Composition for activating estrogen receptor signal transduction, and method for activating estrogen receptor signal transduction

Abstract

To provide a formulation that is effective in treating symptoms caused by disruption of hormonal balance.SOLUTION: The present invention provides a composition for activating estrogen receptor signal transduction via estrogen receptor 1 (ESR1), the composition containing allantoin or a salt thereof as an active ingredient.SELECTED DRAWING: None

Description

[Technical field]

[0001] The present invention relates to a composition for activating estrogen receptor signaling and a method for activating estrogen receptor signaling. [Background technology]

[0002] Hormones are chemical substances secreted from endocrine glands and act via the blood on target sites in the body to control their activity. Receptors exist at hormone target sites, and when hormones bind to them, they transmit information to cause a specific action. Hormones affect growth, development, reproduction, etc., and are also involved in homeostasis, such as maintaining energy metabolism and the concentration of blood components, and control the function of organs throughout the body. Furthermore, because even minute amounts of hormones can cause very large reactions, various symptoms and diseases can appear when the balance is disturbed.

[0003] Estrogen is a hormone produced mainly by granulosa cells in the ovary, and its secretion is controlled by the gonadotropic hormones follicle-stimulating hormone (FSH) and luteinizing hormone (LH) secreted by the anterior pituitary gland. A decrease in the blood level of estrogen promotes the secretion of FSH, while an increase in the blood level of estrogen inhibits the secretion of FSH, so estrogen plays a central role in the ingenious feedback mechanism that regulates ovarian function. Furthermore, estrogen has receptors not only in the target tissues of the diencephalon, anterior pituitary gland, genital organs, and mammary glands, but also in various other tissues, and is known to affect not only reproductive functions but also the skin, brain, cardiovascular system, skeleton, and other tissues throughout the body.

[0004] Menopausal disorders in women occur when ovarian function declines and estrogen secretion decreases, causing hormonal imbalance. Symptoms caused by menopausal disorders include vasomotor symptoms (hot flashes, sweating, chills, palpitations, shortness of breath, swelling), mental and neurological symptoms (headaches, dizziness, insomnia, anxiety, irritability, depression, tinnitus, hearing loss, dizziness, stress), locomotor symptoms (lower back pain, stiffness, joint pain, muscle pain, abnormal bone metabolism), digestive symptoms (loss of appetite, nausea, diarrhea, constipation, thirst, stomach upset, heartburn), general symptoms (fatigue), urinary symptoms (frequent urination, residual urine, pain during urination), reproductive symptoms (menstrual irregularities, decreased libido), sensory symptoms (numbness, hypoesthesia, hypersensitivity, decreased vision), and skin symptoms (dry skin, itching, wrinkles, dullness). It has also been revealed that after menopause, women are more likely to develop hypertension, diabetes, obesity, osteoporosis, autonomic nervous system imbalance, vague symptoms, dementia, etc. (Non-Patent Document 1). It is also known that even young women may experience imbalance of sex hormones, which can lead to premenstrual syndrome (PMS), which includes skin symptoms (acne), vasomotor nervous system symptoms (palpitations, swelling), mental and neurological symptoms (headache, dizziness, slight fever, anxiety, depression, insomnia, drowsiness, stress), locomotor system symptoms (lower back pain, joint pain), digestive system symptoms (constipation, diarrhea, abdominal pain, increased appetite, thirst), and systemic symptoms (fatigue). Although the severity of these symptoms varies greatly from person to person, severe symptoms often interfere with work and daily life.

[0005] On the other hand, an increase in menopausal disorders has been reported in men, and many of these symptoms are believed to be caused by a decrease in blood testosterone concentration, which is associated with various neuropsychiatric symptoms. It has also been reported that in men, a decrease in estrogen occurs along with testosterone, leading to the development of osteoporosis (Non-Patent Document 2).

[0006] For various symptoms caused by the decline of estrogen as described above, so-called phytoestrogens, such as plants containing estrogen-like substances and extracts obtained from them, have been used. For example, soy isoflavones (Patent Document 1, Patent Document 2) and Eucommia ulmoides (Patent Document 3) have been reported as phytoestrogens. Although these phytoestrogens are said to have relatively mild effects and almost no side effects, their effects are not sufficient, and there is an undeniable possibility of a risk of developing breast cancer or uterine cancer, so caution is advised when taking supplements, etc. [Prior art documents] [Patent documents]

[0007] [Patent Document 1] JP 2007-186483 A [Patent Document 2] JP 2005-229855 A [Patent Document 3] Special Publication No. 2012-77012 [Non-patent literature]

[0008] [Non-Patent Document 1] Hormones and Clinical Practice, Medical World Press, Vol. 57, p. 949-954, 2009 [Non-Patent Document 2] Basic aging research, Japan Society for Basic Aging, Vol. 34, p. 13-17, 2010 Summary of the Invention [Problem to be solved by the invention]

[0009] In view of the above-mentioned circumstances, the object of the present invention is to provide a novel preparation which is highly effective against symptoms caused by hormone imbalance. [Means for solving the problem]

[0010] As a result of intensive research to solve the above problems, the present inventors have found that allantoin or its salt, which is known to have anti-inflammatory and tissue repair effects, activates estrogen receptor signaling via estrogen receptor 1 (ESR1). It is suggested that allantoin or its salt normalizes the hormone imbalance caused by estrogen decline by activating estrogen receptor signaling via ESR1. That is, the gist of the present invention is as follows.

