JP7203519B2 - Claudin expression promoter and tight junction formation promoter - Google Patents

Description

TECHNICAL FIELD The present invention relates to claudin expression promoters and tight junction formation promoters.

A tight junction is a cell adhesion apparatus that exists in a belt-like shape around cells and allows adjacent cells to adhere to each other. Tight junctions are well developed in the inner walls of the skin, intestinal tract, kidney tubules, blood vessels, and the like, and control the permeation of liquids and substances in intercellular spaces. Here, it is said that harmful substances such as allergens are likely to enter the body when the function of tight junctions is weakened for some reason. Tight junctions are also known to be involved in the maintenance of the cytoskeleton, the transduction of cell information, the synthesis and metabolism of intercorneocyte lipids, etc., in addition to the function of regulating intercellular permeability. Proteins that constitute such tight junctions include the integral membrane proteins claudin and occludin. .

In recent years, due to the importance of tight junctions in living organisms as described above, the development of substances capable of promoting the formation of tight junctions has been desired. For example, Patent Document 1 discloses that synephrine was found to be a substance that promotes tight junction formation.

JP 2016-222557 A

An object of the present invention is to provide an agent that can be used to promote the formation of tight junctions.

The present inventors have conducted intensive studies to achieve the above-mentioned object, and unexpectedly discovered that glucooligosaccharides themselves have the effect of promoting the expression of claudins, which are major proteins constituting tight junctions. Found it. Then, the present inventors used the glucooligosaccharide as an active ingredient as an agent used to promote the expression of claudin or an agent used to promote the formation of tight junctions, thereby completing the present invention. Arrived.

That is, the gist of the present invention is as follows.

[1] A claudin expression promoter containing glucooligosaccharide as an active ingredient.
[2] A tight junction formation promoter containing glucooligosaccharide as an active ingredient.
[3] A composition for promoting tight junction formation, containing the agent according to [1] or [2].
[4] Promote tight junction formation according to [3], further comprising at least one medicinal ingredient selected from the group consisting of anti-inflammatory agents, vitamins, moisturizers, antibacterial agents, whitening agents and local anesthetics. composition.
[5] The composition for promoting tight junction formation according to [3] or [4], which is an external preparation for skin or a skin cosmetic.
[6] The composition for promoting tight junction formation according to any one of [3] to [5], which is used in the presence of Staphylococcus epidermidis.
[7] A method for promoting expression of claudin, which comprises using glucooligosaccharide.
[8] A method for promoting tight junction formation, characterized by using glucooligosaccharide.
[9] The method of [7] or [8], wherein glucooligosaccharide is used in the presence of S. epidermidis.
[10] A cosmetic method for improving skin, characterized by promoting expression of claudin using glucooligosaccharide.
[11] A cosmetic method for improving skin, characterized by promoting the formation of tight junctions using glucooligosaccharides.
[12] The cosmetic method of [10] or [11], wherein glucooligosaccharide is used in the presence of S. epidermidis.

According to the present invention, an agent that can be used to promote claudin expression and an agent that can be used to promote tight junction formation can be provided.

FIG. 1 shows claudin-1 fluorescent staining images of each test sample in the claudin-1 expression enhancement test. FIG. 2 is a bar graph showing the relative expression level (%) of claudin-1 in each test sample in the claudin-1 expression enhancement test.

The present invention will be described in detail below. In addition, the terms used in this specification are interpreted as meanings commonly used in the technical field unless otherwise specified.

[Claudin expression promoter]
The claudin expression promoter of the present invention is characterized by containing glucooligosaccharide as an active ingredient. As described above, the glucooligosaccharide itself has the effect of promoting the expression of claudin, which is a major protein that constitutes tight junctions. This finding was first discovered by the authors. Claudin expression promoters are used by being blended in, for example, pharmaceuticals, quasi-drugs, cosmetics, food and drink, etc., and are preferably blended in skin pharmaceuticals, quasi-drugs, cosmetics, food and drink, and the like. More preferably, it is incorporated into a composition for external use on the skin and used as a pharmaceutical, quasi-drug, cosmetic, or the like. By using it in this way, it is possible to impart the desired claudin expression-enhancing effect to these pharmaceuticals, quasi-drugs, cosmetics, food and drink, and the like.

Claudins are the predominant proteins responsible for tight junction strand formation and have about 24 subtypes, including claudin-1. It is believed that claudin-1 also contributes to the formation of tight junctions by binding to actin filaments, which are the cytoskeleton, through membrane-lining proteins such as ZO-1. Increased expression levels are thought to promote the formation of tight junctions.

