Reagent for structure maintenance of hyaluronic acid

Abstract

Provided are a reagent and the like for maintaining the structure of hyaluronic acid. The reagent according to the present invention is a reagent for maintaining the structure of hyaluronic acid having a molecular weight of 600 kDa to 2000 kDa, the reagent comprising a compound represented by formula (1) or a salt thereof. [chemical formula 1] # imgabs0 #

Description

Technical Field

[0001] The present invention relates to a reagent for maintaining the structure of hyaluronic acid, etc. Background Art

[0002] The skin, as the forefront directly affected by the external environment, undertakes the important function of maintaining the internal environment of the living body. Therefore, the functions of the skin do not completely stop, but with aging, ultraviolet radiation, exposure of the skin to chemical substances, etc., the functions of the skin gradually decline, and signs of aging such as wrinkles, spots, dullness, and sagging become obvious.

[0003] Hyaluronic acid (HA) is a linear mucopolysaccharide containing a repeating structure of D-glucuronic acid and N-acetyl-D-glucosamine. In vivo, hyaluronic acid is widely present in the skin, eyes, cartilage, synovium, synovial fluid, etc. In the skin, hyaluronic acid is present in the dermis and contributes to skin moisturization and elasticity. Conventionally, hyaluronic acid and raw materials that inhibit its decomposition have been used in cosmetics and the like for skin moisturization and anti-aging (Patent Document 1, Non-Patent Document 1).

[0004] In organisms such as humans (including the dermis of the skin, etc.), as a method of decomposing hyaluronic acid, for example, a KIAA1199 (HYBID)-dependent hyaluronic acid decomposition method can be cited (Non-Patent Document 2).

[0005] Prior Art Documents Patent Documents Patent Document 1: Japanese Unexamined Patent Application Publication No. 2018-135272 Non-Patent Documents Non-Patent Document 1: Shintaro Inoue, Glucosamine Research 5: 4-10, 2009 Non-Patent Document 2: Chemistry and Biology 55(2): 119-127 (2017) Summary of the Invention

[0006] Technical Problem to be Solved by the Invention Here, an object of the present invention is to provide a reagent for maintaining the structure of hyaluronic acid, etc.

[0007] To achieve the above object, the reagent of the present invention is a reagent for maintaining the structure of hyaluronic acid, and the reagent contains a compound represented by the following formula (1) or a salt thereof, the molecular weight of the hyaluronic acid is 600 kDa to 2000 kDa, [Chemical Formula 1]

[0008] In the formula (1), R 1 and R2 They may be the same or different and are each independently a hydrogen atom, a hydroxyl group, or a linear or branched alkoxy group having 1 to 5 carbon atoms. R 3 is a linear or branched hydroxyalkyl group having 1 to 7 carbon atoms, a linear or branched alkyl group having 1 to 7 carbon atoms, a cyclic alkyl group having 3 to 10 carbon atoms which may be substituted, or a hydrogen atom.

[0009] The reagent of the present invention is a reagent for inhibiting the expression of the HYBID gene. The reagent contains a compound represented by the following formula (1) or a salt thereof. [Chemical formula 1]

[0010] In the formula (1), R 1 and R 2 They may be the same or different and are each independently a hydrogen atom, a hydroxyl group, or a linear or branched alkoxy group having 1 to 5 carbon atoms. R 3 is a linear or branched hydroxyalkyl group having 1 to 7 carbon atoms, a linear or branched alkyl group having 1 to 7 carbon atoms, a cyclic alkyl group having 3 to 10 carbon atoms which may be substituted, or a hydrogen atom.

[0011] The method for maintaining the structure of hyaluronic acid of the present invention uses the reagent of the present invention.

[0012] The method for inhibiting the expression of the HYBID gene of the present invention uses the reagent of the present invention.

[0013] The compound or its salt of the present invention is represented by the following formula (2) or formula (3).

[0014] [Chemical formula 2]

[0015] [Chemical formula 3]

[0016] According to the present invention, for example, a reagent for maintaining the structure of hyaluronic acid can be provided. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] Figure 1 It is a diagram of the structural formula of the compound.

[0018] Figure 2 It is a schematic diagram of the skin monitoring test in Example 3.

[0019] Figure 3 It is a photograph showing the states of the gels in Example 6.

[0020] Figure 4 It is a graph showing the results of HPLC analysis in Example 9.

[0021] Figure 5 It is a diagram showing the fractionation scheme of the extract of the capitulum of Artemisia capillaris in Example 10.

[0022] Figure 6 It is a graph showing the gene expression level of HYBID upon addition of histamine in Example 10.

[0023] Figure 7 It is a graph showing the gene expression level of HYBID upon addition of histamine in Example 10.

[0024] Figure 8 It is a graph showing the gene expression level of HYBID upon addition of histamine in Example 10.

[0025] Figure 9 It is a graph showing the gene expression level of HYBID upon addition of histamine in Example 10.

[0026] Figure 10 It is a graph showing the gene expression level of HYBID in Example 10.

[0027] Figure 11 It is a graph showing the expression level of miR-486-5p in Example 11.

[0028] Figure 12 It is a graph showing the expression level of miR-486-5p in Example 11. Detailed implementation mode

[0029] Hereinafter, examples are given to specifically illustrate the present invention. Hereinafter, unless otherwise specifically mentioned, each invention can cite the description of other inventions.

[0030] <Reagent or composition for maintaining the structure of hyaluronic acid> As described above, the reagent or composition for maintaining the structure of hyaluronic acid of the present invention contains a compound represented by the following formula (1) or a salt thereof, and the molecular weight of the hyaluronic acid is 600 kDa to 2000 kDa.

[0031] [Chemical formula 1]

[0032] In the formula (1), R 1 and R 2 may be the same or different and are each independently a hydrogen atom, a hydroxyl group, or a linear or branched alkoxy group having 1 to 5 carbon atoms, R 3 is a straight-chain or branched-chain hydroxyalkyl group having 1 to 7 carbon atoms, a straight-chain or branched-chain alkyl group having 1 to 7 carbon atoms, a cyclic alkyl group having 3 to 10 carbon atoms which may be substituted, or a hydrogen atom.

[0033] The reagent or composition for maintaining the structure of hyaluronic acid according to the present invention is characterized by containing the compound represented by the formula (1) or a salt thereof, and other constitutions and conditions are not particularly limited. Since the reagent or composition for maintaining the structure of hyaluronic acid according to the present invention contains the compound represented by the formula (1) or a salt thereof, it can maintain the structure of hyaluronic acid in the skin. Therefore, according to the reagent or composition for maintaining the structure of hyaluronic acid of the present invention, for example, it can be expected to contribute to the moisturization of the epidermis.

[0034] Hereinafter, each substituent in the compound represented by the formula (1) will be illustrated by way of example. In the description of each substituent, in the case where no particular mention is made, the specific examples in the description of other substituents can be cited. In addition, in the following description, in the case where no particular mention is made, the description of the compound represented by the formula (1) can be cited, for example, to the description of the salt of the compound represented by the formula (1).

[0035] R 1 and R 2 are each a hydrogen atom, a hydroxyl group, or a straight-chain or branched-chain alkoxy group having 1 to 5 carbon atoms. Specifically, examples of the alkoxy group include: methoxy group (methyl oxy, methoxy group), ethoxy group (ethyl oxy, ethoxy group), propoxy group (propyl oxy, propoxy group), isopropoxy group (isopropyl oxy, isopropoxy group), butoxy group (butyl oxy, butoxy group), etc. The butyl group can be any of n-butyl, sec-butyl, isobutyl, and tert-butyl (hereinafter, the same). The R 1 is preferably a hydrogen atom or a hydroxyl group, for example. The R 2 is preferably a hydrogen atom, a hydroxyl group, or a methoxy group.

[0036] R 3is a linear or branched hydroxyalkyl group having 1 to 7 carbon atoms, a linear or branched alkyl group having 1 to 7 carbon atoms, a cyclic alkyl group having 3 to 10 carbon atoms which may be substituted, or a hydrogen atom. As specific examples, the hydroxyalkyl group may include, for example: hydroxymethyl; hydroxyethyl such as 1-hydroxyethyl, 2-hydroxyethyl; hydroxypropyl such as 1-hydroxypropyl, 2-hydroxypropyl or 3-hydroxypropyl; hydroxybutyl such as 1-hydroxybutyl, 2-hydroxybutyl, 3-hydroxybutyl, 4-hydroxybutyl; hydroxypentyl such as 1-hydroxypentyl, 2-hydroxypentyl, 3-hydroxypentyl, 4-hydroxypentyl, 5-hydroxypentyl; hydroxyhexyl such as 1-hydroxyhexyl, 2-hydroxyhexyl, 3-hydroxyhexyl, 4-hydroxyhexyl, 5-hydroxyhexyl, 6-hydroxyhexyl; hydroxyheptyl such as 1-hydroxyheptyl, 2-hydroxyheptyl, 3-hydroxyheptyl, 4-hydroxyheptyl, 5-hydroxyheptyl, 6-hydroxyheptyl, 7-hydroxyheptyl; etc. The alkyl group may include, for example: methyl; ethyl; propyl such as n-propyl, isopropyl (i-propyl); butyl such as n-butyl, isobutyl, tert-butyl, sec-butyl; pentyl such as n-pentyl, isopentyl, tert-pentyl; hexyl such as n-hexyl, isohexyl, tert-hexyl; heptyl such as n-heptyl, isoheptyl, tert-heptyl; etc. The cyclic alkyl group may include, for example: cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl, cyclodecyl, etc. In the cyclic alkyl group, one or more hydrogen atoms may be substituted with a hydroxyl group or a carboxyl group. The R 3 is preferably, for example, a linear or branched hydroxyalkyl group having 1 to 5 carbon atoms, or a linear or branched alkyl group having 1 to 5 carbon atoms, more preferably 1-hydroxypropyl, 2-hydroxypropyl, 3-hydroxypropyl, 1-hydroxybutyl, 2-hydroxybutyl, 3-hydroxybutyl, or 4-hydroxybutyl, or n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, or sec-butyl.

[0037] The compound represented by the formula (1) may be, for example, caffeic acid or a derivative thereof. The caffeic acid is a compound represented by the following formula (4). The compound of the following formula (4) can also be referred to as caffeic acid, (E)-3-(3,4-dihydroxyphenyl)prop-2-enoic acid, for example. The derivative of caffeic acid is a compound obtained by modifying the caffeic acid with an arbitrary substituent, for example. The derivatives of caffeic acid may include, for example, chlorogenic acids. The chlorogenic acids are compounds formed by ester-bonding the caffeic acid or ferulic acid with quinic acid.

[0038] [Chemical formula 4]

[0039] As a specific example, the caffeic acid or its derivative is, for example: in the compound of the formula (1), the R 1 is a hydroxyl group, the R 2 is a hydroxyl group, the R 3is a linear or branched hydroxyalkyl group having 1 to 7 carbon atoms, a linear or branched alkyl group having 1 to 7 carbon atoms, or an optionally substituted cyclic alkyl group having 3 to 10 carbon atoms, preferably hydroxypropyl, hydroxybutyl, propyl, butyl, or cyclohexyl. The substituents of the cyclic alkyl group can refer to the foregoing description, for example.

[0040] As a specific example, the derivative of caffeic acid is preferably a compound represented by the following formula (2) or a salt thereof. The compound represented by the following formula (2) can also be referred to as 3-hydroxybutyl (2E)-3-(3,4-dihydroxyphenyl)prop-2-enoate, for example.

[0041] [Chemical formula 2]

[0042] As a specific example, the derivative of caffeic acid is preferably a compound represented by the following formula (3) or a salt thereof. The compound represented by the following formula (3) can also be referred to as 4-hydroxybut-2-yl (2E)-3-(3,4-dihydroxyphenyl)prop-2-enoate, for example.

[0043] [Chemical formula 3]

[0044] In the present invention, the caffeic acid or its derivative can be in the form of a salt, hydrate or solvate. The caffeic acid or its derivative can be an isomer. Examples of the isomer include a tautomer or a stereoisomer. The tautomer or the stereoisomer includes all theoretically possible tautomers or stereoisomers, for example.

[0045] The salt of caffeic acid or its derivative is not particularly limited, and for example, it is a pharmaceutically acceptable salt. The caffeic acid or its derivative forms an acid addition salt or a salt with a base, for example, according to the type of substituent. The pharmaceutically acceptable salt is not particularly limited, and examples thereof include: alkali metal salts such as sodium salt and potassium salt; alkaline earth metal salts such as calcium salt and magnesium salt; ammonium salt; aliphatic amine salts such as trimethylamine salt, triethylamine salt, dicyclohexylamine salt, ethanolamine salt, diethanolamine salt, and triethanolamine salt; aralkylamine salts such as N,N-dibenzylethylenediamine; heterocyclic aromatic amine salts such as pyridine salt, methylpyridine salt, quinoline salt, and isoquinoline salt; quaternary ammonium salts such as tetramethylammonium salt, tetraethylammonium salt, benzyltrimethylammonium salt, benzyltributylammonium salt, methyltrioctylammonium salt, and tetrabutylammonium salt; amino acid salts such as arginine salt, lysine salt, aspartate salt, and glutamate salt; inorganic acid salts such as hydrochloride, sulfate, nitrate, phosphate, carbonate, hydrogencarbonate, and perchlorate; aliphatic organic acid or aromatic organic acid salts such as acetate, propionate, succinate, glycolate, lactate, maleate, fumarate, tartrate, malate, citrate, ascorbate, hydroxymaleate, pyruvate, phenylacetate, benzoate, 4-aminobenzoate, o-aminobenzoate, 4-hydroxybenzoate, salicylate, 4-aminosalicylate, pamoate, gluconate, and nicotinate; sulfonates such as methanesulfonate, hydroxyethanesulfonate, ethanesulfonate, benzenesulfonate, halogenated benzenesulfonate, p-toluenesulfonate, toluenesulfonate, naphthalenesulfonate, p-aminobenzenesulfonate, and cyclohexylaminosulfonate; and the like.

