Agent, skin application agent, and cosmetic for use in inhibition of binding between ages and rage

Abstract

To provide a new agent capable of inhibiting binding of AGEs and RAGE, and mainly applicable to human skin or the like.SOLUTION: Agents for use in inhibiting the binding of advancedglycationendproducts (AGEs) and receptorforAGEs (RAGE) include Chinese peony extract, butcher's broom extract, common mallow extract, multiflora rose extract, and / or horsetail extract.SELECTED DRAWING: Figure 1

Description

[Technical Field]

[0001] The present disclosure relates to, for example, agents for use in inhibiting the binding of AGEs and RAGE, skin application agents, cosmetics, and the like. [Background technology]

[0002] Advanced glycation end products (AGEs) are a general term for substances that are produced when proteins or lipids combine with reducing sugars such as glucose or fructose through various non-enzymatic reactions, such as the Maillard reaction (Non-Patent Document 1). This process is called glycation, and proteins that have been glycated in the body become less susceptible to degradation due to changes in their structure, and therefore deviate from the normal degradation pathway and become more likely to accumulate. The accumulation of AGEs has been reported to be associated with diseases such as diabetes and atherosclerosis (Non-Patent Document 2).

[0003] In the skin, collagen, elastin, and other substances with long half-lives are glycated in the dermis, while keratin and other substances are glycated in the epidermis. This glycation reduces the quality of these substances, leading to wrinkles, thickened keratin, and other problems. In addition, some AGEs have a yellowish tinge, which is believed to cause dull, yellow skin. Therefore, cosmetic ingredients that suppress the production of AGEs are being researched with the aim of improving beauty (Patent Document 1).

[0004] AGEs not only reduce the quality of proteins, but also bind to receptors for AGEs (RAGE, hereinafter also referred to as "RAGE"), and are reported to cause biological reactions such as diabetic complications, chronic inflammation (Non-Patent Document 3), melanin production (Non-Patent Document 4), inhibition of cell proliferation, and inhibition of adhesion (Non-Patent Documents 5 and 6). For this reason, research into cosmetic ingredients that have the effect of inhibiting AGE production is progressing. [Prior art documents] [Patent documents]

[0005] [Patent Document 1] Patent Publication No. 2020-193182 [Non-patent literature]

[0006] [Non-Patent Document 1] Ott C, Jacobs K, Haucke E, Navarrete Santos A, Grune T, Simm A. Role of advanced glycation end products in cellular signaling. Redox Biol. 2014;2(1):411-429. doi:10.1016 / j.redox.2013.12.016 [Non-patent document 2] Reddy VP, Aryal P, Darkwah EK. Advanced Glycation End Products in Health and Disease. Microorganisms. 2022;10(9). doi:10.3390 / microorganisms10091848 [Non-patent document 3] Park HJ, Boyington JC. The 1.5 A crystal structure of human receptor for advanced glycation endproducts (RAGE) ectodomains reveals unique features determining ligand binding. J Biol Chem. 2010;285(52):40762-40770. doi:10.1074 / jbc.M110.169276 [Non-patent document 4] Eun Jung Lee, Ji Young Kim & Sang Ho Oh. Advanced glycation end products (AGEs) promote melanogenesis through receptor for AGEs. Sci. Rep. 2016; 6, doi: 10.1038 / srep27848 [Non-Patent Document 5] Huijuan Liao, MD / Ph.D., Julia Zakhaleva, MD, and Weiliam Chen, Ph.D., R.Ph., Cells and Tissue Interactions with Glycated Collagen and their Relevance to Delayed Diabetic Wound Healing. Biomaterials. 2009 March; 30(9): 1689-1696. doi:10.1016 / j.biomaterials.2008.11.038. [Non-patent document 6] Lennert Van Putte, Sofie De Schrijver1 and Peter Moortgat, The effects of advanced glycation end products (AGEs) on dermal wound healing and scar formation: a systematic review., Scars, Burns & Healing, 2016;2 doi: 10.1177 / 2059513116676828 Summary of the Invention [Problem to be solved by the invention]

[0007] However, there are few materials that focus on signal transduction through the binding of AGEs and RAGE.

[0008] Therefore, an object of the present disclosure is to provide a new agent that can inhibit the binding of AGEs and RAGE, and that can be applied primarily to human skin. [Means for solving the problem]

[0009] To achieve the above-mentioned objectives, the present disclosure provides an agent for use in inhibiting the binding of AGEs (advanced glycation end products) and RAGE (receptor for AGEs), which comprises peony extract, kaede extract, mallow extract, multiflora rose extract, and / or horsetail extract.

[0010] Agents for application to the skin of the present disclosure include agents of the present disclosure.

[0011] The cosmetic preparation of the present disclosure contains the agent of the present disclosure. [Effects of the Invention]

[0012] According to the present disclosure, for example, it is possible to provide a new agent that can inhibit the binding of AGEs and RAGE and that can be applied mainly to human skin. [Brief explanation of the drawings]

[0013]

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[0014] <Definition> As used herein, "peony" ( Paeonia lactiflora ) is, for example, the Paeoniaceae ( Paeoniaceae ), Paeonia suffruticosa ( Paeonia ) plants or their relatives.

[0015] In this specification, "nagi ikada" ( Ruscus aculeatus ), also known as butcher's broom, is a member of the Asparagaceae family ( Asparagaceae ), Liliaceae ( Nolinoideae ), genus Acanthodes ( Ruscus ) plants or their relatives.

[0016] As used herein, "Mallvasia sylvestris" ( Malva sylvestris ), also known as blue mallow, is a member of the Malvaceae family ( Malvaceae ), mallow ( Malva ) or its related plants. The related plants include, for example, mallow ( Malva mauritiana ) etc.

[0017] In this specification, "Rosa multiflora" ( Rosa multiflora ) is, for example, a member of the Rosaceae family ( Rosaceae), Rosa genus ( Rosa ) or its related plants. The fruit of the above-mentioned Rosa multiflora is also called Rosa multiflora.

[0018] In the present specification, "horsetail" ( Equisetum arvense ) is, for example, Equisetaceae ( Equisetaceae ), Equisetum genus ( Equisetum ) plants or their relatives.

[0019] As used herein, "epidermal keratinocytes" refers to cells capable of differentiating into basal cells, spinous cells, granular cells, and keratinocytes that make up the epidermis. Through this differentiation, epidermal keratinocytes ultimately differentiate into corneocytes that form the stratum corneum, a process known as keratinization.

[0020] As used herein, "dermal fibroblast" refers to a type of cell that differentiates from cells known as mesenchymal stem cells, and has the ability to produce components such as collagen, hyaluronic acid, and elastin in the dermis of the skin.

[0021] As used herein, "melanocyte" refers to a type of cell derived from the neural crest, also known as a pigment cell, which is present in the epidermal basement membrane and hair matrix in the skin and produces melanin pigment.

[0022] As used herein, "TNF (tumor necrosis factor)-α" refers to a multifunctional inflammatory cytokine belonging to the TNF superfamily. TNF-α is produced by macrophages, lymphocytes, fibroblasts, epidermal keratinocytes, and the like during inflammation.

[0023] As used herein, "IL-8" (Interleukin-8) is a type of cytokine (chemokine) that induces neutrophil migration. IL-8 is known to cause inflammatory responses. IL-8 is also known as CXCL8.

[0024] In this specification, "TGM1" (Transglutaminase 1) is a kind of epidermal enzyme that acts on protein cross-linking. TGM1 is expressed at the final differentiation of keratinized squamous epithelium and forms a keratinocyte envelope in differentiated keratinocytes.

[0025] In this specification, "K10 (KRT10)" (Keratin 10) is a kind of keratin that is one of the intermediate diameter filaments (about 10 nm in thickness). K10 is expressed in the differentiated spinous layer and granular layer.

[0026] Hereinafter, the present disclosure will be described with examples, but the present disclosure is not limited to the following examples and can be arbitrarily changed and implemented. In addition, each description in the present disclosure and each embodiment can be mutually incorporated unless otherwise specified. In this specification, when the expression "~" is used, it is used in the sense of including the numerical values or physical values before and after it. In this specification, the expression "A and / or B" includes "only A", "only B", and "both A and B".

[0027] <Agents or compositions for use in inhibiting the binding of AGEs and RAGE> In one aspect, the present disclosure provides an agent or composition for use in inhibiting the binding of AGEs (advanced glycation end products) and RAGE (receptor for AGEs). The agent for use in inhibiting the binding of AGEs and RAGE of the present disclosure includes an extract of Paeonia lactiflora, an extract of Sargassum thunbergii, an extract of Petasites japonicus, an extract of Rosa multiflora, and / or an extract of Stellaria media. In addition, the composition for use in inhibiting the binding of AGEs and RAGE of the present disclosure includes an extract of Paeonia lactiflora, an extract of Sargassum thunbergii, an extract of Petasites japonicus, an extract of Rosa multiflora, and / or an extract of Stellaria media. In the following description, unless otherwise specified, the description of each agent of the present disclosure can be incorporated into the description of the corresponding composition.

[0028] As a result of extensive research, the present inventors have discovered that peony extract, kaede extract, mallow extract, multiflora rose extract, and / or equisetum extract inhibit the binding of AGEs and RAGE in epidermal keratinocytes and fibroblasts, thereby establishing the present disclosure. Therefore, the agent or composition for use in inhibiting the binding of AGEs and RAGE of the present disclosure is expected to have effects such as suppression of TNF-α gene expression, suppression of IL-8 gene expression, promotion of TGM1 gene expression, promotion of K10 gene expression, suppression of melanin production, anti-inflammation, promotion of cell proliferation and adhesion, normalization of cell turnover, and / or moisturizing, which are caused by signal transduction due to the binding of AGEs and RAGE.

[0029] The peony extract, burdock extract, mallow extract, multiflora rose extract, and / or horsetail extract (hereinafter collectively referred to as "plant extract") can be produced by solvent extraction of the respective plants. For example, one or more types of plants may be used. The plant material to be extracted may be, for example, an entire plant or a part of the plant. Examples of plant parts include roots, rhizomes, leaves, stems, flowers, fruits, pericarp, seeds, or mixtures thereof. The material may be the harvested plant itself, or a processed product obtained by freezing, drying, and / or pulverization. When the plant is peony, the material is preferably, for example, the root. When the plant is burdock, the material is preferably, for example, the root and / or rhizome. When the plant is multiflora rose, the material is preferably, for example, the fruit (bark fruit). When the plant is horsetail, the material is preferably, for example, all parts of the plant (whole plant).

[0030] Examples of the solvent include aqueous solvents such as water and buffer solutions; lower alcohols or hydrous lower alcohols such as methanol, ethanol, propyl alcohol, isopropyl alcohol, butanol, and isobutanol; polyhydric alcohols or hydrous polyhydric alcohols such as propylene glycol, 1,3-butylene glycol, 1,2-butylene glycol, 1,4-butylene glycol, 1,5-pentanediol, 1,2-pentanediol, 1,3-pentanediol, 1,4-pentanediol, 1,3,5-pentanetriol, glycerin, and polyethylene glycol (for example, molecular weight of 100 to 100,000); organic solvents such as acetone, ethyl acetate, diethyl ether, dimethyl ether, ethyl methyl ether, dioxane, hexane, acetonitrile, xylene, benzene, chloroform, carbon tetrachloride, phenol, and toluene; and acids (hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid, formic acid, acetic acid, etc.) or alkalis (sodium hydroxide, potassium hydroxide, calcium hydroxide, ammonia, etc.) adjusted to an appropriate normality. The solvent is preferably ethanol, 1,3-butylene glycol, or a mixture thereof. The solvent may be used alone or in combination. The extraction may be carried out, for example, by immersing the plant in the solvent at room temperature (about 25°C) for 1 to 20 days, or 7 to 14 days.

