Improves the balance of the skin's resident flora

Abstract

The present invention addresses the problem of providing a composition for external use on the skin that suppresses a reduction in diversity of cutaneous resident flora and maintains a good skin environment. The present invention solves the problem by providing an agent for improving the balance of cutaneous resident flora, the agent containing lactosucrose as an active ingredient.

Description

[Technical field]

[0001] The present invention relates to an agent for improving the balance of resident skin flora, and more particularly, to an agent for improving the balance of resident skin flora, which contains lactosucrose as an active ingredient and is characterized by maintaining and / or improving the diversity of resident skin flora. [Background technology]

[0002] A state where various kinds of bacteria gather and grow in a certain environment in the body is called the resident flora. In humans, intestinal flora, oral flora, skin flora, etc. are known, and it is said that the bacterial species and their ratios that make up the resident flora change depending on various factors such as the growth environment of the resident flora, external stimuli, and individual differences. In recent years, it has become clear that the state of this resident flora affects various physiological functions of the body, and research on the intestinal flora in particular has progressed.

[0003] On the other hand, it has been revealed that the human skin flora has an effect on skin function. For example, it is known that it improves the skin barrier function that prevents the invasion of pathogenic bacteria (Non-Patent Document 1), that Streptococcus thermophilus produces ceramide (Non-Patent Document 2), and that Staphylococcus epidermidis has a moisturizing effect (Non-Patent Document 3). In addition, Propionibacterium acnes breaks down sebum with lipase to produce free fatty acids, and these free fatty acids keep the skin weakly acidic, suppressing the growth of bad bacteria such as Staphylococcus aureus, which causes food poisoning, but also causes acne, so it is said that an appropriate ratio is desirable (Non-Patent Document 4).

[0004] It is not enough for the skin flora to simply exist; the conditions for maintaining healthy skin are to adjust the composition and number of skin flora, to give dominance to good bacteria such as Staphylococcus epidermidis, to suppress bad bacteria such as Staphylococcus aureus, and to enhance the skin's barrier function. In the field of cosmetics, there are two approaches to skin care that utilize the skin flora: "bacterial cultivation" and "bacterial supplementation." "Bacterial cultivation" is an approach in which good bacteria such as Staphylococcus epidermidis are given nutrients that are favored by the skin and the environment is changed to one in which they can grow easily, giving the good bacteria an advantage. "Bacterial supplementation" is an approach in which good bacteria are taken from the skin, artificially cultivated and multiplied, and then returned to the skin.

[0005] As described above, the effect of the skin flora on the skin indicates that the skin flora and skin condition are closely related, and maintaining the skin flora in good condition is considered to be important for maintaining good skin condition. The skin flora is composed of many types of bacteria, and in recent years, technological advances have made it possible to comprehensively analyze a large number of bacteria, and the diversity of the skin flora is evaluated by calculating a diversity index (such as the Simpson Index or the Shannon Index) from the number of bacterial species and the proportion of each species. Recent studies have revealed that a decrease in the diversity of the skin flora is related to many diseases, including atopic dermatitis, acne, and psoriasis. In addition, it is said that the diversity of the skin flora is also decreased by excessive washing and friction. In addition, there has been an increase in research on the relationship between the diversity index and skin condition, and it has been shown that human skin with good erythema index and skin oxygen saturation index (skin condition considered to be good), which are indices related to skin color, has a high Simpson index. From these results, it is believed that the more a substance or cosmetic method improves the diversity of bacterial species, the more it can improve the erythema index and skin oxygenation index (Patent Document 1). Based on these research results and findings, it is important to maintain and improve the diversity of resident skin bacteria in order to improve skin condition, and appropriate skin care and selection of topical skin preparations and cosmetics are required.

[0006] There have been several reports on skin topical preparations for maintaining the skin flora in good condition. For example, Patent Document 2 discloses a cosmetic composition containing isomaltooligosaccharide and / or its reduced sugar alcohol as a skin flora improving agent using carbohydrates. However, this disclosure only confirms the change in the bacterial mass on the skin before and after continuous application of carbohydrates for two types of skin flora (Staphylococcus epidermidis and Propionibacterium acnes), but does not verify or disclose the action and effect based on the change, and does not disclose any change in the diversity of skin flora.

[0007] In addition, in skin external preparations and cosmetics, when contamination and proliferation by microorganisms occur, the product undergoes changes such as deterioration, discoloration, and odor, and loses its commercial value, so preservatives are used to stabilize the quality. In general, preservatives are often used in combination with a component having antibacterial properties against gram-positive bacteria and a component having antibacterial properties against gram-negative bacteria. In general, preservatives added to products for human use to prevent the deterioration of the product itself are thought to have no adverse effect on skin health, but it has been reported that preservatives affect the skin flora due to their bactericidal and bacteriostatic effects, and that depending on the amount of preservatives added, they can affect the skin flora in healthy skin (Non-Patent Document 5). In addition, bactericides are used to disinfect and keep the skin surface clean, thereby suppressing acne, dandruff, body odor, etc.

[0008] Under these circumstances, it would be extremely useful to prevent a decline in the diversity of resident skin bacteria, maintain a healthy balance of the resident skin flora, and restore the decline in the diversity of resident skin bacteria caused by preservatives and bactericides contained in topical skin preparations and cosmetics, as well as the imbalance of the resident skin flora, to a healthy state. [Prior art documents] [Patent documents]

[0009] [Patent Document 1] JP 2018-99184 A [Patent Document 2] JP 2005-89355 A [Non-patent literature]

[0010] [Non-Patent Document 1] Journal of the Japanese Dermatological Association, Vol. 115, No. 7, pp. 977-984, (2005) [Non-Patent Document 2] Journal of InvestigativeDermatology (1999) Vol.113, No.1 p.98-106 [Non-Patent Document 3] Journal of Dermatologica Science August 2015,Volume 79,Issue2,Pages 119-126 [Non-Patent Document 4] Nature Reviews Microbiology,9(4)p.244-253,2011 [Non-Patent Document 5] Journal of Cosmetic Dermatology 2019,Volume 18,Issue 2,p.652-658 Summary of the Invention [Problem to be solved by the invention]

[0011] The present invention aims to provide a means for suppressing a decrease in the diversity of human resident skin flora and improving the balance of the resident skin flora, thereby maintaining a good skin environment, and in particular for improving disturbance or imbalance of the resident skin flora caused by preservatives and bactericides that are often incorporated into topical skin preparations and cosmetics. [Means for solving the problem]

[0012] In order to solve the above problems, the inventors have conducted extensive research and have unexpectedly discovered that lactosucrose is effective in improving the balance of the normal skin flora, and in particular, inhibits the disruption of the normal skin flora caused by preservatives and disinfectants, thereby maintaining a good skin environment, thus completing the present invention.

[0013] That is, the present invention solves the above-mentioned problems by providing an agent for improving the balance of normal skin flora, which contains lactosucrose as an active ingredient. Effect of the Invention

[0014] The agent for improving the balance of resident skin flora of the present invention can improve the balance of resident skin flora to a favorable state, and can suppress the decrease in the diversity of resident skin flora caused by preservatives and bactericides, and the decrease in resident skin bacteria that are useful for maintaining a healthy skin environment, thereby suppressing the disruption of resident skin flora and maintaining the skin in a healthy state. [Brief description of the drawings]

[0015] [Figure 1] FIG. 1 shows the areas on the face where a test sample and a control sample were applied in a lactosucrose-containing carbohydrate application test. [Diagram 2] FIG. 1 shows the diversity index of all subjects before application of the test sample and the control sample in a lactosucrose-containing carbohydrate application test. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0016] The present invention relates to a skin flora balance improving agent containing lactosucrose as an active ingredient. Lactosucrose is a trisaccharide consisting of galactose, glucose and fructose, and its chemical structure is represented by O-β-Gal-(1→4)-O-α-D-Glc-(1←2)-β-D-Fru (wherein "Gal" represents a galactose residue, "Glc" represents a glucose residue and "Fru" represents a fructose residue). Lactosucrose is a naturally occurring carbohydrate with a mild sweetness, and is a safe and useful carbohydrate that is widely used in the fields of various foods and beverages as a sweetener, excipient, stabilizer, thickener, moisturizer, gloss-imparting agent, etc., and further as a sugar source selected by bifidobacteria.