[0046] In the present invention, as the caffeic acid or its derivative, the reagent may contain Artemisia capillaris extract and / or Lonicera japonica extract.

[0047] In the present invention, "Artemisia capillaris" refers to the plant of the species Artemisia capillaris Thunb. belonging to the genus Artemisia Asteraceae (Compositae). Artemisia Regarding the crude drug name of Artemisia capillaris, the capitulum is also called Artemisia capillaris. The crude drug of Artemisia capillaris is used as a folk medicine and is used as a drug for anti-inflammatory, cholagogic, antipyretic, and diuretic effects. Artemisia capillaris Thunb .( Compositae ))

[0048] The extract of Artemisia capillaris Thunb. can be, for example, a plant extract. The plant extract can be produced, for example, by subjecting the flowers of Artemisia capillaris Thunb. to solvent extraction. The flowers of Artemisia capillaris Thunb. contain, for example, capitula, flower spikes, and / or flower-bearing branches and leaves. The extract of Artemisia capillaris Thunb. can be prepared by oneself or a commercially available product can be used. When the extract of Artemisia capillaris Thunb. is an extract of the flowers of Artemisia capillaris Thunb., the flowers can be the collected flowers themselves or processed products such as those that have been dried and / or pulverized. Examples of the commercially available products include Pharcolex Kawarayomogi B (manufactured by Ichimai Natural Beauty Co., Ltd.) and Pharcolex Kawarayomogi E (manufactured by Ichimai Natural Beauty Co., Ltd.). The manufacturing method of the extract of Artemisia capillaris Thunb. can be referred to the examples described later.

[0049] Examples of the solvent for extracting Artemisia capillaris Thunb. include: aqueous solvents such as water and buffer solutions; lower alcohols or water-containing lower alcohols such as methanol, ethanol, propanol, isopropanol, butanol, and isobutanol; polyhydric alcohols or water-containing polyhydric alcohols such as propylene glycol, 1,3-butanediol, 1,2-butanediol, 1,4-butanediol, 1,5-pentanediol, 1,2-pentanediol, 1,3-pentanediol, 1,4-pentanediol, 1,3,5-pentanetriol, glycerin, and polyethylene glycol (molecular weight 100 to 100,000); organic solvents such as acetone, ethyl acetate, diethyl ether, dimethyl ether, ethyl methyl ether, dioxane, acetonitrile, xylene, benzene, chloroform, carbon tetrachloride, phenol, and toluene; acids (hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid, formic acid, acetic acid, etc.) or bases (sodium hydroxide, potassium hydroxide, calcium hydroxide, ammonia, etc.) with appropriately adjusted normal concentration; etc. The solvent is preferably 1,3-butanediol. The solvent can be used alone or two or more kinds can be used in combination. The extraction can be carried out, for example, by immersing the plant in the solvent at room temperature (about 10°C to 30°C) for 5 days to 10 days.

[0050] The extract of Artemisia capillaris Thunb. can be a purified product after extraction. The purified product can be obtained, for example, by the following treatments: decomposition based on the addition of an acid (hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid, organic acid, etc.) or a base (sodium hydroxide, calcium hydroxide, ammonia, etc.); fermentation or metabolic conversion based on microorganisms; component adsorption based on ion exchange resins, activated carbon, diatomaceous earth, etc.; fractionation using chromatography having various separation modes (ion exchange, hydrophilic adsorption, hydrophobic adsorption, size exclusion, ligand exchange, affinity, etc.); filtration using filter paper, membrane filters, ultrafiltration membranes, etc.; pressurization or depressurization; heating or cooling; drying; pH adjustment; deodorization; decolorization; long-term static storage, etc. One of the treatments can be carried out or a combination of treatments can be carried out.

[0051] In the present invention, "hyaluronic acid" refers to a polysaccharide having a structure formed by linking disaccharide units of glucuronic acid and N-acetylglucosamine. The salt of the hyaluronic acid is not particularly limited as long as it is a food or pharmaceutically acceptable salt, and examples thereof include sodium salt, potassium salt, calcium salt, zinc salt, magnesium salt, ammonium salt, and the like.

[0052] The hyaluronic acid can be, for example, a substance obtained by extracting and concentrating or subjecting to enzymatic treatment from cockscombs or tissues of other animals and plants, or a substance produced by fermentation of hyaluronic acid-producing microorganisms such as microorganisms belonging to the genus Streptococcus. The hyaluronic acid can be used, for example, either a crude extract or a purified product, but from the viewpoint of less coloring and less off-odor during storage, hyaluronic acid with a purity of 90% or more is preferably used.

[0053] In the present invention, "maintaining the structure of hyaluronic acid" means maintaining the molecular weight of hyaluronic acid. The lower limit and upper limit of the molecular weight of the hyaluronic acid can be referred to the examples described later. The lower limit of the molecular weight of the hyaluronic acid is preferably 600 kDa, more preferably 1200 kDa. The upper limit of the hyaluronic acid is preferably 2000 kDa, more preferably 1600 kDa. The molecular weight of the hyaluronic acid is, for example, 600 kDa to 2000 kDa, preferably 1200 kDa to 1600 kDa.

[0054] In the present invention, "Lonicera" refers to a plant of the species Lonicera ( Caprifoliaceae ), genus Lonicera ( Lonicera ), family Caprifoliaceae ( Lonicerajaponica ). The crude drug name of the Lonicera also refers to the leaves as Lonicera and the flowers as Flos Lonicerae. The crude drug of the Lonicera is used as a folk medicine and is used as a drug for diuresis and antipyretic, and the decoction of the Lonicera is used as a gargle.

[0055] The extract of the Lonicera can be, for example, a plant extract. The plant extract can be produced, for example, by subjecting the plant of the Lonicera to solvent extraction. The plant of the Lonicera can be, for example, a plant individual or a part of a plant. Examples of the part of the plant include: leaves; flowers including capitula, flower spikes, flower-bearing branches, etc.; or a mixture thereof. The extract of the Lonicera can be prepared by itself or a commercially available product can be used. Examples of the commercially available product are as follows.

[0056] ·Pharcolex Suikazura FB: An extract of the flowers of Lonicera, prepared with a 1,3-butanediol solution as a solvent, manufactured by Ichibei Shizenken Co., Ltd.

[0057] ·Pharcolex Suikazura FE: An extract of the flowers of Lonicera, prepared with an ethanol solution as a solvent, manufactured by Ichibei Shizenken Co., Ltd.

[0058] ·Pharcolex Suikazura SB: An extract of the leaves of Lonicera japonica Thunb., prepared as an extract using a 1,3 - butanediol solution, manufactured by Ichibei Bijutsu Co., Ltd.

[0059] The reagent for maintaining the structure of hyaluronic acid of the present invention may contain, for example, compounds such as cinnamic acid or its derivatives, p - coumaric acid or its derivatives, or ferulic acid or its derivatives instead of the said caffeic acid or its derivatives. Since these compounds are, for example, analogs of the said caffeic acid or its derivatives, they can be expected to function as active ingredients (compounds) for maintaining the structure of hyaluronic acid. These compounds can be in the form of salts, hydrates or solvates, for example, and can also be isomers. As specific examples of these compounds, for example, the compounds shown in Figure 1 are cited.

[0060] The reagent for maintaining the structure of hyaluronic acid of the present invention can maintain the structure of hyaluronic acid, for example, by being used on a subject to be administered. The usage conditions (administration conditions) of the reagent for maintaining the structure of hyaluronic acid of the present invention are not particularly limited, and for example, the administration method, administration time, administration amount, etc. can be appropriately set according to the type of the subject to be administered, etc.

[0061] The reagent for maintaining the structure of hyaluronic acid of the present invention can be used, for example, in in vivo and can also be used in in vitro .

[0062] The subject to be administered with the reagent for maintaining the structure of hyaluronic acid of the present invention is not particularly limited. When in vivo using the reagent for maintaining the structure of hyaluronic acid of the present invention, the subject to be administered can be, for example, a human or a non - human animal other than a human. As the said non - human animal, for example, mammals such as mice, rats, rabbits, dogs, sheep, horses, cats, goats, monkeys, guinea pigs, etc., birds, etc. can be cited. When in vitro using the reagent for maintaining the structure of hyaluronic acid of the present invention, the subject to be administered can be, for example, cells, tissues, organs, etc. The said cells can be, for example, cells collected from a living body, cultured cells, etc., and the said tissues or organs can be, for example, tissues (biological tissues) or organs collected from a living body, etc.

[0063] In the following external skin agents (for example, transdermal administration agents or coating agents or compositions for the skin) or the following oral administration agents or oral administration compositions containing the reagent for maintaining the structure of hyaluronic acid of the present invention, the blending amount of the extract of the flowers of Artemisia capillaris Thunb. only needs to be within the range that exhibits the effect of maintaining the structure of hyaluronic acid, that is, an effective amount.

[0064] Examples of the administration method of the reagent for maintaining the structure of hyaluronic acid of the present invention include oral administration or parenteral administration. Examples of the parenteral administration include transdermal administration, coating (contact) on the skin, etc. The coating on the skin may include coating on the oral mucosa, that is, coating or contact with the epithelial cells in the oral cavity. In addition, the coating on the skin may include the meaning of administration or injection into the skin or subcutaneous tissue via the skin surface in addition to or instead of coating on the skin surface. Administration or injection into the skin via the skin surface can be carried out using, for example, microneedles.

[0065] The dosage form of the reagent for maintaining the structure of hyaluronic acid of the present invention is not particularly limited and can be appropriately determined according to the above-mentioned administration method, for example. Examples of the dosage form include liquid form and solid form. In the case where the administration method is oral administration, examples of the dosage form include tablets, pills, capsules, granules, powders, liquids, etc.

[0066] The reagent for maintaining the structure of hyaluronic acid of the present invention may contain additives as needed, and when used as a composition, the additives preferably include pharmaceutically acceptable additives or pharmaceutically acceptable carriers. The additives are not particularly limited and examples thereof include base materials, excipients, coloring agents, lubricants, binders, disintegrants, stabilizers, coating agents, preservatives, flavoring and odor-masking agents such as fragrances. In the present invention, the blending amount of the additives is not particularly limited as long as it does not interfere with the effect of maintaining the structure of hyaluronic acid.

[0067] Examples of the excipient include: sugar derivatives such as lactose, lactose hydrate, sucrose, glucose, mannitol, sorbitol; corn starch, potato starch, αStarch, starch derivatives such as dextrin; cellulose derivatives such as crystalline cellulose; gum arabic; dextran; organic excipients such as pullulan; silicate derivatives such as light anhydrous silicic acid, synthetic aluminum silicate, calcium silicate, magnesium metasilicate aluminate; phosphates such as calcium hydrogen phosphate; carbonates such as calcium carbonate; inorganic excipients such as sulfates such as calcium sulfate. Examples of the coloring agent include yellow ferric oxide and the like. Examples of the lubricant include: metal stearates such as stearic acid, calcium stearate, magnesium stearate; talc; polyethylene glycol; silicon dioxide; hydrogenated vegetable oil and the like. Examples of the taste and odor correctives include: spices such as cocoa powder, menthol, aromatic powder, peppermint oil, borneol, cinnamon powder, sweeteners, acidulants and the like. Examples of the binder include: hydroxypropyl cellulose, hydroxypropyl methylcellulose, polyvinylpyrrolidone, polyethylene glycol and the like. Examples of the disintegrant include: cellulose derivatives such as carboxymethyl cellulose, carboxymethyl cellulose calcium; chemically modified starches and chemically modified celluloses such as carboxymethyl starch, sodium carboxymethyl starch, crosslinked polyvinylpyrrolidone, sodium starch glycolate and the like. Examples of the stabilizer include: parabens such as methyl p-hydroxybenzoate, propyl p-hydroxybenzoate; alcohols such as chlorobutanol, benzyl alcohol, phenethyl alcohol; benzalkonium chloride; phenols such as phenol, cresol; thimerosal; dehydroacetic acid; sorbic acid and the like. Examples of the coating agent include: hydroxypropyl methylcellulose, polyethylene glycol such as polyethylene glycol 6000, talc, titanium oxide and the like.

[0068] In the case where the composition for maintaining the structure of hyaluronic acid of the present invention is an oral administration composition, specific examples of the oral administration composition include beverages, foods, pharmaceuticals (categories), quasi-pharmaceuticals (categories) and the like.

