Anti-photoaging agents for the skin

Abstract

The present invention aims to provide a novel anti- and anti-photoaging agent. [Solution] The features of the present invention for solving the above problems are as follows. 1. A UVA-induced cell atrophy inhibitor for fibroblasts, containing purple tea (scientific name: Camellia sinensis, variety name TRFK306) extract as the active ingredient. 2. A cell atrophy inhibitor for fibroblasts induced by UVA irradiation, containing GHG (1,2-di-O-galloyl-4,6-O-(S)-hexahydroxydiphenoyl-β-D-glucose) as the active ingredient. 3. A ferulic acid-containing agent that inhibits UVA-induced cell atrophy in fibroblasts. 4. A photoaging prevention and improvement agent containing one of the agents described in item 1 to 3 above as an active ingredient.

Description

Technical Field

[0001] The present invention relates to an anti-photoaging agent and a photoaging prevention / improvement agent using the same. The present invention is widely used in foods (including health foods such as foods with functional claims and foods for specified health uses), pharmaceuticals, quasi-drugs, cosmetics, external skin preparations, etc.

Background Art

[0002] Aging biologically refers to changes that occur in an organism's individual over time, and among them, it can be said that it is the decline in function that occurs until the organism dies and the process thereof. In particular, since the skin is exposed to ultraviolet rays, which are an external environment, in addition to physiological aging, photoaging, which is an environmental factor, also inevitably occurs simultaneously. That is, "aging" includes physiological aging caused by natural physiological phenomena and photoaging caused by light such as ultraviolet rays.

[0003] Physiological aging is the aging of the skin caused by aging. Macroscopically, it is characterized by fine wrinkles and dryness. The skin becomes thinner, metabolism decreases, and the skin turnover rate slows down. This is due to a decrease in the amount of collagen in the dermis and a decrease in the elasticity of elastin, which is an elastic fiber. Proteins such as collagen and elastin bind to reactive sugars and cause the Maillard reaction (glycation reaction), resulting in a decrease in water retention capacity and skin elasticity. In addition, the epidermal barrier function decreases due to a decrease in sweat and natural moisturizing factors. This leads to a decrease in the water content of the stratum corneum and results in dryness.

[0004] In contrast, photoaging is primarily caused by changes in the skin due to ultraviolet (UV) radiation, and is characterized by deeper wrinkles compared to physiological aging. This is due to changes in the epidermal cells themselves, degeneration of elastic fibers, and delamination due to a decrease in adhesion between the epidermis and dermis. The dermis is composed of the extracellular matrix (collagen, elastin, hyaluronic acid, etc.) and its producers, fibroblasts. It has been confirmed that these substances decrease or disappear due to UV radiation (UVA) that reaches the dermis. Pigmentation, commonly known as age spots, is also mainly caused by the accumulation of melanin pigment in the epidermis due to UV radiation.

[0005] Furthermore, wrinkles can be categorized into deep wrinkles originating from the dermis and shallow wrinkles originating from the epidermis. Of these, deep wrinkles and sagging originating from the dermis are caused by the influence of the extracellular matrix and fibroblasts in the dermis. In addition, it has been reported that irradiating fibroblasts with UVA causes atrophy of the cytoskeleton, leading to cell inactivation (decreased collagen production) and activation of MMP1 (collagen-degrading enzyme) (Non-patent Literature 1). As a result, cytoskeletal atrophy is said to be one of the causes of deep wrinkles originating from the dermis. Therefore, deep wrinkles are primarily caused by photoaging rather than physiological aging. [Non-Patent Document 1] Yamada H., et al, Experimental dermatology, 25 (Suppl. 3), P45-51 (2016). [Disclosure of the Invention] [Problems that the invention aims to solve]

[0006] Against this backdrop, the inventors discovered that purple tea extract, ferulic acid, and GHG suppress cell atrophy induced by UVA irradiation in fibroblasts, thus completing the present invention. In other words, the present invention aims to provide a novel photoaging prevention and improvement agent. [Means for solving the problem]

[0007] The features of the present invention for solving the above problems are as follows. 1. A UVA-induced cell atrophy inhibitor for fibroblasts, containing purple tea (scientific name: Camellia sinensis, variety name TRFK306) extract as the active ingredient. 2. An inhibitor of UVA-induced cell atrophy in fibroblasts, containing GHG (1,2-di-O-galloyl-4,6-O-(S)-hexahydroxydiphenoyl-β-D-glucose) as the active ingredient. 3. A ferulic acid-containing agent that inhibits UVA-induced cell atrophy in fibroblasts. 4. A photoaging prevention and improvement agent containing one of the agents described in item 1 to 3 above as an active ingredient. [Brief explanation of the drawing]

