total RNA of transfected HASMC cells was extracted using TRIzol reagent (Invitrogen, USA), and cDNA was synthesized using oligo dT primer using RT-PreMIX kit (Takara Shuzo, Japan). cDNA was amplified with a PCR device using the following primers.

MMP-9: forward 5'-TCCCTGGAGACCTGAGAACC-3' (SEQ ID NO: 1), reverse 5'-CGGCAAGTCTTCCGAGTAGTT-3' (SEQ ID NO: 2)

MMP-2: forward 5'-TGAAGGTCGGTGTGAACGGA-3' (SEQ ID NO: 3) reverse 5'-CATGTAGCCATGAGGTCCACCAC-3' (SEQ ID NO: 4)

β-actin: forward 5'-TCCTTCTGCATCCTGTCGGC-3' (SEQ ID NO: 5), reverse 5'-CAAGAGATGGCCACGGCTGC-3' (SEQ ID NO: 6)

western blot

Proteins were extracted from the transfected HASMC cells using 1% NP ₄ 0 protein lysis buffer containing 100 mM Na-orthovanadate, 100 mM NaF, and 100 mM Na-pyrophosphate, and the extract was centrifuged at 4 °C and 13000 rpm. . After quantifying the extracted protein by Bio-Rad protein assay (Bio-Rad Laboratories Hercules, CA, USA), the same amount of protein was separated by size through SDS-PAGE (SDS-polyacrylamide). Then, it was transferred to a PVDF (Polyvinylidene difluoride) membrane, the membrane was reacted with a primary antibody and a secondary antibody, and the result was confirmed through ECL (Enhanced chemiluminescence).

Trans-well cell migration assay

HASMC cells were added to the top of the trans-well, and a medium treated with TNF-α was added to the bottom and cultured for 24 hours. Cells migrated through an 8 mm pore size membrane were fixed with methanol, stained with Giemsa, and then stained with an optical microscope.

Wound healing analysis

HASMC cells were cultured in 6-well culture plates. The wounds were artificially scraped with a yellow pipette tip (200 μl) and then washed medium three times. After treatment with avenanthramide or avenanthramide + TNF-α for 24 hours, each well was observed under a microscope.

Gelatin Zymography Analysis

The prepared avenanthramide sample was mixed with 5% tri-glycine SDS sample buffer and loaded onto a 10% polyacrylamide gel containing 1 mg/ml gelatin. The gel was operated at 110 V for 80 minutes, washed with 2.5% Triton X-100 for 20 minutes, and incubated with 10 mM Tris base and 40 mM Tris. Then, it was incubated again with HCl, 200 mM NaCl, 10 mM CaCl ₂ culture buffer at room temperature for 12 hours. Then, the gel was stained with 10% acetic acid in 0.5% Coomassie blue dye in 50% methanol for 2 hours. Except for staining, the MMP-9 active site, which appeared as a transparent band, was detected on a blue background.

Cytokine secretion measurement analysis

The level of inflammatory cytokines such as TNF-α, IL-6 and IL-1β in the culture medium was measured using an ELISA kit (Affymetrix, eBioscience) together with an antibody for each cytokine according to the protocol. The capture antibody was attached to a 96-well plate and incubated for at least 5 hours. Washed 3 to 5 times with the washing liquid. Incubation was performed for 30 minutes using a blocking solution. Washing was performed 3 to 5 times with the same washing solution. The prepared medium (the medium extracted after 24 hours of drug treatment) was added and incubated for 2 hours. And the detection antibody was additionally attached for 1 hour. After washing, the fluorescent tag-attached substrate was put in for 15 minutes, and then a stop solution was added. Thereafter, the absorbance was measured at 450 nm in a micro-reader.

immunofluorescence

After treating HASMC with TNF-α and fixing it using 4% paraformaldehyde (PFA) 24 hours later, the permeability is increased with 0.2% Triton-100, and after sequentially staining with primary antibody, secondary antibody, and DAPI, fluorescence It was confirmed through a microscope.