[0069] When the reagent or composition for maintaining the structure of hyaluronic acid in the present invention is a reagent for transdermal administration or application to the skin (hereinafter, also referred to as "skin external preparation"), the form of the skin external preparation, depending on the usage form, may include ampoules, capsules, powders, granules, liquids, gels, foams, emulsions, tablets, mists, sprays, etc. The usage forms may include, for example: pharmaceuticals (categories); quasi-pharmaceuticals (categories); topical or systemic skin external preparations; pharmaceutical and / or cosmetic preparations applied to the scalp and hair; bath agents used by adding to bath water; other preparations; etc. The topical or systemic skin external preparations may include, for example: basic cosmetics such as lotions, emulsions, creams, ointments, lotions, oils, face masks, solid soaps, liquid soaps, hand sanitizers or skin cleansers, massage agents, cleaning agents, hair removal agents, depilatory agents, shaving treatment agents, aftershave lotions, pre-shave lotions, shaving creams, foundations, lipsticks, blushes, eyeshadows, eyeliner creams, mascaras, etc. for makeup, perfumes, nail agents, nail polishes, nail polish removers, cataplasms, plaster agents, tape agents, sheet agents, patches, aerosol agents, mouthwashes such as toothpastes and mouthwashes, etc. The pharmaceutical and / or cosmetic preparations applied to the scalp and hair may include, for example: shampoos, hair conditioners, hair treatment agents, hair pretreatment agents, permanent wave solutions, hair dyes, hair styling agents, hot oil treatments, hair growth and nourishing agents, cataplasms, plaster agents, tape agents, sheet agents, aerosol agents, etc. The other preparations may include, for example: antiperspirants or deodorants, antiperspirants, sanitary products, sanitary napkins, wet wipes, etc.

[0070] The skin external preparation can be arbitrarily selected and / or used in combination with the following exemplified components and / or additives within the range that does not interfere with the effect of maintaining the structure of hyaluronic acid as needed.

[0071] (1) Various oils Avocado oil, almond oil, anise oil, perilla oil, olive oil, orange oil, orange roughy oil, sesame oil, cocoa butter, chamomile oil, carrot oil, cucumber oil, beef tallow fatty acid, macadamia nut oil, safflower oil, shea butter, liquid shea butter, soybean oil, camellia oil, corn oil, rapeseed oil, peach kernel oil, castor oil, cottonseed oil, peanut oil, turtle oil, mink oil, egg yolk oil, palm oil, palm kernel oil, candelilla wax, coconut oil, beef tallow, lard, squalene, squalane, shark liver alkane or hydrogenated products (such as hardened oils) of these oils, etc.

[0072] (2) Waxes Beeswax, carnauba wax, cetyl wax, lanolin, liquid lanolin, reduced lanolin, hard lanolin, candelilla wax, lignite wax, shellac wax, rice bran wax, etc.

[0073] (3) Mineral oils Liquid paraffin, petrolatum, paraffin wax, ozokerite, pure ozokerite, microcrystalline wax, etc.

[0074] (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-hydroxy stearic acid, undecylenic acid, tall oil fatty acid, lanolin fatty acid, etc.; synthetic fatty acids such as isononanoic acid, caproic acid, 2-ethylbutyric acid, isovaleric acid, 2-methylvaleric acid, 2-ethylhexanoic acid, isovaleric acid, etc.

[0075] (5)Alcohols Natural alcohols such as ethanol, isopropanol, lauryl alcohol, cetyl alcohol, stearyl alcohol, oleyl alcohol, lanolin alcohol, cholesterol, phytosterol, phenoxyethanol, etc.; synthetic alcohols such as 2-hexyldecanol, isostearyl alcohol, 2-octyldodecanol, etc.

[0076] (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-butanediol, pentanediol, glycerol, pentaerythritol, threitol, arabitol, xylitol, ribitol, galactitol, sorbitol, mannitol, lactitol, maltitol, etc.

[0077] (7)Esters Isopropyl myristate, isopropyl palmitate, butyl stearate, hexyl laurate, myristyl myristate, oleyl oleate, decyl oleate, octyldodecyl myristate, di-methyl octanoate hexyl decyl ester, cetyl lactate, myristyl lactate, diethyl phthalate, dibutyl phthalate, lanolin acetate, ethylene glycol monostearate, propylene glycol monostearate, propylene glycol dioleate, etc.

[0078] (8)Metal soaps Aluminum stearate, magnesium stearate, zinc stearate, calcium stearate, zinc palmitate, magnesium myristate, zinc laurate, zinc undecylenate, etc.

[0079] (9)Gums, saccharides or water-soluble high molecular compounds Gum arabic, benzoin gum, dammar gum, guaiac resin, Irish moss, karaya gum, tragacanth gum, carob 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 with an additional alkylene (C2-C4) oxide such as ethylene oxide, low molecular weight chitin or chitosan, chitosan salt, chitin or chitosan sulfate, phosphorylated chitin or chitosan, alginic acid or its salt, hyaluronic acid or its salt, chondroitin sulfate or its salt, 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, polyacrylate, polyalkylene oxide such as polyethylene oxide or polypropylene oxide or its cross-linked polymer, carboxyvinyl polymer, polyethyleneimine, etc.

[0080] (10) Surfactants Anionic surfactants (alkyl carboxylates, alkyl sulfonates, alkyl sulfates, alkyl phosphates), cationic surfactants (alkyl amine salts, alkyl quaternary ammonium salts), amphoteric surfactants: carboxylic acid type amphoteric surfactants (amino type, betaine type), sulfate type amphoteric surfactants, sulfonic acid type amphoteric surfactants, phosphate type 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, derivatives of protein hydrolysates, polymeric surfactants, titanium- and silicon-containing surfactants, fluorocarbon surfactants), etc.

[0081] (11) Various vitamins Vitamin A group: retinol, retinal (vitamin A1), dehydroretinal (vitamin A2), carotenoids, lycopene (provitamin A); Vitamin B group: thiamine hydrochloride, thiamine sulfate (vitamin B1), riboflavin (vitamin B2), pyridoxine (vitamin B6), cyanocobalamin (vitamin B12), folic acid, niacin, pantothenic acid, biotin, choline, inositol; Vitamin C group: ascorbic acid or its derivatives; Vitamin D group: ergocalciferol (vitamin D2), cholecalciferol (vitamin D3), dihydrotachysterol; Vitamin E group: vitamin E or its derivatives, ubiquinones; Vitamin K group: phylloquinone (vitamin K1), menaquinones (vitamin K2), menadione (vitamin K3), hydroquinone (vitamin K4); Other essential fatty acids (vitamin F), carnitine, ferulic acid, γ-oryzanol, orotic acid, vitamin P (rutin, eriocitrin, hesperidin), vitamin U, etc.

[0082] (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, etc., and their amino acid derivatives such as sulfates, phosphates, nitrates, citrates, or pyrrolidonecarboxylic acid.

[0083] (13) Additives The topical skin preparation may further contain various additives derived from animals or plants. The additives can be processed by conventional methods according to the type and form of the product to be added, and can be arbitrarily selected from various raw materials. The processing can be, for example, arbitrarily selecting and / or combining treatments such as crushing, powdering, washing, hydrolysis, fermentation, purification, pressing, extraction, fractionation, filtration, drying, granulation, dissolution, sterilization, pH adjustment, deodorization, and decolorization.

[0084] The solvent used in the extraction can be selected based on the usage purpose, type of the provided product, or subsequent processing. The extraction solvent is preferably, for example, a mixture of one or more selected from lower alcohols such as water, methanol, ethanol, propanol, isopropanol, butanol, isobutanol or water-containing lower alcohols, polyols such as propylene glycol, 1,3-butanediol, glycerol or water-containing polyols, and various organic solvents such as acetone and ethyl acetate. However, when the extraction solvent is not preferably contained with organic solvents according to the usage, water alone can be used, or ethanol that is easily removed after extraction can be used alone or in any mixture with water, or the solvent after squeezing extraction can also be used.

[0085] When the additive derived from plant or animal raw materials is used in external preparations for whole body or local use, or in cosmetics, the external skin preparation can be expected to have, for example, beauty effects such as moisturizing, improvement of touch and feel, imparting softness, alleviating irritation, alleviating stress caused by fragrance, cell activation (preventing cell aging), suppressing inflammation, improving skin and hair quality, preventing and improving skin roughness, hair growth, hair nourishment, preventing hair loss, imparting luster, cleaning effect, alleviating fatigue, promoting blood flow, and warm bath effect, in addition to protecting the skin and hair. In addition, effects such as fragrance imparting, deodorization, thickening, preservation, and buffering can also be expected.

[0086] In addition to the above, the external skin preparation can also be expected to have various beauty and pharmaceutical effects of various raw materials known so far. By combining them, the enhancement of the effects for the purpose of the present invention can be achieved, and a product with multifunctional effects can be produced.

[0087] <Expression inhibitor or composition of HYBID gene> In another aspect, the present invention provides a reagent or composition capable of inhibiting the expression of the HYBID gene. The expression inhibitor or composition of the HYBID gene of the present invention contains a compound represented by the following formula (1) or a salt thereof, and the molecular weight of the hyaluronic acid is 600 kDa to 2000 kDa: [Chemical formula 1]

[0088] In the formula (1), R 1 and R 2 may be the same or different and are each independently a hydrogen atom, a hydroxyl group, or a linear or branched alkoxy group having 1 to 5 carbon atoms, R 3 is a linear or branched hydroxyalkyl group having 1 to 7 carbon atoms, a linear or branched alkyl group having 1 to 7 carbon atoms, a cyclic alkyl group having 3 to 10 carbon atoms which may be substituted, or a hydrogen atom.

[0089] The expression inhibitor of the HYBID gene of the present invention or its composition is characterized by containing the compound represented by the formula (1) or its salt, and other constitutions and conditions are not particularly limited. Since the expression inhibitor or composition of the HYBID gene of the present invention contains the compound represented by the formula (1) or its salt, it can hinder the hyaluronic acid decomposition caused by HYBID and can maintain the structure of hyaluronic acid in the skin. Therefore, according to the expression inhibitor or composition of the HYBID gene of the present invention, for example, it can be expected to contribute to the moisturization of the epidermis. The description of the compound in the expression inhibitor or composition of the HYBID gene of the present invention can refer to the description of the reagent or composition for maintaining the structure of hyaluronic acid of the present invention.

[0090] In the present invention, "HYBID" (hyaluronic acid-binding protein involved in hyaluronic acid depolymerization, KIAA1199) refers to a hyaluronic acid-binding protein. It is known that the HYBID contributes to the decomposition of hyaluronic acid. The HYBID binds to hyaluronic acid of high molecular size (greater than 1000 kDa) and decomposes the hyaluronic acid into intermediate-sized fragments with a molecular weight of about 10 kDa.

[0091] <Method for maintaining the structure of hyaluronic acid> In another aspect, the present invention provides a method capable of maintaining the structure of hyaluronic acid. The method for maintaining the structure of hyaluronic acid of the present invention uses the reagent or composition for maintaining the structure of hyaluronic acid of the present invention. Since the method for maintaining the structure of hyaluronic acid of the present invention uses the reagent or composition for maintaining the structure of hyaluronic acid of the present invention, it can maintain the structure of hyaluronic acid in the skin. Therefore, according to the method for maintaining the structure of hyaluronic acid of the present invention, for example, it can be expected to contribute to the moisturization of the epidermis.

[0092] The method for maintaining the structure of hyaluronic acid of the present invention includes an administration step of administering the reagent and / or composition for maintaining the structure of hyaluronic acid of the present invention to a subject.

[0093] In the method for maintaining the structure of hyaluronic acid of the present invention, the administration step can be carried out, for example, in in vitro or in vivo The subject (administration subject) and administration conditions of the method for maintaining the structure of hyaluronic acid of the present invention can refer to the description of the administration subject and administration conditions in the reagent or composition for maintaining the structure of hyaluronic acid of the present invention.

[0094] <Method for inhibiting the expression of HYBID gene> In another aspect, the present invention provides a method for inhibiting the expression of the HYBID gene. The method for inhibiting the expression of the HYBID gene of the present invention uses an inhibitor or composition for inhibiting the expression of the HYBID gene of the present invention. Since the method for inhibiting the expression of the HYBID gene of the present invention uses the inhibitor or composition for inhibiting the expression of the HYBID gene of the present invention, it can hinder the decomposition of hyaluronic acid caused by HYBID and maintain the structure of hyaluronic acid in the skin. Therefore, according to the method for inhibiting the expression of the HYBID gene of the present invention, for example, it can be expected to contribute to the moisturization of the epidermis.

[0095] The method for inhibiting the expression of the HYBID gene of the present invention includes an administration step of administering an inhibitor or composition for inhibiting the expression of the HYBID gene of the present invention to a subject.

[0096] In the method for inhibiting the expression of the HYBID gene of the present invention, the administration step can be carried out, for example, at in vitro or in vivo . The subject (administration subject) and administration conditions of the method for inhibiting the expression of the HYBID gene of the present invention can, for example, refer to the description of the administration subject and administration conditions in the reagent or composition for maintaining the structure of hyaluronic acid of the present invention.