[0011] [1] A composition for activating estrogen receptor signaling mediated by estrogen receptor 1 (ESR1), characterized in that it contains allantoin or a salt thereof as an active ingredient. [2] The composition for activating estrogen receptor signaling described in [1], wherein the activation of estrogen receptor signaling is promotion of gene expression of estrogen receptor 1 (ESR1) and / or promotion of nuclear translocation of estrogen receptor 1 (ESR1). [3] The composition for activating estrogen receptor signaling described in [1] or [2], which is a composition for activating estrogen receptor signaling mediated by estrogen receptor 1 (ESR1) in epithelial cells. [4] The composition for activating estrogen receptor signaling described in [3], wherein the epithelial cells are epithelial cells in an inflammatory state. [5] A method for activating estrogen receptor signaling mediated by estrogen receptor 1 (ESR1), comprising using allantoin or a salt thereof. Effect of the Invention

[0012] According to the present invention, by using a composition containing allantoin or a salt thereof as an active ingredient, it is possible to activate estrogen receptor signaling mediated by estrogen receptor 1 (ESR1), thereby making it possible to normalize hormone imbalance caused by a decrease in estrogen, etc. [Brief description of the drawings]

[0013] [Figure 1] FIG. 1 is a diagram showing the test system for obtaining RNA-seq and ATAC-seq samples. [Diagram 2] Figure 2 shows the results of RNA-seq principal component analysis. [Figure 3-1] Figure 3-1 shows the results of RNA-seq differential expression gene extraction. [Figure 3-2] Figure 3-2 shows the results of GO (Gene Ontology) analysis performed following the extraction of differentially expressed genes by RNA-seq. [Figure 4] FIG. 4 is a schematic diagram showing that ESR1 acts as a transcription factor after receiving estrogen in the nucleus. [Diagram 5] FIG. 5 shows changes in chromatin opening and closing due to the addition of allantoin. [Figure 6] FIG. 6 is a schematic diagram showing changes in chromatin opening and closing caused by the addition of allantoin. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0014] The composition for activating estrogen receptor signaling and the method for activating estrogen receptor signaling of the present invention are described in detail below.

[0015] [Composition for activating estrogen receptor signaling] The composition for activating estrogen receptor signaling of the present invention is a composition for activating estrogen receptor signaling mediated by estrogen receptor 1 (ESR1), and is characterized by containing allantoin or a salt thereof as an active ingredient.

[0016] Estrogen refers to three types of molecules: estrone (E1), estradiol (E2), and estriol (E3), all of which have the ability to bind to estrogen receptors (ER). Estrogen is a type of steroid hormone that promotes the formation of reproductive functions and cell proliferation. In order to exert its physiological effects, it must bind to the ER present in the target tissue. When a ligand binds to the ER, the ER is activated and its binding to DNA is promoted, and it functions as a transcription factor that controls gene transcription. In addition to estrogen, there are other ER ligands, such as molecules such as isoflavones contained in plants (phytoestrogens) and endocrine disrupting substances that are also among the ligands that have the ability to bind to the ER and are known to exert their effects.

[0017] Estrogen receptors (ER) are one of the molecules belonging to the steroid receptor superfamily. They are also called estrogen receptors. In vivo, they exist in the nuclei of estrogen target cells, and when they bind to estrogen, they are activated, function as transcription factors, and mediate the activation of protein synthesis that depends on specific genes. There are two isoforms of ER, called ERα (NR3A1, 595 amino acid residues) and ERβ (NR3A2, 530 residues). These are produced by independent genes (ESR1, ESR2). ESR1 is located at 6q25.1, and ESR2 is located at 14q21-22. ER proteins activated by ligand binding form homodimers (αα, ββ) or heterodimers (αβ). In addition, both ERα and ERβ are composed of six domains (AF regions), and the amino acid sequences of the two types of receptor proteins are highly similar. The composition of the present invention activates signal transduction mediated by ERα (NR3A1, 595 amino acid residues) derived from the ESR1 gene, one of the two isoforms of the estrogen receptor.

[0018] In the present invention, estrogen receptor signal transduction refers to signal transduction in a series of pathways in which the above-mentioned ligand binds to estrogen receptor 1 (ESR1, ERα) and functions as a transcription factor to activate a protein synthesis pathway that depends on a specific gene. And, estrogen receptor signal transduction activation refers to promoting a part or all of the above-mentioned series of estrogen receptor signal transduction pathways, and activating a specific gene or a protein synthesis pathway that depends on it. In the present invention, estrogen receptor signal transduction activation may promote any step related to the above-mentioned series of estrogen receptor signal transduction pathways, and is preferably the promotion of gene expression of estrogen receptor 1 (ESR1, ERα) and / or the promotion of nuclear translocation of estrogen receptor 1 (ESR1). That is, the composition containing allantoin or a salt thereof of the present invention can activate a protein synthesis pathway that is specifically activated by estrogen receptor 1 functioning as a transcription factor by promoting gene expression of estrogen receptor 1 (ESR1, ERα) and / or promoting nuclear translocation of estrogen receptor 1 (ESR1).

[0019] According to the composition of the present invention containing allantoin or a salt thereof, the activation of estrogen receptor signaling enhances gene expression involved in epidermal differentiation and proliferation, intermediate filament cytoskeleton organization, intermediate filament-based pathways, keratinocyte differentiation, intermediate filament organization, cell migration, epithelial cell migration, epithelial migration, and tissue migration, thereby exerting various effects.

[0020] The estrogen receptor signal transduction mediated by estrogen receptor 1 (ESR1) activated by the composition of the present invention may be in any cell type, but is preferably in epithelial cells. As the above-mentioned epithelial cells, from the viewpoint of significantly achieving the effects of the present invention, epithelial cells of the skin (including scalp), eyes, oral cavity, intestinal tract, and vagina are preferable. In addition, the above-mentioned various epithelial cells are preferably epithelial cells in an inflammatory state, from the viewpoint of similarly significantly achieving the effects of the present invention. The above-mentioned inflammatory state is not particularly limited, but examples thereof include inflammation caused by bacteria, viruses, ultraviolet rays, aging, photoaging, stress, injury, etc.

[0021] Allantoin is a known compound also called 5-ureidohydantoin. Allantoin is contained in the rhizome of Symphytum officinale and is known to have anti-inflammatory, granulation, and tissue repair effects. In the composition of the present invention, Symphytum officinale or an extract thereof containing allantoin may be used as allantoin. Also, it may be a salt of allantoin, and is not limited as long as it is a pharmacopoeiacally acceptable salt, and examples thereof include allantoin β-glycyrrhetinic acid, allantoin dihydroxyaluminum (Aldioxa), allantoin chlorohydroxyaluminum (Alcloxa), allantoin polygalacturonic acid, allantoin ascorbic acid, allantoin acetyl-DL-methionine, allantoin DL-pantothenyl alcohol, and sodium DL-pyrrolidone carboxylate·allantoin. Of these, allantoin is preferred as a component contained in the composition of the present invention. The composition of the present invention may contain one or more components selected from the group consisting of allantoin and salts of allantoin.