[0017] The lactosucrose used in the balance improving agent of the present invention may be a natural product, may be chemically synthesized, or may be produced by fermentation or enzymatic method. For example, as a method for producing lactosucrose or lactosucrose-containing carbohydrates by enzymatic method, a method is known in which an aqueous solution containing sucrose as a donor of fructosyl group and lactose as an acceptor is treated with fructosyltransferase to produce lactosucrose, and glucose and unreacted substrates are removed and purified. As long as it does not interfere with the effect of the present invention, it is not necessary to use purified or isolated lactosucrose, and it may be in the form of an unseparated composition, or a partially purified or highly purified composition, or it may be in any form of liquid, powder, or solid, and commercially available lactosucrose-containing carbohydrates may be used. Commercially available products include, for example, the liquid "NycOligo (registered trademark) 400" (lactosucrose content per solid: approximately 42-47% by mass), "NycOligo (registered trademark) 550" (lactosucrose content per solid: approximately 55-60% by mass), and "NycOligo (registered trademark) 700" (lactosucrose content per solid: approximately 68-73% by mass), all sold by Hayashibara Co., Ltd., and the powdered "NycOligo (registered trademark) 700 Powder" (lactosucrose content per solid: approximately 68-73% by mass). Pearl Ace Co., Ltd. also sells the liquid "LS-40L" (lactosucrose content per solid: approximately 42-47% by mass), "LS-55L" (lactosucrose content per solid: approximately 55-60% by mass), and the powdered "LS-55P" (lactosucrose content per solid: approximately 55-60% by mass), and "LS-90P" (lactosucrose content per solid: approximately 88-93% by mass).

[0018] In the present invention, the "resident skin flora" refers to the flora growing on human skin, and although there are individual differences or differences depending on the site, it is known to be composed of bacteria belonging to the genus Propionibacterium, such as acne bacteria, bacteria belonging to the genus Staphylococcus, such as Staphylococcus aureus, bacteria belonging to the genus Streptococcus, such as Streptococcus thermophilus, bacteria belonging to the genus Corynebacterium, which are considered to be harmful bacteria, and bacteria belonging to the genus Anaerococcus.

[0019] In the present invention, "improving the balance of the resident skin flora" means maintaining the appropriate state of the resident skin flora, that is, maintaining the diversity of the resident skin flora as described above. It is known that an abnormal increase or decrease in a specific resident skin flora can have various adverse effects on the skin. Therefore, in order to maintain the health of the skin, it is very important to maintain the appropriate state of the resident skin flora.

[0020] Whether or not the skin resident bacteria are in a proper balance can be examined by measuring the skin resident flora using the measurement method described below. The following will explain. The skin resident flora can be collected by using a wet cotton swab, tape stripping, etc. Since the skin resident flora is constantly changing due to external stress such as washing the face or washing the body, for example, it is desirable to analyze the bacteria collected after adjusting the conditions such as the face washing method and frequency, the collection site, the collection method, and the collection time for the face, which is the target site for skin care. In addition, since the bacterial species of the skin resident flora are alive at the time of collection, depending on the storage condition, the composition of the resident flora may change due to the influence of bacterial growth, etc., so it is desirable to perform the analysis promptly after collection. For the analysis of the skin resident flora, it is possible to analyze all the bacterial species that make up the skin resident flora and their abundance ratio by using metagenomic analysis using a next-generation sequencer. Contract analysis services for resident skin flora are provided by, for example, World Fusion, Inc. and Takara Bio Inc., but there are no particular limitations as long as the method can clarify the state of resident skin flora and the proportion of each bacterial species present.

[0021] In the present invention, "improving the balance of skin flora" means improving and / or maintaining the diversity of skin flora. The diversity of skin flora can be quantified by the Simpson index, which is calculated from the number of species constituting the flora and the ratio of each species to the total. That is, there are two indices of the diversity of skin flora: the abundance of the species constituting the flora and the evenness of the species. The abundance of the species indicates the number of species present in the flora, and the flora is more diverse as the number of species increases. However, even if the number of species contained in the flora is about the same, if there are many bacteria of a specific species and few bacteria of other species, the diversity is low. In addition, the equality of the number of bacteria among the various species present in the flora is called the evenness of species, and a diversity index that takes into account both the abundance and the evenness of species is used to express the diversity of species. The diversity index can be obtained as the Simpson index (D) calculated by the following formula 1.

[0022] formula 1

number

[0023] In formula 1, S represents the number of species contained in the flora. ni represents the number of bacteria of species i, N represents the total number of bacteria, and ni / N represents the proportion of species i in the total number of bacteria in the flora. The higher the diversity, the closer the Simpson index is to 1, and the lower the diversity, the closer it is to 0. In other words, in the case of skin flora, the closer the Simpson index is to 1, the better the flora is.

[0024] One of the factors that disrupt the balance of the skin resident flora is the preservatives and bactericides contained in various skin external preparations and cosmetics that are applied to the skin, and the "improvement of the balance of the skin resident flora" in this specification also includes the suppression of the decrease in the diversity of the skin resident bacteria caused by the preservatives and bactericides contained in the skin external preparations. In general, preservatives and bactericides are blended in skin external preparations and cosmetics to prevent quality deterioration due to bacterial contamination and decay. Preservatives and bactericides contain bactericidal components, and when a skin external preparation containing a preservative or bactericide is applied to the skin of the face, neck, arms, armpits, legs, etc., the balance of the skin resident flora is disrupted by the influence of the preservative and bactericidal components, such as the death or reduction of specific bacteria. The skin resident flora balance improving agent of the present invention has the effect of suppressing the disruption of the balance of the skin resident flora by blending it with a preservative or bactericide in a skin external preparation or cosmetics. Examples of preservatives and disinfectants commonly used in general skin topical preparations include paraoxybenzoic acid alkyl esters such as phenol, parachlorophenol, parachlorometacresol, parachlorometaxylenol, resorcin, resorcin monoacetate, orthophenylphenol, isobutylparaben, ethylparaben, ethylparaben sodium, butylparaben, butylparaben sodium, propylparaben, propylparaben sodium, methylparaben, and methylparaben sodium, phenoxyethanol, thymol, cresol, hinokitiol, and hydroxybenzathiol, and other phenols, benzoic acid and its salts, salicylic acid and its salts, sorbic acid and its salts, dehydroacetic acid and its salts, 2,4,4'-trichloro-2'-hydroxydiphenyl ether, hexachlorophene, bithionol, and dichlorophene, and other halogenated bisphenols, 3,4,4'-trichlorocarbanilide, 3-trifluoromethyl 4,4'-dichlorocarbanilide, 3,4',Amides such as 5-tribromosalicylanilide, p-aminobenzenesulfonamide, and undecylenic acid monoethanolamide; quaternary ammonium compounds such as benzalkonium chloride, alkylisoquinolinium bromide, benzethonium chloride, cetylpyridinium chloride, dequalinium chloride, and alkyltrimethylammonium chloride; zwitterionic surfactants such as alkylaminoethylglycine chloride and sodium stearyl hydroxyethyl betaine chloride; photosensitizers such as chlorhexidine gluconate, tetramethylthiuram disulfide, photosensitizer No. 101, photosensitizer No. 201, photosensitizer No. 301, photosensitizer No. 401, and photosensitizer No. NK-143; 1-hydroxypyridine-2-thione (zinc pyrithione), imidazolidinyl urea, and N-trichloromethyl-mercapto-4-cyclohexene 1 ,2-dicarboximide, lysozyme chloride, chlorphenesin, chlorobutanol, 2-bromo-2-nitro-1,3-propanediol, anise oil, sea lion oil, citronellol, eugenol, benzyl alcohol, ethanol, 1,3-butylene glycol, hexylene glycol, dipropylene glycol, glycerin, 1,2-propanediol, 1,2-butanediol, 1,2-pentanediol (pentylene glycol), 1,2-hexanediol, 1,2-heptanediol, 1,2-octanediol, 1,2-nonanediol, 1,2-decanediol and other alcohols, parachlormetacresol, chlorhexidine chloride, bis(2-pyridylthio-1-oxide) zinc, thianthol, isopropyl methylphenol, 6-acetoxy-2,Examples of such compounds include 4-dimethyl-m-dioxane, 5-chloro-2-methyl-4-isothiazolin-3-one, 2-methyl-4-isothiazolin-3-one, chlorine, sodium hypochlorite, bleaching powder, iodine, formaldehyde, glutaraldehyde, diethyl pyrocarbonate, ethylene oxide, β-propiolactone, hydrogen peroxide, sulfur, potassium gluconate, DMDM ​​hydantoin, alkyl (C12-14) diethylaminoethyl glycine hydrochloride, quaternium-80, quaternium-87, chloramine T, cymene-5-ol, propyl iodide butylcarbamate, polyaminopropyl biguanide, methylisothiazolinone, methylchloroisothiazolinone, and dimethylaminostyrylheptylmethylthiazolium iodide.