[0097] <Usage> In another aspect, the present invention is the use of a reagent and / or composition for maintaining the structure of hyaluronic acid in maintaining the structure of hyaluronic acid. The present invention is the use of an inhibitor and / or composition of the HYBID gene in inhibiting the expression of the HYBID gene.

[0098] The present invention is the use of a reagent and / or composition for maintaining the structure of hyaluronic acid in the manufacture of a reagent and / or composition for maintaining the structure of hyaluronic acid. The present invention is the use of an inhibitor and / or composition of the HYBID gene in the manufacture of an inhibitor and / or composition of the HYBID gene.

[0099] <Compound> In another aspect, the compound of the present invention is a compound represented by the following formula (2) or formula (3) or a salt thereof. The description of the compound of the present invention can refer to the description of the compound in the reagent or composition for maintaining the structure of hyaluronic acid of the present invention.

[0100] [Chemical formula 2]

[0101] [Chemical formula 3]

[0102] The compound of formula (2) or formula (3) can be produced, for example, by esterifying caffeic acid with 1,3-butanediol.

[0103] Examples Next, examples of the present invention will be described. However, the present invention is not limited to the following examples. Commercially available reagents are used based on their experimental protocols unless otherwise specified. In addition, percentages indicating the addition amounts of active ingredients, etc. refer to % by weight unless otherwise specified.

[0104] <Materials> · Normal adult human dermal fibroblasts: KF-4109 (manufactured by Kurabo Industries Ltd.), cells from humans aged 40 or older, hereinafter referred to as "AD cells".

[0105] · Normal neonatal foreskin dermal fibroblasts: KF-4009 (manufactured by Kurabo Industries Ltd.), hereinafter referred to as "NB cells".

[0106] · Extract of the flowers of Artemisia capillaris: Pharcolex Kawarayomogi B (manufactured by Ichimaru Pharcos Co., Ltd.) [Reference Example 1] For the structural maintenance of hyaluronic acid, a study was conducted using the RT-PCR method.

[0107] (1) NB cells Prepare NB cells. DMEM containing 5% FBS was used for pre-culture, and DMEM containing 0.25% FBS was used in this Reference Example 1. Culturing was carried out under the conditions of 5% CO2 and 37 °C.

[0108] (2) Preparation of hyaluronic acid (HA) Prepare the following hyaluronic acids.

[0109] · H2: HA with a molecular weight of 1200 kDa to 1600 kDa · M2: HA with a molecular weight of 600 kDa to 1120 kDa · S2: HA with a molecular weight of 40 kDa to 80 kDa · U2: HA with a molecular weight of 5 kDa to 10 kDa (3) Confirmation of the structural maintenance of hyaluronic acid (RT-PCR) 6×10 4NB cells were seeded in 6-well plates and cultured until they reached 75% confluence. After the above-mentioned culture, the medium was replaced with DMEM containing 0.25% FBS. After 24 hours of culture, the medium was replaced with fresh DMEM containing 0.25% FBS. For the NB cells after the replacement, the HA was added at a final concentration of 10 μ μg / mL to prepare the following experimental groups.

[0110] (Experimental groups) · Control group: The group without HA addition · H2 addition group: The group with H2 addition · M2 addition group: The group with M2 addition · S2 addition group: The group with S2 addition · U2 addition group: The group with U2 addition After preparing the above-mentioned experimental groups, they were further cultured in a CO2 incubator at 37°C for 1 to 2 hours. After the culture for 1 to 2 hours, in order to make the cells in an inflammatory state, for the above-mentioned experimental groups (including the control group), TNF- was added at a final concentration of 1 ng / mL α . After the addition, the cells were cultured in a CO2 incubator at 37°C for 6 hours. The mRNA was purified from the cultured cells. The purification of the mRNA was performed using QIAshreder and RNeasy Mini Kit (manufactured by QIAGEN). Using the purified mRNA as a template, reverse transcription reaction was performed using PrimeScript RT master Mix (manufactured by Takara Bio Inc.) to synthesize cDNA. Then, RT-PCR was performed using the primer pairs corresponding to each target factor shown in Table 1 below, and the change in expression level was analyzed by relative quantification. The RT-PCR was performed using TB Green Premix Ex Taq (manufactured by TakaraBio Inc.). In addition, the amplification result of RPS18 (ribosomal protein S18) was used as a reference for the analysis of relative quantity change. In addition, "MMP" in Table 1 below is matrix metalloproteinase.

[0111] [Table 1]

[0112] The quantitative results of the mRNA are shown in Table 2 below. Table 2 below shows the values when the value of the control group (the change in expression level shown by this relative quantification) is set to 1. The values are the values after rounding the second decimal place. In addition, in Table 2 below, the asterisk indicates a significant difference (p < 0.01) based on the comparison with the value of the control group in the Dunnett test, The label indicates a significant difference (p < 0.001) based on comparison with the values of the control group in the Dunnett test.

[0113] [Table 2]

[0114] As shown in the said Table 2, it can be seen that the lower limit of the molecular weight for maintaining the structure of hyaluronic acid is preferably 600 kDa, and the upper limit of the molecular weight is preferably 2000 kDa.

[0115] [Reference Example 2] Regarding the expression of HYBID induced by histamine in NB cells, it was studied using the RT-PCR method.

[0116] (1) NB cells Prepare NB cells. For pre-culture, use DMEM containing 5% FBS, and in this Reference Example 2, use DMEM containing 0.25% FBS. Culture under the conditions of 5% CO2 and 37 °C. After the culture, inoculate 6 × 10 4 NB cells into a 6-well plate and culture until 75% confluent. Then, replace it with DMEM containing 0.25% FBS. After culturing for 24 hours, replace it with new DMEM containing 0.25% FBS.

[0117] (2) Experimental groups For the replaced NB cells, use histamine, which is a substance that enhances hyaluronic acid degrading activity, to prepare the following experimental groups.

[0118] (Experimental groups) · Unadded group: The group without added histamine · 0.1 addition group: The group added with histamine at a final concentration of 0.1 μ mol / l · 1.0 addition group: The group added with histamine at a final concentration of 1.0 μ mol / l · 10 addition group: The group added with histamine at a final concentration of 10 μ mol / l (3) Confirmation of gene expression of HYBID (RT-PCR) After preparing the experimental group, it was further cultured in a CO2 incubator at 37 °C for 24 hours. After the culturing, mRNA was purified from the cells. The purification of mRNA was performed using QIAshreder and RNeasy Mini Kit (manufactured by QIAGEN). Using the purified mRNA as a template, reverse transcription reaction was performed using PrimeScript RT master Mix (manufactured by Takara Bio Inc.) to synthesize cDNA. Thereafter, RT-PCR was performed using the primer pairs corresponding to each target factor shown in Table 3 below, and the change in expression level was analyzed by relative quantification. The RT-PCR was performed using TB Green Premix Ex Taq (manufactured by TakaraBio Inc.). In addition, the amplification result of RPS18 (ribosomal protein S18) was used as a reference for the analysis of relative quantity change.

[0119] [Table 3]

[0120] These results are shown in Table 4 below. Table 4 below shows the values when the value of the non-added group (the change in expression level shown by this relative quantification) is set to 1. The values are those obtained by rounding off the third decimal place. In addition, in Table 4 below, The mark indicates a significant difference (p < 0.01) based on comparison with the value of the non-added group in the Dunnett test, The mark indicates a significant difference (p < 0.001) based on comparison with the value of the non-added group in the Dunnett test.

[0121] [Table 4]

[0122] As shown in Table 4 above, it was found that in NB cells, the expression of HYBID was enhanced by the addition of histamine (a substance that enhances hyaluronic acid-degrading activity).

[0123] [Example 1] The expression of HYBID induced by histamine in NB cells when an extract of the flower of Artemisia capillaris was added was studied using the RT-PCR method.

[0124] (1)NB cells NB cells were prepared. For pre-culturing, DMEM containing 5% FBS was used, and in this Example 1, DMEM containing 0.25% FBS was used. The cells were cultured under the conditions of 5% CO2 and 37 °C. After the culturing, 6×10 4NB cells were seeded in 6-well plates and cultured until 75% confluence. Subsequently, the medium was replaced with DMEM containing 0.25% FBS, and after 24 hours of culture, the medium was replaced with fresh DMEM containing 0.25% FBS.

[0125] (2)Experimental groups For the replaced NB cells, the following experimental groups were prepared using the extract of the flowers of Artemisia capillaris.

[0126] (Experimental groups) · Non-added group: The group without the addition of the extract and histamine · Control group: The group without the addition of the extract but with the addition of histamine · 0.125-added group: The group with the addition of the extract at a final concentration of 0.125% and the addition of histamine · 0.25-added group: The group with the addition of the extract at a final concentration of 0.25% and the addition of histamine · 0.5-added group: The group with the addition of the extract at a final concentration of 0.5% and the addition of histamine · 1.0-added group: The group with the addition of the extract at a final concentration of 1.0% and the addition of histamine (3)Confirmation of HYBID gene expression (RT-PCR) After preparing the experimental groups, they were further cultured in a CO2 incubator at 37°C for 3 hours. After the culture, in order to make the cells in an inflammatory state, for the experimental groups (except the non-added group), histamine was added at a final concentration of 10 μ mol / l. After the addition, the cells were cultured in a CO2 incubator at 37°C for 24 hours. In addition, the addition of histamine was carried out to enhance the expression of HYBID as in Reference Example 2. mRNA was purified from the cultured cells. The purification of mRNA was performed using QIAshreder and RNeasy Mini Kit (manufactured by QIAGEN). Using the purified mRNA as a template, reverse transcription reaction was carried out using PrimeScript RT master Mix (manufactured by Takara Bio Inc.) to synthesize cDNA. Subsequently, RT-PCR was performed using the primer pairs corresponding to each target factor shown in Table 5 below, and the change in expression level was analyzed by relative quantitative analysis. The RT-PCR was performed using TB Green Premix Ex Taq (manufactured by Takara Bio Inc.). In addition, the amplification result of RPS18 (ribosomal protein S18) was used as a reference for the analysis of relative quantity change.

[0127] [Table 5]

[0128] These results are shown in Table 6 below. Table 6 below shows the values when the values of the non-added group (the change in expression level shown by the relative quantification) are set to 1. The values are those obtained by rounding the third decimal place. In addition, in Table 6 below, The [mark] indicates a significant difference (p < 0.01) based on the comparison with the values of the control group in the Dunnett test, The [mark] indicates a significant difference (p < 0.001) based on the comparison with the values of the control group in the Dunnett test.

[0129] [Table 6]

[0130] As shown in Table 6 above, it can be seen that in NB cells in an inflammatory state, the expression of HYBID is inhibited by adding an extract of the flower of Artemisia capillaris.

[0131] [Example 2] The expression of miR-600 in NB cells and AD cells when an extract of the flower of Artemisia capillaris was added was studied. Specifically, regarding whether the addition of an extract of the flower of Artemisia capillaris would enhance the expression of miR-600, the RT-PCR method was used for the study. MicroRNA (miRNA) is a short-chain (20 bases to 25 bases) RNA (non-coding RNA) that does not produce proteins. The microRNA binds to the messenger RNA (mRNA) of a gene having the same sequence as itself, and inhibits the expression of the gene by degrading the mRNA or inhibiting translation into a protein. miR-600, one of the microRNAs, is an inhibitory substance for HYBID (Reference 1).

[0132] Reference 1: Sun, Junfeng et al. “LncRNA TUG1 promoted KIAA1199 expression via miR-600 to accelerate cell metastasis and epithelial-mesenchymal transition in colorectal cancer.” Journal of experimental & clinical cancer research: CR vol. 37, 1 106. 18 May. 2018, doi:10.1186 / s13046-018-0771-x (1) Cells 6×10 4NB cells or AD cells were seeded in 6-well plates and cultured until 75% confluence. After the culture, the medium was replaced with DMEM containing 0.25% FBS and cultured for 24 hours. Then, the medium was replaced with fresh DMEM containing 0.25% FBS.

[0133] (2)Experimental group For the NB cells or AD cells after the replacement, the following experimental groups were prepared using the extract of the flowers of Artemisia capillaris.

[0134] (Experimental group) · NB cell non-addition group: The group in which the extract was not added to NB cells · AD cell non-addition group: The group in which the extract was not added to NB cells · NB cell 1.0 addition group: The group in which the extract with a final concentration of 1.0% was added to NB cells · AD cell 1.0 addition group: The group in which the extract with a final concentration of 1.0% was added to NB cells (3)Study on the expression of miR-600 After preparing the experimental groups, they were further cultured in a CO2 incubator at 37°C for 6 hours. After the culture, total RNA was purified and extracted using miRNeasy Tissue / Cells Advanced Mini Kit (manufactured by QIAGEN, Cat.No / ID: 217684). After the purification and extraction, the expression level of miRNA (miR-600) was measured using real-time PCR. Complementary DNA (cDNA) was synthesized from the total RNA using Mir-X (trademark) miRNA First-Strand Synthesis Kit (manufactured by Clontech Laboratories, Inc. (Takara Bio Inc.), product code: 638313, Takara code: Z8313N). After the synthesis, RT-PCR was performed using Mir-X (trademark) miRNA qRT-PCR SYBR Kit (manufactured by Clontech Laboratories, Inc.), the primers listed in Table 7 below, and Thermal Cycler Dice Real Time System TP800 (manufactured by Clontech Laboratories, Inc. (Takara Bio Inc.)). Then, the miRNA expression between the experimental groups was compared by the Delta-delta-CT method.