Measurement of Tyrosinase Inhibitory Activity

Tyrosinase activity was measured using a spectrophotometer. Mushroom tyrosinase (EC 1.14.18.1) having an activity of 110 U/ml at a volume of 0.1 ml in the reaction tube was put together with 0.1 ml of 10 mM L-DOPA. In the reaction tube, 0.05 ml of the measurement sample and 0.05 ml of the measurement sample dissolved in 0.25 ml of 175 mM sodium phosphate buffer (pH 6.8) were mixed. This tube was kept warm at 37°C for 2 minutes. The reaction product, DOPAchrome, was measured at a wavelength of 475 nm. The rate of inhibition of tyrosinase activity was expressed as a rate of decrease in absorbance of the sample solution. SK-MEL-2 human melanoma cells were seeded in a 24-well plate at the number of 2 X 10 ⁵ cells per well, and avenanthramide was added at each concentration, followed by incubation for 24 hours. After 24 hours, each well was washed with 10 mM PBS and suspended by adding 100 μl of 10 mM PBS (1% Triton X-100). The solution was homogeneously suspended with a vortex and centrifuged to obtain the supernatant as an enzyme solution for measurement. 40 μl of the solution thus obtained was transferred to a 96-well microplate (BD Falcon, Bedford, MA, USA), and 100 μl L-dopa (2 mg/ml) as an enzyme substrate was further added thereto. After warming at 37°C for 1 hour, absorbance was measured at 405 nm using a microplate reader.

Enzyme inhibitory activity was determined by the following formula.

Inhibition (%) = [(A - B) - (C - D)]/(A - B) X 100

(A is the absorbance after reaction of the control group, B is the absorbance before reaction of the control group, C is the absorbance after reaction of the sample solution, D is the absorbance of the sample solution before reaction).

To investigate the mechanism of kinetic activity of tyrosinase, various concentrations of L-DOPA (range 0.625 to 5 mM), mushroom tyrosinase solution (1000 U/ml) and 50 mM potassium phosphate buffer (pH 6.5) was injected into a 96-well plate, and the avenanthramide (Ave A, B, C) solution was added to adjust the total solution to 200 μl. The Michaelis constant (Km) of tyrosinase was determined after measuring the enzyme activity according to various L-DOPA concentrations using a Lineweaver-Burk plot.

Measurement of melanin production

SK-MEL-2 cells were inoculated with 2 X 10 ⁵ cells per well in a 24-well plate, and avenanthramides (A, B, C) were prepared at each concentration and pre-treated. After 1 hour, it was treated with α-MSH (100 nM) and kept warm for 48 hours. Thereafter, each well was washed with PBS, and 400 μl of 0.2 N NaOH was added to disperse the cells, and after incubation at 60° C. for 1 hour, absorbance was measured at 405 nm.

Computer simulation of docking of tyrosinase enzymes and compounds

Tyrosinase, a target protein of the compound, was generated through SWISS-MODEL (https://swissmodel.expasy.org/) by tyrosinase related protein 1 (PDB ID: 5M8S). To make docking simulations of avenanthramides A (Avn A), B (Avn B) and C (Avn C), 2D structures of the compounds were generated and converted into 3D structures. The energy minimization process was created with the ChemOffice program (http://www.cambridgesoft.com). Docking simulations with Avn A, Avn B and Avn C with tyrosinase were performed using Autodock Vina (O. Trott, AJ Olson, AutoDock Vina: improving the speed and accuracy of docking with a new scoring function, efficient optimization , and multithreading, J. Comput. Chem . 31 (2010) 455-461). The binding pocket of tyrosinase was predicted using the predicted active site obtained from the tyrosinase-kojic acid (PDB ID: 5M8M) complex. The docking simulation result is represented by Chimera (JE Mills, PM Dean, Three-dimensional hydrogen-bond geometry and probability information from a crystal survey, J. Comput. Aided Mol. Des . 10 (1996) 607-622), and the obtained docking simulation Based on the results, possible hydrogen bonding was analyzed using the default values of 0.4 angstrom and 20.0 degree bonding relaxation constants in Chimera (EF Pettersen, TD Goddard, CC Huang, GS Couch, DM Greenblatt, ECMeng, TE Ferrin, UCSF). Chimera-a visualization system for exploratory research and analysis, J. Comput. Chem. 25 (2004) 1605-1612). The skin irritation of Avn A, B, and C was predicted through quantitative structure-activity relationship (QSAR) analysis.

Example 2. Cytotoxicity evaluation of oat extract and indicator ingredient avenanthramide

The molecular weight of avenanthramide, a major component of oats, is 315.28 g/mol, and the chemical structure is shown in FIG. 1A. In order to confirm the cytotoxicity of 20% alcoholic oat extract and avenanthramide, which is an indicator component of oats, an MTT assay experiment was performed. Typically, human smooth muscle cells (HASMC) were used as model cells. The none group treated with TNF-α (TNF-α only group) showed no significance compared to the cell viability of the control group. When 20% alcoholic oat extract and avenanthramide were treated with 0, 50 and 100 μM, there was no significant difference in cell viability compared to the none group, and there was no cytotoxicity to oat extract and avenanthramide. was confirmed (Fig. 1b, Table 1)

Example 3. The effect of improving blood vessel stenosis of oat extract and index component avenanthramide

It has been reported that TNF-α is involved in vascular stenosis and metastasis (Morisaki N, Xu QP, Koshikawa T, Saito Y, Yoshida S, Ueda S. Tumor necrosis factor-alpha can modulate the phenotype of aortic smooth muscle cells. Scand J Clin Lab Invest . 1993 Jul;53(4):347-52). In HASMC activated with TNF-α, 20% alcohol oat extract and avenanthramide, an indicator component of oats, were tested to determine what effect they had on wound healing and cell migration. .