In the present invention, promotion of claudin expression also means promotion of claudin gene expression and promotion of claudin protein expression. In the present invention, claudin is, for example, claudin-1.

The claudin expression promoter of the present invention can improve claudin expression, for example, compared to when the claudin expression promoter of the present invention is not applied. Here, claudin expression can be confirmed by, for example, the expression level of claudin mRNA and the expression level of claudin protein. The mRNA expression level can be measured by known methods such as real-time PCR and microarray, and the protein expression level can be measured by known methods such as immunostaining and Western blotting.

Gluco-oligosaccharides, which are the active ingredients of the agent for promoting claudin expression of the present invention, are obtained from natural sugars maltose and sucrose by an immobilized enzymatic method. 2)) -(O-α-D-glucopyranosyl-(1→6))-(O-α-D-glucopyranosyl-(1→4))-(O-α-D-glucopyranose) things are mentioned. Such glucooligosaccharides are commercially available, for example, as BIOECOLIA (registered trademark: Soravia).

The claudin expression-enhancing agent of the present invention may contain, for example, various components (eg, known bases, carriers, etc., described below) within a range that does not impair the effects of the present invention. In addition, the form of the agent may be, for example, a liquid system (including liquid, liquid, paste, etc.) or a solid system (including solid, solid, etc.). These agents can be produced, for example, by conventional methods.

[Tight junction formation accelerator]
The tight junction formation promoter of the present invention is characterized by containing glucooligosaccharide as an active ingredient. As described above, the glucooligosaccharide itself has the effect of promoting the expression of claudin, which is a major protein that constitutes tight junctions. This finding was first discovered by the authors. Tight junction formation accelerators are used by being blended in, for example, pharmaceuticals, quasi-drugs, cosmetics, food and drink, etc., and are preferably blended in skin pharmaceuticals, quasi-drugs, cosmetics, food and drink, and the like. More preferably, it is incorporated into a composition for external use on the skin and used as a pharmaceutical, quasi-drug, cosmetic, or the like. By using it in this way, it is possible to impart the desired claudin expression-enhancing effect to these pharmaceuticals, quasi-drugs, cosmetics, food and drink, and the like. As the glucooligosaccharide, those described above can be used. The above description can also be referred to for the form of the agent and the like.

Another aspect of the present invention is an agent for normalizing and/or enhancing stratum corneum function, containing glucooligosaccharide as an active ingredient. That is, the present invention can be used as an agent that normalizes and/or strengthens stratum corneum function by promoting claudin expression or tight junction formation. Here, the function of the stratum corneum refers to the function of the stratum corneum controlled by tight junctions.

Another aspect of the present invention is an agent for normalizing and/or enhancing tight junction function, containing glucooligosaccharide as an active ingredient. That is, the present invention can be used as an agent that normalizes and/or enhances tight junction function by promoting claudin expression or tight junction formation.

Another aspect of the present invention is an agent for normalizing and/or enhancing epidermal barrier function, containing glucooligosaccharide as an active ingredient. That is, the present invention can be used as an agent that normalizes and/or strengthens the epidermal barrier function by promoting the expression of claudins or the formation of tight junctions. Here, the epidermal barrier function refers to the epidermal barrier function controlled by tight junctions.

Another aspect of the present invention is a permeation barrier function normalizing and/or reinforcing agent containing glucooligosaccharide as an active ingredient. That is, the present invention can be used as an agent that normalizes and/or strengthens the permeation barrier function by promoting the expression of claudins or the formation of tight junctions. Here, the permeation barrier function refers to the permeation barrier function controlled by tight junctions.

Another aspect of the present invention is an intercellular barrier function normalizing and/or enhancing agent containing glucooligosaccharide as an active ingredient. That is, the present invention can be used as an agent that normalizes and/or strengthens the intercellular barrier function by promoting claudin expression or tight junction formation. Here, the intercellular barrier function refers to the intercellular barrier function controlled by tight junctions.

[Composition for Promoting Tight Junction Formation]
The composition for promoting tight junction formation of the present invention is characterized by containing a claudin expression promoter or a tight junction formation promoter as an active ingredient. As the claudin expression promoter and the tight junction formation promoter, those described above can be used.

The content of glucooligosaccharide, which is an active ingredient of the claudin expression promoter and tight junction formation promoter, in the composition of the present invention is, for example, 0.00001% by weight or more, 0.0001% by weight or more, and 0.0001% by weight or more, based on the total amount of the composition. 001% by weight or more. As a result, the composition has a sufficient effect of promoting tight junction formation. Also, for example, it can be 30% by weight or less, or 5% by weight or less. Thereby, the other components can be sufficiently blended. The range can be, for example, 0.00001 to 30% by weight, 0.001 to 5% by weight.