[0135] [Table 7]

[0136] It should be noted that the thermal cycling conditions of the RT-PCR are as follows.

[0137] · Step 1: Perform 40 cycles of two-stage PCR with 30 seconds at 95°C, 5 seconds at 95°C, and 30 seconds at 60°C.

[0138] · Step 2: After Step 1, perform 1 cycle of dissociation steps with 15 seconds at 95°C, 30 seconds at 60°C, and 15 seconds at 95°C.

[0139] These results are shown in Table 8 below. Table 8 below shows the values when the group without NB cells (the change in expression level shown by this relative quantification) is set to 1. The values are those after rounding the third decimal place. Additionally, in Table 8 below, The * mark indicates a significant difference (p < 0.05) based on comparison with the value of the group without NB cells in the Student's t-test, and the † mark indicates a significant difference (p < 0.05) based on comparison with the value of the group without AD cells in the Student's t-test.

[0140] [Table 8]

[0141] As shown in Table 8 above, compared with the group without NB cells, the expression level of miR-600 decreased in the group without AD cells. Additionally, compared with the group without AD cells or NB cells, the expression level of miR-600 increased in the group with AD cells or NB cells, respectively. From these results, it can be seen that in AD cells and NB cells, the expression of miR-600 was enhanced by adding the extract of the flower of Artemisia capillaris.

[0142] [Example 3] For the composition containing the extract of the flower of Artemisia capillaris, perform human skin monitoring tests, etc.

[0143] (1) Preparation of lotion First, for use in the skin monitoring test, prepare a lotion containing the components shown in Table 9 below (hereinafter also referred to as "the lotion described in Table 9") and a lotion containing the components shown in Table 10 below (hereinafter also referred to as "the lotion described in Table 10"). As shown in Table 9 below, the lotion described in Table 9 contains 1% of the extract of the flower of Artemisia capillaris.

[0144] [Table 9]

[0145] [Table 10]

[0146] (2)Human skin monitoring test (topical skin agent) For the lotion prepared in Example 3(1) above, a human skin monitoring test was conducted. Specifically, the lotion described in Table 9 and the lotion described in Table 10 (placebo) were applied to the specified parts of the faces of normal human subjects (13 men and women with an average age of 43.6 years) between January 31, 2022 and March 4, 2022. The lotion described in Table 9 was applied to the specified parts of half of the subjects' faces at regular intervals, and the lotion described in Table 10 was applied to the specified parts of the other half of the subjects' faces at regular intervals. The regular interval was twice a day (morning and evening) for 4 weeks. The specified part was "the whole face".

[0147] Figure 2 A schematic diagram showing the outline of the skin monitoring test. As Figure 2 described, the relaxation of the skin was measured. Specifically, under the lower side of an adhesive tape with a circular opening at the center (adhesive tape for elastic probe of DermaLab TM Combo (manufactured by Cotex Technologies)) on the cheeks of the subjects, a staple (No. 11-1M (manufactured by MAX Co., Ltd.)) was hooked. As Figure 2 shown, the pasting position of the adhesive tape was set such that the intersection of the line drawn horizontally from under the nose of the face and the line drawn vertically from the outer corner of the eye overlapped with the lower part inside the circular circle of the adhesive tape.

[0148] Figure 2 The weights were weights (5 g to 20 g). The weights and the fixture were fixed with a rope, and the fixture side was hooked to the needle of the stapler. In addition, the weights were applied to each of the subjects in advance, and the weights used could move the cheeks by 1 mm to 2 mm under the action of gravity.

[0149] To confirm the facial conditions before and after loading the heavy object, left and right face photos were taken using VISIA™ Evolution (manufactured by Canfield Scientific). The photos were taken before the test and 4 weeks after the test. Using photo processing software (Photoshop), the images taken before and after loading the heavy object were overlapped. Then, using image analysis software (Digital Microscope VHX-5000 (manufactured by KEYENCE CORPORATION)), the moving distance of the adhesive tape and the length of the adhesive tape after loading the heavy object were measured before the test and 4 weeks after the test. Regarding the moving distance of the adhesive tape after loading the heavy object, an estimated conversion value was calculated from the length of the adhesive tape. The measurement results are shown in Table 11 and Table 12 below.

[0150] [Table 11]

[0151] Table 11 shows the results of measuring the moving distance. In Table 11, the value before the test (0 week) is represented as 1. In Table 11, The mark indicates a significant difference (p < 0.05) based on the comparison with the value at 0 week (lotion coating group in Table 9) in the Wilcoxon Rank-Sum Test. As shown in Table 11, the measured value 4 weeks after the test for the lotion described in Table 10 was 1.13, while the measured value 4 weeks after the test for the lotion described in Table 9 was 0.71. From these results, it can be seen that the lotion described in Table 9 containing the extract of the flower of Artemisia capillaris Thunb. can inhibit the relaxation of facial skin.

[0152] [Table 12]

[0153] Table 12 shows the results of measuring the length of the adhesive tape 4 weeks after the test. In Table 12, The mark indicates a significant difference (p < 0.05) based on the comparison with the value of the lotion coating group described in Table 10 (100) in the Wilcoxon Rank-Sum Test. As shown in Table 12, when the value of the lotion coating group described in Table 10 is set to 100, the value of the lotion coating group described in Table 9 is 71.20. From these results, it can be seen that the lotion described in Table 9 containing the extract of the flower of Artemisia capillaris Thunb. can inhibit the relaxation of facial skin.

[0154] (3) Human skin monitoring test 2 (skin topical agent) For the lotion prepared in Example 3(1) above, a human skin monitoring test was conducted. Specifically, the lotions described in Table 9 and the lotion described in Table 10 (placebo) were applied to specified parts of the faces of normal human subjects (13 men and women with an average age of 43.6 years), and the test was conducted between January 31, 2022 and March 4, 2022. The lotion described in Table 9 was applied to specified parts of one half of the subjects' faces at specified intervals, and the lotion described in Table 10 was applied to specified parts of the other half of the subjects' faces at specified intervals. The specified interval was twice a day (morning and evening) for 4 weeks. The specified part was "the entire face".

[0155] Before the application of the lotion (week 0) and 4 weeks after the application of the lotion, the size (overall size) and depth of the wrinkles on the eyelids were measured. The eyelids include the upper eyelids and the outer corners of the eyes. The measurement of whether there are wrinkles on the eyelids was performed using ANTERA3D (registered trademark, Galelius Medical Co., Ltd.).

[0156] The measurement was conducted as follows: The average value of the measurement results of each group of subjects before the application of the lotion was set as "100", and the measurement results of each group of subjects 4 weeks after the application of the lotion were calculated as relative values. In the measurement, the value before the application of the lotion (100) was obtained by measuring each person three times at one part, calculating the average value of the above three measurements, collecting and summing up the average values calculated for 13 persons, and setting the average value of these values as 100. The measurement result 4 weeks after the application of the lotion was the value of the change rate calculated compared with the value before the application of the lotion. These results are shown in Table 13 below.

[0157] [Table 13]

[0158] Table 13 shows the measurement results of whether there are wrinkles on the eyelids 4 weeks after the application of the lotion. In Table 13, The markers indicate significant differences (p < 0.05) based on the comparison with the value (100) of the lotion application group described in Table 10 in the Wilcoxon Rank-Sum Test. As shown in Table 13, when the value of the lotion application group described in Table 10 was set as 100, the value of the lotion application group described in Table 9 was 93.51. From these results, it can be seen that with the lotion described in Table 9 containing the extract of the flowers of Artemisia capillaris, the size of the wrinkles becomes smaller and the depth becomes shallower.

[0159] (4) Human skin monitoring test 3 (topical skin agent) For the lotion prepared in Example 3(1), a human skin monitoring test was conducted. Specifically, the lotion described in Table 9 and the lotion described in Table 10 (placebo) were applied to the specified parts of the faces of normal human subjects (13 men and women with an average age of 43.6 years), and the test was conducted between January 31, 2022 and March 4, 2022. The lotion described in Table 9 was applied to the specified parts of half of the subjects' faces at specified intervals, and the lotion described in Table 10 was applied to the specified parts of the other half of the subjects' faces at specified intervals. The specified interval was twice a day (morning and evening) for 4 weeks. The specified part was "the whole face".

[0160] Before the lotion application (week 0) and 4 weeks after the lotion application, the moisture distribution of the skin was measured. The measurement was performed using a Corneometer (CM825, manufactured by Courage+Khazaka). When the average value of the moisture distribution measurements of the 13 people in the lotion application group described in Table 9 or the lotion application group described in Table 10 at week 0 (before the application) was set to 100, the lotion application group described in Table 9 4 weeks after the application was 107.73, and the lotion application group described in Table 10 was 98.98. From these results, it can be seen that the lotion described in Table 9 containing the extract of the flower of Artemisia capillaris has the effect of maintaining a specified amount of skin moisture.

[0161] [Example 4] Regarding the expression of HYBID induced by histamine in NB cells when the extract of the flower of Artemisia capillaris or the extract of the flower of Lonicera japonica was added, a study was conducted using the RT-PCR method.

[0162] (1) NB cells NB cells were prepared. DMEM containing 5% FBS was used for pre-culture, and DMEM containing 0.25% FBS was used in this Example 4. The cells were cultured under the conditions of 5% CO2 and 37 °C. After the culture, 6×10 4 NB cells were seeded in a 6-well plate and cultured until 75% confluent. Then, the medium was replaced with DMEM containing 0.25% FBS. After culturing for 24 hours, the medium was replaced with fresh DMEM containing 0.25% FBS.

[0163] (2) Experimental groups For the NB cells after the replacement, the following experimental groups were prepared using the extract of the flower of Artemisia capillaris or the extract of Lonicera japonica. In addition, the following extracts were used for the extract of the flower of Artemisia capillaris and the extract of Lonicera japonica.

[0164] · Extract of the flower of Artemisia capillaris: Pharcolex Kawarayomogi B, manufactured by Ichimaru Pharcos Co., Ltd. · Extract of the flower of Lonicera japonica: Pharcolex Suikazura FB, manufactured by Ichimaru Natural Beauty Co., Ltd. (Experimental group) · Non-added group: The group without adding the extract and histamine · Control group: The group without adding the extract but adding histamine · 0.125 Suikazura FB added group: The group adding Pharcolex Suikazura FB at a final concentration of 0.125% and adding histamine · 0.25 Suikazura FB added group: The group adding Pharcolex Suikazura FB at a final concentration of 0.25% · 0.5 Suikazura FB added group: The group adding Pharcolex Suikazura FB at a final concentration of 0.5% and adding histamine · 1.0 Suikazura FB added group: The group adding Pharcolex Suikazura FB at a final concentration of 1.0% and adding histamine · 0.125 Kawarayomogi B added group: The group adding Pharcolex Kawarayomogi B at a final concentration of 0.125% and adding histamine · 0.25 Kawarayomogi B added group: The group adding Pharcolex Kawarayomogi B at a final concentration of 0.25% and adding histamine · 0.5 Kawarayomogi B added group: The group adding Pharcolex Kawarayomogi B at a final concentration of 0.5% and adding histamine · 1.0 Kawarayomogi B added group: The group adding Pharcolex Kawarayomogi B at a final concentration of 1.0% and adding histamine (3) Confirmation of gene expression of HYBID (RT-PCR) After preparing the experimental groups, further culture them in a CO2 incubator at 37°C for 3 hours. After the culture, in order to make the cells in an inflammatory state, for the experimental groups (except the non-added group), at a final concentration of 10 μHistamine was added in a manner of mol / l. After the addition, the cells were cultured at 37 °C in a CO2 incubator for 24 hours. In addition, the addition of histamine was carried out to enhance the expression of HYBID. mRNA was purified from the cultured cells. The purification of the mRNA was performed using QIAshreder and RNeasy Mini Kit (manufactured by QIAGEN). Using the purified mRNA as a template, reverse transcription reaction was carried out using PrimeScript RT master Mix (manufactured by Takara Bio Inc.) to synthesize cDNA. Then, using the primer pairs corresponding to each target factor shown in Table 14 below, RT-PCR was performed, and the change in expression level was analyzed by relative quantification. The RT-PCR was carried out using TB Green Premix Ex Taq (manufactured by Takara Bio Inc.). In addition, the amplification result of RPS18 (ribosomal protein S18) was used as a reference for the analysis of relative quantity change.

[0165] [Table 14]

[0166] These results are shown in Table 15 below. Table 15 below shows the values when the value of the non-added group (the change in expression level shown by the relative quantification) is set to 1. In addition, in Table 15 below, The mark indicates the significant difference (p < 0.05) based on the comparison with the value of the non-added group in the Dunnett test, The mark indicates the significant difference (p < 0.001) based on the comparison with the value of the non-added group in the Dunnett test.

[0167] [Table 15]

[0168] As shown in Table 15 above, it was found that in NB cells in an inflammatory state, the expression of HYBID was inhibited by the addition of the extract of the flower of Artemisia capillaris or the addition of the extract of the flower of Lonicera japonica.