[0025] In particular, the skin resident flora balance improving agent of the present invention is excellent in suppressing fluctuations in the abundance ratio of bacteria belonging to the genera Staphylococcus, Anaerococcus, Propionibacterium, and Corynebacterium, which account for the majority of the skin resident flora.

[0026] When the agent for improving the balance of normal skin flora of the present invention is incorporated into skin external preparations or cosmetics for the purpose of improving the balance of normal skin flora, the content of lactosucrose is preferably adjusted to be 0.1% by mass or more and 10% by mass or less, more preferably 0.5% by mass or more and 8% by mass or less, and even more preferably 1% by mass or more and 6% by mass or less, based on the total mass of the skin external preparation or cosmetics. If the content of lactosucrose is less than 0.1% by mass, the effect of improving the balance of normal skin flora may be insufficient, and even if it is used at more than 10% by mass, the effect is not significantly different from that when it is incorporated at 10% by mass, so it is not economically preferable.

[0027] The form of the agent for improving the balance of resident skin flora of the present invention is not particularly limited, and it may be in any appropriate form such as a liquid, ointment, gel, or cream.

[0028] In a suitable case, the agent for improving the balance of normal skin flora of the present invention not only improves the balance of normal skin flora of a person to whom it is applied, but also has the effect of improving skin properties, such as reducing the dryness of the skin, improving the moist feeling, and increasing the moisture content of the stratum corneum.

[0029] As described above, a skin topical preparation or cosmetic comprising the agent for improving the balance of resident skin flora of the present invention may contain a preservative or a bactericide, and further may contain optional ingredients such as the following depending on the dosage form and purpose, to the extent that the effect of improving the balance of resident skin flora is not hindered: surfactants, alcohols, polyhydric alcohols, liquid oil, silicone oil, waxes, oils and fats and other oily ingredients, powders, thickeners, pigments, dyes, fragrances, ultraviolet absorbers, vitamins, pH adjusters, etc.

[0030] Surfactants include anionic surfactants (alkyl carboxylates, alkyl sulfonates, alkyl sulfates, alkyl phosphates), cationic surfactants (alkylamine salts, alkyl quaternary ammonium salts), amphoteric surfactants: carboxylate type amphoteric surfactants (amino type, betaine type), sulfate ester type amphoteric surfactants, sulfonate type amphoteric surfactants, phosphate ester 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), and other surfactants (natural surfactants, derivatives of protein hydrolysates, polymeric surfactants, surfactants containing titanium and silicon, fluorocarbon surfactants).

[0031] Examples of alcohols include natural alcohols such as ethanol, isopropanol, lauryl alcohol, cetanol, stearyl alcohol, oleyl alcohol, lanolin alcohol, cholesterol, phytosterol, and phenoxyethanol, and synthetic alcohols such as 2-hexyldecanol, isostearyl alcohol, and 2-octyldodecanol.

[0032] Examples of polyhydric alcohols include ethylene oxide, ethylene glycol, diethylene glycol, triethylene glycol, ethylene glycol monoethyl ether, ethylene glycol monobutyl ether, diethylene glycol monomethyl ether, diethylene glycol monoethyl ether, polyethylene glycol, propylene oxide, propylene glycol, polypropylene glycol, 1,3-butylene glycol, pentyl glycol, glycerin, pentaerythritol, threitol, arabitol, xylitol, ribitol, galactitol, sorbitol, mannitol, lactitol, and maltitol.

[0033] Examples of liquid vegetable oils include macadamia nut oil, olive oil, avocado oil, rosehip oil, camellia oil, liquid shea butter, almond oil, tea seed oil, sasanqua oil, safflower oil, sunflower oil, soybean oil, cottonseed oil, sesame oil, corn oil, peanut oil, rapeseed oil, rice bran oil, rice germ oil, wheat germ oil, pearl barley oil, grape seed oil, carrot oil, castor oil, linseed oil, and coconut oil. Examples of liquid paraffins include light isoparaffin, light liquid isoparaffin, liquid isoparaffin, and liquid paraffin. Furthermore, examples of saturated fatty acid esters and triglycerides that are liquid oils include isopropyl myristate, octyldodecyl myristate, myristyl myristate, isostearyl myristate, isopropyl palmitate, 2-ethylhexyl palmitate, diisopropyl adipate, diisobutyl adipate, caprylic / capric triglyceride, and glyceryl tricaprylate.

[0034] Examples of silicone oils include dimethylpolysiloxane, methylphenylpolysiloxane, diphenylpolysiloxane, octamethylcyclotetrasiloxane, decamethylcyclopentasiloxane, dodecamethylcyclohexasiloxane, silicone rubber, amino-modified polysiloxane, polyether-modified polysiloxane, alkyl-modified polysiloxane, and fluorine-modified polysiloxane.

[0035] Examples of waxes include beeswax, carnauba wax, spermaceti, lanolin, liquid lanolin, reduced lanolin, hard lanolin, candelilla wax, montan wax, shellac wax, and rice wax.

[0036] Examples of fats and oils include avocado oil, almond oil, fennel oil, perilla oil, olive oil, orange oil, orange roughage oil, sesame oil, cacao butter, chamomile oil, carrot oil, cucumber oil, beef tallow fatty acid, kukui nut oil, safflower oil, shea butter, liquid shea butter, soybean oil, camellia oil, corn oil, rapeseed oil, persic oil, castor oil, cottonseed oil, peanut oil, turtle oil, mink oil, egg yolk oil, palm oil, palm kernel oil, Japan wax, coconut oil, beef tallow, lard, squalene, squalane, and pristane.

[0037] Examples of powders include inorganic powders such as aluminum oxide, magnesium oxide, zirconium oxide, magnesium carbonate, calcium carbonate, aluminum silicate, magnesium silicate, aluminum magnesium silicate, mica, synthetic mica, synthetic sericite, talc, kaolin, silica, silicon carbide, and boron nitride; and organic powders such as magnesium stearate, zinc stearate, N-acyl lysine, and nylon.