The composition of the present invention can be used either orally or parenterally in the form of, for example, pharmaceuticals, quasi-drugs, cosmetics, food and drink, etc., and preferably external skin preparations or skin cosmetics (cosmetics). can be used as In these forms, in addition to the claudin expression promoter and the tight junction formation promoter, in addition to known bases or carriers added to pharmaceuticals, quasi-drugs, cosmetics, etc., to the extent that the effects of the present invention are not impaired. Can be mixed.

Examples of bases or carriers include hydrocarbons such as liquid paraffin, squalane, vaseline, gelling hydrocarbons (plastibase, etc.), ozokerite, α-olefin oligomers, and light liquid paraffin; methylpolysiloxane, highly polymerized methylpolysiloxane, Cyclic silicone, alkyl-modified silicone, amino-modified silicone, polyether-modified silicone, polyglycerin-modified silicone, silicone/alkyl chain co-modified polyether-modified silicone, silicone/alkyl chain co-modified polyglycerin-modified silicone, polyether-modified branched silicone, poly Silicone oils such as glycerin-modified branched silicone, acrylic silicone, phenyl-modified silicone, and silicone resin; fats and oils such as meadowfoam oil, coconut oil, olive oil, rice bran oil, and shea butter; waxes such as jojoba oil, miura, candelilla wax, and lanolin higher alcohols such as cetanol, cetostearyl alcohol, stearyl alcohol, behenyl alcohol, octyldodecanol, isostearyl alcohol, phytosterol, and cholesterol; cellulose derivatives such as ethylcellulose, hydroxypropylcellulose, and hydroxypropylmethylcellulose; polyvinylpyrrolidone; carrageenan; polyethylene glycol; dioxane; butylene glycol adipate polyester; Polysaccharides such as dextrin and maltodextrin; Vinyl polymers such as carboxyvinyl polymer and alkyl-modified carboxyvinyl polymer; Lower alcohols such as ethanol and isopropanol; Ethylene glycol monomethyl ether, ethylene glycol monoethyl ether, ethylene glycol monopropyl Glycol ether such as ether, diethylene glycol monomethyl ether, diethylene glycol monoethyl ether, diethylene glycol monopropyl ether, diethylene glycol monobutyl ether, propylene glycol monoethyl ether, propylene glycol monopropyl ether, dipropylene glycol monoethyl ether, dipropylene glycol monopropyl ether, etc. water;

When the composition of the present invention contains a base or carrier other than water and a lower alcohol, examples of the base or carrier include higher alcohols, hydrocarbons, fats and oils, esters, silicone oils, waxes, vinyl-based high Molecules are preferred, and higher alcohols, ester oils, silicone oils and vinyl polymers are more preferred. Among these components, cetanol, cetostearyl alcohol, stearyl alcohol, behenyl alcohol, glyceryl tri-2-ethylhexanoate, dimethicone, cyclomethicone, polyether-modified silicone, polyglycerin-modified silicone, and carboxyvinyl polymer are more preferred.

In the present invention, the base or carrier may be used singly or in combination of two or more. In addition, the amount of these to be used is appropriately selected from the range known to those skilled in the art.

The composition of the present invention can be a liquid, fluid, or semi-solid formulation. Examples of such dosage forms include liquids, suspensions, emulsions (milk), creams, ointments, gels (gels), liniments, lotions, aerosols, sticks, and non-woven fabric impregnated with a drug solution. A sheet agent (pack agent) and the like can be mentioned, and it can also be provided in the form of a food or drink. Among them, preferred are liquids, emulsions, creams, gels, and lotions. These formulations can be produced according to a conventional method, for example, the method described in the Japanese Pharmacopoeia General Rules for Pharmaceutical Preparations, 17th Edition.

In addition, the composition of the present invention can be blended with other medicinal ingredients within a range that does not impair the effects of the present invention, thereby enhancing the effects of the present invention or imparting further effects. Such medicinal ingredients are, for example, ingredients that exert medicinal effects on the skin and mucous membranes, and include ingredients that bring about all favorable effects such as healing and improvement of inflammation/damage occurring in the skin and mucous membranes and protection of the skin. Specific examples of such medicinal ingredients include anti-inflammatory agents, vitamins, moisturizing agents, antibacterial agents, whitening agents, and local anesthetics. In the composition of the present invention, these medicinal ingredients may be used singly or in combination of two or more.