[0169] [Example 5] Regarding the expression of HYBID induced by IL-1 in NB cells when the extract of the flower of Artemisia capillaris or the extract of the flower of Lonicera japonica was added, it was studied using the RT-PCR method. β The induction of HYBID expression was studied using the RT-PCR method.

[0170] (1)NB cells NB cells were prepared. DMEM containing 5% FBS was used for pre-culture, and DMEM containing 0.25% FBS was used in Example 5. The cells were cultured under the conditions of 5% CO2 and 37 °C. After the culture, 6×104 NB cells were seeded in 6-well plates and cultured until they reached 75% confluence. After that, the medium was replaced with DMEM containing 0.25% FBS. After 24 hours of culture, the medium was replaced with fresh DMEM containing 0.25% FBS.

[0171] (2)Experimental groups For the replaced NB cells, the following experimental groups were prepared using the extract of the flowers of Artemisia capillaris or the extract of Lonicera japonica. In addition, the extracts of the flowers of Artemisia capillaris and the extract of Lonicera japonica were used as follows.

[0172] · Extract of the flowers of Artemisia capillaris: Pharcolex Kawarayomogi B, manufactured by Ichimaru Pharcos Co., Ltd. · Extract of the flowers of Lonicera japonica: Pharcolex Suikazura FB, manufactured by Ichimaru Pharcos Co., Ltd. · Extract of the leaves of Lonicera japonica: Pharcolex Suikazura SB, manufactured by Ichimaru Pharcos Co., Ltd. (Experimental groups) · Unadded group: The extract and IL-1 were not added. β group · Control group: The extract was not added, but IL-1 was added. β group · 1.0 Suikazura FB added group: Pharcolex Suikazura FB was added to a final concentration of 1.0%, and IL-1 was added. β group · 1.0 Suikazura SB added group: Pharcolex Suikazura SB was added to a final concentration of 1.0%, and IL-1 was added. β group · 1.0 Kawarayomogi B added group: Pharcolex Kawarayomogi B was added to a final concentration of 1.0%, and IL-1 was added. β group (3)Confirmation of HYBID gene expression (RT-PCR) After preparing the experimental groups, they were further cultured in a CO2 incubator at 37°C for 3 hours. After the culture, in order to make the cells in an inflammatory state, for the experimental groups (except the unadded group), IL-1 was added to a final concentration of 50 ng / ml. β . After this addition, they were cultured in a CO2 incubator at 37°C for 24 hours. In addition, IL-1 βThe addition was carried out to enhance the expression of HYBID. mRNA was purified from the cultured cells. The purification of the mRNA was performed using QIAshreder and RNeasy Mini Kit (manufactured by QIAGEN). Using the purified mRNA as a template, a reverse transcription reaction was carried out using PrimeScript RT master Mix (manufactured by Takara Bio Inc.) to synthesize cDNA. Subsequently, RT-PCR was performed using the primer pairs corresponding to each target factor shown in Table 16 below, and the change in expression level was analyzed by relative quantification. The RT-PCR was carried out using TB Green Premix Ex Taq (manufactured by Takara Bio Inc.). In addition, the amplification result of RPS18 (ribosomal protein S18) was used as a reference for the analysis of relative quantity change.

[0173] [Table 16]

[0174] These results are shown in Table 17 below. Table 17 below shows the values when the value of the non-addition group (the change in expression level shown by the relative quantification) is set to 1. In addition, in Table 17 below, The mark indicates a significant difference (p < 0.01) based on the comparison with the value of the non-addition group in the Dunnett test, The mark indicates a significant difference (p < 0.001) based on the comparison with the value of the non-addition group in the Dunnett test.

[0175] [Table 17]

[0176] As shown in Table 17, it was found that in NB cells in an inflammatory state, the expression of HYBID was inhibited by the addition of the extract of the flower of Artemisia capillaris, the addition of the extract of the flower of Lonicera japonica, or the addition of the extract of the leaf of Lonicera japonica.

[0177] [Example 6] It was investigated whether the maintenance of the structure of the water-soluble gel differed depending on the molecular size of hyaluronic acid using a collagen gel.

[0178] (1) Gel Using a collagen gel culturing kit (Collagen Gel Culturing Kit, manufactured by Nitta Gelatin Inc., 638-00781), a gel was prepared according to the instructions included in the kit. For clarification, experimental examples using a collagen gel can be referred to the following Reference 2.

[0179] Reference 2: Takahashi, Yu et al. “Drug cytotoxicity screening using human intestinal organoids propagated with extensive cost-reduction strategies.” Scientific reports vol. 13,1 5407. 3 Apr. 2023, doi:10.1038 / s41598-023-32438-2 (2)Hyaluronic acid Prepare the following HA.

[0180] · H2: HA with a molecular weight of 1200 kDa to 1600 kDa · U2: HA with a molecular weight of 5 kDa to 10 kDa (3)Experimental groups For the gel, add the HA so that the final concentration becomes 50 μ g / ml to prepare the following experimental groups.

[0181] (Experimental groups) · H2 addition group: The group to which H2 was added · U2 addition group: The group to which U2 was added (4)Measurement of the height of the gel After preparing the experimental groups, leave them standing at 37 °C for 30 minutes. After the standing, visually confirm the state of the gel in the experimental groups and measure the height of the gel. Show these results in Figure 3 .

[0182] Figure 3 are photos showing the state of each gel. If the height value of the gel in the H2 addition group is set to 100, the height value of the gel in the U2 addition group is 90. As Figure 3 shown, compared with the H2 addition group, in the U2 addition group, the height of the gel decreased by the extent of the arrow. From these results, it can be seen that the U2 addition group has a lower ability to retain water molecules compared with the H2 addition group and cannot maintain the structure of the water-soluble gel. In addition, it is suggested that the molecular weight of HA is important for maintaining the structure of the gel.

[0183] [Example 7] Regarding the expression of HYBID in NB cells when an extract of the flowers of Artemisia capillaris is added, it was studied using the RT-PCR method.

[0184] (1)NB cells Prepare NB cells. For pre-culture, use DMEM containing 10% FBS, and in Example 7 herein, use DMEM containing 0.25% FBS. Culture under the conditions of 5% CO2 and 37 °C. After the culture, inoculate 6×10 4 NB cells into a 6-well plate and culture until 75% confluent. Then, replace with DMEM containing 0.25% FBS. After culturing for 24 hours, replace with fresh DMEM containing 0.25% FBS.

[0185] (2)Experimental groups For the NB cells after the replacement, use the extract of the flowers of Artemisia capillaris Thunb. to prepare the following experimental groups.

[0186] (Experimental groups) · Non-added group: The group without adding the extract · 1.0 added group: The group added with the extract of the flowers of Artemisia capillaris Thunb. at a final concentration of 1.0 μ mol / l (3)Confirmation of the gene expression of HYBID (RT-PCR) After preparing the experimental groups, further culture in a CO2 incubator at 37 °C for 3 hours. After the culture, replace with fresh DMEM containing 0.25% FBS. After this replacement, culture in a CO2 incubator at 37 °C for 24 hours. Purify mRNA from the cultured cells. The purification of the mRNA uses QIAshreder and RNeasy Mini Kit (manufactured by QIAGEN). Using the purified mRNA as a template, perform reverse transcription reaction using PrimeScript RT master Mix (manufactured by Takara Bio Inc.) to synthesize cDNA. Then, perform RT-PCR using the primer pairs corresponding to each target factor shown in Table 18 below, and analyze the change in the expression level by relative quantitative analysis. The RT-PCR is performed using TB Green Premix Ex Taq (manufactured by Takara Bio Inc.). In addition, the amplification result of RPS18 (ribosomal protein S18) is used as a reference for the analysis of the relative quantity change.

[0187] [Table 18]

[0188] Show these results in Table 19 below. Table 19 below shows the values when the value of the non-added group (the change in the expression level shown by the relative quantification) is set to 1. The values are the values after rounding the third decimal place. In addition, in Table 19 below, The markers indicate significant differences (p < 0.001) based on comparison with the values of the non-added group in the Student's t-test.

[0189] [Table 19]

[0190] As shown in Table 19, it can be seen that in NB cells, the expression of HYBID is inhibited by the addition of the extract from the flowers of Artemisia capillaris.

[0191] [Example 8] The expression of HYBID in NB cells or AD cells when the extract from the flowers of Artemisia capillaris was added was studied using the RT-PCR method.

[0192] (1)Cells NB cells and AD cells were prepared. For pre-culture, DMEM containing 10% FBS was used, and in this Example 8, DMEM containing 0.25% FBS was used. The cells were cultured under the conditions of 5% CO2 and 37 °C. After the culture, 6×10 4 NB cells or AD cells were seeded in a 6-well plate and cultured until 75% confluent. After the culture, the medium was replaced with DMEM containing 0.25% FBS and cultured for 24 hours. Then, the medium was replaced with fresh DMEM containing 0.25% FBS.

[0193] (2)Experimental groups For the NB cells or AD cells after the replacement, the following experimental groups were prepared using the extract from the flowers of Artemisia capillaris.

[0194] (Experimental groups) · NB non-added group: The group of NB cells without the addition of the extract · AD non-added group: The group of AD cells without the addition of the extract · AD 1.0 added group: For AD cells, the extract from the flowers of Artemisia capillaris was added at a final concentration of 1.0 μ mol / l (3)Confirmation of the gene expression of HYBID (RT-PCR) After preparing the experimental group, it was further cultured in a CO2 incubator at 37 °C for 3 hours. After the culturing, it was replaced with fresh DMEM containing 0.25% FBS. After this replacement, it was cultured in a CO2 incubator at 37 °C for 24 hours. mRNA was purified from the cultured cells. The purification of the mRNA was performed using QIAshreder and RNeasy Mini Kit (manufactured by QIAGEN). Using the purified mRNA as a template, a reverse transcription reaction was performed using PrimeScript RT master Mix (manufactured by Takara Bio Inc.) to synthesize cDNA. Thereafter, RT-PCR was performed using the primer pairs corresponding to each target factor shown in Table 20 below, and the change in the expression level was analyzed by relative quantification. The RT-PCR was performed using TB Green Premix Ex Taq (manufactured by Takara Bio Inc.). In addition, the amplification result of RPS18 (ribosomal protein S18) was used as a reference for the analysis of the relative quantity change.

[0195] [Table 20]

[0196] These results are shown in Table 21 below. Table 21 below shows the values when the value of the NB non-added group (the change in the expression level shown by the relative quantification) is set to 1. The values are the values after rounding off the second decimal place. In addition, in Table 21 below, The mark indicates a significant difference (p < 0.001) based on the comparison with the value of the AD non-added group in the Dunnett test.

[0197] [Table 21]

[0198] As shown in Table 21 above, it was found that in AD cells, the expression of HYBID was inhibited by the addition of the extract of Artemisia capillaris flowers. In addition, it was found that in AD cells, the addition of the extract of Artemisia capillaris flowers resulted in an expression level of HYBID similar to that of NB cells.

[0199] [Example 9] The distribution of histamine in NB cells when the extract of Artemisia capillaris flowers was added was investigated.

[0200] (1)NB cells NB cells were prepared. For pre-culturing, DMEM containing 10% FBS was used, and in Example 9, DMEM containing 0.25% FBS was used. The cells were cultured under the conditions of 5% CO2 and 37 °C. After the culturing, 6×10 4NB cells were seeded in 6-well plates and cultured until they reached 75% confluence. Then, the medium was replaced with DMEM containing 0.25% FBS. After 24 hours of culture, the medium was replaced with fresh DMEM containing 0.25% FBS.

[0201] (2)Experimental groups For the NB cells after the above replacement, the following experimental groups were prepared using the extract of the flowers of Artemisia capillaris.

[0202] (Experimental groups) · Non-added group: The group without the addition of the extract and histamine · Control group: The group without the addition of the extract but with the addition of histamine · 0.5-added group: The group with the extract added to a final concentration of 0.5% · 1.0-added group: The group with the extract added to a final concentration of 1.0% (3)Study on the distribution of hyaluronic acid (HPLC) After preparing the above experimental groups, they were further cultured in a CO2 incubator at 37°C for 3 hours. After the culture, to make the cells in an inflammatory state, for the experimental groups (except the non-added group), histamine was added to a final concentration of 10 μ mol / l. After this addition, they were cultured in a CO2 incubator at 37°C for 2 hours. In addition, the addition of histamine was carried out to enhance the expression of HYBID. For the cultured cells, fluoresceinamine-labeled sodium hyaluronate (H2, average molecular weight 1.2 million - 1.6 million, FAHA-H2, Iwai Chemical Co., Ltd.) was added. After this addition, they were cultured in a CO2 incubator at 37°C for 48 hours. After the culture, the culture medium was collected. In addition, for experiments using the above FAHA, reference can be made to the following Reference 3 ( Figure 4 ), the following Reference 4 ( Figure 1 , supplementary Figure 1 ). Using the above culture medium, the distribution of hyaluronic acid was studied by HPLC. The conditions of HPLC are as follows. These results are shown in Figure 4 .

