HSF1 expression enhancer, composition containing the same, and screening method

Abstract

The objective of this invention is to provide a novel ingredient that can prevent and improve the deterioration of the skin surface condition. [Solution] The present invention relates to an HSF1 expression enhancer, etc., containing Geranium thunbergii extract.

Description

Technical Field

[0001] The present invention relates to an HSF1 expression enhancer, a composition containing the same, a screening method, and the like.

Background Art

[0002] When skin turnover is disrupted and the skin barrier function is reduced, deterioration of the skin surface condition such as dryness, exfoliation of stratum corneum cells, and itching occurs. Deterioration of the skin surface condition can also be rephrased as rough skin, which is known to be easily caused by accumulation of physical stimuli such as ultraviolet exposure and aging. However, since the onset factors are complicated, it is not easy to deal with, and conventional ingredients that can prevent or improve deterioration of the skin surface condition have not always provided sufficient effects. Therefore, development of a new ingredient capable of preventing and improving deterioration of the skin surface condition has been desired.

[0003] On the other hand, HSF1 (Heat shock transcription factor 1; heat shock transcription factor 1) is a factor identified as a transcription factor that controls the expression of HSP (Heat shock protein; heat shock protein). When cells are exposed to stress conditions such as heat and UV, it regulates various heat shock proteins and contributes to cell protection. It has also been revealed that HSF1 regulates proteins other than heat shock proteins and is involved in various life phenomena such as development, metabolism, aging, neurodegenerative diseases associated with aging, and cancer. Non-Patent Document 1 reports that the expression level of HSF1 decreases with aging in dermal fibroblasts. HSP functions in the dermis such as normalizing collagen structure and protecting fibroblasts, and it is considered that decreased expression of HSF1 reduces these functions, leading to wrinkles and sagging. However, the relationship between HSF1 and deterioration of skin surface condition has not been known.

Prior Art Literature

Non-Patent Literature

[0004] [Non-Patent Document 1] Gutsmann-Conrad A, Heydari AR, You S, Richardson A. The expression of heat shock protein 70 decreases with cellular senescence in vitro and in cells derived from young and old human subjects. Exp Cell Res. 1998 Jun 15;241(2):404-13. [Overview of the project] [Problems that the invention aims to solve]

[0005] In view of these circumstances, the object of the present invention is to provide a new ingredient that can prevent and improve the deterioration of the skin surface condition. [Means for solving the problem]

[0006] In their investigation to solve the above problems, the inventors focused on HSF1. They discovered that the expression level of the HSF1 gene is reduced in aging epidermal keratinocytes. Furthermore, they found that intentionally reducing the expression level of the HSF1 gene leads to a decrease in factors related to epidermal turnover and barrier function. This led them to discover that a decrease in HSF1 contributes to the deterioration of the skin surface condition. Based on this finding, they searched for components that increase the expression level of the HSF1 gene and, among various components, found that Geranium thunbergii extract has this effect, thus completing the present invention. In other words, the gist of this invention is as follows:

[0007] [1] A HSF1 (Heat shock transcription factor 1) expression enhancer containing Geranium thunbergii extract. [2] The agent wherein the enhanced expression is an enhanced expression in the epidermis. [3] The agent wherein the enhancement of expression is an enhancement of gene expression. [4] A composition containing the aforementioned agent for preventing and / or improving the deterioration of the skin surface condition. [5] A composition for improving the appearance of the face, containing the aforementioned agent. [6] The composition is a cosmetic. [7] A method for screening ingredients that prevent and / or improve the deterioration of skin surface conditions, comprising a selection step of selecting active ingredients based on the expression level of the HSF1 gene in cells. [Effects of the Invention]

[0008] According to the present invention, we can provide Geranium thunbergii extract as a novel ingredient that can prevent and improve the deterioration of the skin surface condition. Geranium thunbergii extract can increase the expression level of the HSF1 gene in epidermal keratinocytes, thereby preventing and improving the deterioration of the skin surface condition. Furthermore, by improving the roughness of the skin surface, it can also improve the appearance. In addition, the expression level of the HSF1 gene in cells can be used as an indicator to screen for ingredients that prevent and / or improve the deterioration of the skin surface condition. [Brief explanation of the drawing]

[0009] [Figure 1] This graph shows the expression levels of the HSF1 gene in young and aged epidermal keratinocytes. The expression levels of the HSF1 gene were analyzed in epidermal keratinocytes from three donors each in their 20s (young) and 50s (aged). n=9, mean ± standard deviation. *: p<0.05 (t-test) [Figure 2]This is a graph showing the effect of reduced HSF1 gene expression on factors involved in skin barrier function and turnover. The HSF1 gene expression level was artificially reduced in epidermal keratinocytes, and the gene expression levels of Loricrin (LOR), Hyaluronate synthase 3 (HAS3), and Keratin-1 (KRT1) were analyzed. n=6~8, mean ± standard deviation *:p<0.05, ***:p<0.001 (t-test) [Figure 3] This is a graph showing the effect of Geranium thunbergii extract on HSF1 gene expression level in epidermal keratinocytes. The extract was added to epidermal keratinocytes for culture, and the expression level of HSF1 gene was analyzed. n=8, mean ± standard deviation *:p<0.05 (t-test) [Figure 4] This is a graph showing the effect of Geranium thunbergii extract on HSF1 gene expression level in dermal fibroblasts. Geranium thunbergii extract was added to dermal fibroblasts for culture, and the expression level of HSF1 gene was analyzed. n=4, mean ± standard deviation *:p<0.05 (t-test) [Mode for Carrying Out the Invention]

[0010] The present invention is described below. However, the present invention is not limited to the following embodiments, and can be freely modified within the scope of the present invention.