[0031] The plant extract may be, for example, a crude product of the plant extract, a dried product of the plant extract, a freeze-dried product of the plant extract, or a processed product of the extract such as spray drying.

[0032] Examples of treatments for the treated product include decomposition by adding an acid (hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid, organic acid, etc.) or an alkali (sodium hydroxide, calcium hydroxide, ammonia, etc.); fermentation or metabolic conversion by a microorganism; component adsorption using an ion exchange resin, activated carbon, diatomaceous earth, etc.; fractionation using chromatography with various separation modes (ion exchange, hydrophilic adsorption, hydrophobic adsorption, size exclusion, ligand exchange, affinity, etc.); filtration using filter paper, membrane filter, ultrafiltration membrane, etc.; pressurization or decompression; heating or cooling; drying or freeze-drying; pH adjustment; deodorization; decolorization; prolonged static storage; etc. One type of treatment may be performed alone, or two or more types may be performed.

[0033] The "inhibition of the binding of AGEs and RAGE" means inhibiting, reducing, or suppressing the binding of AGEs and RAGE, and may refer to a state in which the binding of AGEs and RAGE is completely eliminated, or a state in which the binding rate of AGEs and RAGE is reduced. The decrease in the binding rate of AGEs and RAGE can be evaluated by determining whether the binding rate of AGEs and RAGE is reduced when the agent for inhibiting the binding of AGEs and RAGE of the present disclosure is used, compared to when the agent for inhibiting the binding of AGEs and RAGE of the present disclosure is not present. In the evaluation, the agent for inhibiting the binding of AGEs and RAGE can be evaluated as having an AGEs and RAGE binding inhibitory effect when the binding rate of the AGEs and RAGE relative to a control group not using the agent for inhibiting the binding of AGEs and RAGE of the present disclosure is 5% or less, 10% or less, 15% or less, 20% or less, 30% or less, 40% or less, 50% or less, 60% or less, 70% or less, 75% or less, 80% or less, 85% or less, 90% or less, 95% or less, or 99% or less. The inhibition of the binding of AGEs and RAGE can be evaluated, for example, by measuring the binding between AGEs and RAGE in a cell-free assay using recombinant proteins using ELISA or the like in accordance with Example 1 described below.

[0034] The agent for use in inhibiting the binding of AGEs and RAGE of the present disclosure can inhibit downstream signaling of RAGE. Therefore, the inhibition of the binding of AGEs and RAGE of the present disclosure can be evaluated, for example, by evaluating downstream signaling of RAGE. Specifically, the evaluation can be performed by measuring the degree of suppression of TNF-α gene expression, the degree of suppression of IL-8 gene expression, the degree of promotion of TGM1 gene expression, the degree of promotion of K10 gene expression, the degree of suppression of melanin production, the degree of anti-inflammatory effect, the degree of promotion of cell proliferation, the degree of promotion of cell adhesion, etc.

[0035] An example of the RAGE gene is the gene identified in NCBI Gene as Gene ID: 177. Examples of the RAGE gene include mRNA encoded by the human RAGE gene, such as polynucleotides consisting of nucleotide sequences registered in Genbank under accession numbers NM_001136.5, NM_001206929.2, NM_001206932.2, NM_001206934.2, NM_001206936.2, NM_001206940.2, NM_172197.3, NM_001206954.2, etc. The expression level of the RAGE gene may be measured by measuring the expression levels of one or more isoforms of the RAGE gene, or by measuring the expression of all isoforms.

[0036] An example of the TNF-α gene is mRNA encoded by the human TNF-α gene, such as a polynucleotide consisting of the nucleotide sequence registered in Genbank under accession number NM_000594.4. The expression level of the TNF-α gene may be measured by measuring the expression levels of one or more isoforms of the TNF-α gene, or by measuring the expression of all isoforms.

[0037] An example of the IL-8 gene is mRNA encoded by the human IL-8 gene, such as a polynucleotide consisting of the nucleotide sequence registered in Genbank under accession number NM_001354840.3. The expression level of the IL-8 gene may be measured by measuring the expression levels of one or more isoforms of the IL-8 gene, or by measuring the expression of all isoforms.

[0038] An example of the TGM1 gene is mRNA encoded by the human TGM1 gene, such as a polynucleotide consisting of the nucleotide sequence registered in Genbank under accession number NM_000359.3. The expression level of the TGM1 gene may be measured by measuring the expression levels of one or more isoforms of the TGM1 gene, or by measuring the expression of all isoforms.

[0039] An example of the K10 gene is mRNA encoded by the human K10 gene (KRT10 gene), such as a polynucleotide consisting of the nucleotide sequence registered in Genbank under accession number NM_000421.5. The expression level of the K10 gene may be measured by measuring the expression levels of one or more isoforms of the K10 gene, or by measuring the expression of all isoforms.

[0040] The amount of melanin produced can be evaluated, for example, by extracting melanin contained in cells and measuring the absorbance according to Example 3 described below.

[0041] The anti-inflammatory activity can be evaluated, for example, by measuring the expression levels of the TNF-α, IL-8, and the like.

[0042] The agent for inhibiting the binding of AGEs and RAGE of the present disclosure can inhibit the binding of AGEs and RAGE, for example, by administering it to a subject. The conditions for use (administration conditions) of the agent for inhibiting the binding of AGEs and RAGE of the present disclosure are not particularly limited, and the administration form, administration time, dosage, etc. can be appropriately set depending on, for example, the type of subject to be administered.

[0043] The agent for use in inhibiting the binding of AGEs and RAGE according to the present disclosure is, for example, in vivo You can also use in vitro It may also be used in

[0044] There are no particular limitations on the subjects to which the agent for inhibiting the binding of AGEs and RAGE of the present disclosure is administered. in vivo When used in the above, the subject of administration can be, for example, a human or a non-human animal other than a human. Examples of the non-human animal include mammals such as mice, rats, rabbits, dogs, sheep, horses, cats, goats, monkeys, and guinea pigs, as well as birds. The agent for use in inhibiting AGE-RAGE binding of the present disclosure can be used in the following manner: in vitro When used in the above, the subject to be administered can be, for example, a cell, a tissue, an organ, etc., and the cells can be, for example, a cell collected from a living body or a cultured cell, and the tissue or organ can be, for example, a tissue (living tissue) or an organ collected from a living body.

[0045] The administration form of the agent for use in inhibiting the binding of AGEs and RAGE of the present disclosure can be oral or parenteral. Examples of parenteral administration include transdermal administration and application (contact) to the skin. Application to the skin can also mean application to the oral mucosa, i.e., application to or contact with epithelial cells in the oral cavity. Furthermore, application to the skin can also mean administration or injection into the skin or subcutaneously via the skin surface, in addition to or instead of application to the skin surface. Administration or injection into the skin via the skin surface can be performed, for example, using a microneedle.

[0046] The dosage form of the agent for use in inhibiting the binding of AGEs and RAGE according to the present disclosure is not particularly limited and can be appropriately determined depending on, for example, the administration form. Examples of the dosage form include liquid and solid. When the administration form is oral administration, examples of the dosage form include tablets, pills, capsules, granules, powders, and liquids.

[0047] The agent for use in inhibiting the binding of AGEs and RAGE of the present disclosure may contain, for example, additives as needed. When used as a composition, the additives preferably include pharmaceutically acceptable additives or pharmaceutically acceptable carriers. The additives are not particularly limited, and examples include base materials, excipients, colorants, lubricants, binders, disintegrants, stabilizers, coating agents, preservatives, and flavoring agents such as fragrances. In the present disclosure, the amount of the additives added is not particularly limited, as long as they do not interfere with the function of one or more extracts of peony, kaede, mallow, multiflora rose, and horsetail.

[0048] Examples of the excipient include sugar derivatives such as lactose, lactose hydrate, sucrose, glucose, mannitol, and sorbitol; starch derivatives such as corn starch, potato starch, alpha starch, and dextrin; cellulose derivatives such as crystalline cellulose; organic excipients such as gum arabic, dextran, and pullulan; silicate derivatives such as light anhydrous silicic acid, synthetic aluminum silicate, calcium silicate, and magnesium aluminometasilicate; phosphates such as calcium hydrogen phosphate; carbonates such as calcium carbonate; and sulfates such as calcium sulfate. Examples of the colorant include yellow ferric oxide. Examples of the lubricant include metal stearates such as stearic acid, calcium stearate, and magnesium stearate; talc; polyethylene glycol; silica; and hydrogenated vegetable oil. Examples of the flavoring agent include flavorings such as cocoa powder, peppermint, aromatic powder, peppermint oil, borneol, and cinnamon powder, as well as sweeteners and acidulants. Examples of the binder include hydroxypropyl cellulose, hydroxypropylmethyl cellulose, polyvinylpyrrolidone, macrogol, etc. Examples of the disintegrant include cellulose derivatives such as carboxymethyl cellulose and carboxymethyl cellulose calcium; chemically modified starches and chemically modified celluloses such as carboxymethyl starch, carboxymethyl starch sodium, cross-linked polyvinylpyrrolidone, and sodium starch glycolate; examples of the stabilizer include parahydroxybenzoic acid esters such as methylparaben and propylparaben; alcohols such as chlorobutanol, benzyl alcohol, and phenylethyl alcohol; benzalkonium chloride; phenols such as phenol and cresol; thimerosal; dehydroacetic acid; sorbic acid; and examples of the coating agent include hypromellose, macrogols such as Macrogol 6000, talc, titanium oxide, etc.

[0049] When the composition for use in inhibiting the binding of AGEs and RAGE according to the present disclosure is an orally administered composition, specific examples of the orally administered composition include beverages, foods, pharmaceutical products (drugs), and quasi-drug products (quasi-drugs).

[0050] When the agent or composition for inhibiting the binding of AGEs and RAGE disclosed herein is used for transdermal administration or application to the skin (hereinafter also referred to as "topical skin preparation"), the form of the topical skin preparation may be, depending on the form of use, an ampoule, capsule, powder, granule, liquid, gel, foam, emulsion, sheet, mist, spray, etc. Examples of the form of use include: pharmaceutical(s); quasi-drug(s); topical or systemic skin preparations; medicinal and / or cosmetic preparations applied to the scalp and hair; bath additives used by adding them to bathwater; other preparations; etc. Examples of the topical or systemic skin preparations include basic cosmetics such as lotions, milky lotions, creams, ointments, lotions, oils, and packs; face washes or skin cleansers such as solid soaps, liquid soaps, and hand washes; massage agents, cleansing agents, hair removers, depilatories, shaving treatments, aftershave lotions, pre-shave lotions, shaving creams; makeup cosmetics such as foundations, lipsticks, blushers, eye shadows, eyeliners, and mascaras; perfumes; nail polish, nail enamel, nail enamel removers; poultices, plasters, tapes, sheets, patches, aerosols, toothpaste, and mouthwashes. Examples of the medicinal and / or cosmetic preparations to be applied to the scalp and hair include shampoos, rinses, hair treatments, pre-hair treatments, permanent solutions, hair dyes, hair styling products, hair tonics, hair growth and care products, poultices, plasters, tapes, sheets, aerosols, etc. Examples of the other preparations include underarm odor prevention or deodorants, antiperspirants, sanitary products, sanitary cotton, wet tissues, etc.