Examples of anti-inflammatory agents include glycyrrhizic acid, dipotassium glycyrrhizinate, glycyrrhizic acid derivatives such as monoammonium glycyrrhizinate, glycyrrhizic acid or derivatives thereof, allantoin or derivatives thereof, tranexamic acid or salts thereof, ε-aminocaproic acid, licorice extract. substances, menthol, camphor, etc. can be exemplified. Preferred are licorice extract, dipotassium glycyrrhizinate, monoammonium glycyrrhizinate, glycyrrhetinic acid, allantoin, tranexamic acid, ε-aminocaproic acid, menthol and camphor.

These components can be used individually by 1 type or in combination of 2 or more types. The amount of these anti-inflammatory agents in the composition of the present invention is not particularly limited, but is usually 0.01 to 5% by weight, preferably 0.01 to 3% by weight, more preferably 0.01 to 2% by weight. can be appropriately selected and adjusted according to the purpose from the range of

Vitamins include, for example, retinol, retinol acetate, retinol derivatives (vitamin A) such as retinol palmitate, retinal, retinoic acid, methyl retinoate, ethyl retinoate, retinol retinoate, vitamin A oil, vitamin A fatty acid ester β-carotene, α-carotene, γ-carotene, δ-carotene, lycopene, zeaxanthin, cryptoxanthin, provitamin A such as echinenone, dl-α-tocopherol, d-δ-tocopherol, acetic acid Vitamin E such as dl-α-tocopherol, dl-α-tocopherol succinate, dl-α-tocopherol calcium succinate, riboflavin, flavin mononucleotide, flavin adenine dinucleotide, riboflavin butyrate, riboflavin tetrabutyrate, riboflavin 5 '-sodium phosphate, vitamin B2s such as riboflavin tetranicotinic acid ester, dl-α-tocopherol nicotinate, benzyl nicotinate, methyl nicotinate, β-butoxyethyl nicotinate, 1-(4-methylphenyl nicotinate) ) Nicotinic acids such as ethyl, vitamin Cs such as ascorbigen-A, ascorbyl stearate, ascorbyl palmitate, and L-ascorbyl dipalmitate, vitamin Ds such as methylhesperidin, ergocalciferol, and cholecalciferol , vitamin Ks such as phylloquinone and farnoquinone, and γ-oryzanol. Also dibenzoylthiamine, dibenzoylthiamine hydrochloride, thiamine hydrochloride, thiamine cetyl hydrochloride, thiamine thiocyanate, thiamine lauryl hydrochloride, thiamine nitrate, thiamine monophosphate, thiamine lysine salt, thiamine triphosphate, thiamine monoline Vitamin B1s such as acid ester phosphate, thiamine monophosphate, thiamine diphosphate, thiamine diphosphate hydrochloride, thiamine triphosphate, thiamine triphosphate monophosphate, pyridoxine hydrochloride, pyridoxine acetate, pyridoxal hydrochloride, Vitamin B6s such as pyridoxal 5'-phosphate and pyridoxamine hydrochloride; vitamin B12s such as cyanocobalamin, hydroxocobalamin and deoxyadenosylcobalamin; folic acids such as folic acid and pteroylglutamic acid; nicotinic acids such as nicotinic acid and nicotinamide , pantothenic acid, calcium pantothenate, pantothenyl alcohol (panthenol), D-panthesaine, D-pantethine, coenzyme A, pantothenic acids such as pantothenyl ethyl ether, biotin, biotins such as biotycin, ascorbic acid, ascorbic acid Ascorbic acid derivatives such as sodium acid, dehydroascorbic acid, 3-0-ethylascorbic acid, sodium ascorbic acid phosphate, magnesium ascorbic acid phosphate, and other ascorbic acid derivatives, carnitine, ferulic acid, α-lipoic acid , vitamin-like acting factors such as orotic acid and the like can be exemplified.

Preferably, retinol, retinol acetate, retinol palmitate, retinal, retinoic acid, ascorbic acid, sodium ascorbate, 3-0-ethylascorbic acid, sodium ascorbate phosphate, magnesium ascorbate phosphate, ascorbate stearic acid ester, ascorbyl palmitate, riboflavin, riboflavin butyrate, riboflavin 5'-phosphate sodium, pantothenic acid, calcium pantothenate, retinol, vitamin A oil, vitamin A fatty acid ester, methyl hesperidin, folic acid, dibenzoylthiamine, dibenzoylthiamine hydrochloride, thiamine hydrochloride, thiamine cetyl hydrochloride, thiamine thiocyanate, thiamine lauryl hydrochloride, nicotinic acid, nicotinamide, beta-carotene, ergocalciferol, cholecalciferol, pyridoxine hydrochloride, panthenol, dl-α-tocopherol, d-δ-tocopherol and dl-α-tocopherol acetate. More preferably retinol, retinol acetate, retinol palmitate, retinal, retinoic acid, ascorbic acid, magnesium ascorbyl phosphate, ascorbyl stearate, nicotinic acid, nicotinamide, β-carotene, ergocalciferol, cholecalci Ferrol, pyridoxine hydrochloride.