[0011] In the present specification, for numerical ranges expressed as "lower limit to upper limit", the upper limit may be "or less" or "less than", and the lower limit may be "or more" or "more than".

[0012] <HSF1 expression enhancer> One aspect of the present invention relates to an HSF1 expression enhancer containing a Geranium thunbergii extract (hereinafter sometimes referred to as "the HSF1 expression enhancer of the present invention"). The HSF1 expression enhancer of the present invention has an HSF1 expression enhancing effect. Examples of subjects in which the expression of HSF1 is enhanced by the HSF1 expression enhancer of the present invention include the epidermis, and examples of subject cells include keratinocytes, horny layer cells, and the like. Furthermore, the HSF1 expression enhancer of the present invention further enhances the expression of HSF1 in the dermis, and particularly enhances the expression of HSF1 in dermal fibroblasts.

[0013] As used herein, "enhanced expression" means that the expression level and / or expression rate of HSF1 protein or its gene is increased compared to when the expression enhancer of the present invention is not applied, and also refers to suppressing a decrease in said expression level and / or expression rate. The degree of such increase or suppression is not particularly limited. Hereinafter, in the present specification, "extract" is also referred to as "extract", which is synonymous.

[0014] ≪Geranium thunbergii Extract≫ The HSF1 expression enhancer of the present invention contains Geranium thunbergii extract as an active ingredient and increases the expression level of HSF1. Geranium thunbergii is a plant belonging to the genus Geranium of the family Geraniaceae. The Geranium thunbergii extract in the present invention can be prepared by using products sold as naturally grown or cultivated plants, crude Chinese herbal medicine raw materials, etc., or commercially available extracts sold by companies handling plant extracts can be purchased and used.

[0015] The term plant extract is a general term that includes not only the plant extract itself, but also fractions of the extract, purified fractions, solvent-removed products of the extract, fraction, or purified product. Examples of plant-derived extracts include extracts prepared from products sold as naturally grown or cultivated plants, crude Chinese herbal medicine raw materials, etc., and commercially available extracts. The extraction process can use the whole plant, or parts such as the plant body, above-ground parts, rhizomes, trunks, leaves, stems, inflorescences, and flower buds. However, it is preferable to pre-grind or finely chop these parts to improve extraction efficiency. Suitable extraction solvents include one or more polar solvents selected from water, alcohols such as ethanol, isopropyl alcohol, and butanol; polyhydric alcohols such as 1,3-butanediol and polypropylene glycol; ketones such as acetone and methyl ethyl ketone; and ethers such as diethyl ether and tetrahydrofuran. A specific extraction method may involve adding 1 to 30 parts by mass of the solvent to 1 mass of the plant body or its dried form, immersing it for several days at room temperature or for several hours at a temperature near its boiling point, cooling it to room temperature, removing insoluble matter and / or solvent as desired, and then fractionating and purifying it by column chromatography or the like.

[0016] The content of Geranium thunbergii extract in the HSF1 expression enhancer is preferably 0.001% by mass or more, 0.01% by mass or more, 0.1% by mass or more, or 1% by mass or more, based on the total amount of dry solids, and may also be 100% by mass or less, 90% by mass or less, 80% by mass or less, 50% by mass or less, or 10% by mass or less, and may be any non-consistent combination thereof. The content may also be, for example, 0.001 to 100% by mass. Examples of pharmaceutical raw materials to be combined with Geranium thunbergii extract include known pharmaceutical raw materials, excipients, stabilizers, etc. The HSF1 expression enhancer of the present invention can be manufactured by referring to known manufacturing techniques commonly used in the pharmaceutical field.

[0017] <Composition> Another aspect of the present invention relates to a composition for preventing and / or improving the deterioration of skin surface conditions (hereinafter sometimes referred to as "the composition of the present invention"), which contains the HSF1 expression enhancer of the present invention. Here, "prevention and / or improvement of the deterioration of skin surface conditions" specifically means, for example, This may mean preventing and / or improving skin roughness, disorder of the stratum corneum (roughness of the skin surface), decreased barrier function, and decreased cell turnover. The HSF1 expression enhancer of the present invention has an HSF1 expression-enhancing effect. Furthermore, as described below, it has become clear that HSF1 is related to the expression of factors associated with epidermal turnover and barrier function. Therefore, the composition of the present invention, by having an HSF1 expression-enhancing effect, provides a preventive and / or ameliorative effect on the deterioration of skin surface conditions caused by a decrease in HSF1 in the epidermis.

[0018] Furthermore, the inventors have previously shown that the roughness of the skin surface affects various aspects of appearance, such as sagging. Therefore, it is believed that improving the roughness of the skin surface can also improve the overall appearance. Therefore, another aspect of the present invention relates to a composition for improving the appearance of the face, which contains the HSF1 expression enhancer of the present invention (this aspect may also be referred to hereinafter as "the composition of the present invention").

[0019] Furthermore, it has been reported that HSF1 expression decreases in dermal fibroblasts due to aging and other factors. HSPs play a role in normalizing collagen structure and protecting fibroblasts in the dermis, and it is thought that a decrease in HSF1 expression leads to a decline in these functions, resulting in wrinkles and sagging. However, since the composition of the present invention also has an effect of enhancing HSF1 expression in the dermis, it is thought to have a preventive and / or ameliorative effect on phenomena caused by HSF1 reduction in the dermis, such as wrinkles and sagging. In other words, it is thought that the composition of the present invention improves the appearance of the face through both mechanisms: improvement of skin surface roughness in the epidermis and improvement of wrinkles and sagging in the dermis.