[0051] The topical skin preparation can be prepared by arbitrarily selecting and / or combining the following ingredients and / or additives as needed, within the scope that does not interfere with the action of one or more extracts of peony, kaede, mallow, multiflora rose, and horsetail.

[0052] (1) Various oils and fats Avocado oil, almond oil, fennel oil, perilla oil, olive oil, orange oil, orange roughage oil, sesame oil, cacao butter, chamomile oil, carrot oil, cucumber oil, beef tallow fatty acids, kukui nut oil, safflower oil, shea butter, liquid shea butter, soybean oil, camellia oil, corn oil, rapeseed oil, persic oil, castor oil, cottonseed oil, peanut oil, turtle oil, mink oil, egg yolk oil, palm oil, palm kernel oil, Japan wax, coconut oil, beef tallow, lard, squalene, squalane, pristane, and hydrogenated products of these oils and fats (hardened oils, etc.).

[0053] (2) Waxes Beeswax, carnauba wax, spermaceti, lanolin, liquid lanolin, reduced lanolin, hard lanolin, candelilla wax, montan wax, shellac wax, rice wax, etc.

[0054] (3) Mineral oil Liquid paraffin, Vaseline, paraffin, ozokeride, ceresin, microcrystalline wax, etc.

[0055] (4) Fatty acids Natural fatty acids such as lauric acid, myristic acid, palmitic acid, stearic acid, behenic acid, oleic acid, linoleic acid, linolenic acid, docosahexaenoic acid, eicosapentaenoic acid, 12-hydroxystearic acid, undecylenic acid, tall oil, and lanolin fatty acids; synthetic fatty acids such as isononanoic acid, caproic acid, 2-ethylbutanoic acid, isopentanoic acid, 2-methylpentanoic acid, 2-ethylhexanoic acid, and isopentanoic acid; and the like.

[0056] (5) Alcohol Natural alcohols such as ethanol, isopropanol, lauryl alcohol, cetanol, stearyl alcohol, oleyl alcohol, lanolin alcohol, cholesterol, phytosterol, and phenoxyethanol; synthetic alcohols such as 2-hexyldecanol, isostearyl alcohol, and 2-octyldodecanol; and the like.

[0057] (6) Polyhydric alcohols Ethylene oxide, ethylene glycol, diethylene glycol, triethylene glycol, ethylene glycol monoethyl ether, ethylene glycol monobutyl ether, diethylene glycol monomethyl ether, diethylene glycol monoethyl ether, polyethylene glycol, propylene oxide, propylene glycol, polypropylene glycol, 1,3-butylene glycol, pentyl glycol, glycerin, pentaerythritol, threitol, arabitol, xylitol, ribitol, galactitol, sorbitol, mannitol, lactitol, maltitol, etc.

[0058] (7) Esters Isopropyl myristate, isopropyl palmitate, butyl stearate, hexyl laurate, myristyl myristate, oleyl oleate, decyl oleate, octyldodecyl myristate, hexyldecyl dimethyloctanoate, cetyl lactate, myristyl lactate, diethyl phthalate, dibutyl phthalate, lanolin acetate, ethylene glycol monostearate, propylene glycol monostearate, propylene glycol dioleate, etc.

[0059] (8) Metal soaps Aluminum stearate, magnesium stearate, zinc stearate, calcium stearate, zinc palmitate, magnesium myristate, zinc laurate, zinc undecylenate, etc.

[0060] (9) Gums, sugars, or water-soluble polymeric compounds Gum arabic, gum benzoin, gum dammar, guaiac butter, Irish moss, gum karaya, gum tragacanth, carob gum, quince seed, agar, casein, lactose, fructose, sucrose or its ester, trehalose or its derivative, dextrin, gelatin, pectin, starch, carrageenan, carboxymethyl chitin or chitosan, hydroxyalkyl (C2-C4) chitin or chitosan to which alkylene (C2-C4) oxide such as ethylene oxide is added, low molecular weight chitin or chitosan, chitosan salt, sulfated chitin or chitosan, phosphorylated chitin or chitosan, alginic acid or its salt, Hyaluronic acid or a salt thereof, chondroitin sulfate or a salt thereof, heparin, ethyl cellulose, methyl cellulose, carboxymethyl cellulose, carboxyethyl cellulose, sodium carboxyethyl cellulose, hydroxyethyl cellulose, hydroxypropyl cellulose, nitrocellulose, crystalline cellulose, polyvinyl alcohol, polyvinyl methyl ether, polyvinylpyrrolidone, polyvinyl methacrylate, polyacrylates, polyalkylene oxides such as polyethylene oxide and polypropylene oxide or crosslinked polymers thereof, carboxyvinyl polymers, polyethyleneimine, etc.

[0061] (10) Surfactants Anionic surfactants (alkyl carboxylates, alkyl sulfonates, alkyl sulfates, alkyl phosphates), cationic surfactants (alkylamine salts, alkyl quaternary ammonium salts), amphoteric surfactants: carboxylic acid amphoteric surfactants (amino type, betaine type), sulfate ester amphoteric surfactants, sulfonate amphoteric surfactants, phosphate ester amphoteric surfactants, nonionic surfactants (ether type nonionic surfactants, ether ester type nonionic surfactants, ester type nonionic surfactants, block polymer type nonionic surfactants, nitrogen-containing nonionic surfactants), other surfactants (natural surfactants, protein hydrolysate derivatives, polymer surfactants, surfactants containing titanium and silicon, fluorocarbon surfactants), etc.

[0062] (11) Various vitamins Vitamin B complex: thiamine hydrochloride, thiamine sulfate (vitamin B1), riboflavin (vitamin B2), pyridoxine (vitamin B6), cyanocobalamin (vitamin B12), folic acid, nicotinic acid, pantothenic acid, biotin, choline, inositol, vitamin C complex: vitamin C acid or its derivatives, vitamin D complex: ergocalciferol (vitamin D2), cholecalciferol (vitamin D3), dihydrotachysterol, vitamin E complex: vitamin E or its derivatives, ubiquinones, vitamin K complex: phytonadione (vitamin K1), menaquinone (vitamin K2), menadione (vitamin K3), menadiol (vitamin K4), other essential fatty acids (vitamin F), carnitine, ferulic acid, gamma-oryzanol, orotic acid, vitamin P complex (rutin, eriocitrin, hesperidin), vitamin U, etc.

[0063] (12) Various amino acids Valine, leucine, isoleucine, threonine, methionine, phenylalanine, tryptophan, lysine, glycine, alanine, asparagine, glutamine, serine, cysteine, cystine, tyrosine, proline, hydroxyproline, aspartic acid, glutamic acid, hydroxylysine, arginine, ornithine, histidine, and the like, and amino acid derivatives thereof such as sulfates, phosphates, nitrates, citrates, or pyrrolidone carboxylic acid.

[0064] (13) Additives The topical skin preparation may further contain various additives derived from animals or plants. The additives can be selected from a variety of materials and added by conventional processing depending on the type and form of the product to which they are to be added. The processing can be, for example, any of the following processes selected and / or combined: crushing, milling, washing, hydrolysis, fermentation, refining, squeezing, extraction, fractionation, filtration, drying, powdering, granulation, dissolution, sterilization, pH adjustment, deodorization, bleaching, etc.

[0065] The solvent used for the extraction can be selected taking into consideration the intended use and type of the product, as well as subsequent processing. The extraction solvent is preferably one or a mixture of two or more selected from the following: water; lower alcohols or hydrous lower alcohols such as methanol, ethanol, propyl alcohol, isopropyl alcohol, butanol, and isobutanol; polyhydric alcohols or hydrous polyhydric alcohols such as propylene glycol, 1,3-butylene glycol, and glycerin; and various organic solvents such as acetone and ethyl acetate. However, when the use makes it undesirable to include an organic solvent, water alone or ethanol, which is easily removed after extraction, may be used alone or in any mixture with water, or a product obtained by squeezing and extracting the extract.

[0066] When the additives derived from plant or animal raw materials are used in external preparations or cosmetics for systemic or local use, the external preparations for skin can be expected to have cosmetic effects such as protection of the skin and hair, moisturizing, improving feel and texture, imparting softness, easing irritation, relieving stress through fragrance, activating cells (preventing cell aging), suppressing inflammation, improving skin and hair quality, preventing and improving rough skin, promoting hair growth, preventing hair loss, imparting shine, cleansing effects, relieving fatigue, promoting blood flow, and providing a warm bath effect, as well as fragrance, deodorizing, thickening, antiseptic, buffering, and other effects.

[0067] For example, the topical skin preparation can be made into a product that is expected to have multifunctional effects by combining the various cosmetic and pharmaceutical effects of each raw material material that has been known up to now, thereby enhancing the effects aimed at by the present disclosure.

[0068] The agent for use in inhibiting the binding of AGEs and RAGE of the present disclosure can inhibit, for example, the binding of AGEs and RAGE. Therefore, the agent for use in inhibiting the binding of AGEs and RAGE of the present disclosure can also be said to be an agent for use in suppressing downstream signals of RAGE, for example.

[0069] <Agent or composition for use in suppressing the expression of TNF-α gene> In another aspect, the present disclosure provides an agent or composition for use in suppressing the expression of TNF-α gene. The agent for use in suppressing the expression of TNF-α gene of the present disclosure includes Paeonia lactiflora extract, Cissus rotundifolia extract, Plantago asiatica extract, Rosa multiflora extract, and / or Stellaria media extract. The composition for use in suppressing the expression of TNF-α gene of the present disclosure includes Paeonia lactiflora extract, Cissus rotundifolia extract, Plantago asiatica extract, Rosa multiflora extract, and / or Stellaria media extract. According to the agent or composition for use in suppressing the expression of TNF-α gene of the present disclosure, an anti-inflammatory effect on fibroblasts can be obtained.

[0070] The "suppression of TNF-α gene expression" means that the expression level of the TNF-α gene is suppressed or decreased, and may also mean the change of the TNF-α gene expression from a state with expression to a state without expression. The expression of the TNF-α gene can be evaluated, for example, by measuring the expression level of the mRNA of the TNF-α gene according to Example 4 described below.

[0071] The agent for use in suppressing the expression of TNF-α gene of the present disclosure can suppress the expression of the TNF-α gene, for example, by being used for an administration subject. Thereby, the agent for use in suppressing the expression of TNF-α gene of the present disclosure can obtain, for example, an anti-inflammatory effect on dermal fibroblasts. The usage conditions (administration conditions) of the agent for use in suppressing the expression of TNF-α gene of the present disclosure can incorporate the description of the usage conditions of the agent for use in inhibiting the binding of AGEs and RAGE of the present disclosure.

[0072] <Agent or composition for use in suppressing the expression of IL-8 gene> In another aspect, the present disclosure provides an agent or composition for use in suppressing the expression of the IL-8 gene. The agent for use in suppressing the expression of the IL-8 gene of the present disclosure contains Paeonia lactiflora extract, Laminaria japonica extract, Arnebia euchroma extract, Rosa multiflora extract, and / or Eclipta prostrata extract. The composition for use in suppressing the expression of the IL-8 gene of the present disclosure contains Paeonia lactiflora extract, Laminaria japonica extract, Arnebia euchroma extract, Rosa multiflora extract, and / or Eclipta prostrata extract. According to the agent or composition for use in suppressing the expression of the IL-8 gene of the present disclosure, an anti-inflammatory effect on fibroblasts can be obtained.