[0022] The content of allantoin or its salt in the composition of the present invention is not particularly limited, and is appropriately set according to the type and content of other blended components, formulation form, method of use, etc., but from the viewpoint of significantly achieving the effects of the present invention, it is preferably 0.0005% by mass or more, more preferably 0.001% by mass or more, even more preferably 0.005% by mass or more, even more preferably 0.01% by mass or more, and particularly preferably 0.1% by mass or more, based on the total amount of the composition.In addition, from the same viewpoint, the content of allantoin or its salt in the composition of the present invention is preferably 3% by mass or less, more preferably 1% by mass or less, even more preferably 0.75% by mass or less, even more preferably 0.5% by mass or less, and particularly preferably 0.3% by mass or less.

[0023] From the viewpoint of significantly achieving the effects of the present invention, the content of allantoin or a salt thereof in the composition of the present invention is preferably 0.0005% by mass or more and 3% by mass or less, more preferably 0.001% by mass or more and 1% by mass or less, even more preferably 0.005% by mass or more and 0.75% by mass or less, still more preferably 0.01% by mass or more and 0.5% by mass or less, and particularly preferably 0.1% by mass or more and 0.3% by mass or less.

[0024] (Other Ingredients) The composition for activating estrogen receptor signaling of the present invention has an excellent effect of activating estrogen receptor signaling on epithelial cells, and is therefore used on cells of the skin (including the scalp), eyes, oral cavity, intestinal tract, and vagina, and is specifically suitable for use as a composition for external use, a composition for promoting hair growth, an ophthalmic composition, an oral composition, an oral composition, an oral composition, a vaginal composition, etc.

[0025] The composition of the present invention may contain other components in accordance with the purpose and use in addition to allantoin or its salt, which is an essential component.The composition of the present invention may contain, for example, solvent, anti-inflammatory agent other than allantoin or its salt, cooling agent, bactericide, preservative, vitamins, organic acid, moisturizing component, scrub agent, ultraviolet absorbing component, ultraviolet scattering component, astringent component, peptide or its derivative, amino acid or its derivative, polyhydric alcohol, thickener, cleaning component, keratin softening component, cell activation component, anti-aging component, blood circulation promoting component, whitening component, powder, sebum adsorption component, hair growth component, antihistamine component, anti-inflammatory analgesic component, antipruritic component, local anesthetic component, etc.In addition, in the composition of the present invention, one or more of these components may be used alone, or two or more may be used in combination.

[0026] The solvent is preferably one that functions as a base or carrier in the composition, and examples of the solvent include aqueous solvents such as water, hydrocarbons such as liquid paraffin, squalane, petrolatum, gelling hydrocarbons (Plastibase, etc.), ozokerite, α-olefin oligomers, and light liquid paraffin; methyl polysiloxanes such as polymethylsilsesquioxane, crosslinked methyl polysiloxanes, highly polymerized methyl polysiloxanes, cyclic silicones, alkyl-modified silicones, crosslinked alkyl-modified silicones, amino-modified silicones, polyether-modified silicones, polyglycerin-modified silicones such as lauryl dimethicone polyglycerin-3 crosspolymer, crosslinked polyether-modified silicones, crosslinked alkyl polyether-modified silicones, silicone / alkyl chain co-modified polyether-modified silicones, silicone / alkyl chain co-modified polyglycerin-modified silicones, polyether-modified branched silicones, and polyglycerin-modified branched silicones. Examples of the silicone oils include silicone oils such as acrylic silicone, phenyl-modified silicone, and silicone resin; higher alcohols such as cetanol, cetostearyl alcohol, stearyl alcohol, and behenyl alcohol; cellulose derivatives such as ethyl cellulose, hydroxypropyl cellulose, and hydroxypropyl methylcellulose; polyvinylpyrrolidone; carrageenan; polyvinyl butyrate; polyethylene glycol; dioxane; butylene glycol adipate polyester; esters such as isopropyl myristate, octyldodecyl myristate, isopropyl palmitate, cetyl palmitate, octyl palmitate, isononyl isononanoate, pentaerythritol tetra-2-ethylhexanoate, glyceryl tri-2-ethylhexanoate, and jojoba oil; polysaccharides such as dextrin and maltodextrin; and alcohols such as ethanol and isopropanol.

[0027] Examples of anti-inflammatory agents include glycyrrhetinic acid and its derivatives, glycyrrhizic acid and its derivatives, salicylic acid derivatives, aminocaproic acid, azulene and its derivatives, zinc oxide, tocopherol acetate, hydrocortisone, prednisolone, and salts thereof.Specific examples include glycyrrhetinic acid, glycyrrhizic acid, stearyl glycyrrhetinate, glycol salicylate, methyl salicylate, epsilon aminocaproic acid, azulene, guaiazulene, and salts thereof.Note that the term "derivative" refers to esters, ethers, alkylated products, glycosides, etc. of the compounds described herein.Note that the term "salt" refers to, for example, salts of mineral acids such as sulfuric acid, hydrochloric acid, or phosphoric acid, salts of organic acids such as maleic acid or methanesulfonic acid, alkali metal salts such as sodium or potassium, alkaline earth metal salts, ammonium salts, etc.The same applies to the optional components described below.

[0028] Examples of the cooling agents include terpenes such as camphor, 1-menthol, borneol, geraniol, cineole, anethole, limonene, and eugenol (which may be in any of the d-, l-, or dl-forms); and essential oils such as eucalyptus oil, bergamot oil, peppermint oil, cool mint oil, spearmint oil, fennel oil, peppermint oil, cinnamon oil, rose oil, and turpentine.