[0020] The content of the Geranium thunbergii extract, which is the active ingredient in the composition of the present invention, is preferably 0.0001% by mass or more, 0.001% by mass or more, 0.01% by mass or more, 0.1% by mass or more, or 0.5% by mass or more, as dry solid content, and may also be 30% by mass or less, 10% by mass or less, 5% by mass or less, 3% by mass or less, 2% by mass or less, or 1% by mass or less, and may be any non-consistent combination of these. Furthermore, the content may be, for example, 0.0001 to 30% by mass.

[0021] The compositions of the present invention can be specifically applied to cosmetics, quasi-drugs, pharmaceuticals, foods, etc., and may contain the necessary components as appropriate depending on the application. Examples of dosage forms include topical preparations and oral preparations. Topical preparations are preferably in the form of skin preparations, while oral preparations are preferably in the form of supplements such as tablets, granules, or drinks. The compositions of the present invention can be manufactured by referring to known manufacturing techniques commonly used in each respective field.

[0022] The composition of the present invention is particularly preferably applied to cosmetics. When applied to cosmetics, it is possible to incorporate a wide range of ingredients commonly used in cosmetics, and there are no limitations on the dosage form or application. Examples of formulations include liquid, emulsion, semi-solid (gel, ointment, paste, cream, etc.), solid, or foam. Cosmetics include facial cleansers, makeup removers, skincare products (lotions, serums, emulsions, creams, etc.), sunscreens, and makeup products.

[0023] The following describes other ingredients that may be included in cosmetics when applicable.

[0024] ≪Optional ingredients≫ The cosmetic composition of the present invention may optionally contain other ingredients commonly used in cosmetics, as long as they do not impair the effects of the present invention. Examples of such optional ingredients include oils, alcohols, ethers, powders, clay minerals, humectants, surfactants, metal ion chelating agents, pearlescent agents, amino acids, organic amines, polymer emulsions, pH adjusters, vitamins, antioxidants, preservatives, water-soluble polymers, fragrances, solvents such as water, and various active ingredients. Note that only one optional ingredient may be included, or two or more may be included in combination.

[0025] Examples of oils include polar oils, natural oils, and hydrocarbon oils.

[0026] As polar oils, synthetic ester oils include isopropyl myristate, cetyl octanoate, octyldodecyl myristate, isopropyl palmitate, butyl stearate, hexyl laurate, myristyl myristate, decyl oleate, hexyldecyl dimethyloctanoate, cetyl lactate, myristyl lactate, lanolin acetate, isocetyl stearate, isocetyl isostearate, cholesteryl 12-hydroxystearylate, ethylene glycol di-2-ethylhexylate, dipentaerythritol fatty acid ester, N-alkyl glycol monoisostearate, neopentyl glycol dicaprate, diisostearyl malate, glyceryl di-2-heptylundecanoate, trimethylolpropane tri-2-ethylhexylate, trimethylolpropane triisostearate, pentaneerythritol tetra-2-ethylhexylate, glyceryl tri-2-ethylhexanoate, triisoste Trimethylolpropane arate, cetyl 2-ethylhexanoate, 2-ethylhexyl palmitate, glyceryl trimiristic acid, glyceride tri-2-heptyl undecanoate, methyl castor oil fatty acid ester, oleic acid oil, cetostearyl alcohol, acetoglyceride, 2-heptyl undecyl palmitate, diisobutyl adipate, 2-octyldodecyl N-lauroyl-L-glutamate, di-2-heptyl adipate Examples include ethyl undecyl, ethyl laurate, di-2-ethylhexyl sebatate, 2-hexyldecyl myristate, 2-hexyldecyl palmitate, 2-hexyldecyl adipic acid, diisopropyl sebatate, 2-ethylhexyl succinate, ethyl acetate, butyl acetate, amyl acetate, triethyl citrate, octyl methoxycinnamate, 2-ethylhexyl paramethoxycinnamate, and diethylamino hydroxybenzoyl hexyl benzoate.

[0027] Examples of natural oils include avocado oil, camellia oil, turtle oil, macadamia nut oil, corn oil, mink oil, olive oil, rapeseed oil, egg yolk oil, sesame oil, peach kernel oil, wheat germ oil, sasanqua oil, castor oil, linseed oil, safflower oil, cottonseed oil, elm oil, soybean oil, peanut oil, tea seed oil, kaya oil, rice bran oil, cinnamon oil, Japanese tung oil, jojoba oil, wheat germ oil, sunflower oil, triglycerin, glyceryl trioctanoate, and glyceryl triisopalmitate. Examples of hydrocarbon oils include isododecane, isohexadecane, squalane, hydrogenated poly(C6-12) olefin, and hydrogenated polyisobutene.