[0073] The "suppression of IL-8 gene expression" means that the expression level of the IL-8 gene is suppressed or decreased, and may also mean a change in the expression of the IL-8 gene from a state of expression to a state of no expression. The expression of the IL-8 gene can be evaluated, for example, by measuring the expression level of the mRNA of the IL-8 gene according to Example 4 described below.

[0074] The agent for use in suppressing the expression of the IL-8 gene of the present disclosure can suppress the expression of the IL-8 gene, for example, by being used for an administration subject. Thereby, the agent for use in suppressing the expression of the IL-8 gene of the present disclosure can obtain, for example, an anti-inflammatory effect on dermal fibroblasts. The usage conditions (administration conditions) of the agent for use in suppressing the expression of the IL-8 gene of the present disclosure can refer to the description of the usage conditions of the agent for use in inhibiting the binding of AGEs and RAGE of the present disclosure.

[0075] <Agent or composition for use in promoting the expression of the TGM1 gene> In another aspect, the present disclosure provides an agent or composition for promoting the expression of the TGM1 gene. The agent for promoting the expression of the TGM1 gene of the present disclosure contains a peony extract, a snake gourd extract, a dwarf morning glory extract, a rose extract, and / or a shepherd's purse extract. The composition for promoting the expression of the TGM1 gene of the present disclosure contains a peony extract, a snake gourd extract, a dwarf morning glory extract, a rose extract, and / or a shepherd's purse extract. According to the agent or composition for promoting the expression of the TGM1 gene of the present disclosure, normalization of turnover and / or a moisturizing effect can be obtained.

[0076] The "promotion of the expression of the TGM1 gene" means that the expression level of the TGM1 gene is promoted or increased, and may also mean a change from a state where the TGM1 gene is not expressed to a state where it is expressed. The expression of the TGM1 gene can be evaluated, for example, by measuring the expression level of the mRNA of the TGM1 gene according to Example 7 described below.

[0077] The agent for promoting the expression of the TGM1 gene of the present disclosure can promote the expression of the TGM1 gene, for example, by being used for an administration subject. Thereby, the agent for promoting the expression of the TGM1 gene of the present disclosure can obtain, for example, normalization of turnover and / or a moisturizing effect. The usage conditions (administration conditions) of the agent for promoting the expression of the TGM1 gene of the present disclosure can refer to the description of the usage conditions of the agent for inhibiting the binding of AGEs and RAGE of the present disclosure.

[0078] <Agent or composition for promoting the expression of the K10 gene> In another embodiment, the present disclosure provides an agent or composition for use in promoting the expression of K10 gene.The agent for use in promoting the expression of K10 gene of the present disclosure comprises peony extract, raft extract, mallow extract, multiflora rose extract, and / or horsetail extract.The composition for use in promoting the expression of K10 gene of the present disclosure comprises peony extract, raft extract, mallow extract, multiflora rose extract, and / or horsetail extract.The agent or composition for use in promoting the expression of K10 gene of the present disclosure can achieve the effect of normalizing cell turnover and / or moisturizing.

[0079] The "promotion of K10 gene expression" means that the expression level of the K10 gene is promoted or increased, and may also mean that the expression level of the K10 gene changes from an unexpressed state to an expressed state. The expression of the K10 gene can be evaluated, for example, by measuring the expression level of K10 gene mRNA according to Example 7 described below.

[0080] The agent for promoting K10 gene expression of the present disclosure can promote K10 gene expression, for example, by administering it to a subject. As a result, the agent for promoting K10 gene expression of the present disclosure can achieve, for example, the effects of normalizing cell turnover and / or moisturizing. The conditions for use (administration conditions) of the agent for promoting K10 gene expression of the present disclosure can be the same as those described above for the agent for inhibiting the binding of AGEs and RAGE of the present disclosure.

[0081] <Melanin production inhibitor or composition> In another aspect, the present disclosure provides an agent or composition capable of inhibiting melanin production in melanocytes. The melanin production inhibitor of the present disclosure comprises peony extract, kaede extract, malva sylvestris extract, multiflora rose extract, and / or horsetail extract. The melanin production inhibitor of the present disclosure comprises peony extract, kaede extract, malva sylvestris extract, multiflora rose extract, and / or horsetail extract. The melanin production inhibitor or composition of the present disclosure can inhibit melanin production in melanocytes, thereby achieving a whitening effect.

[0082] The melanin production inhibitor of the present disclosure can, for example, inhibit melanin production in melanocytes by administering it to a subject. The conditions for use (administration conditions) of the melanin production inhibitor of the present disclosure can be the same as those described above for the agent used to inhibit the binding of AGEs and RAGE of the present disclosure.

[0083] <Inflammation suppressant or composition> In another aspect, the present disclosure provides an agent or composition capable of suppressing inflammation of epidermal keratinocytes. The inflammation suppressing agent of the present disclosure comprises peony extract, raft extract, mallow extract, multiflora extract, and / or horsetail extract. The inflammation suppressing composition of the present disclosure comprises peony extract, raft extract, mallow extract, multiflora extract, and / or horsetail extract. The inflammation suppressing agent or composition of the present disclosure can suppress inflammation of epidermal keratinocytes.

[0084] The inflammation inhibitor of the present disclosure can suppress inflammation of epidermal keratinocytes, for example, by administering it to a subject. The conditions for use (administration conditions) of the inflammation inhibitor of the present disclosure can be the same as those described above for the agent used to inhibit the binding of AGEs and RAGE of the present disclosure.

[0085] <Cell proliferation promoter or composition> In another aspect, the present disclosure provides an agent or composition capable of promoting cell growth. The cell growth promoting agent of the present disclosure includes Paeonia extract, Sargassum extract, Abelmoschus manihot extract, Rosa multiflora extract, and / or Stellaria media extract. The cell growth promoting composition of the present disclosure includes Paeonia extract, Sargassum extract, Abelmoschus manihot extract, Rosa multiflora extract, and / or Stellaria media extract. According to the cell growth promoting agent or composition of the present disclosure, for example, the growth of cells such as epidermal keratinocytes and dermal fibroblasts can be promoted.

[0086] The cell growth promoting agent of the present disclosure can promote the growth of cells such as epidermal keratinocytes and dermal fibroblasts, for example, by being used for an administration subject. The usage conditions (administration conditions) of the cell growth promoting agent of the present disclosure can incorporate the description of the usage conditions of the agent for inhibiting the binding of AGEs and RAGE in the present disclosure.

[0087] <Cell adhesion promoting agent or composition> In another aspect, the present disclosure provides an agent or composition capable of promoting cell adhesion. The cell adhesion promoting agent of the present disclosure includes Paeonia extract, Sargassum extract, Abelmoschus manihot extract, Rosa multiflora extract, and / or Stellaria media extract. The cell adhesion promoting composition of the present disclosure includes Paeonia extract, Sargassum extract, Abelmoschus manihot extract, Rosa multiflora extract, and / or Stellaria media extract. According to the cell adhesion promoting agent or composition of the present disclosure, for example, the adhesion of cells such as epidermal keratinocytes and dermal fibroblasts can be promoted.

[0088] The cell adhesion promoting agent of the present disclosure can promote the adhesion of cells such as epidermal keratinocytes and dermal fibroblasts, for example, by being used for an administration subject. The usage conditions (administration conditions) of the cell growth adhesion agent of the present disclosure can incorporate the description of the usage conditions of the agent for inhibiting the binding of AGEs and RAGE in the present disclosure.

[0089] <Method for inhibiting the binding of AGEs and RAGE> In another aspect, the present disclosure discloses a method capable of inhibiting the binding of AGEs and RAGE. The method for inhibiting the binding of AGEs and RAGE of the present disclosure uses the agent and / or composition for inhibiting the binding of AGEs and RAGE of the present disclosure. According to the method for inhibiting the binding of AGEs and RAGE of the present disclosure, for example, it is expected that the binding of AGEs and RAGE can be inhibited and the signal suppression effect downstream of RAGE can be obtained.

[0090] The method for inhibiting the binding of AGEs and RAGE of the present disclosure includes a use step of using, for a subject, the agent and / or composition for inhibiting the binding of AGEs and RAGE of the present disclosure. The use may be, for example, contact with the skin or the like, or administration.

[0091] In the method for inhibiting the binding of AGEs and RAGE of the present disclosure, the use step may be performed, for example, in vitro or in vivo The subject (administration subject) and administration conditions of the method for inhibiting the binding of AGEs and RAGE of the present disclosure can, for example, refer to the description of the administration subject and administration conditions in the agent and / or composition for inhibiting the binding of AGEs and RAGE of the present disclosure.

[0092] <Method for suppressing the expression of TNF-α gene> In another aspect, the present disclosure discloses a method capable of suppressing the expression of TNF-α gene. The method for suppressing the expression of TNF-α gene of the present disclosure uses the agent and / or composition for suppressing the expression of TNF-α gene of the present disclosure. According to the method for suppressing the expression of TNF-α gene of the present disclosure, for example, an anti-inflammatory effect on fibroblasts can be obtained.

[0093] The method for suppressing the expression of TNF-α gene of the present disclosure includes a use step of using, for a subject, the agent and / or composition for suppressing the expression of TNF-α gene of the present disclosure. The use may be, for example, contact with the skin or the like, or administration.

[0094] In the method for suppressing the expression of the TNF-α gene of the present disclosure, the usage step is, for example, in vitro or in vivo It may be carried out. The subject (administration subject) and administration conditions of the method for suppressing the expression of the TNF-α gene of the present disclosure can, for example, refer to the description of the administration subject and administration conditions in the agent and / or composition used for suppressing the expression of the TNF-α gene of the present disclosure.

[0095] <Method for Suppressing the Expression of the IL-8 Gene> In another aspect, the present disclosure discloses a method capable of suppressing the expression of the IL-8 gene. The method for suppressing the expression of the IL-8 gene of the present disclosure uses the agent and / or composition used for suppressing the expression of the IL-8 gene of the present disclosure. According to the method for suppressing the expression of the IL-8 gene of the present disclosure, for example, an anti-inflammatory effect on fibroblasts can be obtained.

[0096] The method for suppressing the expression of the IL-8 gene of the present disclosure includes a usage step of using the agent and / or composition used for suppressing the expression of the IL-8 gene of the present disclosure on a subject. The usage may be, for example, contact with the skin or the like, or administration.

[0097] In the method for suppressing the expression of the IL-8 gene of the present disclosure, the usage step is, for example, in vitro or in vivo It may be carried out. The subject (administration subject) and administration conditions of the method for suppressing the expression of the IL-8 gene of the present disclosure can, for example, refer to the description of the administration subject and administration conditions in the agent and / or composition used for suppressing the expression of the IL-8 gene of the present disclosure.

[0098] <Method for Promoting the Expression of the TGM1 Gene> In another aspect, the present disclosure discloses a method capable of promoting the expression of the TGM1 gene. The method for promoting the expression of the TGM1 gene of the present disclosure uses an agent and / or composition for use in promoting the expression of the TGM1 gene of the present disclosure. According to the method for promoting the expression of the TGM1 gene of the present disclosure, for example, normalization of turnover and / or the effect of moisturizing can be obtained.

[0099] The method for promoting the expression of the TGM1 gene of the present disclosure includes a use step of using, for a subject, an agent and / or composition for use in promoting the expression of the TGM1 gene of the present disclosure. The use may be, for example, contact with the skin or the like, or administration.