[0029] Examples of the disinfectants include isopropylmethylphenol, chlorhexidine, acrinol, gluconic acid and its derivatives, povidone-iodine, potassium iodide, iodine, triclocarban, triclosan, photosensitizer No. 101, photosensitizer No. 201, alkyldiaminoglycine hydrochloride, piroctoolamine, isoconazole, oxiconazole nitrate, miconazole nitrate, sulconazole nitrate, luliconazole, amorolfine hydrochloride, clotrimazole, ketoconazole, bifonazole, neticonazole hydrochloride, lanoconazole, liranaftate, efinaconazole, dequalinium chloride, chlorhexidine hydrochloride, chlorhexidine gluconate, alkyldiaminoglycine hydrochloride ... Examples of the antioxidants include dimethylaminoethylglycine, cetylpyridinium chloride, sodium benzoate, chlorobutanol, salicylic acid, gluconic acid, thymol, hexachlorophene, berberine, terbinafine hydrochloride, butenafine hydrochloride, lysozyme chloride, salicylic acid, salicylate salts, sulfur or sulfur compounds, hinokitiol, trichlorocarbanilide, halocarban, chlorophenesin, sorbic acid, potassium sorbate, sodium dehydroacetate, methyl parahydroxybenzoate, ethyl parahydroxybenzoate, propyl parahydroxybenzoate, butyl parahydroxybenzoate, oxyquinoline sulfate, phenethyl alcohol, benzyl alcohol, and biguanide compounds.

[0030] Examples of the preservatives include methyl benzoate, ethyl benzoate, propyl benzoate, sodium benzoate, chlorphenesin, and caprylhydroxamic acid.

[0031] The vitamins may be either water-soluble or oil-soluble, and examples thereof include vitamin B6 such as pyridoxine, pyridoxal, pyridoxamine, 5'-pyridoxal phosphate, and salts thereof (e.g., pyridoxine hydrochloride, pyridoxine acetate, pyridoxal hydrochloride, pyridoxamine hydrochloride); pantothenic acids such as pantothenic acid, calcium pantothenate, pantothenyl alcohol (panthenol), D-panthesine, D-pantethine, coenzyme A, pantothenyl ethyl ether, and salts thereof; nicotinic acid, nicotinic nicotinic acids such as dl-α-tocopherol nicotinate, benzyl nicotinate, methyl nicotinate, β-butoxyethyl nicotinate, 1-(4-methylphenyl)ethyl nicotinate, nicotinamide, and salts thereof; gamma-oryzanol, thiamine, dibenzoylthiamine, thiamine cetyl, thiamine monophosphate, thiamine diphosphate, thiamine triphosphate, and salts thereof (e.g., dibenzoylthiamine hydrochloride, thiamine hydrochloride, thiamine cetyl hydrochloride, thiamine thiocyanate, thiamine lauryl hydrochloride, thiamine Vitamin B1 such as nitrate, thiamine monophosphate, thiamine lysine salt, thiamine triphosphate, thiamine monophosphate phosphate, thiamine diphosphate hydrochloride, and thiamine triphosphate monophosphate; Vitamin B2 such as riboflavin, flavin mononucleotide, flavin adenine dinucleotide, riboflavin butyrate, riboflavin tetrabutyrate, riboflavin 5'-phosphate sodium, riboflavin tetranicotinate, and salts thereof; Vitamin B3 such as biotin, biocytin, and salts thereof. tins; folates such as folic acid, pteroylglutamic acid, and their salts; vitamin B12 such as cyanocobalamin, hydroxocobalamin, deoxyadenosylcobalamin, and their salts; water-soluble vitamin C such as ascorbic acid, dehydroascorbic acid, ascorbic acid phosphate, ascorbic acid-2-glucoside, 3-O-ethyl ascorbic acid, and their salts (e.g., sodium ascorbate, sodium ascorbyl phosphate, magnesium ascorbyl phosphate);Vitamin E such as dl-α-tocopherol, dl-α-tocopherol acetate, dl-α-tocopherol succinate, and dl-α-tocopherol calcium succinate; oil-soluble vitamin C such as ascorbigen-A, ascorbyl stearate, ascorbyl palmitate, L-ascorbyl dipalmitate, and ascorbyl tetra-2-hexyldecanoate; vitamin D such as ergocalciferol and cholecalciferol; vitamin K such as phylloquinone and farnoquinone; and vitamin-like factors such as ferulic acid.

[0032] Examples of the organic acids include gluconic acid, aspartic acid, aminoethylsulfonic acid, citric acid, glutamic acid, succinic acid, oxalic acid, fumaric acid, propionic acid, malic acid, salicylic acid, glycolic acid, phytic acid, tartaric acid, acetic acid, lactic acid, and salts thereof. Examples of the salts include salts of mineral acids such as sulfuric acid, hydrochloric acid, and phosphoric acid, salts of organic acids such as maleic acid and methanesulfonic acid, salts of alkali metals such as sodium and potassium, alkaline earth metal salts, and ammonium salts.

[0033] Examples of the moisturizing components include diglycerin trehalose; polymeric compounds such as sodium hyaluronate, heparinoids, sodium chondroitin sulfate, collagen, elastin, keratin, chitin, and chitosan; amino acids such as glycine, aspartic acid, and arginine; natural moisturizing factors such as sodium lactate, urea, and sodium pyrrolidone carboxylate; lipids such as ceramide, cholesterol, and phospholipids; and plant extracts such as chamomile extract, witch hazel extract, tea extract, and perilla extract.

[0034] Examples of the scrubbing agent include apricot kernel powder, almond shell powder, apricot kernel powder, sodium chloride granules, olive kernel powder, seawater dried granules, candelilla wax, walnut shell powder, cherry kernel powder, coral powder, charcoal powder, hazel shell powder, polyethylene powder, and anhydrous silicic acid.