[0028] Examples of alcohols include monohydric alcohols such as ethanol; dihydric alcohols such as ethylene glycol, 1,3-butylene glycol, trimethylene glycol, 1,2-butylene glycol, propylene glycol, dipropylene glycol, propanediol, 1,2-pentanediol, 3-methyl-1,3-butanediol, tetramethylene glycol, 2,3-butylene glycol, pentamethylene glycol, 2-butene-1,4-diol, hexylene glycol, octylene glycol; trihydric alcohols such as trimethylolpropane; tetrahydric alcohols such as pentaerythritol; pentahydric alcohols such as xylitol; polyhydric alcohol polymers such as triglycerin, tetraglycerin, and polyglycerin; and sugar alcohols such as starch-degrading sugar-reducing alcohols. Ethylene glycol monomethyl ether, ethylene glycol monoethyl ether, Alcohol alkyl ethers such as ethylene glycol monobutyl ether, ethylene glycol monophenyl ether, ethylene glycol monohexyl ether, ethylene glycol mono-2-methylhexyl ether, ethylene glycol isoamyl ether, ethylene glycol benzyl ether, and ethylene glycol isopropyl ether; Alcohol alkyl ethers such as diethylene glycol monomethyl ether, diethylene glycol monoethyl ether, diethylene glycol monobutyl ether, triethylene glycol monomethyl ether, triethylene glycol monoethyl ether, propylene glycol monomethyl ether, propylene glycol monoethyl ether, propylene glycol monobutyl ether, propylene glycol isopropyl ether, dipropylene glycol methyl ether, dipropylene glycol ethyl ether, and dipropylene glycol butyl ether; Glycerin monoalkyl ethers such as xyl alcohol, ceracyl alcohol, and batyl alcohol; Alcohol polymers such as diethylene glycol, triethylene glycol, polypropylene glycol, tetraethylene glycol, and polyethylene glycol; Examples include glycolid; tetrahydrofurfuryl alcohol; POE-tetrahydrofurfuryl alcohol; POP-butyl ether; POP·POE-butyl ether; tripolyoxypropylene glycerin ether; POP-glycerin ether; POP-glycerin ether phosphate; POP·POE-pentaneerythritol ether, etc.

[0029] Ethers include those that fall under the category of alcohols mentioned above, Examples include ethylene glycol dimethyl ether, ethylene glycol diethyl ether, ethylene glycol dibutyl ether, diethylene glycol dimethyl ether, diethylene glycol diethyl ether, diethylene glycol methyl ethyl ether, ethylene glycol monomethyl ether acetate, ethylene glycol monoethyl ether acetate, ethylene glycol monobutyl ether acetate, ethylene glycol monophenyl ether acetate, ethylene glycol diazibate, ethylene glycol disuccinate, diethylene glycol monoethyl ether acetate, diethylene glycol monobutyl ether acetate, propylene glycol monomethyl ether acetate, propylene glycol monoethyl ether acetate, propylene glycol monopropyl ether acetate, and propylene glycol monophenyl ether acetate.

[0030] The shape of the powder is not particularly limited, and can be spherical, needle-shaped, plate-shaped, etc. Examples include inorganic powders (e.g., talc, kaolin, mica, sericite, muscovite, phlogopite, synthetic mica, red mica, biotite, vermiculite, magnesium carbonate, calcium carbonate, aluminum silicate, barium silicate, calcium silicate, magnesium silicate, strontium silicate, tungstate metal salts, magnesium, silica, alumina, zeolite, barium sulfate, calcined calcium sulfate (calcined gypsum), calcium phosphate, fluorine apatite, hydroxyapatite, ceramic powder, boron nitride, etc.); organic powders (e.g., polyamide resin powder (nylon powder), polyethylene powder, poly(meth)acrylate powder, polystyrene powder, styrene-acrylic acid copolymer resin powder, silicone resin powder, benzoguanamine resin powder, polytetrafluoroethylene powder, cellulose powder, etc.); metal soaps (e.g., Zinc lysitate, calcium palmitate, aluminum stearate; inorganic white pigment (e.g., titanium dioxide, zinc oxide, etc.); inorganic red pigments (e.g., iron oxide (red iron oxide), iron titanate, etc.); inorganic brown pigments (e.g., γ-iron oxide, etc.); inorganic yellow pigments (e.g., yellow iron oxide, ochre, etc.); inorganic black pigments (e.g., black iron oxide, lower titanium oxide, etc.); inorganic purple pigments (e.g., mango violet, cobalt violet, etc.); inorganic green pigments (e.g., chromium oxide, chromium hydroxide, cobalt titanate, etc.); inorganic blue pigments (e.g., ultramarine, Prussian blue, etc.); pearl pigments (e.g., titanium dioxide-coated mica, titanium dioxide-coated bismuth oxychloride, titanium dioxide-coated talc, colored titanium dioxide-coated mica, bismuth oxychloride, fish scale foil, etc.); gold Examples include powdered pigments (e.g., aluminum powder, copper powder); organic pigments such as zirconium, barium, or aluminum lake (e.g., organic pigments such as Red 201, Red 202, Red 204, Red 205, Red 220, Red 226, Red 228, Red 405, Orange 203, Orange 204, Yellow 205, Yellow 401, and Blue 404; Red 3, Red 104, Red 106, Red 227, Red 230, Red 401, Red 505, Orange 205, Yellow 4, Yellow 5, Yellow 202, Yellow 203, Green 3, and Blue 1); natural pigments (e.g., chlorophyll, β-carotene); and organically modified clay minerals.

[0031] Examples of organically modified clay minerals include hectorite, montmorillonite, saponite, bentonite, and smectite. Alternatively, organically modified clay minerals treated with organic cations are also acceptable, such as dimethyldistearylammonium hectorite (quaternium-18 hectorite, etc.), dimethyldistearylammonium bentonite (quaternium-18 bentonite, quaternium-90 bentonite, etc.), benzyldimethylstearylammonium hectorite, dioctadecyldimethylammonium salt-modified montmorillonite, octadecyldimethylbenzylammonium salt-modified montmorillonite, and dihexadecyldimethylammonium salt-modified montmorillonite, and dimethyldistearylammonium smectite.