[0100] In the method for promoting the expression of the TGM1 gene of the present disclosure, the use step may be, for example, in vitro or in vivo and may be carried out. The subject (administration subject) and administration conditions of the method for promoting the expression of the TGM1 gene of the present disclosure can, for example, refer to the description of the administration subject and administration conditions in the agent and / or composition for use in promoting the expression of the TGM1 gene of the present disclosure.

[0101] <Method for promoting the expression of the K10 gene> In another aspect, the present disclosure discloses a method capable of promoting the expression of the K10 gene. The method for promoting the expression of the K10 gene of the present disclosure uses an agent and / or composition for use in promoting the expression of the K10 gene of the present disclosure. According to the method for promoting the expression of the K10 gene of the present disclosure, for example, normalization of turnover and / or the effect of moisturizing can be obtained.

[0102] The method for promoting the expression of the K10 gene of the present disclosure includes a use step of using, for a subject, an agent and / or composition for use in promoting the expression of the K10 gene of the present disclosure. The use may be, for example, contact with the skin or the like, or administration.

[0103] In the method for promoting the expression of the K10 gene of the present disclosure, the use step may be, for example, in vitroor in vivo The subject (administration subject) and administration conditions of the method for promoting expression of the K10 gene of the present disclosure can be, for example, the description of the administration subject and administration conditions of the agent and / or composition used for promoting expression of the K10 gene of the present disclosure.

[0104] <Melanin production inhibition method> In another aspect, the present disclosure discloses a method for inhibiting melanin production. The method for inhibiting melanin production of the present disclosure uses the melanin production inhibitor and / or melanin production inhibitor composition of the present disclosure. The method for inhibiting melanin production of the present disclosure can inhibit melanin production in melanocytes, thereby achieving a whitening effect.

[0105] The method for inhibiting melanin production according to the present disclosure includes a step of administering the melanin production inhibitor and / or composition according to the present disclosure to a subject. The use may be, for example, by contact with the skin or the like or by administration.

[0106] <Inflammation suppression method> In another aspect, the present disclosure discloses a method for suppressing inflammation. The method for suppressing inflammation of the present disclosure uses the inflammation-suppressing agent and / or inflammation-suppressing composition of the present disclosure. The method for suppressing inflammation of the present disclosure can suppress, for example, inflammation of fibroblasts.

[0107] The method for suppressing inflammation of the present disclosure includes a step of using the inflammation suppressing agent and / or composition of the present disclosure in a subject. The use may be, for example, by contact with the skin or the like or by administration.

[0108] In the inflammation suppression method of the present disclosure, the using step may include, for example, in vitro or in vivo The subject (administration subject) and administration conditions of the inflammation suppression method of the present disclosure can be determined by reference to, for example, the explanation of the administration subject and administration conditions of the agent and / or composition used for inhibiting the binding of AGEs and RAGE of the present disclosure.

[0109] <Method for promoting cell proliferation> In another aspect, the present disclosure discloses a method for promoting cell proliferation. The cell proliferation promoting method of the present disclosure uses the cell proliferation promoting agent and / or cell proliferation promoting composition of the present disclosure. The cell proliferation promoting method of the present disclosure can promote the proliferation of cells such as epidermal keratinocytes and dermal fibroblasts.

[0110] The cell proliferation promoting method of the present disclosure includes a step of using the cell proliferation promoting agent and / or composition of the present disclosure in a subject. The use may be, for example, by contact with the skin or the like, or by administration.

[0111] In the cell proliferation promoting method of the present disclosure, the using step may include, for example, in vitro or in vivo The subject (administration subject) and administration conditions of the cell proliferation-promoting method of the present disclosure can be determined by reference to, for example, the explanation of the administration subject and administration conditions of the agent and / or composition used for inhibiting the binding of AGEs and RAGE of the present disclosure.

[0112] <Method for promoting cell adhesion> In another aspect, the present disclosure discloses a method for promoting cell adhesion. The cell adhesion promoting method of the present disclosure uses the cell adhesion promoter and / or cell adhesion promoting composition of the present disclosure. The cell adhesion promoting method of the present disclosure can promote the adhesion of, for example, epidermal keratinocytes, dermal fibroblasts, and the like.

[0113] The cell adhesion promoting method of the present disclosure includes a step of using the cell adhesion promoting agent and / or composition of the present disclosure in a subject. The use may be, for example, by contact with the skin or the like, or by administration.

[0114] In the cell adhesion promoting method of the present disclosure, the using step may include, for example, in vitro or in vivoThe subject (administration subject) and administration conditions of the cell adhesion-promoting method of the present disclosure can be determined by reference to, for example, the explanation of the administration subject and administration conditions of the agent and / or composition used for inhibiting the binding of AGEs and RAGE of the present disclosure.

[0115] <Cosmetics> In another aspect, the present disclosure provides cosmetics. The cosmetics of the present disclosure include agents and / or compositions for use in inhibiting the binding of AGEs and RAGE, agents and / or compositions for use in suppressing TNF-α gene expression, agents and / or compositions for use in suppressing IL-8 gene expression, agents and / or compositions for use in promoting TGM1 gene expression, agents and / or compositions for use in promoting K10 gene expression, melanin production inhibitors and / or compositions, inflammation inhibitors and / or compositions, cell proliferation promoters and / or compositions, and cell adhesion promoters and / or compositions. Specifically, the cosmetics of the present disclosure include peony extract, kaede extract, mallow extract, multiflora rose extract, and / or horsetail extract. Because the cosmetics of the present disclosure can inhibit melanin production resulting from the inhibition of the binding of AGEs and RAGE, they can also be referred to as whitening cosmetics. Therefore, the cosmetics of the present disclosure can achieve whitening effects.

[0116] <Use> In another aspect, the present disclosure relates to the use of an agent and / or composition for use in inhibiting the binding of AGEs and RAGE. The present disclosure relates to the use of an agent and / or composition for use in inhibiting the expression of the TNF-α gene. The present disclosure relates to the use of an agent and / or composition for use in inhibiting the expression of the IL-8 gene. The present disclosure relates to the use of an agent and / or composition for use in promoting the expression of the TGM1 gene. The present disclosure relates to the use of an agent and / or composition for use in promoting the expression of the K10 gene. The present disclosure relates to the use of a melanin production inhibitor and / or composition for use in inhibiting melanin production. The present disclosure relates to the use of an inflammation inhibitor and / or composition for use in inhibiting inflammation. The present disclosure relates to the use of a cell proliferation promoter and / or composition for use in promoting cell proliferation. The present disclosure is the use of cell adhesion promoting agents and / or compositions for use in promoting cell adhesion.

[0117] The present disclosure relates to the use of an agent and / or composition for inhibiting the binding of AGEs and RAGE, for producing an agent and / or composition for use in inhibiting the binding of AGEs and RAGE. The present disclosure relates to the use of an agent and / or composition for inhibiting TNF-α gene expression, for producing an agent and / or composition for use in inhibiting TNF-α gene expression. The present disclosure relates to the use of an agent and / or composition for inhibiting IL-8 gene expression, for producing an agent and / or composition for use in inhibiting IL-8 gene expression. The present disclosure relates to the use of an agent and / or composition for inhibiting TGM1 gene expression, for producing an agent and / or composition for use in promoting TGM1 gene expression. The present disclosure relates to the use of an agent and / or composition for inhibiting K10 gene expression, for producing an agent and / or composition for use in promoting K10 gene expression. The present disclosure relates to the use of an agent and / or composition for inhibiting melanin production, for producing an agent and / or composition for use in inhibiting melanin production. The present disclosure relates to the use of an inflammation suppressing agent and / or composition for producing an inflammation suppressing agent and / or composition.The present disclosure relates to the use of a cell growth promoting agent and / or composition for producing an agent and / or composition for use in a cell growth promoting agent.The present disclosure relates to the use of a cell adhesion promoting agent and / or composition for producing an agent and / or composition for use in a cell adhesion promoting agent. [Example]

[0118] Next, examples of the present invention will be described. However, the present invention is not limited to the following examples. Commercially available reagents were used according to their protocols unless otherwise specified. The unit % used to indicate the amount of each extract used in the examples means % (w / v).

[0119] <Production Example 1 of Peony Extract> To 100 g of dried peony root, 1.5 kg of 50% 1,3-butylene glycol solution was added. After the addition, the mixture was soaked at 10 to 30°C for 7 to 14 days. After the soaking, the mixture was filtered using a filter material. After the filtration, the mixture was left to stand at -30 to 10°C for 7 to 14 days. After the standing, the mixture was filtered using a filter material.

[0120] <Production Example 1 of Nagiikada Extract> To 100 g of dried underground parts (roots and / or rhizomes) of the raft was added 1 kg of a 50% 1,3-butylene glycol solution. After the addition, the mixture was soaked at 10°C to 30°C for 7 to 14 days. After the soaking, the mixture was filtered using a filter material. After the filtration, the mixture was left to stand at -30°C to 10°C for 7 to 14 days. After the standing, the mixture was filtered using a filter material.

[0121] <Production Example 1 of Malva Sylvestris Extract> To 100 g of dried mallow flowers, 2 kg of a 50% 1,3-butylene glycol solution was added. After the addition, the mixture was soaked at 10°C to 30°C for 7 to 14 days. After the soaking, the mixture was filtered using a filter material. After the filtration, the mixture was left to stand at -30°C to 10°C for 7 to 14 days. After the standing, the mixture was filtered using a filter material.

[0122] <Production Example 1 of Rose Multiflora Extract> 1 kg of a 50% 1,3-butylene glycol solution was added to 100 g of dried wild rose fruit (Rosa multiflora). After the addition, the mixture was immersed at 10°C to 30°C for 7 to 14 days. After the immersion, filtration was carried out using a filter material. After the filtration, the mixture was left to stand at -30°C to 10°C for 7 to 14 days. After the standing, filtration was carried out using a filter material.

[0123] <Equisetum arvense extract production example 1> To 100 g of dried whole horsetail plant, 1 kg of 50% 1,3-butylene glycol solution was added. After the addition, the mixture was soaked at 10°C to 30°C for 7 to 14 days. After the soaking, the mixture was filtered using a filter material. After the filtration, the mixture was left to stand at -30°C to 10°C for 7 to 14 days. After the standing, the mixture was filtered using a filter material.

[0124] The dry solid content of each extract obtained in Production Example 1 was 0.1 to 5%.

[0125] <Production Example 2 of Peony Extract> 1.5 kg of 50% ethanol solution was added to 100 g of dried peony root. After the addition, the mixture was soaked at 10 to 30°C for 7 to 14 days. After the soaking, filtration was carried out using a filter material. After the filtration, the mixture was left to stand at -30 to 10°C for 7 to 14 days. After the standing, filtration was carried out using a filter material.

[0126] <Production Example 2 of Nagiikada Extract> 1 kg of 50% ethanol solution was added to 100 g of dried underground parts (roots and / or rhizomes) of the raft. After the addition, the mixture was soaked at 10°C to 30°C for 7 to 14 days. After the soaking, the mixture was filtered using a filter material. After the filtration, the mixture was left to stand at -30°C to 10°C for 7 to 14 days. After the standing, the mixture was filtered using a filter material.

[0127] <Production Example 2 of Mallorca Extract> 2 kg of a 50% ethanol solution was added to 100 g of dried mallow flowers. After the addition, the mixture was soaked at 10 to 30°C for 7 to 14 days. After the soaking, the mixture was filtered using a filter material. After the filtration, the mixture was left to stand at -30 to 10°C for 7 to 14 days. After the standing, the mixture was filtered using a filter material.