[0008] [Figure 1] This is a micrograph (fluorescent immunostaining of β-actin) showing the state of cell atrophy in normal diploid fibroblasts (NB1RGB cells) induced by UVA irradiation. [Figure 2] These are micrographs (fluorescent immunostaining of β-actin, 100x magnification) and graphs showing the evaluation results of the inhibitory effect of purple tea extract and ferulic acid on UVA irradiation-induced cell atrophy. [Figure 3] This image shows a micrograph (fluorescent immunostaining of β-actin, 100x magnification) and graph illustrating the evaluation results of the inhibitory effect of UVA irradiation on cell atrophy in GHG cells. [Figure 4] The image above is a magnified micrograph (fluorescent immunostaining of β-actin) showing the evaluation results of the cell atrophy inhibitory effect shown in Figure 3. [Figure 5] This is the HPLC chromatogram of the purple tea (scientific name: Camellia sinensis, variety name TRFK306) extract used in this example. [Modes for carrying out the invention]

[0009] The present invention will be described in detail below. This invention is characterized by containing purple tea (scientific name: Camellia sinensis, variety name TRFK306) extract, GHG, and ferulic acid as active ingredients.

[0010] The purple tea used as the raw material for purple tea extract (scientific name: Camellia sinensis, variety name TRFK306, hereinafter simply referred to as "purple tea") is a tea plant developed by the Kenyan government through crossbreeding, and is named variety TRFK306. Kenyan purple tea leaves contain anthocyanins, which give them their purple color, hence the name "purple tea." In addition to TRFK306, other purple teas such as Sunrouge, developed by the National Agriculture and Food Research Organization (NARO), are also known, but TRFK306 contains a high concentration of GHG (1,2-di-O-galloyl-4,6-O-(S)-hexahydroxydiphenoyl-β-D-glucose), a unique component not found in other purple teas.

[0011] Here, the above GHG is the compound represented by the following chemical formula (1). [ka]

[0012] The part of the Kenyan purple tea used in this invention is not particularly limited; leaves, stems, roots, flowers, seeds, etc., can be used, but it is especially preferable to use leaves, as this allows for obtaining a higher concentration of GHG.

[0013] The purple tea extract of the present invention can preferably be obtained, for example, by crushing fresh or dried purple tea leaves (hereinafter referred to as "purple tea leaves") and extracting them using a polar solvent (including water; the same applies hereinafter). To improve the extraction efficiency, the purple tea leaves may be subjected to appropriate chemical treatments such as acid or alkali decomposition or enzymatic decomposition before extraction.

[0014] Specifically, purple tea extract can be produced by the method described below. That is, first, as the above-mentioned purple tea leaves, the raw or dried purple tea leaves are subjected to chemical treatments such as acid or alkali decomposition, enzymatic decomposition, etc.

[0015] Then, a polar solvent is added to the above-mentioned purple tea leaves, and shaken or heated under reflux to extract GHG into the solvent.

[0016] At this time, the above-mentioned polar solvent is not particularly limited, and water, alcohols, and ketones can be used. In addition, these may be used alone or in combination of two or more. Furthermore, it is particularly preferable to use a hydrous alcohol or a hydrous ketone among these.

[0017] As the hydrous alcohol-based solvent, hydrous solvents such as ethanol, methanol, and propanol can be used, and particularly, hydrous ethanol is preferable. Also, as the hydrous ketone-based solvent, hydrous solvents such as acetone, methyl ethyl ketone, diethyl ketone, and chloroacetone can be used, and particularly, hydrous acetone is preferable.

[0018] Regarding the water content rate, in the case of hydrous ethanol, ethanol is 1 to 99.9% by mass, preferably 30 to 99% by mass, more preferably 40 to 80% by mass, and most preferably 40 to 60% by mass. In the case of hydrous acetone, it is preferable that it contains 20 to 99.9% by mass of acetone. This is because the extraction efficiency of GHG is excellent in the above range. In the following, regarding the notation of the water content rate of the hydrous solvent, for the sake of simplicity, for example, 80% ethanol hydrated with 20% by mass of water is denoted as "80% hydrous ethanol".

[0019] In the method for producing the purple tea extract of the present invention, the heating under reflux can be carried out by a well-known method using the above-mentioned hydrous alcohol-based solvent or hydrous ketone-based solvent. The heating temperature is 30 to 95°C, more preferably about 30 to 50°C, and the reflux time is preferably about 1 to 4 hours.

[0020] Furthermore, in the method for producing purple tea extract according to the present invention, shaking, stirring, etc. may be performed as appropriate, if necessary.

[0021] Furthermore, in the method for producing purple tea extract of the present invention, it is preferable to remove the solvent under reduced pressure after extraction. This makes it possible to obtain a composition that does not contain organic solvents, and it is possible to meet safety standards for use as a food ingredient in functional foods, health foods, and other food and beverage products.

[0022] Furthermore, in the method for producing purple tea extract according to the present invention, stepwise extraction can be performed using multiple solvents. This makes it possible to produce purple tea extract containing a high concentration of GHG in a higher yield.