[0038] Examples of thickening agents include guar gum, locust bean gum, queen seed, carrageenan, galactan, gum arabic, tara gum, tamarind, furcellan, karaya gum, aoi, caragam, tragacanth gum, pectin, pectinic acid and its salts such as sodium salt, alginic acid and its salts such as sodium salt, mannan, starch such as rice, corn, potato, wheat, etc., dextrin, xanthan gum, dextran, succinoglucan, curdlan, hyaluronic acid and its salts, xanthan gum, pullulan, gellan gum, chitin, chitosan, agar. , cassou extract, chondroitin sulfate, casein, collagen, gelatin, albumin, methyl cellulose, ethyl cellulose, hydroxypropyl cellulose, hydroxyethyl cellulose, carboxymethyl cellulose and its sodium salts, methylhydroxypropyl cellulose, sodium cellulose sulfate, dialkyldimethylammonium cellulose sulfate, crystalline cellulose, cellulose powder and other cellulose and derivatives thereof, soluble starch, carboxymethyl starch, methylhydroxypropyl starch, methyl starch and other starches. Polymers, starch derivatives such as hydroxypropyltrimonium starch chloride and corn starch aluminum octenylsuccinate, alginic acid derivatives such as propylene glycol alginate, polyvinylpyrrolidone (PVP), polyvinyl alcohol (PVA), vinylpyrrolidone-vinyl alcohol copolymer, vinyl alcohol-vinyl acetate copolymer, polyvinyl methyl ether, polyethylene glycol, polypropylene glycol, polyoxyethylene-polyoxypropylene copolymer, amphoteric methacrylate ester copolymers such as (methacryloyloxyethyl carboxybetaine / alkyl methacrylate) copolymer, (acrylates / stearyl acrylate / ethylamine oxide methacrylate) copolymer, (dimethicone / vinyl dimethicone) crosspolymer, (alkyl acrylate / diacetone acrylamide) copolymer, (alkyl acrylate / diacetone acrylamide) copolymer AMP, polyvinyl acetate partially saponified product, maleic acid copolymer, polyvinylpyrrolidone, vinylpyrrolidone-dialkylaminoalkyl methacrylate copolymer; acrylic resin alkanolamine,Polyester, water-dispersible polyester, polyacrylamide, polyacrylic acid ester copolymers such as polyethyl acrylate, carboxyvinyl polymer, polyacrylic acid and its salts such as sodium salt, acrylic acid-methacrylic acid ester copolymer, acrylic acid-alkyl methacrylate copolymer, cationic cellulose such as polyquaternium-10, diallyldimethylammonium chloride-acrylamide copolymer such as polyquaternium-7, acrylic acid-diallyldimethylammonium chloride copolymer such as polyquaternium-22, acrylic acid-diallyldimethylammonium chloride-acrylamide copolymer such as polyquaternium-39, acrylic acid-cationized methacrylic acid ester copolymer, acrylic acid-cationized methacrylic acid amide copolymer, acrylic acid-methyl acrylate-methacrylamidopropyltrimethylammonium chloride copolymer such as polyquaternium-47, methacrylic acid choline ester chloride polymer, cationic oligosaccharide, cationic dextran, cationic polysaccharide such as guar hydroxypropyltrimonium chloride, polyethyleneimine, cationic polymer, Polymers of 2-methacryloyloxyethyl phosphorylcholine such as polyquaternium-51 and copolymers with butyl methacrylate copolymers, etc., polymer emulsions such as acrylic resin emulsions, polyethyl acrylate emulsions, polyacrylic alkyl ester emulsions, polyvinyl acetate resin emulsions, natural rubber latex, synthetic latex, and other polymer emulsions, nitrocellulose, polyurethanes and various copolymers, various silicones, various silicone copolymers such as acrylic-silicone graft copolymers, various fluorine-based polymers, 12-hydroxystearic acid and its salts, dextrin fatty acid esters such as dextrin palmitate and dextrin myristate, cholesteryl hydroxystearate, anhydrous silicic acid, fumed silica (ultrafine silicic acid anhydrous), magnesium aluminum silicate, magnesium sodium silicate, metal soaps, dialkyl phosphate metal salts, bentonite, hectorite, organically modified clay minerals, sucrose fatty acid esters, fructooligosaccharide fatty acid esters, quince seed extract, hydroxyethyl guar gum, carboxymethyl guar gum, starch, carboxymethyl chitin, and other natural polymeric substances,Semisynthetic polymer substances and synthetic polymer substances are included.

[0039] Examples of pigments include extender pigments such as talc, kaolin, mica, sericite, calcium carbonate, magnesium carbonate, magnesium silicate, and anhydrous silicic acid; white pigments such as titanium oxide and zinc oxide; colored pigments such as yellow iron oxide, red ocher, black iron oxide, gunjo, konjo, chromium oxide, chromium hydroxide, and carbon black; pearl pigments such as fish scale foil, bismuth oxychloride, titanium mica, and titanium colored mica; powder pigments such as magnesium, calcium, and aluminum salts of stearic acid, zinc salts of lauric acid, palmitic acid, and myristic acid, starch powder, silk powder, nylon powder, powdered polymethyl methacrylate, and powdered polyethylene.

[0040] Dyes include 5-amino orthocresol, 2-amino-4-nitrophenol, 2-amino-5-nitrophenol, 1-amino-4-methylaminoanthraquinone, 3,3'-iminodiphenol, 2,4-diaminophenoxyethanol hydrochloride, 2,4-diaminophenol hydrochloride, toluene-2,5-diamine hydrochloride, nitroparaphenylenediamine hydrochloride, paraphenylenediamine hydrochloride, N-phenylparaphenylenediamine hydrochloride, metaphenylenediamine hydrochloride, and orthoaminophenon. , N-phenyl-p-phenylenediamine acetate, 1,4-diaminoanthraquinone, 2,6-diaminopyridine, 1,5-dihydroxynaphthalene, toluene-2,5-diamine, toluene-3,4-diamine, nitro-p-phenylenediamine, para-aminophenol, para-nitro-ortho-phenylenediamine, para-phenylenediamine, para-methylaminophenol, picramic acid, sodium picramate, N,N´-bis(4-aminophenyl)-2,5-diamino-1,4-quinone diamine amine, 5-(2-hydroxyethylamino)-2-methylphenol, N-phenyl paraphenylenediamine, meta aminophenol, meta phenylenediamine, 5-amino orthocresol sulfate, 2-amino-5-nitrophenol sulfate, ortho aminophenol sulfate, ortho chloro paraphenylenediamine sulfate, 4,4'-diaminodiphenylamine sulfate, 2,4-diaminophenol sulfate, toluene-2,5-diamine sulfate, nitro paraphenylenediamine sulfate, para aminophenol sulfate, para nitro ortho phenylenediamine sulfate, para nitro meta phenylenediamine sulfate, para phenylenediamine sulfate, para methyl aminophenol sulfate, meta aminophenol sulfate, meta phenylenediamine sulfate, catechol, diphenylamine, α-naphthol, hydroquinone, pyrogallol, phloroglucin, gallic acid, resorcin, tannic acid, sodium 2-hydroxy-5-nitro-2',4'-diaminoazobenzene-5'-sulfonate, and hematein.

[0041] Fragrances include natural animal fragrances such as musk, civet, castoreum, and ambergris, anise essential oil, angelica essential oil, ylang-ylang essential oil, iris essential oil, fennel essential oil, orange essential oil, cananga essential oil, caraway essential oil, cardamom essential oil, guaiac wood essential oil, cumin essential oil, black letter essential oil, cassia bark essential oil, cinnamon essential oil, geranium essential oil, copaiba balsam essential oil, coriander essential oil, shiso essential oil, cedarwood essential oil, citronella essential oil, jasmine essential oil, ginger grass essential oil, cedar essential oil, spearmint essential oil, Western hawthorn essential oil, and other essential oils. Examples of plant-based fragrances include kaolin essential oil, aniseed essential oil, tuberose essential oil, clove essential oil, orange flower essential oil, wintergreen essential oil, tolu balsam essential oil, butterbur essential oil, rose essential oil, palmarosa essential oil, cypress essential oil, Japanese cedar essential oil, sandalwood essential oil, petitgrain essential oil, bay essential oil, vetiver essential oil, bergamot essential oil, Peru balsam essential oil, bois de rose essential oil, camphor essential oil, mandarin essential oil, eucalyptus essential oil, lime essential oil, lavender essential oil, linaloe essential oil, lemongrass essential oil, lemon essential oil, rosemary (rosemary) essential oil, and Japanese peppermint essential oil, as well as other synthetic fragrances.