These components can be used individually by 1 type or in combination of 2 or more types. The amount of these vitamins in the composition of the present invention is not particularly limited, but is usually 0.001 to 10% by weight, preferably 0.005 to 5% by weight, more preferably 0.01 to 2% by weight. It can be appropriately selected and adjusted from the range according to the purpose.

Examples of moisturizing agents include polyethylene glycol, propylene glycol, glycerin, diglycerin, 1,3-butylene glycol, dipropylene glycol, sorbitol, mannitol, glucose, sucrose, fructose, xylitol, lactose, maltose, maltitol, and the like. Polyhydric alcohols, sodium hyaluronate, heparinoids, sodium chondroitin sulfate, biopolymers such as collagen, elastin, and keratin, amino acids such as glycine, aspartic acid, and arginine, natural moisturizing agents such as sodium lactate, urea, and sodium pyrrolidonecarboxylate Factors include lipids such as ceramide, cholesterol, and phospholipids. Also, plant extracts such as lavender extract, eucalyptus extract, peppermint extract, and the like can be exemplified. Preferably, glycerin, diglycerin, 1,3-butylene glycol, dipropylene glycol, sodium hyaluronate, heparinoid, sodium chondroitin sulfate, collagen, keratin, glycine, aspartic acid, arginine, sodium lactate, urea, pyrrolidone carboxylic acid Sodium, Ceramides, Cholesterol, Lavender Extract, Eucalyptus Extract, Peppermint Extract.

These components can be used individually by 1 type or in combination of 2 or more types. The amount of these moisturizing agents in the composition of the present invention is not particularly limited, but is usually 0.005 to 50% by weight, preferably 0.01 to 30% by weight, more preferably 0.01 to 10% by weight. It can be appropriately selected and adjusted from the range.

Examples of antibacterial agents include isopropylmethylphenol, chlorhexidine hydrochloride, benzalkonium chloride, cetylpyridinium chloride, ethanol, parabens, phenoxyethanol, glycerin fatty acid esters, and the like.

These components can be used individually by 1 type or in combination of 2 or more types. The amount of these antibacterial agents in the composition of the present invention is not particularly limited, but is usually in the range of 0.001 to 10% by weight, preferably 0.001 to 5% by weight, more preferably 0.001 to 1% by weight. can be appropriately selected and adjusted from

Whitening agents include, for example, hydroquinone, α-arbutin, β-arbutin, ascorbic acid, 2-O-ethylascorbic acid, 3-O-ethylascorbic acid, 3-O-cetylascorbic acid, tetra-2-hexyldecyldecane Ascorbyl Acid, Methylsilanol Ascorbate, Sulfuric Ascorbate, Ascorbyl Glucoside, Potassium (Ascorbyl/Tocopheryl) Phosphate, Ascorbyl Phosphate, Ascorbyl Palmitate, Ascorbyl Dipalmitate, Trisodium Ascorbyl Palmitate Phosphate, Ascorbyl Aminopropyl Phosphate , Ascorbyl Stearate, Dehydroascorbic Acid, Disodium Ascorbyl Sulfate, Calcium Ascorbate, Magnesium Ascorbate, Sodium Ascorbate, Zinc Ascorbate, Chitosan Ascorbate, Sodium Ascorbyl Phosphate, Magnesium Ascorbyl Phosphate, Trisodium Ascorbyl Phosphate , kojic acid, rucinol, tranexamic acid and the like.

These components can be used individually by 1 type or in combination of 2 or more types. The amount of these whitening agents in the composition of the present invention is not particularly limited, but is usually in the range of 0.00001 to 50% by weight, preferably 0.00001 to 30% by weight, more preferably 0.0001 to 4% by weight. can be appropriately selected and adjusted from

Examples of local anesthetics include eucalyptus oil, eugenol, menthol, camphor, peppermint oil and the like.

These components can be used individually by 1 type or in combination of 2 or more types. The amount of these local anesthetics in the composition of the present invention is not particularly limited, but can be appropriately selected and adjusted from the range of usually 0.01 to 20% by weight, preferably 0.1 to 15% by weight.

Further, the composition of the present invention may contain other additives as long as the effects of the present invention are not impaired. Specific examples of such additives include pigments, fragrances, preservatives , surfactants, plant extracts, pH adjusters, sequestering agents, oils, gelling agents, higher alcohols, hydrocarbons, penetration enhancers, pigments, antioxidants, and the like. In addition, in the composition of the present invention, these additives may be used singly or in combination of two or more.