[0023] Specifically, for example, purple tea leaves are mixed with either the aqueous alcohol-based solvent or the aqueous ketone-based solvent, shaken or heated under reflux, and GHG is extracted into the solvent to obtain a first extract. The extract is separated from the residue that was not recovered as an extract by centrifugation or the like, and the other solvent that was not selected is added to the residue, shaken or heated under reflux, and GHG is extracted into the solvent to obtain a second extract. The first extract and the second extract are then mixed. It goes without saying that this second extract can also be used as an extract of purple tea leaves on its own.

[0024] Thus, by performing stepwise extraction with multiple solvents, it is conceivable that the physical properties and other characteristics of the purple tea leaves will change to those suitable for extraction after undergoing the first extraction treatment with the aqueous alcohol-based solvent or the aqueous ketone-based solvent. Therefore, in the subsequent second extraction treatment, it can be expected that the extraction efficiency will be improved, not only when using the aqueous alcohol-based solvent or the aqueous ketone-based solvent, but also when using other solvents.

[0025] Furthermore, the present invention is characterized by using GHG as an active ingredient. This GHG can be obtained by purification from the purple tea extract described above.

[0026] Ferulic acid is a compound shown in the following chemical formula (2). [ka]

[0027] There are no particular limitations on the method of producing ferulic acid; it can be extracted from natural sources or manufactured synthetically. Alternatively, commercially available products may be used; for example, "ferulic acid" (product name) manufactured by Oryza Oil & Fat Chemical Co., Ltd. may be used.

[0028] The photoaging prevention and improvement agent 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 confectionery (gum, candy, caramel, chocolate, cookies, snacks, jelly, gummies, tablets, etc.), noodles (soba, udon, ramen, etc.), dairy products (milk, ice cream, yogurt, etc.), seasonings (miso, soy sauce, etc.), soups, and beverages (juice, coffee, 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.

[0029] These foods and beverages can be formulated with various ingredients depending on their type. For example, they can contain food ingredients 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 ester, polyglycerin fatty acid ester, sucrose fatty acid ester, sorbitan fatty acid ester, propylene glycol fatty acid ester, gum arabic, carrageenan, casein, gelatin, pectin, agar, B vitamins, nicotinamide, calcium pantothenate, amino acids, calcium salts, colorants, flavorings, and preservatives.

[0030] Specifically, the agent of the present invention can be spray-dried or freeze-dried together with powdered cellulose, and this can be easily incorporated into food and beverages (such as instant foods) by processing it into a powder, granules, tablets, or solution. Furthermore, the agent of the present invention can be dissolved in, for example, oils and fats, ethanol, glycerin, or mixtures thereof to form a liquid, which can then be added to beverages or solid foods. If necessary, it 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.

[0031] When applying the agent of the present invention to food and beverages, the amount added is preferably such that the total content of the active ingredient relative to the food and beverage is 1 to 20 wt%, since the main purpose is disease prevention and health maintenance.

[0032] The photoaging prevention and improvement agent of the present invention may be used as a material for pharmaceuticals (including pharmaceuticals and quasi-drugs). It can be manufactured by appropriately blending the agent of the present invention with raw materials for pharmaceutical formulations. Examples of formulation ingredients 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, hydrogenated vegetable oil, kaolin, talc, etc.), binders (distilled water, physiological 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, monoglyceride stearate, starch, lactose, acacia 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.), and lubricants (purified talc, stearate, polyethylene glycol, etc.).

[0033] The present invention's photoaging prevention and improvement agent can generally be administered orally in the form of tablets, pills, soft or hard capsules, granules, powders, granules, or liquids, but it may also be administered parenterally. When administered parenterally, it can be administered in solution form, or with the addition of dispersants, suspensions, stabilizers, etc., by local tissue administration, intradermal, subcutaneous, intramuscular, or intravenous injection. It may also be administered in the form of suppositories or other preparations.

[0034] The dosage may vary depending on the method of administration, the patient's condition, and the patient's age, but typically, adults can be administered 0.5 to 5000 mg of the active ingredient per day, and children can usually be administered 0.5 to 3000 mg. The ratio of photoaging prevention and improvement agents can be adjusted as appropriate depending on the dosage form, but generally, when administered orally or via mucosal absorption, it is appropriate to use approximately 0.3 to 15.0 wt%, and when administered parenterally, approximately 0.01 to 10 wt%. Note that the dosage will vary depending on various conditions, so in some cases a smaller amount than the above may be sufficient, and in other cases it may be necessary to administer a larger amount. The photoaging prevention and improvement agent of the present invention can also be expected to exhibit photoaging prevention and improvement effects when used as a topical skin preparation (including cosmetics, pharmaceuticals, and quasi-drugs). Examples of external skin preparations that may contain the photoaging prevention and improvement agent of the present invention include lotions, soaps, facial cleansers, bath additives, creams, lotions, toners, colognes, shaving creams, shaving lotions, cosmetic oils, sunscreens, face powders, foundations, perfumes, face masks, nail creams, nail polish, nail polish removers, eyebrow pencils, blushes, eye creams, eyeshadows, mascaras, eyeliners, lipsticks, lip balms, shampoos, conditioners, hair dyes, dispersions, and cleansing agents. Examples of pharmaceuticals or quasi-drugs that may contain the photoaging prevention and improvement agent of the present invention include ointments, creams, and topical solutions.