[0032] Examples of moisturizing agents include chondroitin sulfate, hyaluronic acid, mucoitin sulfate, carotenoid acid, atelocollagen, cholesteryl-12-hydroxystearate, sodium lactate, dl-pyrrolidone carboxylate, short-chain soluble collagen, diglycerin (EO)PO adduct, Rosa rugosa extract, Achillea millefolium extract, and Melilotus extract.

[0033] When a surfactant is used in combination with the composition of the present invention, examples of surfactants include anionic surfactants, cationic surfactants, amphoteric surfactants, and nonionic surfactants.

[0034] Examples of anionic surfactants include fatty acid soaps (e.g., sodium laurate, sodium palmitate, etc.); higher alkyl sulfate salts (e.g., sodium lauryl sulfate, potassium lauryl sulfate, etc.); alkyl ether sulfate salts (e.g., POE lauryl sulfate triethanolamine, POE lauryl sulfate sodium, etc.); N-acyl sarcosinate (e.g., sodium lauroyl sarcosinate, etc.); higher fatty acid amide sulfonates (e.g., sodium N-myristoyl-N-methyl taurate, sodium coconut oil fatty acid methyl taulide, sodium lauryl methyl taulide, etc.); phosphate salts (e.g., sodium POE oleyl ether phosphate, POE stearyl ether phosphate, etc.); sulfosuccinates (e.g., sodium di-2-ethylhexyl sulfosuccinate, sodium monolauroyl monoethanolamide polyoxyethylene sulfosuccinate, lauryl polypropylene glycol sulfosuccinate) Examples include sodium citrate, alkylbenzene sulfonates (e.g., linear dodecylbenzenesulfonate sodium, linear dodecylbenzenesulfonate triethanolamine, linear dodecylbenzenesulfonate, etc.), higher fatty acid ester sulfates (e.g., hydrogenated coconut oil fatty acid glycerin sulfate sodium, etc.), N-acyl glutamates (e.g., N-lauroyl glutamate monosodium, N-stearoyl glutamate disodium, N-myristoyl-L-glutamate monosodium, etc.), sulfated oils (e.g., belladonna oil, etc.), POE alkyl ether carboxylic acids, POE alkyl allyl ether carboxylic acid salts, α-olefin sulfonates, higher fatty acid ester sulfonates, secondary alcohol sulfates, higher fatty acid alkylolamide sulfates, lauroyl monoethanolamide succinate sodium, N-palmitoyl aspartate ditriethanolamine, sodium caseinate, etc.

[0035] Examples of cationic surfactants include alkyltrimethylammonium salts (e.g., stearyltrimethylammonium chloride, lauryltrimethylammonium chloride, etc.); alkylpyridinium salts (e.g., cetylpyridinium chloride, etc.); distearyldimethylammonium chloride dialkyldimethylammonium salt; poly(N,N'-dimethyl-3,5-methylenepiperidinium) chloride; alkylquaternary ammonium salts; alkyldimethylbenzylammonium salts; alkylisoquinolinium salts; dialkylmolyphonium salts; POE alkylamines; alkylamine salts; polyamine fatty acid derivatives; amyl alcohol fatty acid derivatives; benzalkonium chloride; benzethonium chloride, etc.

[0036] Examples of amphoteric surfactants include imidazoline-based amphoteric surfactants (e.g., 2-undecyl-N,N,N-(hydroxyethylcarboxymethyl)-2-imidazoline sodium, 2-cocoyl-2-imidazolinium hydroxide-1-carboxyethyloxy disodium salt, etc.); and betaine-based surfactants (e.g., 2-heptadecyl-N-carboxymethyl-N-hydroxyethylimidazolinium betaine, lauryldimethylaminoacetic acid betaine, alkyl betaine, amide betaine, sulfobetaine, etc.).