[0203] Reference 3: Tobisawa, Yuki et al. “The cell surface hyaluronidase TMEM2 is essential for systemic hyaluronan catabolism and turnover.” The Journal of biological chemistry vol. 297, 5(2021): 101281. doi:10.1016 / j.jbc.2021.101281 Reference 4: Yoshida, Hiroyuki et al. “KIAA1199, a deafness gene of unknown function, is a new hyaluronan binding protein involved in hyaluronan depolymerization.” Proceedings of the National Academy of Sciences of the United States of America vol. 110,14(2013): 5612-7. doi:10.1073 / pnas.1215432110 Using the said culture medium, the distribution of hyaluronic acid was studied by HPLC under the following measurement conditions. The results are shown in Figure 4 and Table 22 below.

[0204] (Conditions for HPLC) Apparatus: SHIMADZU LC-20A Prominence (manufactured by Shimadzu Corporation) Column: TSKgel G5000 PWXL (manufactured by Tosoh Corporation) Eluent: 0.2 mol / L NaCl Flow rate: 0.5 mL / min Detector wavelength: Excitation wavelength 490 nm, Fluorescence wavelength 525 nm [Table 22]

[0205] Figure 4 is a graph showing the results of HPLC analysis. In Figure 4 the vertical axis represents the fluorescence intensity and the horizontal axis represents the time (minutes). Table 22 shows Figure 4The fluorescence intensity values at the peaks of each group shown. Table 22 shows the values when the value of the non - added group (fluorescence intensity) is set to 100. The values are those obtained by rounding the fourth decimal place. As Figure 4 shown in Table 22, compared with the non - added group, in the groups added with histamine (control group, 0.5 - added group, 1.0 - added group), the fluorescence intensity at the peak is lower. These results suggest that through histamine, HYBID is enhanced, and through HYBID, a certain amount of hyaluronic acid is decomposed into fragments.

[0206] [Example 10] It was confirmed that caffeic acid derivatives inhibit the production of HYBID.

[0207] (1) Gene expression level test of Component Fraction 1 and HYBID The components contained in the extract of the flower of Artemisia capillaris were analyzed. Specifically, according to the Figure 5 shown protocol, the extract of the capitulum of Artemisia capillaris (Artemisia capillaris flower extract: ACFE) was fractionated. The raw materials used in this fractionation were purchased from Rippon Tenkaido Co., Ltd. Regarding this fractionation, specifically, water was added to the extract (ACFE) obtained by extracting the capitulum of Artemisia capillaris with 30% butylene glycol (BG). After making a 15% BG aqueous solution through this addition, it was roughly fractionated using a column filled with DIAION HP - 20 (manufactured by Mitsubishi Chemical Corporation) to obtain 4 fractions, namely Fr.1 - Fr.4. Then, for these 4 fractions, the inhibitory effect on HYBID expression was studied. In this study, human NHDF (normal human dermal fibroblasts) cells were used as cells, and these 4 fractions were used instead of the extract. Except for this, the gene expression of HYBID was confirmed by the same method as in Example 1. These results are shown in Figure 6 .

[0208] Figure 6 is a graph showing the gene expression level of HYBID when histamine is added. In Figure 6 , the vertical axis represents the relative value of the expression level of the HYBID gene when the non - added group (NT) is set to 100, and the horizontal axis represents the experimental groups. As Figure 6 shown, compared with the control group, in the fraction of Fr.2, the expression level of the HYBID gene decreased. From these results, it can be known that the fraction of Fr.2 contains components that inhibit the production of HYBID.

[0209] (2) Gene expression level test of Component Fraction 2 and HYBID For the components contained in Fr.2, preparative HPLC was further used for sorting under the following measurement conditions. Through the above sorting, four fractions, namely Fr.2-1, Fr.2-2, Fr.2-3, and Fr.2-4, were obtained. Subsequently, for the above four fractions, the inhibitory effect on HYBID expression was studied. In the above study, human NHDF (normal human dermal fibroblasts) cells were used as cells, and the above four fractions were used instead of the extract. Except for this, the gene expression of HYBID was confirmed by the same method as in Example 1. The results are shown in Figure 7 and Figure 8 .

[0210] Figure 7 is a graph showing the gene expression level of HYBID when histamine is added. In Figure 7 , the vertical axis represents the relative value of the expression level of the HYBID gene with the non-added group (NT) set to 100, and the horizontal axis represents the experimental group. As Figure 7 shows, compared with the control group, the expression level of the HYBID gene decreased in the fractions of Fr.2-2 and Fr.2-3. From these results, it can be known that the fractions of Fr.2-2 and Fr.2-3 contain components that inhibit the production of HYBID.

[0211] Figure 8 is a graph showing the gene expression level of HYBID when histamine is added. In Figure 8 , the vertical axis represents the relative value of the expression level of the HYBID gene with the non-added group (NT) set to 100, and the horizontal axis represents the experimental group. As Figure 8 shows, in the fraction of Fr.2-2, the expression level of the HYBID gene decreased depending on the concentration. From these results, it can be known that the fraction of Fr.2-2 contains components that inhibit the production of HYBID, and the production of HYBID is inhibited depending on the concentration of the fraction of Fr.2-2.

[0212] (3) Structural analysis For the structural analysis of Fr.2-2 and Fr.2-3 containing a single component, NMR analysis was performed using the following analysis conditions. The results of the above analysis identified that Fr.2-2 (Compound 1) is 1-Caffeoyl-3-hydroxybutane represented by Chemical Formula (2), and Fr.2-3 (Compound 2) is 3-Caffeoyl-1-hydroxybutane represented by Chemical Formula (3). The measured values of NMR are as follows. In addition, the above 1-Caffeoyl-3-hydroxybutane was obtained as a yellow solid, and the above 3-Caffeoyl-1-hydroxybutane was obtained as a yellow oil.

[0213] (Conditions for NMR analysis) ·Optical rotation Apparatus: JASCO P-1020 polarimeter (manufactured by JASCO Corporation) Solvent: MeOH ·UV spectrum Apparatus: Shimadzu UV-3100 spectrometer (manufactured by Shimadzu Corporation) Solvent: MeOH ·High-resolution electrospray ionization mass spectrometry (HRESIMS) Apparatus: Shimadzu LCMS-IT-TOF spectrometer (manufactured by Shimadzu Corporation) Ion mode: negative ion mode ·Nuclear magnetic resonance spectroscopy (NMR) Apparatus: JEOL JNM-ECA-500 spectrometer (manufactured by JEOL Ltd.) Solvent: MeOH-d4 (Measured values of nuclear magnetic resonance spectroscopy) 1-caffeoyl-3-hydroxybutane (Compound 1): α D +4.0° (c = 0.1 g / dL, MeOH). UV λ MeOHmax nm (logε): 218 (4.11), 243 (3.97), 329 (4.20). calcd. for C 13 H 1５ O5251.0925(M-H), found 251.0911. 1 H NMR (500 MHz, MeOH-d4): δ =7.53 (d, J =15.8 Hz, 1H),7.03 (d, J =1.9 Hz, 1H), 6.94 (dd, J =8.0, 1.9 Hz, 1H), 6.77 (d, J =8.0 Hz, 1H),6.25 (d, J =15.8 Hz, 1H), 4.27 (br t, J =6.5 Hz, 2H), 3.90 (dqd, J ​=8.0, 6.3, 4.6 Hz, 1H), 1.83 (dtd, 14.2, 7.1, 4.6 Hz, 1H), 1.77 (ddt, 14.2, 8.0, 6.0 Hz,1H), 1.21 (d, J =6.3 Hz, 3H). 13 C NMR (125 MHz, MeOH-d4): δ =169.4, 149.6, 146.9(2C), 127.7, 122.9, 116.5, 115.10, 115.07, 65.5, 62.7, 39.0, 23.8. 3-Caffeoyl-1-hydroxybutane (Compound 2): α D +3.6° (c = 0.1 g / dL, MeOH). UV λ MeOHmax nm (logε): 217 (3.99), 234 (3.80), 243 (3.80), 300 (3.89), 328 (3.98). calcd. for C 13 H 15 O5251.0925 (M-H), found 251.0911. 1 H NMR (500 MHz, MeOH-d4): δ =7.52 (d, J =16.0 Hz, 1H), 7.03 (d, J =1.9 Hz, 1H), 6.94 (dd, J =8.2, 1.9 Hz, 1H), 6.77 (d, J =8.2 Hz, 1H), 6.24 (d, J =16.0 Hz, 1H), 5.12 (dqd, J =8.2, 6.3, 4.9 Hz,1H), 3.62 (br t, J =6.6 Hz, 2H), 1.89 (ddt, 14.0, 8.2, 6.6 Hz, 1H), 1.81 (dtd,14.0, 6.6, 4.9 Hz, 1H), 1.30 (d, J =6.3 Hz, 3H). 13 C NMR (125 MHz, MeOH-d4): δ ​=169.0, 149.6, 146.8, 146.7, 127.7, 122.9, 116.5, 115.5, 115.1, 69.5, 59.3, 39.9, 20.6. (4) HYBID gene expression assay based on the identified compounds Furthermore, for the separated Compound 1 and Compound 2, the HYBID expression inhibitory effect was studied. In this study, human NHDF (normal human dermal fibroblasts) cells were used, and Compound 1, Compound 2, and Fr.2 fraction were used instead of the extract. Except for this, the gene expression of HYBID was confirmed in the same manner as in Example 1. These results are shown in Figure 9 .

[0214] Figure 9 is a graph showing the gene expression level of HYBID when histamine is added. In Figure 9 , the vertical axis represents the relative value of the gene expression level of HYBID with the non-added group (NT) set to 100, and the horizontal axis represents the experimental groups. As Figure 9 shown, compared with the control group, the gene expression level of HYBID decreased in Compound 1, Compound 2, and Fr.2 fraction. From these results, it can be seen that in NHDF cells in the inflammatory state, Compound 1 and Compound 2 are compounds that inhibit the production of HYBID.

[0215] (5) HYBID gene expression assay For the extract (ACFE) of the capitulum of Artemisia capillaris, Fr.2 fraction, and the separated Compound 1, the HYBID expression inhibitory effect was studied. Specifically, human NHDF (normal human dermal fibroblasts) cells were used, and ACFE, Fr.2 fraction, and Compound 1 were used instead of the extract. Except for this, the gene expression of HYBID was confirmed in the same manner as in Example 7. These results are shown in Figure 10 .

[0216] Figure 10 is a graph showing the gene expression level of HYBID. In Figure 10 , the vertical axis represents the relative value of the gene expression level of HYBID with the control group (extract non-added group, control) set to 100, and the horizontal axis represents the experimental groups. As Figure 10 shown, compared with the control group, the gene expression level of HYBID decreased in ACFE, Fr.2 fraction, and Compound 1. From these results, it can be seen that ACFE, Fr.2 fraction, and Compound 1 inhibit the production of HYBID.

[0217] [Example 11] It was confirmed that the Artemisia capillaris Thunb. extract inhibits the production of HYBID.

[0218] (1)Experiment on the expression level of miR-486p-5p in NB cells based on the extract of the flowers of Artemisia capillaris Thunb. As an upstream microRNA that directly targets the 3'-UTR region of HYBID, miR-486p-5p is known (Reference 5). Therefore, it was investigated whether the extract of the flowers of Artemisia capillaris Thunb. inhibits the expression of miR-486p-5p. Specifically, 6×10 4 NB cells were seeded in a 6-well plate and cultured until 75% confluence. After the culture, it was replaced with DMEM containing 0.25% FBS and cultured for 24 hours. Then, it was replaced with fresh DMEM containing 0.25% FBS. After the replacement, it was cultured for 6 hours in the presence or absence of AFCF. After the culture, to measure the expression of miR-486-5p, total RNA was purified and extracted using the miRNeasy Tissue / Cells Advanced Mini Kit (manufactured by QIAGEN). After the purification and extraction, the expression level of miRNA (miR-486p-5p) was measured using real-time PCR. Complementary DNA (cDNA) was synthesized from the total RNA using the Mir-X (trademark) miRNA First-Strand Synthesis Kit (manufactured by Clontech Laboratories, Inc. (Takara Bio Inc.), product code: 638313, Takara code: Z8313N). After the synthesis, RT-PCR was performed using the Mir-X (registered trademark) miRNA qRT-PCR SYBR Kit (manufactured by Clontech Laboratories, Inc.), the primers listed in Table 23 below, and the Thermal Cycler Dice Real Time System TP800 (manufactured by Clontech Laboratories, Inc. (Takara Bio Inc.)). Then, the miRNA expression between the experimental groups was compared by the Delta-delta-CT method. These results are shown in Figure 11 .

[0219] Reference 5: Jiao, Xuehua et al. “KIAA1199, a Target of MicoRNA-486-5p, Promotes Papillary Thyroid Cancer Invasion by Influencing Epithelial-Mesenchymal Transition (EMT).” Medical science monitor : international medical journal of experimental and clinical research vol. 25 6788-6796. 10 Sep. 2019, doi:10.12659 / MSM.918682 [Table 23]

[0220] Figure 11 is a graph showing the expression level of miR-486-5p. In Figure 11 , the vertical axis represents the relative value of the expression level of miR-486p-5p when the control group (the group without extract added, control) is set to 100, and the horizontal axis represents the experimental groups. As Figure 11 shown, compared with the control group, the expression level of miR-486p-5p increased in the ACFE-added group. From these results, it can be seen that in NB cells, ACFE increases the expression level of miR-486-5p and inhibits the production of HYBID.