The pH of the composition of the present invention is generally pH 3-9, preferably pH 4-8. In addition, this pH can be adjusted by using the pH adjuster.

The composition of the present invention can be usefully used when promotion of tight junction formation is desired. For example, it is useful for inhibiting the entry of harmful substances into the body, and is useful for improving skin and sensitive skin. It is also useful for normalizing and strengthening interstitial barrier function. Furthermore, application to research on claudins themselves or tight junctions themselves is also conceivable, and is useful in a very wide range of fields. For these terms, the above description can be referred to.

The composition of the present invention can be used, for example, when promotion of tight junction formation in the skin is desired, and normalization and enhancement of the stratum corneum function, tight junction function, epidermal barrier, permeation barrier, and intercellular barrier function in the skin. It can be usefully used when desired. At this time, for example, by normalizing the control function of the skin and improving and strengthening the function of the skin itself, the permeation of irritating substances from the external environment is suppressed, and the ions, water, etc. existing inside the skin are removed. It is expected to prevent it from passing outside and improve skin troubles such as rough skin. In addition, for example, it is expected to normalize and strengthen the synthesis and metabolism of intercorneocyte lipids and to normalize and strengthen the expulsion function of keratin.

The composition of the present invention used for the above purposes includes, for example, external skin preparations such as ointments, gels, liquids, and lotions, and skin cosmetics such as body soap cleansers, hand cleansers, and facial cleansers. It can be suitably used in the form of powders, lotions, milky lotions, creams, packs and the like.

The compositions of the invention can be used, for example, in the presence of Staphylococcus epidermidis. By using the composition of the present invention in the presence of Staphylococcus epidermidis, the claudin expression promoting effect and tight junction formation promoting effect of the present invention are remarkably enhanced.

Another aspect of the present invention is a composition for promoting claudin expression, comprising a claudin expression-enhancing agent as an active ingredient. Since the present invention contains a claudin expression promoter as an active ingredient, the expression of claudin can be promoted. The description of the composition for promoting tight junction formation can be referred to as appropriate.

[Method for Promoting Claudin Expression and Method for Promoting Tight Junction Formation]
The method for promoting claudin expression and promoting tight junction formation of the present invention are characterized by using glucooligosaccharides. As described above, the present inventors found that glucooligosaccharides themselves have the effect of promoting the expression of claudins, which are major proteins constituting tight junctions. Based on this finding, the present inventors have completed the present invention by using glucooligosaccharides as a method for promoting claudin expression and tight junction formation. The method for promoting claudin expression and the method for promoting tight junction formation of the present invention do not include medical practice.

In the method of the present invention, the glucooligosaccharides described above can be used. In addition, the glucooligosaccharide-containing agent and composition described above can also be suitably used as the glucooligosaccharide in the method of the present invention.

The method of the invention can be performed, for example, in the presence of Staphylococcus epidermidis. By performing the method of the present invention in the presence of Staphylococcus epidermidis, the claudin expression promoting effect and tight junction formation promoting effect of the present invention are remarkably enhanced. As for Staphylococcus epidermidis, the above description can be referred to.

[Beauty method]
The cosmetic method of the present invention is characterized by promoting the expression of claudins using glucooligosaccharides or promoting the formation of tight junctions using glucooligosaccharides. As described above, the present inventors found that glucooligosaccharides themselves have the effect of promoting the expression of claudins, which are major proteins constituting tight junctions. Based on this finding, the present inventors have come to use glucooligosaccharides to promote the expression of claudins or to promote the formation of tight junctions using glucooligosaccharides as cosmetic methods for skin improvement. , completed the present invention. In the present invention, the term "beauty method" refers not only to a method that is performed individually, but also to a method that is provided as a cosmetic prescription tailored to the customer when providing products related to beauty, and that is used by non-doctors to sell cosmetics. including those provided by staff and estheticians. In addition, it also includes what is provided as a method of using the product in the instruction manual (package insert, etc.) of the product related to beauty. Note that the cosmetic method of the present invention does not include medical practice.

In the cosmetic method of the present invention, the glucooligosaccharides described above can be used. In addition, the above glucooligosaccharide-containing agent and composition can also be suitably used as the glucooligosaccharide in the cosmetic method of the present invention. Such compositions can be used, for example, by applying to the skin of a desired site on a subject. Application to the skin may occur, for example, once or multiple times a day.

The cosmetic method of the present invention can be used, for example, to improve skin conditions for which promotion of tight junction formation is desired. For example, it can be used to improve sensitive skin, and can also be used to normalize and strengthen stratum corneum function, tight junction function, epidermal barrier function, permeation barrier function, and intercellular barrier function in skin. For these terms, the above description can be referred to.