[0037] Nonionic surfactants include polyether-modified silicones (e.g., polyethylene glycol-10 dimethicone, polyethylene glycol-12 dimethicone, PEG-9 polydimethylsiloxyethyl dimethicone, PEG-9 polydimethylsiloxyethyl dimethicone, etc.); polyglycerin-modified silicones (e.g., polyglyceryl-3 disiloxane dimethicone, polyglyceryl-3 polydimethylsiloxyethyl dimethicone, etc.); sorbitan fatty acid esters (e.g., sorbitan monooleate, sorbitan monostearate, sorbitan monoisostearate, sorbitan monolaurate, sorbitan monopalmitate, sorbitan monostearate, sorbitan) Examples of non-hydrophilic nonionic surfactants include: sesquioleates, sorbitan trioleates, diglycerol sorbitan penta-2-ethylhexylate, diglycerol sorbitan tetra-2-ethylhexylate, etc.); glycerin fatty acids (e.g., monocottonseed oil fatty acid glycerin, monoerucate glycerin, sesquioleate glycerin, monostearate glycerin, α,α'-oleate pyroglutamate glycerin, monostearate glycerin malic acid, etc.); propylene glycol fatty acid esters (e.g., propylene glycol monostearate, etc.); POE castor oil and hydrogenated castor oil derivatives (POE castor oil, POE hydrogenated castor oil, etc.); and glycerin alkyl ethers. Furthermore, polyglycerin fatty acid esters (e.g., polyglyceryl monooleate, polyglyceryl monostearate, etc.); POE sorbitan fatty acid esters (e.g., POE sorbitan monooleate, POE sorbitan monostearate, POE sorbitan tetraoleate, etc.); POE sorbitan fatty acid esters (e.g., POE sorbitan monolaurate, POE sorbitan monooleate, POE sorbitan pentaoleate, POE sorbitan monostearate, etc.); POE glycerin fatty acid esters (e.g., POE monooleates such as POE glycerin monostearate, POE glycerin monoisostearate, POE glycerin triisostearate, etc.); POE fatty acid esters (e.g., POE distearate, POE monodioleate, ethylene glycol distearate, etc.);POE alkyl ethers (e.g., POE lauryl ether, POE oleyl ether, POE stearyl ether, POE-behenyl ether, POE-2-octyldodecyl ether, POE cholestanol ether, etc.); POE alkylphenyl ethers (e.g., POE nonylphenyl ether, etc.); Pluronic type ethers (e.g., Pluronic®, etc.); POE·POP alkyl ethers (e.g., POE·POP cetyl ether, POE·POP-2-decyltetradecyl ether, POE·POP monobutyl ether, POE·POP hydrogenated lanolin, POE·POP glycerin ether, etc.); tetra Other examples include POE-tetraPOP ethylenediamine condensates (e.g., Tetronic); POE castor oil hydrogenated castor oil derivatives (e.g., POE castor oil, POE hydrogenated castor oil, POE hydrogenated castor oil monoisostearate, POE hydrogenated castor oil triisostearate, POE hydrogenated castor oil monopyroglutamic acid monoisostearate diester, POE hydrogenated castor oil maleic acid, etc.); POE beeswax-lanolin derivatives (e.g., POE sorbitol beeswax, etc.); alkanolamides (e.g., coconut oil fatty acid diethanolamide, lauric acid monoethanolamide, fatty acid isopropanolamide, etc.); POE propylene glycol fatty acid esters; POE alkylamines; POE fatty acid amides; sucrose fatty acid esters; alkyl glucosides; alkyl ethoxydimethylamine oxides; and hydrophilic nonionic surfactants such as trioleyl phosphate.

[0038] Examples of metal ion chelating agents include 1-hydroxyethane-1,1-diphosphonic acid, tetrasodium 1-hydroxyethane-1,1-diphosphonic acid, disodium edetate, trisodium edetate, tetrasodium edetate, sodium citrate, sodium polyphosphate, sodium metaphosphate, gluconic acid, phosphoric acid, citric acid, ascorbic acid, succinic acid, edetate, and trisodium ethylenediaminehydroxyethyl triacetate.

[0039] Examples of pearlescent agents include glycol distearate and titanium mica.

[0040] Examples of amino acids include neutral amino acids (e.g., threonine, cysteine, etc.) and basic amino acids (e.g., hydroxylysine, etc.). Examples of amino acid derivatives include sodium acylsarcosinate (sodium lauroylsarcosinate), and acylsarcosinate. Examples include glutamate, acyl β-alanine sodium, glutathione, and pyrrolidone carboxylic acid.

[0041] Examples of organic amines include monoethanolamine, diethanolamine, triethanolamine, morpholine, triisopropanolamine, 2-amino-2-methyl-1,3-propanediol, and 2-amino-2-methyl-1-propanol.

[0042] Examples of polymer emulsions include acrylic resin emulsion, ethyl polyacrylate emulsion, acrylic resin liquid, polyacrylic alkyl ester emulsion, polyvinyl acetate resin emulsion, and natural rubber latex.

[0043] Examples of pH adjusting agents include buffers such as lactate-sodium lactate, citrate-sodium citrate, and succinate-sodium succinate, and are usually used to adjust the pH of the internal phase (aqueous component).

[0044] Examples of vitamins include vitamins A, B1, B2, B6, C, E and their derivatives. Examples include phosphate, pantothenic acid and its derivatives, and biotin.

[0045] Examples of antioxidants include tocopherols, dibutylhydroxytoluene (BHT), butylhydroxyanisole, pyrosulfites, gallic acid esters, phosphoric acid, citric acid, ascorbic acid, maleic acid, malonic acid, succinic acid, fumaric acid, kephalin, hexametaphosphorate, phytic acid, and ethylenediaminetetraacetic acid.

[0046] Examples of preservatives include parabens and phenoxyethanol.

[0047] Examples of water-soluble polymers include plant-derived polymers (e.g., gum arabic, tragacanth gum, galactan, guar gum, carob gum, karaya gum, carrageenan, pectin, agar, quince seed (quince), algae colloid (cassia extract), starch (co (Corn, potato, wheat), glycyrrhizic acid); microbial polymers (for example) For example, xanthan gum, dextran, succinoglucan, pullulan, etc.; natural water-soluble polymers such as animal-derived polymers (e.g., collagen, casein, albumin, gelatin, etc.), Semi-synthetic water-soluble polymers such as starch polymers (e.g., carboxymethyl starch, methylhydroxypropyl starch, etc.); cellulose polymers (hydroxypropyl methylcellulose, hydroxyethylcellulose, hydroxypropylcellulose, methylcellulose, ethylcellulose, hydroxypropyl methylcellulose stearoxyl ether, sodium cellulose sulfate, carboxymethylcellulose, sodium carboxymethylcellulose, crystalline cellulose, cellulose powder, etc.); and alginate polymers (e.g., sodium alginate, propylene glycol alginate, etc.). Examples of water-soluble polymers used in the synthesis of vinyl polymers (e.g., polyvinyl alcohol, polyvinyl methyl ether, polyvinylpyrrolidone, carboxyvinyl polymer, etc.); polyoxyethylene polymers (e.g., polyethylene glycol 20,000, 40,000, 60,000, etc.); acrylic polymers (e.g., sodium polyacrylate, polyethyl acrylate, polyacrylamide, etc.); polyethyleneimines; and cationic polymers.