[0042] Examples of ultraviolet absorbents include paraaminobenzoic acid-based ultraviolet absorbents such as paraaminobenzoic acid, paraaminobenzoic acid monoglycerin ester, N,N-dipropoxyparaaminobenzoic acid ethyl ester, N,N-diethoxyparaaminobenzoic acid ethyl ester, N,N-dimethylparaaminobenzoic acid ethyl ester, and N,N-dimethylparaaminobenzoic acid butyl ester; anthranilic acid-based ultraviolet absorbents such as homomenthyl-N-acetylanthranilate; cinnamic acid; and oxybenzone- 3. Benzophenone derivatives such as oxybenzone-4, oxybenzone-5, and oxybenzone-9; sugar-based UV absorbers in which sugar or sugar alcohol is bound to a 1,3-propanedione derivative; salicylic acid and its sodium salts, amyl salicylate, menthyl salicylate, homomenthyl salicylate, octyl salicylate, phenyl salicylate, benzyl salicylate, p-isopropanol phenyl salicylate, dipropylene glycol salicylate, and other salicylic acid-based UV absorbers. Absorbents, Octyl Cinnamate, Ethyl-4-Isopropyl Cinnamate, Methyl-2,5-Diisopropyl Cinnamate, Ethyl-2,4-Diisopropyl Cinnamate, Methyl-2,4-Diisopropyl Cinnamate, Propyl-p-Methoxy Cinnamate, Isopropyl-p-Methoxy Cinnamate, Isoamyl-p-Methoxy Cinnamate, 2-Ethylhexyl-p-Methoxy Cinnamate, 2-Ethoxyethyl-p-Methoxy Cinnamate (Cinoxate), Cyclohexyl Cinnamic acid-based UV absorbers such as sil-p-methoxycinnamate, ethyl-α-cyano-β-phenylcinnamate, 2-ethylhexyl α-cyano-β-phenylcinnamate (Octocrine), glyceryl mono-2-ethylhexanoyl-di-para-methoxycinnamate, ferulic acid and its derivatives, 2,4-dihydroxybenzophenone, 2,2'-dihydroxy-4-methoxybenzophenone, 2,2'-dihydroxy-4,4'-dimethoxybenzophenone, 2,2',4,Benzophenone-based ultraviolet absorbers such as 4'-tetrahydroxybenzophenone, 2-hydroxy-4-methoxybenzophenone, 2-hydroxy-4-methoxy-4'-methylbenzophenone, 2-hydroxy-4-methoxybenzophenone-5-sulfonate, 4-phenylbenzophenone, 2-ethylhexyl-4'-phenyl-benzophenone-2-carboxylate, 2-hydroxy-4-n-octoxybenzophenone, and 4-hydroxy-3-carboxybenzophenone, 3-(4'-methylbenzylidene) -d,l-camphor, 3-benzylidene-d,l-camphor, 2-phenyl-5-methylbenzoxazole, 2,2´-hydroxy-5-methylphenylbenzotriazole, 2-(2´-hydroxy-5´-t-octylphenyl)benzotriazole, 2-(2´-hydroxy-5´-methylphenylbenzotriazole, dibenzalazine, dianisoylmethane, 5-(3,3-dimethyl-2-norbornylidene)-3-pentan-2-one, 4-methoxy-4´-t-butyldibenzoylmethane, 4-t- dibenzoylmethane derivatives such as butyl methoxydibenzoylmethane, octyl triazone, urocanic acid derivatives such as urocanic acid and ethyl urocanate, hydantoin derivatives such as 2-(2'-hydroxy-5'-methylphenyl)benzotriazole, 1-(3,4-dimethoxyphenyl)-4,4-dimethyl-1,3-pentanedione, dimethoxybenzylidene dioxoimidazolidinepropionate octyl, dimethoxybenzylidene dioxoimidazolidinepropionate 2-ethylhexyl, phenylbenzyl Midasozole sulfonic acid, terephthalidene dicamphor sulfonic acid, drometrizole trisiloxane, methyl anthranilate, rutin and its derivatives, oryzanol and its derivatives, octyl dimethyl paraaminobenzoate, titanium oxide, zinc oxide, kaolin, talc, ethylhexyl triazone, octocrylene, dimethyl PABA octyl, homosalate, methylene bisbenzotriazolyl tetramethylbutylphenol, ethylhexyl methoxycinnamate, octyl methoxycinnamate, etc.

[0043] Vitamins include vitamin A group: retinol, retinal (vitamin A1), dehydroretinal (vitamin A2), carotene, lycopene (provitamin A), vitamin B group: thiamine hydrochloride, thiamine sulfate (vitamin B1), riboflavin (vitamin B2), pyridoxine (vitamin B6), cyanocobalamin (vitamin B12), folic acid, nicotinic acid, pantothenic acid, biotin, choline, inositol, vitamin C group: vitamin C acid or its derivatives, vitamin D group: ergocalciferol, These include vitamin D2, cholecalciferol (vitamin D3), dihydrotachysterol, vitamin E group: vitamin E or its derivatives, ubiquinones, vitamin K group: phytonadione (vitamin K1), menaquinone (vitamin K2), menadione (vitamin K3), menadiol (vitamin K4), and others, essential fatty acids (vitamin F), carnitine, ferulic acid, γ-oryzanol, orotic acid, vitamin P group (rutin, eriocitrin, hesperidin), vitamin U, etc.

[0044] Examples of pH adjusters include citric acid, sodium citrate, lactic acid, sodium lactate, glycolic acid, succinic acid, acetic acid, sodium acetate, potassium acetate, malic acid, tartaric acid, fumaric acid, phosphoric acid, hydrochloric acid, sulfuric acid, boric acid, borax, nitrilotriethanol, monoethanolamine, diethanolamine, triethanolamine, isopropanolamine, triisopropanolamine, 2-amino-2-methyl-1,3-propanediol, 2-amino-2-hydroxymethyl-1,3-propanediol, arginine, ammonium hydroxide, sodium hydroxide, potassium hydroxide, calcium hydroxide, magnesium hydroxide, potassium hydrogen phosphate aqueous ammonia, guanidine carbonate, ammonium carbonate, sodium carbonate, potassium carbonate, potassium bicarbonate, sodium bicarbonate, magnesium oxide, calcium oxide, ammonium phosphate, sodium monohydrogen phosphate, sodium dihydrogen phosphate sodium phosphate, potassium phosphate, magnesium phosphate, calcium phosphate, sodium borate, sodium metaborate, and potassium citrate.

[0045] Examples of topical skin preparations containing the skin resident flora balance improving agent of the present invention include topical liquid preparations, topical gel preparations, cream preparations, ointments, liniments, lotions, poultices, plasters, sprays, aerosol preparations, etc.

[0046] Examples of cosmetics containing the skin resident bacterial flora balance improver of the present invention include skin care cosmetics such as lotions, beauty serums, milky lotions, creams, packs, massage products, facial cleansers, cleansing products, and sunscreen products; body care cosmetics such as body powders and body lotions; makeup cosmetics such as bases, foundations, face powders, concealers, eye colors, and lipsticks; ointments, aerosols, and attachments.

[0047] Furthermore, when the above-mentioned topical skin preparations or cosmetics are emulsion compositions, it is possible to incorporate the skin resident flora balance improving agent of the present invention into any of the multi-layer types such as O / W type, W / O type, O / W / O type, W / O / W type, etc.

[0048] As described above, the skin resident flora balance improving agent of the present invention can be suitably used by being incorporated into skin external preparations or cosmetics. Furthermore, the skin resident flora balance improving agent of the present invention can also be used by being impregnated into nonwoven fabrics or the like, that is, in wipe-off sheet-type facial cleansers, sheet-type cleansing agents, and sheet-type wet napkins.

[0049] The present invention will now be described in detail with reference to experiments.

[0050] <Experiment 1: Assimilation test of various carbohydrates by skin flora> Among the resident skin bacteria, Lactobacillus pentosus JCM 1558 and Micrococcus kristinae ATCC 27570, which are considered to be good bacteria, and Propionibacterium acnes JCM 6425 and Staphylococcus aureus ATCC 6538P, which are considered to be bad bacteria, were used to evaluate the assimilation of various carbohydrates. The carbohydrates used were glucose (special reagent grade, Fuji Film Wako Pure Chemical Industries, Ltd.), sucrose (special reagent grade, Fuji Film Wako Pure Chemical Industries, Ltd.), and lactosucrose (purity 96.8% by mass, Ensuiko Sugar Co., Ltd.).

[0051] The previously cultured bacteria were inoculated into 10 mL of a medium suitable for each bacteria to which 1.0% by mass of each carbohydrate had been added, and cultured at either 30 or 37°C for either 2 or 4 days under culture conditions suitable for each bacteria. The amount of residual sugar in the culture solution at the end of the culture was measured using the anthrone-sulfuric acid method, and the sugar assimilation of each bacteria was evaluated on the following 5-point scale based on the residual rate of each sugar calculated based on the sugar content of the medium at the start of the culture. The culture conditions and assimilation results of each skin resident bacteria are shown in Table 1. The residual rate of sugar was calculated by determining the amount of sugar in the culture solution at the start and end of the culture based on D-glucose conversion using the anthrone-sulfuric acid method commonly used in the art, and calculating the percentage of the amount of sugar in the culture solution at the end of the culture relative to the amount of sugar in the culture solution before the start of the culture.

[0052] <Sugar assimilation rate (sugar residual rate)> +++: Residual sugar content is less than 20% ++: Residual sugar content is over 20% and below 50% +: Residual sugar rate is over 50% and below 80% ±: Residual sugar rate is over 80% and below 90% -: Sugar remaining rate is over 90%

[0053] [Table 1]

[0054] As shown in Table 1, glucose was assimilated by all the bacteria used in the test, and sucrose was not assimilated by Propionibacterium acnes, one of the bad bacteria used in the test, but was assimilated by Staphylococcus aureus, another bad bacterium, and two good bacteria. In other words, it was found that glucose and sucrose are assimilated by both good and bad bacteria, although to different degrees. In contrast, lactosucrose was not assimilated by Micrococcus cristinae, a type of good bacterium, but was selectively assimilated by Lactobacillus pentosus, another good bacterium, while it was not assimilated by Propionibacterium acnes and Staphylococcus aureus, which are bad bacteria. From these results, it was expected that lactosucrose is selectively assimilated by the good bacteria that are resident on the skin, and selectively increases the number of good bacteria, thereby improving the resident skin flora.