As a result, as shown in FIG. 2, wound healing and cell migration were increased when human smooth muscle cells were treated with TNF-α, but wounds were treated with 20% alcohol oat extract and avenanthramide. Healing and cell migration were inhibited (Table 2).

Example 4. MMP-9 expression and activity inhibitory effect of oat extract and index component avenanthramide

To evaluate the skin protection, skin regeneration activation and skin elasticity effects of 20% alcoholic oat extract and avenanthramide, an indicator component of oats, the expression levels of MMP-9 and MMP-2 stimulated by TNF-α were RT -PCR and Western blot, enzyme activity was evaluated by gelatin zymography analysis. Human arterial smooth muscle cells were treated with 100 ng/ml TNF-α and 0-100 μM 20% alcoholic oat extract or avenanthramide, and protein and mRNA levels were measured after incubation for 24 hours.

As a result, as shown in FIG. 3a , there was little change in MMP-2 in HASMC treated with 20% alcoholic oat extract and avenanthramide, but the protein level of MMP-9 decreased (Table 3).

In addition, as shown in FIG. 3b, MMP-9 mRNA levels were decreased in cells treated with 20% alcoholic oat extract and avenanthramide at high concentrations in the range of 0-100 μM (Table 4).

In addition, as shown in FIG. 3c , when TNF-α-induced MMP-9 in human smooth muscle cells was treated with 20% alcoholic oat extract and avenanthramide, in control cells (MMP-9 induced only with TNF-α) MMP-9 enzyme activity was inhibited by material treatment at a significantly lower level (Table 5).

The above results show that 20% alcoholic oat extract and avenanthramide can reduce the mRNA and protein levels of MMP-9 and inhibit the enzymatic activity.

Example 5. Cytotoxicity evaluation of oat extract and indicator ingredient avenanthramide in skin cells

A cytotoxicity assay for securing the safety of 20% alcoholic oat extract and avenanthramides (Avn A, Avn B and Avn C) was performed by MTT assay. As a result, as shown in FIG. 4 , the untreated group and the α-MSH treated group did not show cytotoxicity as a control group. In addition, when Avn A, Avn B, Avn C and oat extracts were treated with 0, 50 and 100 μM concentrations and 50 μM and 100 μM, no cell toxicity was observed (Table 6). Specifically, as a result of treatment with 20% alcoholic oat extract and avenanthramide up to 100 μM, it did not show toxicity to the proliferation of SK-MEL-2 cells. Therefore, in subsequent experiments, it was treated at this concentration.

Example 6. Tyrosinase inhibitory activity of oat extract and indicator ingredient avenanthramide

As shown in Figure 5a, in vitro 20% oat extract and Avn A, Avn B, Avn C concentration-dependently inhibited the activity of tyrosinase at concentrations 0, 10, 25, 50 and 100 μM (Table 7) ). The inhibitory activity of Avn C was the best, followed by Avn A and Avn B in that order.

In addition, as shown in FIG. 5b, tyrosinase activity in α-MSH-induced SK-MEL-2 cells was also inhibited in a concentration-dependent manner (0, 50, 100 μM) (Table 8). In SK-MEL-2 cells, Avn C inhibited the enzyme activity the most, and the effect was superior to Avn A or Avn B.

The above results show that oat extract and avenanthramide inhibit the activity of α-MSH-induced tyrosinase in skin cells.

Example 7. Melanin production inhibitory effect of oat extract and index ingredient avenanthramide in skin cells

Tyrosinase is involved in the synthesis of melanin, and tyrosinase converts L-tyrosine to 3,4-dihydroxyphenylalanine (L-DOPA), and L-DOPA is again synthesized into melanin through DOPAquinone (Masaru). Murakami 1, Fumihide Matsuzaki, Masayuki Funaba. Regulation of Melanin Synthesis by the TGF-beta Family in B16 Melanoma Cells. Mol Biol Rep . 2009 Jul;36(6):1247-50).

As shown in FIG. 6 , melanin biosynthesis was inhibited by 20% alcohol oat extract and Avn A, Avn B, Avn C treatment at concentrations of 0, 50 and 100 μM (Table 9). Avn C had the best effect, followed by Avn A and Avn B in order. These results show that oat extract and avenanthramide effectively inhibit melanin biosynthesis in skin cells.