[0035] Examples of the ultraviolet absorbing component include cinnamic acid-based ultraviolet absorbers such as di-paramethoxycinnamate mono-2-ethylhexanoate glyceryl; paramethoxycinnamate 2-ethylhexyl; dimethoxybenzylidene dioxoimidazolidinepropionate 2-ethylhexyl; 2,2'-methylenebis[6-(2H-benzotriazol-2yl)-4-(1,1,3,3-tetramethylbutyl)phenol]; 2,4-bis-[{4-(2-ethylhexyloxy)-2-hydroxy}-phenyl]-6-(4-methoxyphenyl)-1,3,5-triazine, diethylhexylbutamidotriazon, 2,4,6-tris[4-(2-ethylhexyloxycarbonyl)anilino]-1,3,5-triazine, diethylhexylbutamidotriazon ... benzalmalonate derivative ultraviolet absorbers such as dimethicodiethyl benzalmalonate; octocrylene ultraviolet absorbers such as 2-cyano-3,3-diphenylprop-2-enoic acid 2-ethylhexyl ester; imidazole sulfonic acid derivative ultraviolet absorbers such as 2-phenylbenzimidazole-5-sulfonic acid and disodium phenyldibenzimidazole tetrasulfonate; and benzophenone derivative ultraviolet absorbers such as 2-hydroxy-4-methoxybenzophenone, 2-hydroxy-4-methoxybenzophenone-5-sulfonic acid and salts thereof, dihydroxydimethoxybenzophenone, dihydroxybenzophenone, and tetrahydroxybenzophenone.

[0036] Examples of the ultraviolet ray scattering component include inorganic compounds such as hydrous silicic acid, zinc silicate, cerium silicate, titanium silicate, zirconium oxide, cerium oxide, zinc oxide, titanium oxide, iron oxide, and anhydrous silicic acid; these inorganic compounds are coated with inorganic powders such as hydrous silicic acid, aluminum hydroxide, mica, and talc; or are composited with resin powders such as polyamide, polyethylene, polyester, polystyrene, and nylon; and further treated with silicone oil, aluminum salt of a fatty acid, or the like.

[0037] Examples of the astringent ingredient include zinc sulfate, aluminum chloride, zinc sulfocarbonate, tannic acid, and the like.

[0038] Examples of the above peptides or derivatives thereof include keratin hydrolyzed peptides, hydrolyzed keratin, collagen, fish-derived collagen, atelocollagen, gelatin, elastin, elastin hydrolyzed peptides, collagen hydrolyzed peptides, hydrolyzed collagen, hydroxypropylammonium chloride hydrolyzed collagen, elastin hydrolyzed peptides, conchiolin hydrolyzed peptides, hydrolyzed conchiolin, silk proteolytic peptides, hydrolyzed silk, sodium lauroyl hydrolyzed silk, soybean proteolytic peptides, hydrolyzed soybean protein, wheat protein, wheat proteolytic peptides, hydrolyzed wheat protein, casein hydrolyzed peptides, acylated peptides (palmitoyl oligopeptide, palmitoyl pentapeptide, palmitoyl tetrapeptide, etc.), and the like.

[0039] Examples of the above amino acids or derivatives thereof include betaine (trimethylglycine), proline, hydroxyproline, arginine, lysine, serine, glycine, alanine, phenylalanine, β-alanine, threonine, glutamic acid, glutamine, asparagine, aspartic acid, cysteine, cystine, methionine, leucine, isoleucine, valine, histidine, taurine, γ-aminobutyric acid, γ-amino-β-hydroxybutyric acid, carnitine, carnosine, creatine, and the like.

[0040] Examples of the polyhydric alcohol include glycols (ethylene glycol, diethylene glycol, propylene glycol, isopropylene glycol, dipropylene glycol, polypropylene glycol, polyethylene glycol, 1,3-butylene glycol, pentylene glycol, etc.), glycerin, diglycerin, triglycerin, polyglycerin, sorbitol, and alkanediols (propanediol, 3-methyl-1,3-butanediol, pentanediol, etc.).

[0041] Examples of the thickening agent include agar, gellan gum, guar gum, locust bean gum, carrageenan, xanthan gum, dextran, methyl cellulose, ethyl cellulose, carboxymethyl cellulose, hydroxymethyl cellulose, hydroxyethyl cellulose, hydroxypropyl cellulose, hydroxypropyl methyl cellulose, carboxymethyl cellulose, carboxyethyl cellulose, sodium alginate, propylene glycol alginate, dextran, polyvinyl alcohol, polyvinylpyrrolidone, polyvinyl methyl ether, carboxyvinyl polymer, acrylic acid alkyl methacrylate copolymer, sodium polyacrylate, Examples of suitable active ingredients include polyethylene glycol, bentonite, dextrin fatty acid ester, pectin, (hydroxyethyl acrylate / sodium acryloyldimethyltaurate) copolymer, dimethyl distearyl ammonium hectorite, (ammonium acryloyldimethyltaurate / vinylpyrrolidone) copolymer, (ammonium acryloyldimethyltaurate / beheneth-25 methacrylate) cross copolymer, (ammonium acryloyldimethyltaurate / steareth-25 methacrylate) crosspolymer, polyethylene glycol distearate, ethylene glycol triisostearate, and polyoxyethylene (20) methyl glucoside triisostearate.

[0042] Examples of the cleaning components include soaps selected from alkali metal salts, alkanolamide salts, and amino acid salts, such as potassium laurate, potassium myristate, potassium palmitate, and potassium stearate; amino acid surfactants, such as sodium cocoyl glutamate and sodium cocoyl methyl taurate; ether sulfate salts, such as sodium laureth sulfate; ether carboxylates, such as sodium lauryl ether acetate; sulfosuccinate salts, such as sodium alkyl sulfosuccinate; fatty acid alkanolamides, such as coconut oil fatty acid monoethanolamide and coconut oil fatty acid diethanolamide; monoalkyl phosphate salts, such as sodium lauryl phosphate and sodium polyoxyethylene lauryl ether phosphate; betaine-type amphoteric surfactants, such as coconut oil fatty acid amidopropyl dimethylaminoacetate betaine, lauryl dimethylaminoacetate betaine, 2-alkyl-N-carboxymethyl-N-hydroxyethyl imidazolinium betaine, lauryl hydroxysulfobetaine, and sodium lauroyl amidoethyl hydroxyethyl carboxymethyl betaine hydroxypropyl phosphate; and amino acid amphoteric surfactants, such as sodium lauryl aminopropionate.

[0043] Examples of the keratin softening ingredient include lanolin, lactic acid, lactate salts, salicylic acid, gluconic acid, citric acid, malic acid, fruit acid, phytic acid, urea, glycolic acid, and sulfur.