[0055] <Experiment 2: Effect of lactosucrose-containing carbohydrates on skin flora> Based on the results of Experiment 1, we investigated the effect of lactosucrose-containing carbohydrates on the diversity of skin resident bacteria and the abundance ratio of various skin resident bacteria.

[0056] <Experiment 2-1: Preparation of test sample solution> Lactosucrose-containing carbohydrate (product name "Nycto Oligo 700 Powder", lot number: 8G06, lactosucrose purity 72.1% by mass, Hayashibara Sales Co., Ltd.) was dissolved in purified water to a final concentration of 5% by mass, and sterilized by filtering through a filter (Stericup 250 μL durapore 0.45 μm PVDF (manufactured by Merck Millipore Co., Ltd.)) to prepare a test sample solution (lactosucrose-containing carbohydrate solution).

[0057] <Experiment 2-2: Lactosucrose-containing carbohydrate application test> Twenty-three healthy adult women aged 25 to 55 years were used as subjects, and the application test was performed on the skin of the face as the test site. When applying the test sample solution or purified water (control) to the face, the subjects washed their faces before application, and 1 mL of the test sample solution or purified water dispensed in the container was poured onto cotton. As shown in Figure 1, the test sample solution was applied to the right half of the subject's face, and the purified water was applied to the left half of the subject's face. The subjects were asked to apply the test sample solution twice a day, morning and night, for 14 consecutive days. In order to prevent overlapping of the application sites of the test sample solution and purified water, the sample was not applied to a width of about 2 cm in the vertical center of the face. During the test period, the subjects were instructed to continue using the same cosmetics they normally use, except for avoiding the use of skin cleansing sheets containing alcohol that affect the normal bacteria on the skin.

[0058] <Experiment 2-3: Collection of normal skin flora and flora analysis> The collection of the skin flora was carried out before and after continuous application of the test sample solution or purified water (control). Since some bacteria on the skin surface are damaged by face washing, the skin flora was collected with a cotton swab for collecting bacteria before face washing. The collected samples were entrusted to World Fusion Co., Ltd., who performed 16S ribosomal RNA sequence identification by a conventional method, and the bacterial species and their abundance ratio were calculated to analyze the skin flora. As an index for evaluating the diversity of the skin flora, the diversity index (Simpson index) obtained by formula 1 was calculated. Note that the skin flora varies depending on the surface part of the body even in the same individual, and since the change over time in the state of the skin flora at the same site is important, the state of the skin flora before and after application of each sample was compared. The same applies to the subsequent experiments.

[0059] The average diversity index values ​​for all subjects at the test sites where the test sample solution or purified water (control) was applied before and after continuous application are shown in Table 2. Note that a paired t-test was performed for the diversity index before and after application at each test site, and a significance level of p<0.05 was determined to indicate a significant difference, which is indicated by * in the table.

[0060] [Table 2]

[0061] As shown in Table 2, in the left half of the face where purified water (control) was applied, the diversity index (Simpson index) of the resident skin flora increased slightly from 0.76 before application to 0.79 after application, but the change was not significant. In contrast, in the right half of the face where the test sample solution was applied, the diversity index increased significantly from 0.75 before application to 0.81 after application. These results indicate that the diversity of resident skin bacteria increases by continuously applying lactosucrose-containing carbohydrates to the skin.

[0062] Next, in the same experiment, the presence ratio of Streptococcus thermophilus, a normal skin bacterium that has been reported to repair the skin's barrier function and be useful for moisturizing the skin, was analyzed at each time point before and after continuous application. The average values ​​of the abundance ratio of Streptococcus thermophilus at the test sites where the test sample solution or purified water (control) was applied at each time point before and after continuous application for all subjects are shown in Table 3. Note that for the abundance ratio, a paired t-test was performed, and a significance level of p<0.05 was considered to indicate a significant difference, which is indicated by * in the table.

[0063] [Table 3]

[0064] As shown in Table 3, in the left half of the face where purified water (control) was applied, the abundance ratio of Streptococcus thermophilus tended to increase, from 0.10% before application to 0.18% after application. In contrast, in the right half of the face where the test sample solution was applied, the abundance ratio was significantly increased, from 0.08% before application to 0.17% after application. In other words, by applying lactosucrose-containing carbohydrates to the skin, the diversity of skin resident bacteria increased, and the abundance ratio of skin resident bacteria that are considered to be useful for maintaining a good skin environment increased, indicating that lactosucrose-containing carbohydrates are useful as a material for maintaining skin health.

[0065] <Experiment 2-4: Sensory evaluation> Before the continuous application of the test sample solution or purified water (control), during the continuous application period (7 days after the start of application), and after the continuous application (14 days after the start of application), all subjects were asked to complete a sensory questionnaire survey on the two items of skin condition, "dryness" and "moistness." Regarding the "dryness" and "moistness" of the skin condition at each time point, Feel: 5 Somewhat: 4 Don't know: 3 Not much: 2 Not at all: 1 The scores of all subjects at each time point were evaluated on a 5-point scale. Table 4 shows the scores of all subjects at each time point. The average ranking of the scores of all subjects at each time point was also calculated. The results are shown in Table 5. In addition, Friedman's test and Scheffe's pairwise comparison were performed for the skin condition of each test site before and during or after application, and a risk level of p<0.05 was determined to indicate a significant difference, which is indicated by * in the table, and a risk level of p≦0.05 to 0.10 was determined to indicate a trend, which is indicated by + in the table.

[0066] [Table 4]

[0067] [Table 5]

[0068] As shown in Table 5, no significant change in "dryness" or "moisturization" was observed on the left half of the face where purified water (control) was applied, but on the right half of the face where the test sample solution was applied, the "dryness" was no longer felt during or after application compared to before application, and a "moisturization" was felt. This result shows that applying lactosucrose-containing carbohydrates to the skin can provide a moisturizing effect on the skin.

[0069] These results indicate that applying lactosucrose-containing carbohydrates to the skin improves the diversity of resident skin bacteria and increases the proportion of resident skin bacteria that are useful for maintaining a good skin environment. Lactosucrose-containing carbohydrates can be advantageously used as a material to maintain the health of the skin, and can also be incorporated into external skin preparations such as cosmetics to provide a moisturizing effect on the skin.

[0070] <Experiment 2-5: Effects of lactosucrose-containing carbohydrates on subjects with low diversity of skin flora> In the above-mentioned Experiment 2-3, the diversity index of all subjects before application of the test sample solution or purified water (control) is shown in Figure 2. The diversity index of all subjects was distributed widely from 0.31 to 0.98, with the median being 0.83. Here, six subjects (hereinafter referred to as the "low diversity group"), which are the bottom 1 / 4 of all subjects and are located below the line showing the diversity index of 0.60 in Figure 2, were selected and stratified analysis was performed.

[0071] <Experiment 2-5-1: Collection of normal skin flora and flora analysis> The skin resident flora of the low diversity group was analyzed using the same method as in Experiment 2-3. The average diversity index of the test sites to which the test sample solution or purified water (control) was applied at each time point before and after continuous application in the low diversity group is shown in Table 6. Note that a paired t-test was performed for the diversity index of each test site before and after application, and a significance level of p<0.01 was determined to indicate a significant difference, which is indicated by ** in the table.

[0072] [Table 6]

[0073] As shown in Table 6, in the left half of the face where purified water (control) was applied, the diversity index increased slightly (not significant) from 0.46 before application to 0.57 after application, whereas in the right half of the face where the test sample solution was applied, the diversity index increased significantly from 0.45 to 0.60 after application. These results show that the effect of increasing the diversity of skin resident bacteria by applying lactosucrose-containing carbohydrates to the skin is more pronounced in the low diversity group than in all subjects.

[0074] Next, the ratio of resident skin bacteria was analyzed for the low diversity group. For Streptococcus thermophilus, which has been reported to restore the skin barrier function and be useful for skin moisturization, and Propionibacterium acnes, known as the causative bacterium of acne, the average ratio of each resident skin bacteria at the test sites to which the test sample solution or purified water was applied at each time point before and after continuous application in the low diversity group is shown in Tables 7 and 8. Note that a paired t-test was performed for the ratio of each resident skin bacteria at each test site before and after application, and a risk level of p<0.05 was determined to indicate a significant difference, and p<0.05 and p<0.01 were indicated with * and ** in the tables.