[0035] In addition to the photoaging prevention and improvement agent according to the present invention, the above-described form of topical skin preparation may contain ingredients commonly used in cosmetics, quasi-drugs, and other topical skin preparations, such as oils, higher alcohols, fatty acids, UV absorbers, powders, pigments, surfactants, polyhydric alcohols / sugars, polymers, physiologically active ingredients, solvents, antioxidants, fragrances, preservatives, etc., to the extent that they do not impair the photoaging prevention and improvement effect. Examples are listed below, but the present invention is not limited to these examples.

[0036] (1) Examples of oil content Ester-based oil phase components: 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, diethyl adipate Sopropyl, Isoarachil 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, Iso 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, Di(Capryl / Capric Acid)Propylene Glycol, 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 (C12-18) carbonate, triisocetyl citrate, triisoarachil citrate, triisooctyl citrate, lauryl lactate, myristyl lactate, cetyl lactate, octyldecyl lactate, triethyl citrate, acetyl triethyl citrate, acetyl tributyl citrate, trioctyl citrate, diisostearyl malate, 2-ethylhexyl hydroxystearate, di-2-ethylhexyl succinate, diisobutyl adipate, diisopropyl sebacate Examples include ropil, dioctyl sebacate, cholesteryl stearate, cholesteryl isostearate, cholesteryl hydroxystearate, cholesteryl oleate, dihydrocholesteryl oleate, phytosteryl isostearate, phytosteryl oleate, isocetyl 12-stearoylhydroxystearate, stearyl 12-stearoylhydroxystearate, and isostearyl 12-stearoylhydroxystearate. Hydrocarbon-based oil phase components include 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: animal oils and their hydrogenated oils such as beef tallow, hydrogenated beef tallow, pork tallow, hydrogenated pork tallow, horse oil, hydrogenated horse oil, mink oil, orange roughy oil, fish oil, hydrogenated fish oil, egg yolk oil, 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 include 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, meadowfoam oil, cottonseed oil, hydrogenated cottonseed oil, coconut oil, hydrogenated coconut oil, and other vegetable oils and their hydrogenated oils, beeswax, high-acid value beeswax, lanolin, reduced lanolin, hydrogenated lanolin, liquid lanolin, carnauba wax, montan wax, and other waxes. Silicone-based oil phase components include dimethylpolysiloxane, methylphenylpolysiloxane, methylcyclopolysiloxane, octamethylpolysiloxane, decamethylpolysiloxane, dodecamethylcyclosiloxane, methylhydrogenpolysiloxane, polyether-modified organopolysiloxane, dimethylsiloxane-methylcetyloxysiloxane copolymer, dimethylsiloxane-methylstearoxane copolymer, alkyl-modified organopolysiloxane, terminal-modified organopolysiloxane, amino-modified silicone oil, amino-modified organopolysiloxane, dimethiconol, silicone gel, acrylic silicone, trimethylsiloxysilicate, silicone RTV rubber, etc. Examples of fluorine-based oil phase components include perfluoropolyethers, fluorine-modified organopolysiloxanes, fluorinated pitch, fluorocarbons, fluoroalcohols, and fluoroalkyl / polyoxyalkylene copolymerized organopolysiloxanes.

[0037] (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.

[0038] (3) Examples of fatty acids Examples 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.

[0039] (4) Examples of UV absorbers Para-aminobenzoic acid, amyl para-aminobenzoate, ethyl dihydroxypropyl para-aminobenzoate, glyceryl para-aminobenzoate, ethyl para-aminobenzoate, octyl para-aminobenzoate, octyl dimethyl 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 mono-2-ethylhexanoate dipara-methoxycinnamate, isopropyl para-methoxycinnamate, diethanolamine para-methoxyhydrocinnamate salt, diisopropyl / diisopropyl cinnamic acid ester mixture, urocanic acid, ethyl urocanic acid, hydroxymethoxybenzophenone, hydroxymethoxybenzophenone sulfonic acid and its salts, di Examples include hydroxymethoxybenzophenone, 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-diphenyl acrylate, phenylbenzimidazole sulfate, 3-(4-methylbenzylidene)camphor, isopropyldibenzoylmethane, 4-(3,4-dimethoxyphenylmethylene)-2,5-dioxo-1-imidazolidinepropionate 2-ethylhexyl, and their polymer derivatives and silane derivatives.

[0040] (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, 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, ultramarine, Prussian blue, zinc oxide, titanium dioxide, acid Examples include white pigments such as cerium oxide, extender pigments such as talc, mica, sericite, kaolin, and plate-like barium sulfate, pearl pigments such as titanium mica, 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 restrictions on the shape (spherical, rod-shaped, needle-shaped, plate-shaped, irregular shape, flake-shaped, spindle-shaped, etc.) and particle size of these powders. These powders may or may not have been pre-treated by conventional 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.