[0048] Examples of various active ingredients include: anti-inflammatory agents (e.g., glycyrrhizic acid derivatives, glycyrrhetinic acid derivatives, salicylic acid derivatives, hinokitiol, zinc oxide, allantoin, etc.); whitening agents (e.g., placenta extract, saxifrage extract, arbutin, resorcinol derivatives, etc.); various extracts (e.g., Phellodendron bark, Coptis rhizome, Lithospermum root, Peony root, Swertia herb, birch, sage, loquat leaf, carrot, aloe, mallow, iris, grape, coix seed, loofah, lily, saffron, Ligusticum chuanxiong rhizome, Zingiber officinale root, Hypericum perforatum, Ononis spinosa, garlic, capsicum, citrus peel, Angelica acutiloba root, seaweed, etc.); activators (e.g., royal jelly, photosensitizing pigments, cholesterol derivatives, etc.); blood circulation promoters (e.g., nonylic acid vanillylamide, benzyl nicotinate, β-butoxyethyl nicotinate, capsaicin, zingerone, cantharides tincture, ichthammol, tannic acid, α-borneol, tocopherol nicotinate, inositol hexanicotinate, cyclandelate, cinnarizine, tolazoline, acetylcholine, verapamil, cephalanthin, γ-oryzanol, etc.); antiseborrheic agents (e.g., sulfur, thianthol, etc.); anti-inflammatory agents (e.g., tranexamic acid, taurine, hypotaurine, etc.).

[0049] <Screening method> Another aspect of the present invention relates to a method for screening a component that prevents and / or ameliorates deterioration of skin surface conditions (hereinafter sometimes referred to as "the screening method of the present invention"), comprising a selection step of selecting an active ingredient with the expression level of the HSF1 gene in cells as an indicator.

[0050] All matters described in the sections of <HSF1 expression enhancer> and <composition> apply to the description of the screening method of the present invention.

[0051] In the screening method of the present invention, it is preferable to use at least one type of epidermal cell, such as keratinocytes or corneocytes. The inventors have found that the expression level of the HSF1 gene decreases in epidermal keratinocytes due to aging, etc. Therefore, using at least one of these cells is efficient for measuring the expression level of HSF1. It is also preferable to use both of these cells.

[0052] The screening method of the present invention uses the expression level of the HSF1 gene in cells as an indicator. The process includes a selection step for selecting an active ingredient. Specifically, the test substance is added to a cell culture system, the expression level of HSF1 in the cells is measured, and if the expression level of HSF1 in cells cultured with the test substance is statistically significantly higher than the expression level of HSF1 in cells cultured without the test substance, the test substance can be selected as an active ingredient for preventing and / or improving the deterioration of the skin surface condition. Specifically, if the expression level is greater than 1 times that of cells cultured without the test substance, the test substance can be selected as an active ingredient for preventing and / or improving the deterioration of the skin surface condition.

[0053] The expression level of HSF1 can be measured by conventional methods such as protein measurement, mRNA measurement, and immunohistochemical analysis. It can also be measured according to the methods described in the examples below. For example, the expression level of the gene encoding HSF1 can be quantitatively detected by PCR using a DNA fragment having a sequence that specifically binds to the gene sequence as a primer. Since the gene sequence encoding HSF1 is publicly available, those skilled in the art can design appropriate primers. Furthermore, commercially available primers can be used to measure the expression level in cells.

[0054] The test substance targeted by the screening method of the present invention is not particularly limited and may be a pure substance, an extract derived from plants or animals, or a mixture thereof. Extracts derived from animals or plants refer not only to the extracts themselves, but also to fractions of the extracts, purified fractions, and solvent-removed extracts or fractions or purified products. Plant-derived extracts include extracts made from wild or cultivated plants, extracts made from plants sold as raw materials for herbal medicines, and commercially available extracts. [Examples]

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

[0056] <Example 1> Confirmation of the effect of aging on HSF1 gene expression in epidermal keratinocytes The effects of aging on HSF1 gene expression in skin cells remain unclear, particularly in epidermal keratinocytes. Therefore, we conducted a study to clarify the effects of aging on HSF1 expression in healthy epidermal keratinocytes.

[0057] ≪Cells used≫ This is shown in the table below.

[0058] [Table 1]

[0059] ≪Experimental Method≫ Seeds were seeded in groups of n=3 from each donor, and the gene expression level of HSF1 was measured and analyzed by qRT-PCR. The expression levels of young epidermal keratinocytes (No. 4-6) and aged epidermal keratinocytes (No. 1-3) were compared.

[0060] ≪Results≫ The results are shown in Figure 1. Compared to young epidermal keratinocytes, the gene expression level of HSF1 was significantly lower in aged epidermal keratinocytes.

[0061] <Example 2> Confirmation of the effect of HSF1 knockdown on the expression of epidermal-related genes in epidermal keratinocytes Experiments were conducted to investigate whether HSF1 regulates the expression levels of factors related to epidermal differentiation and stratum corneum formation.