[0075] [Table 7]

[0076] As shown in Table 7, in the left half of the face where purified water (control) was applied, the abundance ratio of Streptococcus thermophilus was 0.02% to 0.06%, with only slight changes in the abundance ratio between before and after continuous application. In contrast, in the right half of the face where the test sample solution was applied, a significant increase was observed, from 0.01% to 0.11%.

[0077] [Table 8]

[0078] As shown in Table 8, the presence ratio of Propionibacterium acnes in the left half of the face where purified water (control) was applied decreased from 69.40% to 61.61%, but the change was not significant. In contrast, in the right half of the face where the test sample solution was applied, the presence ratio decreased significantly from 70.01% to 59.59%.

[0079] <Experiment 2-5-2: Measurement of skin condition> During the sensory evaluation in Experiment 2-4, the moisture content of the stratum corneum was measured three times for all subjects before, during, and after the application of the sample using a skin surface moisture content measuring device (SKICON-200EX, manufactured by IBS Co., Ltd.), and the average value at each time point was calculated. This measuring device evaluates the moisture content of the stratum corneum as skin conductance (electrical conductivity, unit: μS). The central part (1.5 cm x 1.5 cm) of both cheeks of each subject's face was used as the measurement site. The change in the average moisture content of the stratum corneum from before the application of the sample in the low diversity group selected in Experiment 2-5-1 is shown in Table 9. The change in the average moisture content of the stratum corneum before, during, and after the application of the sample at each test site was performed using a paired t-test, and a significance level of p≦0.05 to 0.1 was determined to indicate a trend, and is indicated by + in the table.

[0080] [Table 9]

[0081] As seen in Table 9, in the low diversity group, the change in skin conductance (stratum corneum moisture content) in the left half of the face where purified water (control) was applied was -2.06 and 10.56, showing slight fluctuations (not significant), whereas in the right half of the face where the test sample solution was applied, the change in skin conductance was 6.94 and 25.83, showing a tendency for an increase, indicating an increase in the moisture content of the stratum corneum.

[0082] The above results show that the application of lactosucrose-containing carbohydrates to the skin increases the diversity of resident skin bacteria, balances the ratio of good and bad bacteria, and improves the balance of resident skin flora, leading to a better skin environment. In other words, lactosucrose-containing carbohydrates can be advantageously used as an active ingredient in an agent for improving the balance of resident skin flora.

[0083] <Experiment 3: Effects of preservatives on the skin flora and the effect of lactosucrose-containing carbohydrates> Next, we investigated the effect of preservatives on the balance of normal skin flora and the effect of adding carbohydrates containing lactosucrose.

[0084] <Experiment 3-1: Preparation of test sample solution> A test sample solution was prepared by mixing lactosucrose-containing carbohydrates (product name "Nyuka Oligo 700 Powder", lactosucrose purity 72.1% by mass, Hayashibara Sales Co., Ltd.) and general-purpose cosmetic raw materials in the ratios shown in Table 10, and designated as test sample A. A test sample solution prepared in the same manner as test sample A except that lactosucrose-containing carbohydrates were not mixed was designated as placebo sample B. The formulation shown in Table 10 was manufactured in accordance with European Cosmetics Regulation 1223 / 2009, and the amounts of the preservatives phenoxyethanol and chlorphenesin are below the maximum amounts stipulated in the "Cosmetics Standards."

[0085] [Table 10]

[0086] <Experiment 3-2: Lactosucrose-containing carbohydrate application test> Twenty-three healthy men and women aged 18-70 years (10 men and 13 women) were used as subjects and randomly divided into two groups. For each group, either the left or right armpit was used as the test site. After washing both armpits with unscented soap, one double application (approximately 0.3 mL) of test sample A or placebo sample B packed in a roll-on container was applied to the test site for 14 consecutive days.

[0087] <Experiment 3-3: Collection of normal skin flora and flora analysis> The bacterial flora was collected twice, before and 24 hours after the continuous application of test sample A or placebo sample B. The subjects' armpits were wiped 50 times with a cotton swab for collecting bacteria to collect the resident skin flora. The collected samples were entrusted to ProDerm Institute of Applied Dermatology (Federal Republic of Germany), which performed 16S ribosomal RNA sequence identification by standard methods, and the species, number and abundance ratio of bacteria were calculated to analyze the resident skin flora. As in Experiment 2, a diversity index (Simpson index) was used as an index to evaluate the diversity of resident skin bacteria.

[0088] The average diversity index values ​​before and 24 hours after the end of continuous application in the armpits to which test sample A or placebo sample B was applied are shown in Table 11. Note that a paired t-test was performed on the diversity index before and 24 hours after the end of continuous application of each test sample, and a significance level of p<0.05 was determined to indicate a significant difference, which is indicated by * in the table.

[0089] [Table 11]

[0090] As shown in Table 11, in the armpits where placebo sample B was applied, the diversity index of the skin resident flora was 0.44 before application, but was 0.23 24 hours after continuous application, indicating a significant decrease in the diversity index. Since placebo sample B is a blend of purified water and general-purpose cosmetic ingredients, it was speculated that the significant decrease in the diversity index of the skin resident flora at the test site where placebo sample B was applied was due to the influence of phenoxyethanol and chlorphenesin, which are preservatives added to the general-purpose cosmetic ingredients to maintain the quality of the formulation. In contrast, in the armpits where test sample A containing lactosucrose-containing carbohydrates was applied, the diversity index of the skin resident flora was 0.53 before application, but was 0.52 24 hours after continuous application, indicating no change in the diversity index of the skin resident flora.

[0091] The above results show that even within the range of the amount prescribed in the "Cosmetics Standards," when topical skin preparations containing preservatives are applied to the skin, they have a bactericidal effect on some resident skin bacteria and inhibit their growth, thereby reducing the diversity of resident skin bacteria. However, when lactosucrose-containing carbohydrates are further added, the reduction in the diversity of resident skin bacteria caused by preservatives is mitigated, and the diversity of resident skin bacteria is maintained.

[0092] Analysis of the bacterial flora of all subjects revealed that the bacterial flora of the subjects' armpits was dominated by three types of skin resident bacteria: Staphylococcus, Corynebacterium, and Anaerococcus. The average ratios of these three types of skin resident bacteria in the armpits to which test sample A or placebo sample B was applied before and 24 hours after continuous application are shown in Table 12. In addition, a Wilcoxon signed rank test was performed on the abundance ratio of each skin resident flora before and 24 hours after continuous application of each test sample, and a significance level of p<0.01 was determined to indicate a significant difference, which is indicated with ** in the table.

[0093] [Table 12]

[0094] As shown in Table 12, in the armpits where placebo sample B was applied, the abundance ratio of Corynebacterium was 21.38% before application, but it was significantly reduced to 8.39% 24 hours after continuous application. In addition, the abundance ratio of Anaerococcus was 8.99% before application, but it was significantly reduced to 4.07% 24 hours after continuous application. On the other hand, the abundance ratio of Staphylococcus was 68.64% before application, but it was significantly increased to 87.04% 24 hours after continuous application. It is speculated that these changes in the abundance ratio of skin flora are due to the influence of preservatives such as phenoxyethanol and chlorphenesin contained in placebo sample B.

[0095] In contrast, in the armpits where test sample A containing lactosucrose-containing carbohydrates was applied, as shown in Table 12, for all types of skin resident bacteria, the change in the proportion of skin resident bacteria was very slight between before and after continuous application, and no significant change was observed as was seen with placebo sample B.

[0096] Furthermore, the number of bacteria present in the skin was also analyzed before and 24 hours after the continuous application of the test sample A or the placebo sample B in the armpits. The average number of bacteria at each time point in each sample application group is shown in Table 13.

[0097] [Table 13]

[0098] As seen in Table 13, there was no significant change in the number of resident skin bacteria before and 24 hours after continuous application in the armpits to which test sample A or placebo sample B was applied. In other words, it was shown that application of placebo sample B did not affect the number of bacteria, but reduced the diversity of resident skin bacteria and changed the abundance ratio of various resident skin bacteria.