[0041] (6) Examples of surfactants Anionic surfactants include fatty acid soaps, α-acyl sulfonates, alkyl sulfonates, alkylallyl sulfonates, alkylnaphthalene sulfonates, alkyl sulfates, POE alkyl ether sulfates, alkylamide sulfates, alkyl phosphates, POE alkyl phosphates, alkylamide phosphates, alkylylalkyl taurates, N-acyl amino acid salts, POE alkyl ether carboxylates, alkyl sulfosuccinates, alkyl sulfoacetate sodium, acylated hydrolyzed collagen peptides, perfluoroalkyl phosphates, and the like. Cationic surfactants include alkyltrimethylammonium chloride, stearyltrimethylammonium chloride, stearyltrimethylammonium bromide, cetostearyltrimethylammonium chloride, distearyldimethylammonium chloride, stearyldimethylbenzylammonium chloride, behenyltrimethylammonium bromide, benzalkonium chloride, beheninamidepropyldimethylhydroxypropylammonium chloride, diethylaminoethylamide stearate, dimethylaminopropylamide stearate, quaternary ammonium salts of lanolin derivatives, and the like. Amphoteric surfactants include carboxybetaine type, amidebetaine type, sulfobetaine type, hydroxysulfobetaine type, amidesulfobetaine type, phosphobetaine type, aminocarboxylate type, imidazoline derivative type, and amideamine type. Nonionic surfactants include 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 alkanolamides, alkylamine oxides, hydrogenated soybean phospholipids, and the like. Examples of natural surfactants include lecithin, saponin, and sugar-based surfactants.

[0042] (7) Examples of polyhydric alcohols and sugars Examples 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 can also be used.

[0043] (8) Examples of polymers Acrylic acid ester / methacrylic acid ester copolymer (Plussize, manufactured by Go-O Chemical Co., Ltd.), vinyl acetate / crotonic acid copolymer (Resin 28-1310, manufactured by NSC Corporation), vinyl acetate / crotonic acid / vinyl neodecanate copolymer (28-2930, manufactured by NSC Corporation), methyl vinyl ether maleate half ester (Gantlets ES, manufactured by ISP Corporation), T-butyl acrylate / ethyl acrylate / methacrylic acid copolymer (Rubymer, manufactured by BASF Corporation), vinylpyrrolidone / vinyl acetate / vinyl propionate copolymer (Rubiscol VAP, manufactured by BASF Corporation), vinyl acetate β / crotonic acid copolymer (Rubicet CA, BASF), vinyl acetate / crotonic acid / vinylpyrrolidone copolymer (Rubicet CAP, BASF), vinylpyrrolidone / acrylate copolymer (Rubiflex, BASF), acrylate / acrylamide copolymer (Ultrahold, BASF), vinyl acetate / butyl maleate / isobornyl acrylate copolymer (Advantage, ISP), carboxyvinyl polymer (Carbopol, BFGoodrich), acrylic acid / alkyl methacrylate copolymer (Pemulene, BF Examples include anionic polymer compounds such as those manufactured by Goodrich, amphoteric polymer compounds such as the amphoteric acetate of dialkylaminoethyl methacrylate polymer (Yukaformer, manufactured by Mitsubishi Chemical Corporation), octylacrylamide acrylate / hydroxypropyl acrylate / butylaminoethyl methacrylate copolymer (AMPHOMER, manufactured by NSC), cationic polymer compounds such as the quaternary compound of vinylpyrrolidone / dimethylaminoethyl methacrylate (GAFQUAT, manufactured by ISP), methylvinylimidazolium chloride / vinylpyrrolidone copolymer (Rubicort, manufactured by BASF), and nonionic polymer compounds such as polyvinylpyrrolidone (Rubiscol K, manufactured by BASF), vinylpyrrolidone / vinyl acetate copolymer (Rubiscol VA, manufactured by BASF), vinylpyrrolidone / dimethylaminoethyl methacrylate copolymer (Copolymer 937, manufactured by ISP), and vinylcaprolactam / vinylpyrrolidone / dimethylaminoethyl methacrylate copolymer (Copolymer VC713, manufactured by ISP).Furthermore, naturally derived polymer compounds such as cellulose or its derivatives, keratin and collagen or their derivatives, calcium alginate, pullulan, agar, gelatin, tamarind seed polysaccharides, xanthan gum, carrageenan, high methoxyl pectin, low methoxyl pectin, guar gum, acacia gum, crystalline cellulose, arabinogalactan, karaya gum, tragacanth gum, alginic acid, albumin, casein, curdlan, gellan gum, and dextran can also be suitably used.