[0221] (2) Experiment on the expression level of miR-486p-5p in AD cells based on the extract from the flowers of Artemisia capillaris It was studied whether the extract from the flowers of Artemisia capillaris also inhibits the expression of miR-486p-5p in cells other than NHDF cells. Specifically, except for using AD cells instead of NB cells, the same method as in Example 11(1) was used. These results are shown in Figure 12 .

[0222] Figure 12 is a graph showing the expression level of miR-486-5p. In Figure 12 , the vertical axis represents the relative value of the expression level of miR-486p-5p when the control group (the group without extract added, control) is set to 100, and the horizontal axis represents the experimental groups. As Figure 12 shown, compared with the control group, the expression level of miR-486p-5p increased in the ACFE-added group. From these results, it can be seen that in AD cells, ACFE increases the expression level of miR-486-5p and inhibits the production of HYBID.

[0223] The present invention has been described with reference to the embodiments and examples, but the present invention is not limited to the above embodiments and examples. The configuration and details of the present invention can be variously modified within the scope of the present invention that can be understood by those skilled in the art.

[0224] This application claims priority based on Japanese Patent Application No. 2023-214475 filed on December 20, 2023, and Japanese Patent Application No. 2024-016827 filed on February 7, 2024, and incorporates all of their disclosure herein.

[0225] <Supplementary Note> Part or all of the above-described embodiments and examples can be described as in the following supplementary notes, but are not limited to the following.

[0226] <Structural Stabilizer of Hyaluronic Acid> (Supplementary Note 1) A reagent for maintaining the structure of hyaluronic acid, comprising a compound represented by the following formula (1) or a salt thereof, wherein the molecular weight of the hyaluronic acid is 600 kDa to 2000 kDa, [Chemical Formula 1]

[0227] In the formula (1), R 1 and R 2 may be the same or different and are each independently a hydrogen atom, a hydroxyl group, or a linear or branched alkoxy group having 1 to 5 carbon atoms, R 3 is a linear or branched hydroxyalkyl group having 1 to 7 carbon atoms, a linear or branched alkyl group having 1 to 7 carbon atoms, a cyclic alkyl group having 3 to 10 carbon atoms which may be substituted, or a hydrogen atom.

[0228] (Supplementary Note 2) The reagent according to Supplementary Note 1, wherein the R 1 is a hydrogen atom or a hydroxyl group.

[0229] (Supplementary Note 3) The reagent according to Supplementary Note 1 or 2, wherein the R 2 is a hydrogen atom, a hydroxyl group, or a methoxy group.

[0230] (Supplementary Note 4) The reagent according to any one of Supplementary Notes 1 to 3, wherein the R 3It is a straight-chain or branched-chain hydroxyalkyl group having 1 to 5 carbon atoms, a straight-chain or branched-chain alkyl group having 1 to 5 carbon atoms, or a cyclic alkyl group having 3 to 10 carbon atoms which may be substituted.

[0231] (Appendix 5) The reagent according to any one of Appendices 1 to 4, wherein said R 1 is a hydroxyl group, said R 2 is a hydroxyl group, said R 3 is a straight-chain or branched-chain hydroxyalkyl group having 1 to 7 carbon atoms, a straight-chain or branched-chain alkyl group having 1 to 7 carbon atoms, or a cyclic alkyl group having 3 to 10 carbon atoms which may be substituted.

[0232] (Appendix 6) The reagent according to any one of Appendices 1 to 5, wherein the compound or its salt represented by the formula (1) is a compound or its salt represented by the following formula (2) or formula (3).

[0233] [Chemical formula 2]

[0234] [Chemical formula 3]

[0235] (Appendix 7) The reagent according to any one of Appendices 1 to 6, wherein the molecular weight of the hyaluronic acid is 1200 kDa to 1600 kDa.

[0236] (Appendix 8) The reagent according to any one of Appendices 1 to 7, wherein, as the compound or its salt represented by the formula (1), it contains an extract of Artemisia capillaris Thunb. and / or an extract of Lonicera japonica Thunb.

[0237] <Expression inhibitor of HYBID gene> (Appendix 9) A reagent which is a reagent for inhibiting the expression of the HYBID gene, comprising a compound or its salt represented by the following formula (1), [Chemical formula 1]

[0238] In the formula (1), R 1 and R 2 may be the same or different and are each independently a hydrogen atom, a hydroxyl group, or a straight-chain or branched-chain alkoxy group having 1 to 5 carbon atoms, R3 is a linear or branched hydroxyalkyl group having 1 to 7 carbon atoms, a linear or branched alkyl group having 1 to 7 carbon atoms, a cyclic alkyl group having 3 to 10 carbon atoms which may be substituted, or a hydrogen atom.

[0239] (Supplementary Note 10) For the reagent according to Supplementary Note 9, wherein the R 1 is a hydrogen atom or a hydroxyl group.

[0240] (Supplementary Note 11) For the reagent according to Supplementary Note 9 or 10, wherein the R 2 is a hydrogen atom, a hydroxyl group or a methoxy group.

[0241] (Supplementary Note 12) For the reagent according to any one of Supplementary Notes 9 to 11, wherein the R 3 is a linear or branched hydroxyalkyl group having 1 to 5 carbon atoms, a linear or branched alkyl group having 1 to 5 carbon atoms, or a cyclic alkyl group having 3 to 10 carbon atoms which may be substituted.

[0242] (Supplementary Note 13) For the reagent according to any one of Supplementary Notes 9 to 12, wherein the R 1 is a hydroxyl group, the R 2 is a hydroxyl group, the R 3 is a linear or branched hydroxyalkyl group having 1 to 7 carbon atoms, a linear or branched alkyl group having 1 to 7 carbon atoms, or a cyclic alkyl group having 3 to 10 carbon atoms which may be substituted.

[0243] (Supplementary Note 14) For the reagent according to any one of Supplementary Notes 9 to 13, wherein the compound represented by the formula (1) or a salt thereof is a compound represented by the following formula (2) or formula (3) or a salt thereof.

[0244] [Chemical Formula 2]

[0245] [Chemical Formula 3]

[0246] (Supplementary Note 15) For the reagent according to any one of Supplementary Notes 9 to 14, wherein, as the compound represented by the formula (1) or a salt thereof, it contains an extract of Artemisia capillaris and / or an extract of Lonicera japonica.

[0247] <Method for Maintaining the Structure of Hyaluronic Acid> (Supplementary Note 16) A method for maintaining the structure of hyaluronic acid, which uses the reagent described in any one of Supplementary Notes 1 to 8.

[0248] (Supplementary Note 17) The maintenance method according to Supplementary Note 16, wherein the maintenance method includes an administration step of administering the reagent to a subject.

[0249] (Supplementary Note 18) According to the maintenance method described in Supplementary Note 16 or 17, wherein in vitro or in vivo the reagent is used.

[0250] <Method for inhibiting the expression of HYBID gene> (Supplementary Note 19) A method for inhibiting the expression of HYBID gene, which uses the reagent described in any one of Supplementary Notes 9 to 15.

[0251] (Supplementary Note 20) According to the expression inhibition method described in Supplementary Note 19, wherein the expression inhibition method includes an administration step of administering the reagent to a subject.

[0252] (Supplementary Note 21) According to the expression inhibition method described in Supplementary Note 19 or 20, wherein the reagent is used in vitro or in vivo.

[0253] <Use> (Supplementary Note 22) Use of the compound represented by the formula (1) described in any one of Supplementary Notes 1 to 8 or a salt thereof for maintaining the structure of hyaluronic acid.

[0254] (Supplementary Note 23) Use of the compound represented by the formula (1) described in any one of Supplementary Notes 9 to 15 or a salt thereof for inhibiting the expression of HYBID gene.

[0255] <Compound> (Supplementary Note 24) A compound or a salt thereof, which is represented by the following formula (2) or formula (3).

[0256] [Chemical formula 2]

[0257] [Chemical formula 3]

[0258] Industrial applicability As described above, according to the present invention, it is possible to provide a reagent for maintaining the structure of hyaluronic acid and the like. Therefore, it can be said that the present invention is extremely useful in fields such as cosmetics and skin external preparations.

Claims

1. A reagent, characterized in that it is A reagent for maintaining the structure of hyaluronic acid, The reagent contains a compound represented by the following formula (1) or a salt thereof, The molecular weight of the hyaluronic acid is 600 kDa to 2000 kDa, ; In the formula (1), R 1 and R 2 which are the same as or different from each other, and are each independently a hydrogen atom, a hydroxyl group, or a linear or branched alkoxy group having 1 to 5 carbon atoms, R 3 is a straight-chain or branched-chain hydroxyalkyl group having 1 to 7 carbon atoms, a straight-chain or branched-chain alkyl group having 1 to 7 carbon atoms, a substituted or unsubstituted cyclic alkyl group having 3 to 10 carbon atoms, or a hydrogen atom.

2. The reagent according to claim 1, wherein The R 1 is a hydrogen atom or a hydroxyl group.

3. The reagent according to claim 1 or 2, wherein The R 2 is a hydrogen atom, a hydroxyl group or a methoxy group.

4. The reagent according to any one of claims 1 to 3, wherein, The R 3 is a linear or branched hydroxyalkyl group having 1 to 5 carbon atoms, a linear or branched alkyl group having 1 to 5 carbon atoms, or a substituted or unsubstituted cyclic alkyl group having 3 to 10 carbon atoms.

5. The reagent according to any one of claims 1 to 4, wherein, The said R 1 is a hydroxyl group, The said R 2 is a hydroxyl group, The R 3 is a linear or branched hydroxyalkyl group having 1 to 7 carbon atoms, a linear or branched alkyl group having 1 to 7 carbon atoms, or a substituted or unsubstituted cyclic alkyl group having 3 to 10 carbon atoms.

6. The reagent according to any one of claims 1 to 5, wherein The compound represented by the formula (1) or a salt thereof is a compound represented by the following formula (2) or formula (3) or a salt thereof, ; 。 7. The reagent according to any one of claims 1 to 6, wherein The molecular weight of the hyaluronic acid is 1200 kDa to 1600 kDa.

8. The reagent according to any one of claims 1 to 7, wherein As the compound represented by the formula (1) or a salt thereof, it contains an extract of Artemisia capillaris and / or an extract of Lonicera japonica.

9. A reagent, characterized in that it is A reagent for inhibiting the expression of the HYBID gene, The reagent contains a compound represented by the following formula (1) or a salt thereof, ; In the formula (1), R 1 and R 2 are the same as or different from each other, and are each independently a hydrogen atom, a hydroxyl group, or a linear or branched alkoxy group having 1 to 5 carbon atoms, R 3 is a linear or branched hydroxyalkyl group having 1 to 7 carbon atoms, a linear or branched alkyl group having 1 to 7 carbon atoms, a substituted or unsubstituted cyclic alkyl group having 3 to 10 carbon atoms, or a hydrogen atom.

10. The reagent according to claim 9, wherein, The R 1 is a hydrogen atom or a hydroxyl group.

11. The reagent according to claim 9 or 10, wherein The R 2 is a hydrogen atom, a hydroxyl group or a methoxy group.

12. The reagent according to any one of claims 9 to 11, wherein, The R 3 is a straight-chain or branched-chain hydroxyalkyl group having 1 to 5 carbon atoms, a straight-chain or branched-chain alkyl group having 1 to 5 carbon atoms, or a substituted or unsubstituted cyclic alkyl group having 3 to 10 carbon atoms.

13. The reagent according to any one of claims 9 to 12, wherein, The said R 1 is a hydroxyl group, The said R 2 is a hydroxyl group, The R 3 is a linear or branched hydroxyalkyl group having 1 to 7 carbon atoms, a linear or branched alkyl group having 1 to 7 carbon atoms, or a substituted or unsubstituted cyclic alkyl group having 3 to 10 carbon atoms.

14. The reagent according to any one of claims 9 to 13, wherein The compound represented by the formula (1) or a salt thereof is a compound represented by the following formula (2) or formula (3) or a salt thereof, ; 。 15. The reagent according to any one of claims 9 to 14, wherein, As the compound represented by the following formula (1) or a salt thereof, it contains an extract of Artemisia capillaris and / or an extract of Lonicera japonica.

16. A method for maintaining the structure of hyaluronic acid, characterized in that, Use the reagent according to any one of claims 1 to 8.

17. The maintenance method according to claim 16, wherein, The maintaining method includes an administration step of administering the reagent to a subject.

18. The maintenance method according to claim 16 or 17, wherein, In In vitro or In vivo use the reagent(s).

19. A method for inhibiting the expression of a HYBID gene, characterized in that, Use the reagent according to any one of claims 9 to 15.

20. The expression inhibition method according to claim 19, wherein, The expression inhibition method includes an administration step of administering the reagent to a subject.

21. The expression inhibition method according to claim 19 or 20, wherein, In In vitro or In vivo use the reagent.

22. A compound or its salt, characterized in that, Represented by the following formula (2) or formula (3), ; 。

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