[0044] Examples of the cell activation components include amino acids such as γ-aminobutyric acid; vitamins such as retinol, thiamine, riboflavin, pyridoxine hydrochloride, pantothenic acids, etc.; α-hydroxy acids such as glycolic acid, lactic acid, etc.; tannins, flavonoids, saponin, photosensitizer No. 301, etc.

[0045] Examples of the anti-aging ingredients include pangamic acid, kinetin, ursolic acid, turmeric extract, sphingosine derivatives, silicon, silicic acid, N-methyl-L-serine, and mevalonolactone.

[0046] Examples of the blood circulation promoting component include components derived from plants (e.g., ginseng, angelica, arnica, ginkgo, fennel, Japanese oak, Dutch oak, chamomile, Roman chamomile, carrot, gentian, burdock, rice, hawthorn, shiitake mushroom, ginger, European hawthorn, European juniper, Cnidium rhizome, Swertia japonica, thyme, clove, tangerine peel, chili pepper, angelica tree, peach kernel, spruce, carrot, garlic, butcher's broom, grape, peony, horse chestnut, melissa, yuzu, coix seed, ryokucha, rosemary, rose hip, tangerine peel, angelica tree, spruce, peach, apricot, walnut, corn, etc.); acetylcholine, ichthammol, cantharides tincture, gamma oryzanol, cepharanthine, tolazoline, tocopherol nicotinate, glucosyl hesperidin, etc.).

[0047] Examples of the whitening ingredient include vitamins such as tocopherol, tranexamic acid, nicotinamide, ascorbic acid or a salt thereof, an ascorbic acid derivative or a salt thereof, hydroquinone, placenta, arbutin, kojic acid, ellagic acid, phytic acid, 4-n-butylresorcinol, chamomile extract, vitamin A or a derivative thereof, and pantothenic acid or a derivative thereof. In addition, plant ingredients having a whitening effect may be used as the whitening ingredient. Examples of such plant ingredients include iris, almond, and the like. , aloe, ginkgo, oolong tea, ginger, Scutellaria, Coptis, Hypericum, white lamb, seaweed, pueraria, gardenia, solanaceae, chlorella, barley, rice, rice hyacinth, oryzanol, rice bran, saishin, Japanese pepper, perilla, peony, cnidium, soybean, natto, tea, angelica tree, calendula, garlic, witch hazel, safflower, peonies, coix seed, angelica tree, amethyst, astragalus, aceae, bracken, dogwood, enoki, persimmon (Diospyros Examples of ingredients that can be used include those derived from kaki, catalpa, black bean, gentian, gentian, salsa, kidney bean, jūrou, sage, zenko, radish, azalea, horsetail, toshin, bittern, parsley, holly, hops, Japanese bush clover, clove, licorice, and grapefruit.

[0048] Examples of the powder components include aluminum starch octenylsuccinate, talc, corn starch, magnesium hydroxide, anhydrous silicic acid, light anhydrous silicic acid, mica, hydroxyapatite, zinc oxide, titanium oxide, nylon powder, charcoal, kaolin, and calamine.

[0049] Examples of the sebum-adsorbing component include talc, mica, hydroxyapatite, zinc oxide, and aluminum silicate.

[0050] Examples of the hair growth ingredients include procyanidin, dipotassium glycyrrhizinate, capronium chloride, cepharanthine, menthol, hinokitiol, L-hydroxyproline, acetylhydroxyproline, fucoidan, capsicum tincture, cepharanthine, swerthianin, shinhongginisin, flavonosteroids, minoxidil, carpronium chloride, FGF-10, Enmeisou extract, Swertia japonica extract, Mitsuishikonbu extract, Gynostemma pentaphyllum extract, Hypericum perforatum extract, Gentiana extract, sage extract, peppermint extract, hop extract, Coix seed extract, persimmon leaf extract, Rehmannia Root extract, carrot extract, linden extract, Peony extract, and Jiu extract.

[0051] Examples of the antihistamine components include ethanolamine compounds such as diphenhydramine, diphenhydramine hydrochloride, and dimenhydrinate; propylamine compounds such as chlorpheniramine maleate; phenothiazine compounds such as promethazine hydrochloride; piperazine compounds such as hydroxyzine; piperidine compounds such as cyproheptadine hydrochloride, as well as epinastine hydrochloride, loratadine, and fexofenadine hydrochloride.

[0052] Examples of the anti-inflammatory and analgesic ingredients include indomethacin, felbinac, ibuprofen, ibuprofen piconol, bufexamac, butyl flufenamate, bendazac, piroxicam, and ketoprofen.

[0053] Examples of the antipruritic ingredients include crotamiton, chlorpheniramine, chlorpheniramine maleate, diphenhydramine, diphenhydramine hydrochloride, diphenhydramine salicylate, vanillylamide nonylate, mequitazine, camphor, thymol, eugenol, polyoxyethylene lauryl ether, and perilla extract.

[0054] Examples of the local anesthetic component include lidocaine, lidocaine hydrochloride, dibucaine, dibucaine hydrochloride, ethyl aminobenzoate, eucalyptus oil, eugenol, chlorobutanol, and the like.

[0055] In addition to the above-mentioned components, the composition may contain components that are commonly used in the fields of pharmaceuticals, quasi-drugs, cosmetics, etc., depending on the application or dosage form. The components that can be added are not particularly limited, but may include additives such as surfactants, preservatives, pH adjusters, chelating agents, stabilizers, irritation reducers, colorants, dispersants, and fragrances. These components may be added alone or in any combination of two or more.

[0056] Examples of the surfactant include sorbitan esters such as sorbitan stearate, PEG sorbitan stearate, sorbitan monooleate, sorbitan monoisostearate, and sorbitan monolaurate; POE-sorbitan monooleate; polyoxyethylene hydrogenated castor oil (HCO-10); glycerin fatty acid esters such as glycerin monooleate, glycerin monostearate, and glycerin monomyristate; glycerin alkyl ethers such as monoisostearyl glyceryl ether and monomyristyl glyceryl ether; various nonionic surfactants such as polyglycerin fatty acid esters such as polyglyceride stearate, polyglyceryl-10 isostearate, diglyceryl monostearate, decaglyceryl decastearate, decaglyceryl decastearate, and diglyceryl diisostearate; and naturally derived surfactants such as lecithin, hydrogenated lecithin, saponin, sodium surfactin, cholesterol, and bile acid.