[0044] (9) Examples of bioactive ingredients Bioactive ingredients include substances that provide some kind of bioactivity to the skin when applied to the skin. Examples include whitening ingredients, immune stimulants, anti-aging agents, UV protection agents, slimming agents, skin tightening agents, antioxidants, hair growth agents, hair tonics, 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 keiskei extract, avocado extract, Hydrangea macrophylla extract, Althaea officinalis extract, Arnica montana extract, Aloe vera extract, apricot extract, apricot kernel extract, Ginkgo biloba extract, Fennel extract, Turmeric extract, Oolong tea extract, Rosa multiflora extract, Echinacea japonica leaf extract, Scutellaria baicalensis extract, Phellodendron amurense extract, Coptis japonica extract, Barley extract, Hypericum perforatum extract, Lamium album extract, Nasturtium officinale extract, Orange extract, Dried seawater, seaweed extract, hydrolyzed elastin, hydrolyzed wheat powder, hydrolyzed silk, chamomile extract, carrot extract, Artemisia capillaris extract, licorice extract, carrot extract, Arctium moniliforme extract, cinchona extract, cucumber extract, guanosine, gardenia extract, bamboo grass extract, Sophora flavescens extract, walnut extract, grapefruit extract, clematis extract, chlorella extract, mulberry extract, gentian extract, black tea extract, yeast extract, burdock extract, corn Rice bran fermentation extract, rice germ oil, comfrey extract, collagen, lingonberry extract, asarum extract, bupleurum extract, umbilical cord extract, salvia extract, soapwort extract, bamboo extract, hawthorn extract, sansho extract, shiitake mushroom extract, rehmannia extract, lithospermum extract, perilla extract, linden extract, meadowsweet extract, peony extract, calamus root extract, birch extract, horsetail extract, ivy extract, hawthorn extract, ce Elderflower extract, yarrow extract, peppermint extract, sage extract, mallow extract, Cnidium officinale extract, Swertia japonica extract, soybean extract, jujube extract, thyme extract, tea extract, clove extract, cogongrass extract, citrus peel 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 extracts include Ophiopogon japonicus extract, parsley extract, honey, witch hazel extract, Parietaria trifolia extract, Isodon japonicus extract, bisabolol, loquat extract, coltsfoot extract, butterbur 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, lemon balm 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, royal jelly extract, etc. Furthermore, it contains 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 pyrrolidone carboxylate, betaine, whey, and trimethylglycine; oily ingredients such as sphingolipids, ceramides, phytosphingosine, cholesterol, cholesterol derivatives, and phospholipids; ε-aminocaproic acid, glycyrrhizic acid, β-glycyrrhetinic acid, lysozyme chloride, and guaiazulene. Immunostimulants such as hydroxyl, hydrocholtisone, vitamins such as vitamin A, vitamin B2, vitamin B6, vitamin C, vitamin D, vitamin E, calcium pantothenate, biotin, nicotinamide, and vitamin C ester, active ingredients such as allantoin, diisopropylamine dichloroacetate, 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, and blood circulation promoters such as gamma-oryzanol and vitamin E derivatives. Wound-healing agents such as retinol and retinol derivatives, whitening agents such as arbutin, kojic acid, placenta extract, sulfur, ellagic acid, linoleic acid, tranexamic acid, and glutathione, cepharanthine, licorice extract, capsicum tincture, hinokitiol, garlic iodide extract, pyridoxine hydrochloride, DL-α-tocopherol, DL-α-tocopherol acetate, nicotinic acid, nicotinic acid derivatives, calcium pantothenate, D-pantothenyl alcohol, acetyl pantothenyl ethyl ether, biotin, allantoin, isopropylmethylphenol, estradiol All, ethinylestradiol, capronium chloride, benzalkonium chloride, diphenhydramine hydrochloride, tacanal, camphor, salicylic acid, vanillyl nonyl acid, vanillyl nonanoate, piroctone olamine, glyceryl pentadecanoate, L-menthol, mononitroguaiacol, resorcinol, gamma-aminobutyric acid, benzethonium chloride, mexiletine hydrochloride, auxin, female hormones, cantharis tincture, cyclosporine, zinc pyrithione, hydrocoltisone, minoxidil, polyoxyethylene sorbitan monostearate, peppermint oil,Examples include hair growth products containing Sasanishiki rice extract.

[0045] (10) Examples of antioxidants Examples include sodium bisulfite, sodium sulfite, erythorbic acid, sodium erythorbate, dilauryl thiodipropionate, tocopherol, tol biguanide, nordihydroguaiaretinic acid, parahydroxyanisole, butylhydroxyanisole, dibutylhydroxytoluene, ascorbyl stearate, ascorbyl palmitate, octyl gallate, propyl gallate, carotenoids, flavonoids, tannins, lignans, saponins, and plant extracts with antioxidant effects such as apple extract and clove extract.

[0046] (11) Example of Be Examples include purified water, ethanol, lower alcohols, ethers, LPG, fluorocarbons, N-methylpyrrolidone, fluoroalcohols, volatile linear silicones, and next-generation chlorofluorocarbons (CFCs).