[0062] ≪Method≫ Normal human epidermal keratinocytes (neonatal, Kurabo Industries Ltd.) were seeded in 24-well plates and cultured for 24 hours at 37°C in a 5% CO2 environment. Subsequently, siRNA was introduced using a standard method (Lipofectamine). TM Cells were cultured for 72 hours using 3000 Transfection Reagent (Invitrogen). After culturing, cells were harvested and RNA was extracted by a standard method. cDNA was synthesized from the extracted RNA by a standard method, and the epidermal barrier was analyzed by RT-qPCR using Fast SYBR Green Master Mix (Applied Biosystems). We measured and analyzed the expression levels of genes involved in function and turnover. The GAPDH gene was used as an endogenous control.

[0063] ≪Results≫ The results are shown in Figure 2. Knockdown of the HSF1 gene resulted in changes to Loricrin (LOR) and Hyaluronan Significant decrease in the expression of synthase 3 (HAS3) and keratin-1 (KRT1) genes was observed. Loricrin is a protein that constitutes the cornified envelope, which prevents excessive water loss from within the stratum corneum. KRT1 is a component protein of the cytoskeleton and is necessary for the formation of normal barrier function. HAS3 is an enzyme involved in hyaluronic acid synthesis and contributes to the normal turnover of the epidermis.

[0064] <Example 3> Selection of plant extracts that increase HSF1 gene expression in epidermal keratinocytes The gene expression level of HSF1 in the epidermis decreases with age, and the gene expression level of HSF1 It was revealed that a decrease in HSF1 leads to a decrease in the gene expression levels of various epidermal-related factors. In response, plant extract screening was conducted to develop ingredients that increase HSF1 expression and improve skin condition.

[0065] • Extracts used • Geranium thunbergii extract (contains solid components in the extraction solvent)

[0066] ≪Method≫ Using HuMedia-KG2 medium (manufactured by Kurabo Industries Ltd.), normal human-derived epidermal keratinocytes (neonatal) The HSF1 extract (manufactured by Kurabo Industries Ltd.) was seeded in a 24-well plate and cultured overnight at 37°C and 5% CO2. After culture, the medium was removed and replaced with HuMedia-KG2 medium containing 0.1% by mass of the test extract, and cultured for another 24 hours at 37°C and 5% CO2. After culture, the cells were washed with PBS(-) and collected, and RNA was extracted by conventional methods. cDNA was synthesized from the extracted RNA by conventional methods, and the expression levels of each HSF1 gene were analyzed by RT-qPCR using Fast SYBR Green Master Mix (manufactured by Applied Biosystems). The GAPDH gene was used as an endogenous control.

[0067] ≪Results≫ The results are shown in Figure 3. Addition of Geranium thunbergii extract resulted in a significant increase in HSF1 gene expression. This was confirmed.

[0068] <Example 4> Selection of plant extracts that increase HSF1 gene expression in dermal fibroblasts HSF1, a transcription factor for HSPs, shows decreased expression levels in dermal fibroblasts with age. It has been reported that HSP normalizes collagen structure and protects fibroblasts in the dermis. HSF1 plays a role in these functions, and a decrease in HSF1 expression can lead to a decline in these functions, potentially resulting in wrinkles and sagging. We investigated whether Geranium thunbergii extract, which has been found to increase HSF1 expression in the epidermis, also increases HSF1 expression in the dermis.

[0069] ≪Method≫ D-MEM medium (Sigma-A) was used to collect normal human dermal fibroblasts (neonatal, Kurabo Industries Ltd. strain). The test extract was seeded in a 24-well plate and cultured overnight at 37°C and 5% CO2. After culturing, the medium was removed and replaced with HuMedia-KG2 medium containing 0.1% by mass of the test extract, and cultured further at 37°C and 5% CO2. The cells were cultured for 24 hours under a %CO2 environment. After culturing, the cells were washed with PBS(-) and collected, and RNA was extracted by a standard method. cDNA was synthesized from the extracted RNA by a standard method and mixed with Fast SYBR Green Master Mix. Using an Applied Biosystems device, the expression levels of each HSF1 gene were analyzed by RT-qPCR. The GAPDH gene was used as an endogenous control.

[0070] ≪Results≫ The results are shown in Figure 4. The addition of Geranium thunbergii extract resulted in a significant decrease in HSF1 gene expression in dermal fibroblasts.

[0071] As described above, it was confirmed that the expression level of the HSF1 gene is decreased in aging epidermal keratinocytes. Furthermore, it was confirmed that intentionally reducing the expression level of the HSF1 gene leads to a decrease in factors related to epidermal turnover and barrier function. This suggests that a decrease in HSF1 contributes to the disorder of the skin surface. In addition, it was found that Geranium thunbergii extract has the effect of increasing the expression level of the HSF1 gene in epidermal keratinocytes and dermal fibroblasts. Therefore, Geranium thunbergii extract can function as an HSF1 expression enhancer, preventing and improving the deterioration of skin surface condition, and also improving the overall appearance by improving the roughness of the skin surface. [Industrial applicability]

[0072] This invention can be applied to topical skin preparations, cosmetics, and the like.

Claims

1. A HSF1 (Heat shock transcription factor 1) expression enhancer containing Geranium thunbergii extract.

2. The agent according to claim 1, wherein the enhanced expression is an enhanced expression in the epidermis.

3. The agent according to claim 1, wherein the enhancement of expression is an enhancement of gene expression.

4. A composition for preventing and / or improving the deterioration of the skin surface condition, comprising the agent described in claim 1.

5. A composition for improving the appearance of the face, comprising the agent described in claim 1.

6. The composition according to claim 4 or 5, which is a cosmetic.

7. A method for screening ingredients that prevent and / or improve the deterioration of skin surface conditions, comprising a selection step of selecting an active ingredient based on the expression level of the HSF1 gene in cells.

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