[0057] Examples of the preservatives include benzoic acid, sodium benzoate, dehydroacetic acid, sodium dehydroacetate, isobutyl parahydroxybenzoate, isopropyl parahydroxybenzoate, butyl parahydroxybenzoate, ethyl parahydroxybenzoate, propyl parahydroxybenzoate, benzyl parahydroxybenzoate, methyl parahydroxybenzoate, and phenoxyethanol.

[0058] Examples of the pH adjuster include inorganic acids (hydrochloric acid, sulfuric acid, etc.), organic acids (lactic acid, sodium lactate, citric acid, sodium citrate, succinic acid, sodium succinate, etc.), inorganic bases (potassium hydroxide, sodium hydroxide, etc.), and organic bases (triethanolamine, diisopropanolamine, triisopropanolamine, etc.).

[0059] Examples of the chelating agent include ethylenediaminetetraacetic acid (edetic acid), ethylenediaminetetraacetate (sodium salt (sodium edetate: Japanese Pharmacopoeia, EDTA-2Na, etc.), potassium salt, etc.), phytic acid, gluconic acid, polyphosphoric acid, metaphosphoric acid, etc.

[0060] Examples of the stabilizer include magnesium sulfate, sodium polyacrylate, dibutylhydroxytoluene, and butylhydroxyanisole.

[0061] Examples of the irritation reducer include licorice extract, gum arabic, polyvinylpyrrolidone, sodium alginate, and the like.

[0062] Examples of the colorant include inorganic pigments and natural dyes.

[0063] Examples of the dispersant include sodium pyrophosphate, sodium hexametaphosphate, polyvinyl alcohol, polyvinylpyrrolidone, methyl vinyl ether / maleic anhydride crosslinked copolymer, and organic acids.

[0064] (pH) The composition of the present invention may generally have a liquid property of pH 2.0 to 9.0, but from the viewpoints of low irritation to the skin, mucous membranes, etc., and good usability, the pH is preferably 3.0 to 8.5, more preferably 3.5 to 8.0.

[0065] (Properties / Formulation) The properties of the composition of the present invention are not particularly limited, and it can be liquid, fluid, or semi-solid. The formulation form can be, for example, a liquid, suspension, emulsion, cream, milky lotion, ointment, gel, liniment, lotion, or a sheet of nonwoven fabric impregnated with a drug solution.

[0066] The composition of the present invention is preferably used in the fields of cosmetics, quasi-drugs and pharmaceuticals, particularly as a composition for improving, treating and preventing symptoms caused by hormonal imbalance in the tissues and cells of the skin (including the scalp), eyes, oral cavity, intestinal tract and vagina, both in inflammatory and normal states. The composition of the present invention is also preferably used to supplement symptoms caused by a decrease in hormones such as estrogen. The composition of the present invention exhibits collagen production promotion effects, skin and mucous membrane moisturizing effects, wound healing effects, hair cycle normalization effects and the like by activating estrogen receptor signaling via estrogen receptor 1 (ESR1), and therefore can be preferably used in the fields of cosmetics, quasi-drugs and pharmaceuticals, for example, as a composition for external use, a composition for promoting hair growth, an ophthalmic composition, an oral composition, an internal composition, a vaginal composition and the like. Specific examples include external and internal compositions for anti-acne (acne treatment), rough skin treatment, dermatitis treatment, scalp eczema treatment, hair growth, and suppression of body odor; oral compositions for treating periodontal disease, etc.; ophthalmic compositions for treating blurred vision and dry eyes, etc.; and vaginal compositions for treating vaginal candidiasis, etc.

[0067] [Method for activating estrogen receptor signaling] The present invention also includes a method for activating estrogen receptor signaling via estrogen receptor 1 (ESR1), characterized by using allantoin or a salt thereof. According to the method of the present invention, allantoin or a salt thereof can activate signaling via ERα (NR3A1, 595 amino acid residues) derived from the ESR1 gene, one of the two isoforms of the estrogen receptor. Specifically, allantoin or a salt thereof enhances gene expression involved in epidermal differentiation and proliferation, intermediate filament cytoskeleton organization, intermediate filament-based pathway, keratinocyte differentiation, intermediate filament organization, cell migration, epithelial cell migration, epithelial migration, and tissue migration through activation of estrogen receptor signaling, thereby exerting various effects.

[0068] The method of the present invention can be said to be a method for activating estrogen receptor signaling via estrogen receptor 1 (ESR1) using the above-mentioned composition for activating estrogen receptor signaling. Therefore, the description in the section on the composition for activating estrogen receptor signaling can be applied to the specific explanation of the estrogen receptor, estrogen receptor signaling, activation of estrogen receptor signaling, allantoin or a salt thereof, etc. in the method of the present invention. EXAMPLES

[0069] The present invention will be described in detail below with reference to examples and test examples, but the present invention is not limited to these examples.

[0070] 1. Acquiring RNA-seq and ATAC-seq data Using human epidermal keratinocytes (NHEK (NB) cells), we acquired RNA-seq and ATAC-seq data to investigate how cellular behavior changes when allantoin is added to cells in an inflammatory state. RNA-seq is a method to comprehensively and quantitatively examine gene expression in a cell population, and ATAC-seq is a method to investigate which genes are easily regulated by detecting open chromatin regions of DNA in cells. The purpose of this study was to combine these two methods to elucidate what kind of epigenetic control allantoin exerts on skin cells and how cellular gene expression changes as a result.

[0071] To obtain these data, cells were seeded on the first day, the medium was replaced with serum-free medium on the second day, and inflammation was induced on the third day by adding allantoin (three conditions: Alla 0.0% (Ctrl), 0.1%, 0.2%) and Pam3CSK4 (two conditions: 0 μg / mL (Ctrl) and 100 μg / mL). 24 h after stimulation with Pam3CSK4, total RNA was collected for RNA-seq and cells were collected for ATAC-seq, and sequences were obtained after library preparation. The experimental scheme is shown in Figure 1.