[0128] <Production Example 2 of Rose Multiflora Extract> 1 kg of 50% ethanol solution was added to 100 g of dried fruit of Rosa multiflora. After the addition, the mixture was immersed at 10 to 30°C for 7 to 14 days. After the immersion, filtration was carried out using a filter material. After the filtration, the mixture was left to stand at -30 to 10°C for 7 to 14 days. After the standing, filtration was carried out using a filter material.

[0129] <Equisetum arvense extract production example 2> To 100 g of dried whole horsetail plant, 1 kg of 50% ethanol solution was added. After the addition, the mixture was soaked at 10 to 30°C for 7 to 14 days. After the soaking, the mixture was filtered using a filter material. After the filtration, the mixture was left to stand at -30 to 10°C for 7 to 14 days. After the standing, the mixture was filtered using a filter material.

[0130] The dry solid content of each extract obtained in Production Example 2 was 0.1 to 5%.

[0131] <Production Example 3 of Nagiikada Extract> To 100 g of dried underground parts (roots and / or rhizomes) of the raft was added 1 kg of 50% ethanol solution. After the addition, the raft was soaked at 10 to 30°C for 7 to 14 days. After the soaking, the raft was collected, and 1 kg of 50% ethanol solution was added again. After the addition, the raft was soaked at 10 to 30°C for 7 to 14 days. The extracts after each soaking were mixed and adsorbed onto a column packed with Diaion HP20. After the adsorption, the column was washed with 50% ethanol and eluted with 100% ethanol. After the elution, the obtained extract was treated with activated carbon in an amount equal to the solid content and filtered using a filter material.

[0132] The dry solid content of each extract obtained in Production Example 3 was 0.1 to 5%.

[0133] [Example 1] Each plant extract was examined to determine whether it exhibited inhibitory effects on the binding of AGEs and RAGE.

[0134] (1) Sample preparation Recombinant Human RAGE Fc Chimera Protein (R&D Systems, 1145-RG) (hereinafter referred to as RAGE-Fc) was prepared at 2 μg / mL using PBS. Peony extract, Falcorex Peony B (Ichimaru Pharcos, Inc.), was prepared at final concentrations of 0.25%, 0.5%, and 1% using PBS containing 0.1% BSA. The rhubarb extract (Preparation Example 1 above), mallow extract (Falcorex Mallow B, Ichimaru Pharcos, Inc.), rose multiflora extract (Falcorex Eijitsu B, Ichimaru Pharcos, Inc.), and horsetail extract (Preparation Example 1 above) were prepared at final concentrations of 0.125%, 0.25%, and 0.5%, respectively, using PBS containing 0.1% BSA. AGE-BSA Biotinylated (BT4127, manufactured by R&D Systems) (hereinafter referred to as AGEs-Biotin) was adjusted to a final concentration of 20 μg / mL using PBS containing 0.1% BSA.

[0135] (2) Binding inhibition assay of AGEs and RAGE 50 μL of RAGE-Fc prepared in Example 1(1) was added to a 96-well plate at each well and allowed to stand at 4°C to coat the plate with RAGE-Fc. After standing overnight, the plate was returned to room temperature and blocked by adding 200 μL of PBS containing 1% BSA to each well. Two to four hours after blocking, the wells were washed with PBS containing 0.1% Tween (PBS-T). After washing, 50 μL of each of the plant extracts prepared in Example 1(1) was added. For the control group without extract, PBS containing 0.1% BSA and 50% BG was added instead of each plant extract. After the addition, the wells were allowed to react for 1 to 2 hours, and then washed with PBS-T. After washing, 50 μL of AGEs-Biotin prepared in Example 1(1) was added to each well. After the addition, the wells were allowed to react for 30 minutes to 1 hour, and then washed with PBS-T. After the washing, 50 μL of HRP-labeled streptavidin diluted 1 / 1000 with PBS containing 0.1% BSA was added to each well. After the addition, the wells were allowed to react for 30 minutes to 1 hour, and then washed with PBS-T. After the washing, detection was performed using TMB, and the reaction was stopped using sulfuric acid diluted 1 / 10, and the absorbance at 450 nm was measured. Each group was tested with one sample (n=1). These results are shown in Figure 1.

[0136] Figure 1 is a graph showing the inhibitory effect of AGEs and RAGE binding when various plant extracts were added. In Figure 1, (A) shows the results when peony extract was added, (B) shows the results when kabuto extract was added, (C) shows the results when mallow extract was added, (D) shows the results when multiflora extract was added, and (E) shows the results when horsetail extract was added. In Figure 1, the vertical axis shows the AGE and RAGE binding inhibition rate (%), and the horizontal axis shows the concentration of each plant extract. As shown in Figure 1, when various plant extracts were added, the AGE and RAGE binding inhibition rate increased in a concentration-dependent manner. These results demonstrate that peony extract, kabuto extract, mallow extract, multiflora extract, and horsetail extract each exhibit inhibitory effects on AGEs and RAGE binding.

[0137] [Example 2] AGEs are known to promote melanin production via RAGE, so we investigated whether each plant extract exhibits an inhibitory effect on melanin production caused by AGEs.

[0138] (1) Sample preparation Peony extract (Falcorex Peony B, Ichimaru Falcos Co., Ltd.), kaede extract (Preparation Example 1 above), mallow extract (Falcorex Mallow B, Ichimaru Falcos Co., Ltd.), multiflora rose extract (Falcorex Euglena B, Ichimaru Falcos Co., Ltd.), and horsetail extract (Preparation Example 1 above) were each prepared using MGM™-4 Melanocyte Growth Medium-4 BulletKit™ (MGM, manufactured by Lonza) medium so that the final concentrations shown in Table 1 below were achieved when added to the medium. AGEs (AGE-BSA, manufactured by CalbioChem) were prepared using PBS to a concentration of 10 mg / mL.

[0139] [Table 1]

[0140] (2) Melanocyte culture 1.4 x 10 cells in a 12-well plate 5 Melanocytes (normal human neonatal epidermal melanocytes, LIFELINE, Lot: 07965, passage number 5) were seeded at 100 cells / well and cultured for 2 days at 5% CO2 and 37°C. MGM was used as the culture medium. After the culture, the medium in the plate was aspirated and replaced with MGM medium (MGM FBS-) from which fetal bovine serum (FBS) had been removed. After the replacement, the medium was cultured overnight, and then replaced with MGM FBS- containing each plant extract prepared in Example 2(1) above, and the melanocytes were cultured for 1 hour. Note that the medium for the untreated group (nt group) and negative control group (NC group) was replaced with MGM FBS- containing the same amount of solvent as the extract-added group. After the culture, AGEs prepared in Example 2(1) above were added to a final concentration of 400 μg / mL. Note that an equal volume of PBS was added instead of AGEs to the nt group. After the addition, the cells were cultured under conditions of 5% CO2, 37°C, and 72 hours. After the culture, the cells were treated with each plant extract and AGEs were added again, and cultured under conditions of 5% CO2, 37°C, and 72 hours.

[0141] (3) Evaluation of cell number After the culture in Example 2(2), the medium was removed. After removal, Cell Counting Kit-8 (DOJINDO) diluted 1 / 20 with MGM FBS- was added, and the number of viable cells was assessed by measuring absorbance at 450 nm. Each group consisted of three samples (n=3). Statistical analysis was performed using Dunnett's test (*: p<0.05, ***: p<0.001 vs. nt).

[0142] (4) Evaluation of melanin content The cells obtained by the culture in Example 2(2) were detached using trypsin and collected. After collection, a mixture of ethanol and diethyl ether (3:1 by volume) was added, followed by centrifugation and removal of the supernatant to obtain a melanin precipitate. The precipitate was dried overnight, and the resulting dried melanin was dissolved in 25 μL of a 1N NaOH / 10% DMSO mixture. After dissolution, absorbance at 420 nm was measured, and the amount of melanin produced was calculated. Each group was run with three samples (n=3). Statistical analysis was performed using Dunnett's test (*: p<0.05, ***: p<0.001 vs. NC). These results are shown in Figure 2.

[0143] Figure 2 is a graph showing the amount of melanin produced per cell when each plant extract was added, in which the vertical axis indicates the relative value to the nt group, and the horizontal axis indicates the test group. In Figure 2, (A) shows the results when peony extract was added (the value of the nt group in the figure is set to 1; AGEs group: 1.22, 0.3% addition group: 1.19, 1% addition group: 1.08), (B) shows the results when kaede extract was added (the value of the nt group in the figure is set to 1; AGEs group: 1.22, 0.3% addition group: 1.08), (C) shows the results when mallow extract was added (the value of the nt group in the figure is set to 1; AGEs group: 1.27, 0.5% addition group: 0.96), (D) shows the results when wild rose extract was added (the value of the nt group in the figure is set to 1; AGEs group: 1.21, 1% addition group: 1.02), and (E) shows the results when horsetail extract was added (the value of the nt group in the figure is set to 1; AGEs group: 1.18, 0.3% addition group: 1.06). As shown in Figure 2, when each plant extract was added, the relative value decreased compared to the group without the plant extract (negative control group). These results suggest that peony extract, kaede extract, mallow extract, multiflora rose extract, and horsetail extract inhibit the binding of AGEs and RAGE, respectively, thereby suppressing the signal downstream of RAGE and, as a result, suppressing melanin production.

[0144] [Example 3] We investigated whether the extract of the raft obtained during the extraction process of the raft can suppress melanin production caused by AGEs.

[0145] (1) Sample preparation In Production Example 3 of the above-mentioned raft extract, the solution obtained by washing with 50% ethanol was used as raft HP20W, the solution obtained by eluting with 100% ethanol was used as raft HP20E, and the solution obtained by treating HP20E with an amount of activated carbon equal to the solid content and filtering through a filter was used as raft HP20E charcoal. The above-mentioned raft extract was prepared using MGM™-4 Melanocyte Growth Medium-4 BulletKit™ (hereinafter referred to as MGM, manufactured by Lonza) medium so that the final concentrations shown in Table 2 below were obtained upon addition to the medium. Hereinafter, raft HP20W, raft HP20E, and HP20E charcoal will also be referred to as raft extract. AGEs (AGE-BSA, manufactured by CalbioChem) were prepared using PBS to a concentration of 10 mg / mL.

[0146] [Table 2]

[0147] (2) Melanocyte culture 1.4 x 10 cells in a 12-well plate 5Melanocytes (normal human neonatal epidermal melanocytes, LIFELINE, Lot: 07965, passage number 5) were seeded at 100 cells / well and cultured for 2 days at 37°C with 5% CO2. MGM medium was used for the culture. After the culture, the medium in the plate was aspirated and replaced with MGM FBS-. After the replacement, the medium was cultured overnight, and then replaced with MGM FBS- containing Nautical Raft HP20W, Nautical Raft HP20E, or Nautical Raft HP20E charcoal prepared in Example 3(1), and the melanocytes were cultured for 1 hour. The medium for the non-added group (nt group) and negative control group (NC group) was replaced with MGM FBS- containing the same amount of solvent as the Nautical Raft extract-added group. After the culture, AGEs prepared in Example 3(1) were added to a final concentration of 400 μg / mL. An equal volume of PBS was added to the nt group instead of AGEs. After the addition, the cells were cultured under conditions of 5% CO2, 37°C, and 72 hours. After the culture, pretreatment with the early stage Nagi-i-kada extract and addition of AGEs were again performed, and the cells were cultured under conditions of 5% CO2, 37°C, and 72 hours.