[0047] The photoaging prevention and improvement agent of the present invention can be used as a raw material for food and beverage compositions, pharmaceutical compositions, and topical skin preparations. The same raw materials used in the aforementioned agents can be incorporated into these formulations, and the manufacturing and administration methods can be the same as those used for photoaging prevention and improvement agents. [Examples]

[0048] The following describes examples of the present invention. These examples are provided to confirm the various actions and effects of the agent obtained by the present invention, and the scope of the present invention is not limited to these products and manufacturing methods.

[0049] Examples 1. Preparation of purple tea extract and GHG [Example: Preparation of purple tea extract (GHG-containing composition)] 50 g of purple tea leaves were immersed in 500 mL of a 50% ethanol aqueous solution and extracted by heating under reflux at 40 °C for 2 hours while stirring. 400 mL of the extract was obtained by suction filtration. The extract was concentrated and dried to obtain 16.6 g of purple tea extract.

[0050] [Purification of GHG from purple tea extract] When HPLC analysis was performed on the purple tea extract under the following conditions, a peak of a specific component unique to the purple tea extract that is not contained in common teas such as green tea, oolong tea, and black tea was confirmed at 27.5 min (the arrow part in Figure 5). Sample preparation: 350 mg of purple tea extract was dissolved in a 30% methanol aqueous solution and diluted to 20 mL in a volumetric flask. The solution was diluted 2-fold, and after filter filtration, HPLC analysis was performed. The HPLC analysis conditions are as follows. <HPLC analysis conditions> Flow rate: 0.7 mL / min Mobile phase A: 0.3% TFA aqueous solution Mobile phase B: Acetonitrile Gradient: As shown in Table 1 below Column: SunFire C18, 4.6×150 mm (Waters) or equivalent Column temperature: 30 °C Sample injection volume: 10 μL Detection wavelength: 280 nm

[0051]

Table 1

[0052] Separation and purification of the above-mentioned specific component was carried out, and NMR analysis was performed. The results are shown in Table 2. As a result of Table 2, it was identified as the known component 1,2-di-O-galloyl-4,6-O-(S)-hexahydroxydiphenoyl-β-D-glucose (GHG).

[0053]

Table 2

[0054] Quantitative analysis using HPLC with GHG-purified product as the standard substance revealed that the purple tea extract contained 8.70% GHG. Using the same method as described above, purple tea extract was prepared two more times, and the GHG content in the extract was measured using the same method. The results showed that the GHG content was 6.79% and 6.38%, respectively. This confirmed that the purple tea extract produced by the method of this example contains approximately 6-9% GHG.

[0055] 2. About ferulic acid In this example, "ferulic acid" (product name) manufactured by Oryza Oil & Fat Chemical Co., Ltd. was used.

[0056] Test example: Evaluation of the inhibitory effect of UVA irradiation on cell atrophy. 1. Method for culturing normal diploid fibroblasts (NB1RGB cells) derived from human neonatal skin. NB1RGB cells were cultured and used for the experiment. The culture medium used was Dulbecco's Modified Eagles Medium (DMEM) containing low-concentration glucose (1000 mg / L), supplemented with 10% (v / v) FBS (Fetal Bovine Serum), 100 units / mL penicillin G, and 100 mg / mL streptomycin. Cell culture was performed at 75 cm². 2 The procedure was carried out in a culture flask at 37°C under 5% CO2. For subculturing, the cultured cells were washed twice with PBS (-), and then detached using a phenol red-containing 0.05 (w / v) % trypsin-0.53 mmol / l EDTA·4Na solution before being used in the experiment.

[0057] 2. Method for evaluating the inhibitory effect of UVA irradiation on cell atrophy. 3.0 × 10 on an ibidi 8-well plate 4Cells / mL of cell suspension were seeded in 300 μL / well and cultured for 1 day. Then, the medium was changed to serum-free medium containing the test substance and cultured for another day. Finally, the medium was changed again with PBS (-) and irradiated with UVA (10 J / cm²). 2 Subsequently, the culture medium was changed to a serum-free medium and incubated for 3 hours. Immunofluorescence staining was then performed according to standard procedures. The luminescence intensity and cell number (number of nuclei) after immunofluorescence staining were analyzed using ImageJ. The results are shown in Figures 1 and 2. Figure 1 shows the normal (no UV irradiation) and control (no test substance added) conditions, Figure 2 shows the results after adding the test substances (purple tea and ferulic acid), Figure 3 shows the results after adding the test substance (GHG), and Figure 4 shows a magnified micrograph of Figure 3 (immunofluorescence staining of β-actin).

[0058] 3. Results and Effects of the Examples Normally, when fibroblasts are cultured on a monolayer, they expand their cytoskeleton as shown in the upper part of Figure 1. However, it was confirmed that irradiating with UVA caused the cytoskeleton to atrophy, as shown in the lower part of Figure 1 and the control cells in Figures 2-4. We investigated the effects of purple tea extract, ferulic acid, and GHG, and found that all of them suppressed cell atrophy caused by UVA irradiation (Figures 2-4). The graphs reflect the corrected result of dividing the fluorescence intensity of the cytoskeleton (actin) in the photograph by the number of cell nuclei, thus calculating the actin emission intensity per cell. These results revealed that purple tea extract, ferulic acid, and GHG have cell atrophy-inhibiting effects, confirming their usefulness as anti- and anti-aging agents for the skin.