[0072] 2. RNA-seq analysis results – principal component analysis, GO analysis The obtained RNA-seq sequence data was trimmed of adapter sequences, mapped to the reference genome, and counted the number of mapped reads to quantify the expression level of genes in each sample. In order to examine whether the tendency of gene expression changes depending on the presence or absence of inflammation or allantoin stimulation, a principal component analysis was performed using the prcomp function of the statistical analysis programming language "R" (Figure 2). The sample names are [No inflammation: Ctrl, Inflammation: Pam]_[Concentration of added allantoin]_[Sampling time]. In addition to the clusters being separated by the presence or absence of inflammation, it was found that the position of the points changed with increasing allantoin concentration. This revealed that the tendency of gene expression changes with the presence or absence of inflammation and with changes in allantoin concentration. In addition, since the direction of change in the position of the plot with increasing allantoin concentration was the same regardless of the presence or absence of inflammation, it was revealed that the direction of change in gene expression with changes in allantoin concentration was the same regardless of the presence or absence of inflammation.

[0073] Next, genes whose expression was significantly increased in the sample with 0.2% allantoin concentration compared to 0% under Pam3CSK4 stimulation (inducing inflammation) were extracted for each time point. As a result, it was revealed that the expression of 151 genes was increased. To investigate what kind of cell phenotype these genes are related to, GO (Gene Ontology) analysis was performed using ClusterProfiler in "R". GO analysis is an analytical method to extract gene functions that are significantly more frequently observed in a list of genes compared to the entire gene population. The top 5 hits were 1. Epidermals development, 2. Ameboidal-type cell migration, 3. Epithelial cell migration, 4. Epithelium migration, and 5. Tissue migration. From these results, it was found that the induction of keratinocyte differentiation (cornification) was occurring through cell migration. In summary, it was found that the addition of allantoin induces keratinocyte differentiation, that is, cornification. The analysis results are shown in Figure 3.

[0074] 3. RNA-seq analysis results – Upstream analysis We used the biological framework Lisa (epigenetic Landscape In Silico deletion Analysis) (https: / / lisa.cistrome.org / ) to investigate the molecular mechanisms by which allantoin-induced keratinocyte differentiation is controlled. Lisa is a tool that determines transcription factors and chromatin regulators that are directly involved in the perturbation of differentially expressed gene sets. Using this tool, we investigated what transcription factors control the genes whose expression is increased by allantoin, as extracted in 2 above. As a result, it was revealed that ESR1 is active in the analysis based on the database of DNase ChIP-seq, which can examine transcription activation (Table 1), and the analysis based on the database of H3K27ac ChIP-seq, which examines the acetylation of lysine 27 of histone H3 protein (Table 2).

[0075] [Table 1]

[0076] [Table 2]

[0077] ESR1 is known as a gene that encodes an estrogen receptor, and is known to act as a transcription factor after receiving estrogen in the nucleus (Figure 4). From this, it is thought that the addition of allantoin exerts some kind of control, which increases the activity of ESR1 as a transcription factor. Estrogen is a type of female hormone, and its blood concentration is known to change depending on the menstrual cycle and age. In addition, it is known that the action of estrogen on the skin is to promote collagen production in skin fibroblasts and maintain skin moisture, etc. In addition, it has been reported in previous studies that an increase in estrogen levels promotes wound healing (Int J Mol Sci. 2017 Nov; 18(11): 2325), and that a low ratio of estrogen concentration to blood androgen worsens acne in women (Clin Cosmet Investig Dermatol. 2022; 15: 2211-2219), suggesting that estrogen is effective in wound healing and acne treatment in the skin. Therefore, since allantoin promotes the transcriptional activity of ESR1, it was suggested that allantoin or its salt may be effective in healing skin wounds and treating acne by acting like estrogen in skin epidermal cells.

[0078] 4. Results of ATAC-seq analysis To investigate what kind of epigenetic regulation occurs in ESR1 by the addition of allantoin, ATAC-seq was pretreated with the nf-core analysis pipeline (https: / / nf-co.re / atacseq / 2.1.2) and then analyzed. As a result, it was revealed that a part of the chromatin region encoding ESR1 was opened by the addition of allantoin under the condition of inflammation mimicking acne (Figure 5). In Figure 5, the peak indicates that the chromatin is open. The chromatin region in the box is opened by the addition of allantoin. This suggests that the transcriptional activity of ESR1 may be induced by epigenetic regulation of skin epidermal cells by allantoin or its salt.

[0079] When allantoin is added to skin epidermal cells that have been induced to cause inflammation to mimic acne, the chromatin region of ESR1 opens. This increases the activity of ESR1 as a transcription factor and increases the expression of various genes, inducing keratinocyte differentiation (cell keratinization), suggesting the possibility of wound healing and preventing the worsening of acne. These actions of allantoin are shown diagrammatically in Figure 6. From this, it is expected that allantoin or its salt will act like estrogen in skin epidermal cells and normalize the abnormality in the skin hormone balance that causes acne. [Industrial Applicability]

[0080] According to the present invention, by using a composition containing allantoin or a salt thereof as an active ingredient, it is possible to activate estrogen receptor signaling mediated by estrogen receptor 1 (ESR1), thereby making it possible to normalize hormone imbalance caused by a decrease in estrogen, etc.

Claims

1. A composition for activating estrogen receptor signaling mediated by estrogen receptor 1 (ESR1), characterized in that it contains allantoin or a salt thereof as an active ingredient.

2. The composition for activating estrogen receptor signaling according to claim 1, wherein the activation of estrogen receptor signaling is promotion of gene expression of estrogen receptor 1 (ESR1) and / or promotion of nuclear translocation of estrogen receptor 1 (ESR1).

3. The composition for activating estrogen receptor signaling according to claim 1 or 2, which is a composition for activating estrogen receptor signaling mediated by estrogen receptor 1 (ESR1) in epithelial cells.

4. The composition for activating estrogen receptor signaling according to claim 3 , wherein the epithelial cells are epithelial cells in an inflammatory state.

5. A method for activating estrogen receptor signaling mediated by estrogen receptor 1 (ESR1), comprising using allantoin or a salt thereof.

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