[0148] (3) Evaluation of cell number After the culture in Example 3(2), the medium was removed. After removal, Cell Counting Kit-8 (DOJINDO) diluted 1 / 20 with MGM FBS- was added, and the number of viable cells was assessed by measuring the absorbance at 450 nm. Each group consisted of three samples (n=3). Statistical analysis was performed using Dunnett's test (*: p<0.05, ***: p<0.001 vs. nt).

[0149] (4) Evaluation of melanin content The cells obtained by the culture in Example 3(2) were detached using trypsin and collected. After collection, a mixture of ethanol and diethyl ether (3:1 by volume) was added, followed by centrifugation. The supernatant was removed to obtain a melanin precipitate. The precipitate was dried overnight, and the resulting dried melanin was dissolved in 25 μL of a 1N NaOH / 10% DMSO mixture. After dissolution, the absorbance at 420 nm was measured, and the amount of melanin produced was calculated. Each group consisted of three samples (n=3). Statistical analysis was performed using Dunnett's test (*: p<0.05, ***: p<0.001 vs. NC). These results are shown in Figure 3.

[0150] Figure 3 is a graph showing the amount of melanin produced per cell when each of the Naumann's raft extracts was added. In Figure 3, the vertical axis shows the relative value to the nt group, and the horizontal axis shows the test group and the concentration of the Naumann's raft extract added. In Figure 3, (A) shows the results when Nagiikada HP20W was added (the value of the nt group in the figure is set to 1, with the AGEs group being 1.30, the 0.1% addition group being 1.28, the 0.3% addition group being 1.23, and the 1% addition group being 1.32), (B) shows the results when Nagiikada HP20E was added (the value of the nt group in the figure is set to 1, with the AGEs group being 1.30, the 0.001% addition group being 1.43, the 0.003% addition group being 1.07, and the 0.01% addition group being 1.07), and (C) shows the results when HP20E charcoal was added (the value of the nt group in the figure is set to 1, with the AGEs group being 1.30, the 0.001% addition group being 1.18, the 0.003% addition group being 1.36, and the 0.01% addition group being 1.19). As shown in Figure 3, when Nagi-Kaida HP20E or HP20E charcoal was added, the relative values ​​decreased compared to the group without addition of either plant extract (negative control group). These results suggest that Nagi-Kaida HP20E and Nagi-Kaida HP20E charcoal inhibit the binding of AGEs and RAGE, respectively, thereby suppressing the signaling downstream of RAGE, and as a result, are able to suppress melanin production.

[0151] [Example 4] We investigated whether peony extract can suppress inflammation induced by AGEs.

[0152] (1) Sample preparation Peony extract (Falcorex Peony B, Ichimaru Pharcos Co., Ltd.) was prepared using 0.25% FBS-containing D-MEM medium (Fujifilm Wako Pure Chemical Industries, Ltd.) so that the concentration was 0.4% when added to the medium. AGEs were prepared using PBS so that the concentration was 10 mg / mL.

[0153] (2) Culture of normal human dermal fibroblasts (NBNHDF) 1 x 10 in a 35 mm dish 5 NBNHDF (normal human neonatal dermal fibroblasts, KURABO, Lot: 5884, passage number 5) were seeded at 100 cells / dish and cultured for 2 days under conditions of 5% CO2 and 37°C. The culture medium used was D-MEM containing 10% FBS. After the culture, the medium in the dish was aspirated and replaced with D-MEM containing 0.25% FBS. After the replacement, the culture was continued overnight, and the medium was replaced with D-MEM containing 0.25% FBS and containing the peony extract prepared in Example 4(1), and the NBNHDF were cultured for 1 hour. The medium for the non-added group (nt group) and negative control group (NC group) was replaced with D-MEM containing 0.25% FBS containing the same amount of solvent as the peony extract-added group. After the culture, AGEs prepared in Example 4(1) were added to a final concentration of 400 μg / mL. For the nt group, an equal volume of PBS was added instead of AGEs. After the addition, the cells were cultured under conditions of 5% CO2, 37°C, and 16 hours. After the culture, total RNA was extracted using an RNeasy Kit (QIAGEN).

[0154] (3) Expression analysis The total RNA concentration extracted in Example 4(2) was measured, and cDNA was prepared using ReverTra Ace® qPCR RT Master Mix (TOYOBO). After preparation, quantitative analysis of gene expression levels was performed by real-time PCR using THUNDERBIRD® Next SYBR® qPCR (TOYOBO). In the quantitative analysis, interleukin-8 (IL-8), tumor necrosis factor-α (TNF-α), and receptor for AGEs (RAGE) were used as target genes. RPS18 was used as a housekeeping gene. Table 3 below lists the primers used in the quantitative analysis. Each group was analyzed using three samples (n=3). Statistical analysis was performed using Dunnett's test (*: p<0.05, ***: p<0.001 vs. NC). These results are shown in Figure 4.

[0155] [Table 3]

[0156] Figure 4 is a graph showing the expression levels of each gene in NBNHDF cells when peony extract was added. In Figure 4, the vertical axis shows the relative value of each gene expression level to the nt group, and the horizontal axis shows the test group. In Figure 4, (A) shows the results of IL-8 gene expression (the value of the nt group in the figure is set to 1, and the results are AGEs group: 1.27, 0.4% addition group: 0.56), (B) shows the results of TNF-α gene expression (the value of the nt group in the figure is set to 1, and the results are AGEs group: 1.42, 0.4% addition group: 0.95), and (C) shows the results of RAGE gene expression (the value of the nt group in the figure is set to 1, and the results are AGEs group: 1.17, 0.4% addition group: 0.84). As shown in Figure 4, when peony extract was added, the relative values ​​decreased compared to the groups to which each plant extract was not added (negative control groups). These results indicate that peony extract can suppress the gene expression of IL-8, TNF-α, and RAGE. Furthermore, it suggests that peony extract inhibits the binding of AGEs and RAGE, thereby suppressing downstream signals of RAGE, and as a result, suppressing the expression of IL-8 and TNF-α.

[0157] [Example 5] We investigated whether peony extract can suppress the delay in cell proliferation caused by glycation of epidermal basement membrane components.

[0158] (1) Sample preparation Peony extract (Falcorex Peony B, Ichimaru Falcos Co., Ltd.) was added to the medium at a concentration of 0.4%. Among the additives included in KGM™ Gold Keratinocyte Growth Medium BulletKit™, Calcium Free (hereafter referred to as KGM, manufactured by Lonza), BPE and EGF were added at 1 / 5 of the specified amount, and the other additives were added at their specified amounts. The medium used was adjusted to a calcium ion concentration of 0.06 mmol / L (hereafter referred to as test KGM).

[0159] (2) Plate coating and saccharification reaction Epidermal basement membrane components, Cultrex UltiMatrix BME (hereinafter referred to as BME, R&D, 3433-005-01), iMatrix332 (Nippi, 892031), iMatrix511 (Nippi, 892011), and Fibronectin (Corning, 354008), were diluted as shown in Table 4 below. After dilution, they were coated onto a 96-well plate according to the respective product protocols. After coating, D(+)-glucose (Kishida, 000-34105) dissolved in PBS was added to a final concentration of 100 mg / ml. The plates were incubated at 37°C for 7 days to allow glycation of the basement membrane components. An equal volume of PBS was added to the nt group.

[0160] [Table 4]

[0161] (3) Culture of normal human neonatal epidermal keratinocytes (NBNHEK) The 96-well plate prepared in Example 5(2) was washed twice with PBS. After the washing, 1.5 × 10 NBNHEK cells (LIFELINE, Lot: 10232, passage number 4) were added. 3 After seeding, the cells were cultured under conditions of 5% CO2 and 37°C. During the culture, the medium contained all the specified amounts of additives and 0.06 mmol / L Ca 2+ KGM containing 100% ethanol was used. After 24 hours of culturing, the medium was replaced with the medium prepared in Example 5(1) above, and culturing was continued for an additional 72 hours. For the nt and NC groups, the medium was replaced with test KGM containing an equal volume of solvent. After culturing, the cells in each group were photographed using an inverted microscope (Olympus, main body: IX70, camera: DP70). The results are shown in Figure 5.

[0162] Figure 5 shows photographs of the results of observing the proliferation of NBNHEK cells induced by glycated epidermal basement membrane components when peony extract was added. In Figure 5, (A) shows the control results, (B) shows the results of the glycation model, and (C) shows the results when peony extract was added. As shown in Figure 5, cell proliferation was inhibited in the glycation model, whereas no inhibition of cell proliferation was observed when peony extract was added. These results indicate that peony extract inhibits the binding of AGEs and RAGE, thereby suppressing signaling downstream of RAGE, and as a result, is able to ameliorate the inhibition of cell proliferation caused by glycation.

[0163] [Example 6] We investigated whether peony extract has the effect of suppressing uneven skin tone.

[0164] (1) Correlation between color nonuniformity index and human skin age The inventors of the present invention have investigated whether peony extract can suppress uneven skin tone in humans.

[0165] First, we confirmed whether the information processing method could evaluate the correlation with skin tone unevenness caused by aging. Specifically, we calculated a skin nonuniformity index value from images containing skin as an index of skin tone unevenness, and confirmed that the color nonuniformity index value correlated with human skin age. First, we acquired images of approximately the same area of ​​cheek skin from the same subject at predetermined ages (38, 40, 41, 42, 43, 44, and 47 years old) (Figure 6(A)). For the images containing cheek skin at each age, we performed segmentation using SLIC registered in OpenCV so that the number of color types (number of segments) in the image was 100 or less, thereby integrating the colors in the image and dividing the regions (Figure 6(B)).

[0166] For the obtained image (Figure 6(B)), the variation (standard deviation) of the color (Lab) of each region, the area of ​​each region (segment), the color change rate (color gradient) between each region and its adjacent regions, and the average value of the lightness (L value) of each region were calculated. Next, variables and formulas for each index value were set using a large-scale language model so that each obtained index value correlates with the subject's skin age, which is the item being evaluated.

[0167] As a result, the L value (ω L ), a value (ω a ), b value (ω b ), the area of ​​each region, and the color gradient (ω grad The variables for the average L value (correction coefficient) were L value: a value: b value: area of ​​each region: color gradient = 3:2:1.5:1:2:0.03, and the index value of color nonuniformity (corrected hybrid score) calculated using the following formulas (3) to (12) correlated with the age of the subject's skin. The values ​​in the following formulas (6) to (12) were calculated based on the corrected L value (L i '), a value (a i ') and b value (b i The average value of the L value in the following formula (11) is calculated using the corrected L value (L i The results are shown in Figure 7.

number

[0168] FIG. 7 is a graph showing the correlation between the subject's skin age and the color non-uniformity index value. In FIG. 7, the horizontal axis represents the subject's skin age, and the vertical axis represents the color non-uniformity index value. As shown in FIG. 7, the color non-uniformity index value showed a high correlation (r=0.7196) with the subject's skin age (age / score: 38 / 4.68, 40 / 4.78, 41 / 5.12, 42 / 5.66, 43 / 6.85, 44 / 7.42, 47 / 9.06). These results confirmed that the information processing method can be used to evaluate the correlation with skin color unevenness caused by aging.