[0059] Examples of formulations of the photoaging prevention and improvement agent (purple tea extract, ferulic acid, or GHG) according to the present invention are shown below. Note that the following formulation examples are not limiting to the present invention. Example formulation 1: Chewing gum Sugar 53.0 wt% Gum base 20.0 Glucose 10.0 Starch syrup 16.0 Fragrance 0.5 Photoaging prevention and improvement agent 0.5 100.0 wt%

[0060] Example formulation 2: Gummy Reduced starch syrup 40.0 wt% Granulated sugar 20.0 Glucose 20.0 Gelatin 4.7 Wednesday 9.68 Yuzu juice 4.0 Yuzu flavor 0.6 Dye 0.02 Photoaging prevention and improvement agent 1.0 100.0 wt%

[0061] Formula example 3: Candy Sugar 50.0 wt% Starch syrup 33.0 Wednesday 14.4 organic acid 2.0 Fragrance 0.2 Photoaging prevention and improvement agent 0.4 100.0 wt%

[0062] Example of formulation 4: Yogurt (hard / soft) Milk 41.5 wt% Skim milk powder 5.8 Sugar 8.0 Agar 0.15 Gelatin 0.1 Lactic acid bacteria 0.005 Photoaging prevention and improvement agent 0.4 fragrance trace amount water residue 100.0 wt%

[0063] Formulation example 5: Soft drink Fructose-glucose liquid sugar 30.0 wt% Emulsifier 0.5 Photoaging prevention and improvement agent 0.3 Fragrance (appropriate amount) Purified water remainder 100.0 wt%

[0064] Formulation example 6: Tablet confectionery Sugar 76.4 wt% Glucose 19.0 Sucrose fatty acid ester 0.2 Photoaging prevention and improvement agent 0.5 Purified water 3.9 100.0 wt%

[0065] Formulation Example 7: Soft Capsules Brown rice germ oil 47.0 wt% Yuzu seed oil 40.0 Emulsifier 12.0 Photoaging prevention and improvement agent 1.0 100.0 wt%

[0066] Formulation example 8: Tablets Lactose 54.0wt% Crystalline cellulose 30.0 Starch decomposition product 10.0 Glycerin fatty acid ester 5.0 Photoaging prevention and improvement agent 1.0 100.0 wt%

[0067] Example formulation 9: Cosmetic cream Squalane 20.0 wt% Beeswax 5.0 Refined jojoba oil 5.0 Glycerin 5.0 Glycerin monostearate 2.0 Polyoxyethylene (20) sorbitan Monosterate 2.0 Photoaging prevention and improvement agent 2.0 Preservative (appropriate amount) Fragrance (appropriate amount) Purified water remainder 100.0 wt%

[0068] Formula example 10: Lotion Ethanol 5.0 wt% Glycerin 2.0 1,3-Butylene glycol 2.0 Polyethylene oleyl ether 0.5 Sodium citrate 0.1 Citric acid 0.1 Photoaging prevention and improvement agent 0.1 Purified water remainder 100.0 wt%

[0069] Formula Example 11: Body Gel Macadamia nut oil 2.0 wt% 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 Retinyl palmitate 0.1 Preservative (appropriate amount) Centella asiatica extract 1.0 Photoaging prevention and improvement agent 1.0 1,3-Butylene glycol 5.0 Purified water remainder 100.0 wt%

[0070] Formulation example 12: Emulsion Squalane 4.0 wt% Vaseline 2.5 Cetanol 2.0 Glycerin 2.0 Lipophilic glyceryl monostearate 1.0 Stearic acid 1.0 L-arginine 1.0 Photoaging prevention and improvement agent 0.5 Potassium hydroxide 0.1 fragrance trace amount Purified water remainder 100.0 wt%

[0071] Formula Example 13: Bath additive (liquid) Propylene glycol 50.0 wt% Ethanol 20.0 Sodium sulfate 5.0 Photoaging prevention and improvement agent 0.5 Lanolin 0.5 Avocado oil 0.5 Dye 1.5 Fragrance 22.0 100.0 wt% [Industrial applicability]

[0072] Based on the above, the present invention can provide a novel anti-photoaging agent for the skin.

Claims

1. A UVA-induced cell atrophy inhibitor for fibroblasts, containing purple tea (scientific name: Camellia sinensis, variety name TRFK306) extract as the active ingredient.

2. A fibroblast cell atrophy inhibitor containing GHG (1,2-di-O-galloyl-4,6-O-(S)-hexahydroxydiphenoyl-β-D-glucose) as the active ingredient.

3. A ferulic acid-containing agent that inhibits UVA-induced cell atrophy in fibroblasts.

4. A photoaging prevention and improvement agent comprising the agent described in any one of claims 1 to 3 as an active ingredient.

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