Example 8. Prediction of binding sites of avenanthramide and tyrosinase through molecular docking simulation analysis

As a result of the docking simulation, it was confirmed that Avn A, Avn B and Avn C were effectively present at binding sites during the formation of the tyrosinase-kojic acid complex ( FIG. 7 ). It was confirmed that all three phenylpropanoids of these compounds were located in the pocket. The binding affinity of Avn C to tyrosinase was calculated to be -4.0 kcal/mol, and the binding affinity values of Avn A (-3.7 kcal/mol) and Avn B (-2.9 kcal/mol) were calculated using AutoDock Vina assay. was confirmed to be higher.

The description of the present invention described above is for illustration, and those of ordinary skill in the art to which the present invention pertains can understand that it can be easily modified into other specific forms without changing the technical spirit or essential features of the present invention. will be. Therefore, it should be understood that the embodiments described above are illustrative in all respects and not restrictive.

<110> Research & Business Foundation SUNGKYUNKWAN UNIVERSITY <120> Composition for improving skin condition or angiostenosis comprising oat extract or avenanthramide <130> MP20-059 <160> 6 <170> KoPatentIn 3.0 <210> 1 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Forward primer for MMP-9 <400> 1 tccctggaga cctgagaacc 20 <210> 2 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> Reverse primer for MMP-9 <400> 2 cggcaagtct tccgagtagt t 21 <210> 3 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Forward primer for MMP-2 <400> 3 tgaaggtcgg tgtgaacgga 20 <210> 4 <211> 23 <212> DNA <213> Artificial Sequence <220> <223> Reverse primer for MMP-2 <400> 4 catgtagcca tgaggtccac cac 23 <210> 5 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Forward primer for beta-actin <400> 5 tccttctgca tcctgtcggc 20 <210> 6 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Reverse primer for beta-actin <400> 6 caagagatgg ccacggctgc 20

Claims (13)

A cosmetic composition for improving skin condition, comprising an oat extract, or avenanthramide, which is an indicator component of oats, or a cosmetically acceptable salt thereof as an active ingredient. According to claim 1,
The skin condition improvement is skin whitening, skin regeneration, skin elasticity enhancement or skin wrinkle improvement, a cosmetic composition. 3. The method of claim 2,
The skin whitening is caused by blemishes, freckles, black spots, age spots, spots, milk coffee spots, Ota nevus, blue nevus, hyperpigmented spots, hyperpigmentation after drug use, gravidic chloasma, and wounds or dermatitis A cosmetic composition, characterized in that it relieves one or more selected from the group consisting of hyperpigmentation after inflammation. According to claim 1,
The cosmetic composition, characterized in that the avenanthramide is avenanthramide A, avenanthramide B or avenanthramide C. According to claim 1,
The oat extract is alcohol, water, acetone, ether (ether), benzene, chloroform, ethyl acetate, methylene chloride, hexane (hexane), cyclohexane (cyclohexane), petroleum ether (petroleum ether), subcritical having 1 to 6 carbon atoms. A cosmetic composition, characterized in that it is extracted with a solvent selected from the group consisting of a fluid, a supercritical fluid, and a mixed solvent thereof. According to claim 1,
The active ingredient is characterized in that inhibiting the activity of tyrosinase, a cosmetic composition. According to claim 1,
The active ingredient is characterized in that inhibiting melanin production, cosmetic composition. According to claim 1,
The active ingredient is a cosmetic composition, characterized in that it inhibits the expression of MMP-9 (Matrix metallopeptidase 9) or its enzymatic activity. A food composition for improving skin condition, comprising an oat extract, or avenanthramide, which is an indicator component of oats, or a pharmaceutically acceptable salt thereof as an active ingredient. A pharmaceutical composition for preventing or treating skin pigmentation diseases, comprising an oat extract, or an oat index component, Avenanthramide, or a pharmaceutically acceptable salt thereof, as an active ingredient. 11. The method of claim 10,
The skin pigmentation diseases include melasma, freckles, black spots, age spots, spots, milk coffee spots, nevus ota, blue nevus, hyperpigmentation spots, hyperpigmentation after drug use, gravidic chloasma, and wounds or dermatitis A pharmaceutical composition, characterized in that at least one selected from the group consisting of hyperpigmentation after inflammation due to A food composition for preventing or improving vascular stenosis, comprising an oat extract, or avenanthramide, which is an indicator component of oats, or a pharmaceutically acceptable salt thereof as an active ingredient. A pharmaceutical composition for preventing or treating vascular stenosis, comprising as an active ingredient an oat extract, or avenanthramide, which is an indicator component of oats, or a pharmaceutically acceptable salt thereof.

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