[0169] (2) Human skin monitor test Using the above-mentioned information processing method, it was investigated whether peony extract can suppress uneven skin tone in humans. Prior to the investigation, a lotion and a placebo lotion were prepared for use in a human skin monitoring test. The lotion used in the human skin monitoring test (Example 6) was prepared with the ingredients and content ratios shown in Table 5 below. The placebo lotion (Comparative Example 6) was prepared with the ingredients and content ratios shown in Table 6 below.

[0170] [Table 5]

[0171] [Table 6]

[0172] A monitoring test was conducted by applying the lotion listed in Table 5 and the placebo lotion listed in Table 6 to designated areas on the faces of normal human subjects (four women with an average age of 46 years). The monitoring test was conducted between November 18, 2024, and December 24, 2024. The lotion listed in Table 5 was applied to designated areas on one side of the subject's face (Example 6), and the lotion listed in Table 6 was applied to designated areas on the other side of the subject's face (Comparative Example 6). The application was conducted twice daily (morning and evening) at intervals of four weeks. In the monitoring test, photographs of the designated areas were taken using a camera on the first day of application, two weeks after application, and four weeks after application. The color non-uniformity index values ​​of the photographed images were calculated using the same information processing method as in Example 6(1). The color non-uniformity index value on the first day of application (0 w) was set as the reference (100), and the relative color non-uniformity value was calculated. These results are shown in FIG.

[0173] FIG. 8 is a graph showing the results of a human skin monitoring test. In FIG. 8, the vertical axis shows the relative value when the index value for color unevenness on the first day of application (0w) is set to 100, and the horizontal axis shows the number of weeks (w) from the first day of application. In FIG. 8, the value for Comparative Example 6 after two weeks was 107.2, the value for Comparative Example 6 after four weeks was 107.5, the value for Example 6 after two weeks was 88.7, and the value for Example 6 after four weeks was 77.5. As shown in FIG. 8, the index value for color unevenness was lower in Example 6 after four weeks of application than in Comparative Example 6 after four weeks of application. These results demonstrate that peony extract suppresses color unevenness.

[0174] [Example 7] We investigated whether peony extract can suppress the delayed differentiation caused by glycation of epidermal basement membrane components.

[0175] (1) Sample preparation Peony extract (Falcorex Peony B, Ichimaru Pharcos Co., Ltd.) was added to the medium at a concentration of 0.2% or 0.4%. The medium used in this preparation was KGM™ Gold Keratinocyte Growth Medium BulletKit™, Calcium Free (hereafter referred to as KGM, manufactured by Lonza) supplemented with specified amounts of BPE, EGF, and insulin, and adjusted to a calcium ion concentration of 0.15 mmol / L (hereafter referred to as KGM for differentiation tests).

[0176] (2) Plate coating and glycation reaction As an epidermal basement membrane component, iMatrix332 (Nippi, 892031) was used at 4.2 μg / cm 2 After the dilution, the solution was coated onto a 12-well plate according to the protocol. After the coating, D(+)-glucose (Kishida, 000-34105) dissolved in PBS was added to a final concentration of 100 mg / ml, and the plate was incubated at 37°C for 14 days to glycate the basement membrane components. An equal volume of PBS was added to the negative control group.

[0177] (3) Culture of normal human neonatal epidermal keratinocytes (NBNHEK) The 12-well plate prepared in Example 7(2) was washed twice with PBS. After the washing, 1.8 × 10 NBNHEK cells (LIFELINE, Lot: 10232, passage number 4) were added. 5 After seeding, the cells were cultured under conditions of 5% CO2 and 37°C. During the culture, the medium contained all the specified amounts of additives and 0.06 mmol / L Ca 2+ After 24 hours of culturing, the medium was replaced with the medium prepared in Example 7(1) (KGM for differentiation test), and the cells were cultured for another 24 hours. Thereafter, the medium was replaced with KGM for differentiation test, and the cells were cultured for 6 hours under conditions of 5% CO2 and 37°C. After the culturing, total RNA was extracted using an RNeasy Kit (QIAGEN).

[0178] (4) Expression analysis The total RNA concentration extracted in Example 7(3) was measured, and cDNA was prepared using ReverTra Ace® qPCR RT Master Mix (TOYOBO). After preparation, quantitative analysis of gene expression levels was performed by real-time PCR using THUNDERBIRD® Next SYBR® qPCR (TOYOBO). In the quantitative analysis, transglutaminase 1 (TGM1) and keratin 10 (K10) were used as target genes. RPS18 was used as the housekeeping gene. Table 7 below lists the primers used in the quantitative analysis (all manufactured by Takara Bio Inc.). Each group was analyzed using three samples (n=3). Statistical analysis was performed using Dunnett's test (*: p<0.05, ***: p<0.001 vs. NC). These results are shown in Figure 9.

[0179] [Table 7]

[0180] Figure 9 is a graph showing the expression levels of each gene in NBNHEK cells when peony extract was added. In Figure 9, the vertical axis shows the relative value of each gene expression level to the nt group, and the horizontal axis shows the test group. In Figure 9, (A) shows the results of TGM1 gene expression (the value of the nt group in the figure is set to 1, with PC group: 2.7, NC group: 2.0, 0.2% addition group: 2.1, and 0.4% addition group: 0.4), and (B) shows the results of K10 gene expression (the value of the nt group in the figure is set to 1, with PC group: 3.9, NC group: 1.7, 0.2% addition group: 2.3, and 0.4% addition group: 2.6). As shown in Figure 9, the addition of peony extract increased the relative values ​​compared to the group without peony extract (negative control group). These results demonstrate that peony extract can enhance the expression of TGM1 and K10 genes. These results suggest that peony extract may have a normalizing effect on turnover due to epidermal differentiation, normalizing barrier function, and / or moisturizing effects.

[0181] [Example 8] We investigated whether peony extract can suppress melanin production.

[0182] (1) Component fractionation The components contained in peony extract were analyzed. Specifically, 200 g of the peony extract prepared in Preparation Example 1 was diluted 5-fold with water to prepare a sample solution. After this preparation, 200 g of HP-20 gel was added to the sample solution, and adsorption was allowed to proceed overnight with stirring. In parallel, 200 g of HP-20 gel was swollen with 10% ethanol solution and packed into a column. The adsorbed gel and the non-adsorbed solution were added to the column. After this addition, the batch solution was applied to the column, and then elution was performed with 1 L of 10% ethanol solution to remove BG. The ethanol concentration was increased in 10% increments, and 250 mL of eluate was collected (1 L total). After collection, the eluate was divided into five fractions (Fr. 1 to Fr. 5) based on the HPLC analysis results, and 200 g of 50% ethanol solution was used to prepare the eluate for cell testing.

[0183] (2) Study of the inhibition of melanin production Melanocytes were cultured and melanin levels were evaluated in the same manner as in Example 2, except that peony extract (final concentration: 0.4%, Falcorex Peony B, Ichimaru Pharcos Co., Ltd.) or fractions Fr. 1 to Fr. 5 (final concentration: 0.48%) fractionated in Example 8(1) were added to the medium. The results are shown in Figure 10.

[0184] Figure 10 is a graph showing the amount of melanin produced per cell when peony extract was added. In Figure 10, the vertical axis represents the relative value to the AGEs group, and the horizontal axis represents the test group. In Figure 10, the values ​​for the AGEs group are set to 1, and the following values ​​are obtained: nt group: 0.77, 0.4% peony extract group: 0.87, Fr.1 addition group: 1.01, Fr.2 addition group: 1.02, Fr.3 addition group: 0.97, Fr.4 addition group: 0.99, Fr.5 addition group: 0.99. As shown in Figure 10, when peony extract was added, the relative value decreased compared to Fr.1 to Fr.5. These results suggest that melanin production can be suppressed by using peony extract as a whole rather than by using each fraction.

[0185] From the above results, it was found that the extracts of Paeonia lactiflora, Saururus chinensis, Bidens pilosa L., Rosa multiflora Thunb., and Thlaspi arvense L. inhibit the binding of AGEs and RAGE. It was also suggested that these extracts can suppress melanogenesis induced by AGEs by inhibiting the binding of AGEs and RAGE. Furthermore, the Paeonia lactiflora extract was suggested to exhibit effects such as anti-inflammation, improvement of cell growth inhibition due to glycation of the basement membrane, improvement of cell adhesion inhibition, improvement of skin color unevenness, normalization of turnover due to epidermal differentiation, normalization of barrier function, and / or moisturizing, etc. by inhibiting the binding of AGEs and RAGE. For the extracts showing AGEs and RAGE binding inhibitory activity (each extract of Saururus chinensis, Bidens pilosa L., Rosa multiflora Thunb., and Thlaspi arvense L.), it can be expected that, by suppressing the RAGE signal, similar effects to those of the Paeonia lactiflora extract, such as anti-inflammation, improvement of cell growth inhibition due to glycation of the basement membrane, improvement of cell adhesion inhibition, improvement of skin color unevenness, normalization of turnover due to epidermal differentiation, normalization of barrier function, and / or moisturizing, etc. can be shown.

[0186] The present disclosure has been described above by referring to the embodiments and examples, but the present disclosure is not limited to the above embodiments and examples. Various changes that can be understood by those skilled in the art can be made to the configuration and details of the present disclosure within the scope of the present disclosure.

[0187] This application claims priority based on Japanese Patent Application No. 2024-131958 filed on August 8, 2024, and incorporates all of its disclosure herein.

[0188] <Supplementary Note> Some or all of the above embodiments and examples can be described as follows in the supplementary note, but are not limited thereto. <Agent for use in inhibiting the binding of AGEs and RAGE> (Supplementary Note 1) An agent for use in inhibiting the binding of AGEs (advanced glycation end products) and RAGE (receptor for AGEs), comprising peony extract, kaede extract, mallow extract, multiflora rose extract, and / or horsetail extract. (Appendix 2) The agent described in Appendix 1 for use in inhibiting the expression of the RAGE gene, inhibiting the expression of the TNF-α gene, inhibiting the expression of the IL-8 gene, promoting the expression of the TGM1 gene, and / or promoting the expression of the K10 gene. (Appendix 3) The agent described in Appendix 1 for use in inhibiting melanin production, anti-inflammation, promoting cell proliferation, and / or promoting cell adhesion. <Skin application agent> (Appendix 4) 4. An agent for application to the skin, comprising an agent according to any one of appendices 1 to 3. <Cosmetics> (Appendix 5) A cosmetic comprising an agent according to any one of appendices 1 to 4. (Appendix 6) The cosmetic according to claim 5, wherein the cosmetic is a whitening cosmetic. [Industrial Applicability]

[0189] As described above, the present disclosure provides an agent that can inhibit the binding of AGEs and RAGE and that can be applied primarily to human skin, etc. Therefore, the present disclosure can be said to be extremely effective, for example, in the field of cosmetics, etc.

Claims

1. An agent for use in inhibiting the binding of AGEs (advanced glycation end products) and RAGE (receptor for AGEs), comprising peony extract, kaede extract, mallow extract, multiflora rose extract, and / or horsetail extract.

2. The agent according to claim 1, which is used for suppressing the expression of the RAGE gene, the TNF-α gene, the IL-8 gene, promoting the expression of the TGM1 gene, and / or promoting the expression of the K10 gene.

3. The agent according to claim 1 for use in inhibiting melanin production, anti-inflammation, promoting cell proliferation, and / or promoting cell adhesion.

4. An agent for application to the skin, comprising an agent according to any one of claims 1 to 3.

5. A cosmetic preparation comprising the agent according to any one of claims 1 to 4.

6. The cosmetic according to claim 5, wherein the cosmetic is a whitening cosmetic.

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