The cosmetic method of the present invention can be performed, for example, in the presence of Staphylococcus epidermidis. Specifically, for example, in implementing the cosmetic method of the present invention, it can be performed by supplementing the target skin with Staphylococcus epidermidis. can be performed by carrying out the cosmetic method of the present invention on the subject. By performing the cosmetic method of the present invention in the presence of Staphylococcus epidermidis, the claudin expression promoting effect and tight junction formation promoting effect of the present invention are remarkably enhanced. As for Staphylococcus epidermidis, the above description can be referred to.

EXAMPLES The present invention will be described in more detail below with reference to examples, but the present invention is not limited by these examples.

[Test Example 1] Claudin-1 expression promotion test (test sample)
As a test sample, glucooligosaccharide (Nikko Chemical Co., Ltd.) and/or Staphylococcus epidermidis (Microbiologies) were dissolved in PBS (-) (Kohjin Bio Co., Ltd.) to obtain a glucooligosaccharide (1% by mass)-containing solution, A solution containing S. epidermidis (1×10 ⁸ CFU) and a solution containing glucooligosaccharides (1% by weight) and S. epidermidis (1×10 ⁸ CFU) were prepared. As a control, a solution containing only PBS(-) was prepared.

(Test method and results)
A three-dimensional tissue human epidermis model RHE (Nikoderm Research) was cultured in a 12-well culture plate. More specifically, 1 mL of MAINTENANCE MEDIUM (Nikoderm Research), a dedicated medium for human epidermis model RHE without antibacterial agents, was used as a culture medium for each well, and cultured at 37° C. in an environment of 5% carbon dioxide gas and 95% air. cultured for days. After exchanging the culture solution, the test sample was added from the stratum corneum side and cultured for 5 more days. A three-dimensional tissue human epidermis model RHE was dissected from the support, fixed with 4% PFA for 20 hours, and embedded in paraffin. Sections were prepared by slicing at 10 μm, deparaffinized, and subjected to antigen retrieval treatment using an antigen retrieval solution (Target Retrieval Splution, pH 9, Dako). Rabbit-derived anti-Claudin-1 antibody (Rabbit anti-Claudin-1, invitrogen) as primary antibody, Alexa Fluor 594-labeled anti-rabbit IgG antibody (Goat anti-Rabbit IgG (H + L) Highly Cross-Adsorbed Secondary Antibody, Alexa 5 4 , invitrogen) was used as a secondary antibody to stain claudin-1 protein. In addition, cell nuclei were stained with Hoechst 33342 (invitrogen). After staining, the sections were observed and photographed using a fluorescence microscope (Olympus Corporation). Based on each photographed image, the amount of claudin-1 protein was quantified, and the relative expression level (%) of claudin-1 was calculated with respect to the control (no addition) as 100%. The results are shown in FIG. 1 and Tables 1 and 2.

FIG. 1 is a fluorescence-stained image of each post-staining section, claudin-1 is shown in red and cell nuclei in blue. Table 1 shows the relative expression level (%) of claudin-1 calculated based on each fluorescence-stained image, and FIG. 2 shows this as a bar graph. As shown in FIG. 1 and Tables 1 and 2, when glucooligosaccharide was added, the expression level of claudin-1 increased about 8-fold compared to the control (no addition). , the expression level of claudin-1 was significantly enhanced. In addition, when glucooligosaccharide was added together with S. epidermidis, the expression level of claudin-1 was as follows: It was shown that the expression level of claudin-1 was synergistically enhanced by the co-addition of glucooligosaccharides and S. epidermidis. . These test results using human epidermis models revealed that glucooligosaccharides promote the expression of claudins and promote the formation of tight junctions. In particular, in the presence of glucooligosaccharides and Staphylococcus epidermidis, claudin expression was markedly enhanced, showing a synergistic tight junction formation effect.

Examples of formulations of the composition for promoting tight junction formation of the present invention are shown below.