[0099] As described above, topical skin preparations containing preservatives reduced the diversity of resident skin bacteria and significantly changed the abundance ratio of resident skin bacteria, whereas topical skin preparations further containing lactosucrose-containing carbohydrates did not significantly change the diversity or abundance ratio of resident skin bacteria. This indicates that lactosucrose-containing carbohydrates are useful for suppressing the disruption of resident skin flora caused by preservatives such as bactericides and bacteriostatic agents and for improving the balance of resident skin flora to protect the skin.

[0100] The present invention will be described in more detail below with reference to examples. However, these examples are merely illustrative and the present invention is not limited to these examples. EXAMPLES

[0101] <Emulsion> Ingredients Mass% Stearic Acid 2.0 Cetanol 1.0 Vaseline 3.0 Lanolin Alcohol 2.0 Liquid Paraffin 8.0 Squalane 3.0 Glycyrrhetin 0.5 Allantoin 0.5 Hyaluronic Acid 1.0 Lactosucrose-containing carbohydrates 3.0 (Product name: "Nyuko Oligo 700 Powder", manufactured by Hayashibara Co., Ltd.) POE(10) monooleate 2.5 Triethanolamine 1.0 Propylene glycol 5.0 Preservatives (appropriate amount) Fragrance (appropriate amount) Distilled water remaining Total 100.0

[0102] According to the above recipe, a milky lotion was produced by a conventional method. This product contains lactosucrose-containing carbohydrates, which can suppress the change in the diversity of skin flora caused by preservatives. EXAMPLES

[0103] <Astringent lotion> Ingredients Mass% Dipropylene glycol 2.0 Citric acid 0.03 Sodium citrate 0.05 Lactosucrose-containing carbohydrates 3.0 (Product name: "Nyuko Oligo 700 Powder", manufactured by Hayashibara Co., Ltd.) Histidine 5.0 Ethyl alcohol 15.0 Polyoxyethylene (15 moles added) 0.5 Oleyl alcohol ether Preservatives (appropriate amount) Fragrance (appropriate amount) Distilled water remaining Total 100.0

[0104] According to the above recipe, an astringent lotion was produced by a conventional method. This product contains lactosucrose-containing carbohydrates, which can suppress the variation in the diversity of skin flora. EXAMPLES

[0105] <Moisture Cream> Ingredients Mass% (A) Cetyl Phosphate 2.0 Cetanol 4.0 Coconut oil 20.0 Methylheptyl laurate 10.0 Dimethyl silicone 3.0 (B) Glycerin 5.0 Lactosucrose-containing carbohydrates 5.0 (Product name: "Nyuko Oligo 700 Powder", manufactured by Hayashibara Co., Ltd.) Xanthan gum (2% aqueous solution) 5.0 Arginine 1.0 Distilled water remaining Total 100.0

[0106] The above phases A and B were each heated to 80°C to dissolve uniformly, and then while stirring phase A, phase B was added and stirred to make uniform, and the mixture was cooled to 30°C with continued stirring to prepare a moisture cream. This product contains lactosucrose-containing carbohydrates, which can suppress changes in the diversity of resident skin bacteria. EXAMPLES

[0107] <Body cream> Ingredients Mass% (A) Glyceryl Oleate 2.0 Squalane 4.0 Methylheptyl laurate 3.0 Coconut oil 1.5 Fragrance (appropriate amount) Tocopherol 0.05 Preservatives (phenoxyethanol, chlorphenesin) appropriate amount (B) Glycerin 10.0 Lactosucrose-containing carbohydrates 5.0 (Product name: "Nyuko Oligo 700 Powder", manufactured by Hayashibara Co., Ltd.) Magnesium sulfate 0.5 Plant extract 0.3 pH adjuster (appropriate amount) Distilled water remaining Total 100.0

[0108] After mixing phases A and B uniformly at room temperature, phase B was added while stirring phase A, and the mixture was stirred for a while after adding phase B to prepare a body cream. This product contains lactosucrose-containing carbohydrates, which can suppress the change in diversity of resident skin bacteria caused by preservatives. EXAMPLES

[0109] <Moisture Lotion> Ingredients Mass% (A) Chimyl alcohol 2.0 Batyl alcohol 0.5 Palmitic acid 0.1 Stearic acid 0.3 Coconut oil 0.05 Liquid Paraffin 1.0 (B) Sodium stearoyl glutamate 0.3 Lactosucrose-containing carbohydrates 5.0 (Product name: "Nyuko Oligo 700 Powder", manufactured by Hayashibara Co., Ltd.) Arginine 0.2 Glycerin 3.0 Preservatives (phenoxyethanol, chlorphenesin) appropriate amount Distilled water remaining Total 100.0

[0110] The above phases A and B were each uniformly heated to 80°C, and then while stirring phase A, phase B was added and stirred to make it uniform, and the mixture was cooled to 30°C with continued stirring to prepare a moisture lotion. As this product contains lactosucrose-containing carbohydrates, it is able to suppress changes in the diversity of resident skin bacteria caused by preservatives. EXAMPLES

[0111] <Bath additives> Ingredients Mass% (A) PEG-6 Oleate 10.0 Liquid Paraffin 45.0 Squalane 10.0 Coconut oil 5.0 Fragrance (appropriate amount) (B) Glycerin 25.0 Lactosucrose-containing carbohydrates 5.0 (Product name: "Nyuko Oligo 700 Powder", manufactured by Hayashibara Co., Ltd.) Plant extract (appropriate amount) 1,3-butylene glycol (appropriate amount) Hydroxypropyltrimonium Hyaluronate 1.0 Ethanol (appropriate amount) Dye appropriate amount Total 100.0

[0112] Phase A was mixed uniformly, and then Phase B was dissolved by heating. Phase A was then mixed uniformly while stirring to prepare the bath additive. This product contains lactosucrose-containing carbohydrates, which can suppress changes in the diversity of resident skin bacteria. EXAMPLES

[0113] <Ointment> Ingredients Mass% (A) Polyglyceryl oleate 2.0 Coconut oil 10.0 Squalane 2.0 Ceresin 5.0 Vaseline 2.0 Preservatives (phenoxyethanol, chlorphenesin) appropriate amount (B) Dipotassium glycyrrhizinate 1.0 Isostearic acid 0.1 Decaglyceryl Laurate 5.0 Glycerin 20.0 Lactosucrose-containing carbohydrates 5.0 (Product name: "Nyuko Oligo 700 Powder", manufactured by Hayashibara Co., Ltd.) Disodium edetate 0.1 Distilled water remaining Total 100.0

[0114] The above phases A and B were each heated to 80℃ to dissolve uniformly, and then phase B was gradually added while stirring phase A, and the ointment was prepared by cooling to 30℃ with stirring. This product contains lactosucrose-containing carbohydrates, which can suppress the change in diversity of resident skin bacteria caused by preservatives. [Industrial Applicability]

[0115] The agent for improving the balance of normal skin flora of the present invention can suppress the imbalance and bias of normal skin flora that occurs when a topical skin preparation containing a preservative is applied to the skin, and can maintain and improve the normal skin flora in a healthy state. The present invention, which provides an agent for improving the balance of normal skin flora, will contribute to the fields of cosmetics, medicines, etc. that use topical skin preparations, and its industrial significance is extremely great.

Claims

1. An agent for application to the skin that contains lactosucrose as an active ingredient and is used to improve the balance of the skin's resident bacterial flora.

2. The agent according to claim 1, wherein the improvement of balance is an increase in the diversity of normal skin flora.

3. The agent according to claim 1 or 2, wherein the improvement of balance is suppression of a decrease in the diversity of normal skin flora caused by a preservative.

4. 4. The agent according to claim 1, wherein the balance improvement is suppression of fluctuations in the abundance ratio of bacteria belonging to the genera Staphylococcus, Anaerococcus, Propionibacterium, and Corynebacterium.

5. The agent according to any one of claims 1 to 4, comprising lactosucrose in an amount of 0.1% by mass or more and 10% by mass or less in the formulation.

6. 6. The agent according to any one of claims 1 to 5, which is in the form of a liquid, ointment, gel or cream.

7. The agent according to any one of claims 1 to 6, which has an effect of improving skin properties.

8. 8. The agent according to claim 1, wherein the improvement in skin properties is a reduction in dryness, an improvement in moistness, and / or an increase in the moisture content of the stratum corneum.

9. A cosmetic or topical skin preparation comprising the agent according to any one of claims 1 to 8.

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