<Formulation Formulation Example 1: Skin Cosmetic (Lotion)>
1,3-butylene glycol 12.0% by weight
Glycerin 3.0% by weight
Gluco-oligosaccharide 2.0% by weight
Hydroxyethyl cellulose 0.1% by weight
Hydrolyzed collagen 0.01% by weight
Allantoin 0.1% by weight
Polyquaternium-51 0.5% by weight
PEG-1500 0.5% by weight
pH adjuster Appropriate amount Preservative Trace amount
Appropriate amount of purified water
Total 100% by weight

<Formulation Formulation Example 2: Whitening Skin Cosmetic (Lotion)>
Tranexamic acid 2.0% by weight
Gluco-oligosaccharide 1.0% by weight
Glycerin 5.0% by weight
Carboxyvinyl polymer 0.1% by weight
EDTA-2Na 0.05% by weight
Hyaluronic acid Na 0.1% by weight
pH adjuster Appropriate amount Preservative Trace amount
Appropriate amount of purified water
Total 100% by weight

<Formulation Formulation Example 3: External Skin Preparation (Emulsion)>
Gluco-oligosaccharide 3.0% by weight
1,3-butylene glycol 8.0% by weight
Squalane 3.0% by weight
Glycerin 4.0% by weight
Vaseline 3.5% by weight
Polyoxyethylene (40) hydrogenated castor oil 1.5% by weight
Glyceryl stearate 1.2% by weight
Dimethicone 0.5% by weight
Hydrolyzed collagen 0.1% by weight
Carboxyvinyl polymer 0.1% by weight
Xanthan gum 0.1% by weight
Cetanol 0.5% by weight
pH adjuster Appropriate amount Preservative Trace amount
Appropriate amount of purified water
Total 100% by weight

<Formulation Formulation Example 4: External Skin Preparation (Cream)>
1,3-butylene glycol 15.0% by weight
Vaseline 10.0% by weight
Meadowfoam oil 2.0% by weight
Glycerin 7.0% by weight
Gluco-oligosaccharide 5.0% by weight
Behenyl alcohol 1.0% by weight
Shea butter 2.0% by weight
Glyceryl stearate 1.5% by weight
Polyglyceryl-10 stearate 2.0% by weight
Hydrogenated lecithin 0.8% by weight
Hydrolyzed collagen 0.5% by weight
Dipotassium glycyrrhizinate 0.1% by weight
Carboxyvinyl polymer 0.3% by weight
pH adjuster Appropriate amount Preservative Trace amount
Appropriate amount of purified water
Total 100% by weight

<Formulation Formulation Example 5: External Skin Preparation (Gel)>
Gluco-oligosaccharide 0.5% by weight
Alkyl-modified carboxyvinyl polymer 0.4% by weight
1,3-butylene glycol 5.0% by weight
Glycerin 10.0% by weight
Sodium hyaluronate 0.1% by weight
Arginine Appropriate amount Preservative Trace amount
Appropriate amount of purified water
Total 100% by weight

<Formulation Formulation Example 6: Skin Cosmetic (Facial Cleansing Cream)>
Gluco-oligosaccharide 2.0% by weight
Glycerin 15.0% by weight
1,3-butylene glycol 5.0% by weight
Sodium cocoyl glutamate 3.0% by weight
Cocamidopropyl betaine 1.0% by weight
Hydroxypropyl starch phosphate 2.0% by weight
Stearic acid 0.2% by weight
Lauric acid 0.2% by weight
EDTA-2Na 0.1% by weight
Glyceryl stearate (SE) 1.5% by weight
Polyquaternium-7 0.3% by weight
Dipotassium glycyrrhizinate 0.05% by weight
Hydrolyzed collagen 0.1% by weight
Citric acid Appropriate amount Perfume Appropriate amount pH adjuster Appropriate amount
Appropriate amount of purified water
Total 100% by weight

<Formulation Formulation Example 7: Skin Cosmetic (Cleansing Milk)>
Gluco-oligosaccharide 1.0% by weight
Squalane 10.0% by weight
Cetyl ethylhexanoate 10.0% by weight
Tri(caprylic/capric)glyceryl 5.0% by weight
Cetanol 0.5% by weight
Glyceryl stearate 2.0% by weight
Steareth-20 1.0% by weight
PEG-12 diisostearate 2.0% by weight
Alkanediol appropriate amount Acrylates/C10-30 alkyl acrylate crosspolymer
Appropriate amount Xanthan gum Appropriate amount pH adjuster Appropriate amount Perfume Appropriate amount
Appropriate amount of purified water
Total 100% by weight

INDUSTRIAL APPLICABILITY The present invention has industrial applicability in that it can provide an agent that can be used to promote claudin expression and an agent that can be used to promote tight junction formation.

Claims (6)

A claudin expression promoter comprising a glucooligosaccharide . A tight junction formation accelerator comprising a glucooligosaccharide . A method for promoting expression of claudin by glucooligosaccharide . A method for promoting claudin expression by glucooligosaccharides in the presence of Staphylococcus epidermidis. A method for promoting formation of tight junctions with glucooligosaccharides . A method for promoting tight junction formation by glucooligosaccharides in the presence of Staphylococcus epidermidis.

Images