Composition for controlling skin resident flora

Abstract

The present invention relates to a composition for controlling the resident bacterial flora of the skin. A primary object of the present invention is to provide a novel composition for controlling the resident bacterial flora of the skin that can increase or improve the species diversity of the resident bacterial flora of the skin compared to the pre-treatment state. Examples of the present invention include compositions (e.g., ointments or liquids) containing an effective amount of earthworm lipids extracted from Lumbricus rubellus for controlling the species diversity of the resident bacterial flora of the skin. The compositions of the present invention are useful, for example, as raw materials for skin care products and cosmetics.

Description

Technical Field

[0001] The present invention relates to a composition for controlling the resident bacterial flora of the skin, belonging to the technical field of skin care. Specifically, the present invention relates to a composition for controlling the resident bacterial flora of the skin, containing earthworm lipid extracted from earthworms. Background Art

[0002] In addition to the functions of the epidermis and stratum corneum for separating the body from the outside world, the formation of the sebum film also plays an important role in the human skin barrier mechanism. The sebum film is formed by sodium fatty acids. After the sebum secreted by the sebaceous glands is decomposed into free fatty acids by the lipase as a skin microbial source enzyme, it is combined with the sodium contained in the sweat secreted by the sweat glands to generate the sodium fatty acids. The sebum film prevents foreign matter from invading as a physical barrier, and suppresses water evaporation from the epidermis to prevent dry skin. It also has the function of stabilizing the skin surface environment. It is generally believed that when the sebum secretion amount is small and the sebum secretion is sufficient but the sweating amount is small, it is difficult to generate sodium fatty acids, and the formation of the sebum film is insufficient, and the barrier effect is reduced.

[0003] Normally, the skin hosts a balanced community of over 200 microorganisms adapted to this environment, preventing the proliferation of pathogenic bacteria. It is generally believed that some of the free fatty acid isomers produced by the breakdown of sebum by lipases in skin microbes have the effect of inhibiting the proliferation of specific bacterial species, and the abundance ratio of the constituent fatty acids influences the skin microbiome. Furthermore, the composition of the skin microbiome is believed to contribute to differences in resistance to pathogenic microbes.

[0004] Therefore, controlling the composition and diversity of the skin microbiome or resident skin flora to that of healthy skin is crucial for maintaining skin health, preventing the invasion of pathogenic microorganisms, and suppressing inflammation in conditions such as atopic dermatitis. Non-Patent Document 1 reports that the diversity of the resident skin flora is low in sensitive skin.

[0005] As a technology for controlling the resident bacterial flora of the skin, for example, a coating for controlling resident bacteria of the skin containing magnesium lactate and / or calcium lactate as an active ingredient is known (Patent Document 1). Patent Document 1 claims that the coating for controlling resident bacteria of the skin can suppress the decrease in the number of beneficial bacteria among the resident bacteria of the skin and reduce the number of harmful bacteria.

[0006] Meanwhile, earthworms, particularly cultivated earthworms (Lumbricus rubellus), are widely distributed as health supplements and medicines when dried and powdered. These earthworms contain components with antipyretic and analgesic properties, as well as lumbrokinase, an enzyme believed to dissolve blood clots.

[0007] Prior art literature

[0008] Patent Document

[0009] Patent Document 1: Japanese Patent Laid-Open No. 2018-52891

[0010] Non-Patent Document

[0011] Non-Patent Document 1: "Shiseido discovers that the diversity of the skin's resident flora is low in sensitive skin", August 19, 2020, Publisher: Shiseido Company, Limited, Internet (URL: https: / / corp.shiseido.com / jp / news / detail.html?n=00000000002960) Summary of the Invention

[0012] Problems to be solved by the invention

[0013] As described above, controlling the composition or diversity of the skin's resident flora to the same state as that of healthy skin is important for maintaining skin health, preventing the invasion of pathogenic microorganisms, and suppressing inflammation such as atopic dermatitis.

[0014] The main object of the present invention is to provide a new composition for controlling the skin's resident flora, which can increase or improve the species diversity (diversity of microorganisms on the skin) of the skin's resident flora compared to the state before treatment.

[0015] Solutions for solving problems

[0016] The inventors of the present invention conducted in-depth research and found that so-called earthworm lipids extracted from earthworms have the effect of increasing the diversity of skin resident bacteria, etc., thus completing the present invention.

[0017] As the present invention, for example, the following embodiments can be cited.

[0018] [1] A composition for controlling the species diversity of the skin's resident flora, which contains an effective amount of earthworm lipids and a base.

[0019] [2] The composition according to [1] above, wherein the earthworm lipids are obtained from the extraction residue after removing proteins from live earthworms.

[0020] [3] The composition according to [1] or [2] above, wherein the earthworm is Lumbricus rubellus.

[0021] [4] The composition according to any one of [1] to [3] above, which is used for application to an object with a low sebum secretion amount or an object with atopic symptoms on the skin.

[0022] [5] A C16 monounsaturated fatty acid enhancer for the skin, comprising the composition according to any one of [1] to [4].

[0023] [6] The composition according to any one of [1] to [4] above or the C16 monounsaturated fatty acid enhancer on the skin according to [5] above, wherein the dosage form is an ointment, cream, gel, liquid, patch, patch, tablet, aerosol, external powder, spray or bath preparation.

[0024] [7] A cosmetic comprising the composition described in any one of [1] to [4] above or the C16 monounsaturated fatty acid enhancer for the skin described in [5] or [6].

[0025] [8] A composition for controlling resident bacterial flora on the skin, obtained by extracting the residue (earthworm dehydrate) from living earthworms after removing protein using a polar organic solvent; or by saponifying the earthworm dehydrate with an alkali and extracting the saponified product using a non-polar organic solvent.

[0026] [9] An earthworm lipid for controlling the resident bacterial flora of the skin, obtained by extracting the residue (earthworm dehydrate) from living earthworms after removing the protein using a polar organic solvent; or by saponifying the earthworm dehydrate with an alkali and extracting the saponified product using a non-polar organic solvent.

[0027]

[10] The composition for controlling resident bacteria on the skin according to [8] or the earthworm lipid for controlling resident bacteria on the skin according to [9], wherein the polar organic solvent is a mixed solvent of a chlorine-based organic solvent and an alcohol-based organic solvent.

[0028]

[11] The composition for controlling resident bacteria on the skin according to [8] or the earthworm lipid for controlling resident bacteria on the skin according to [9], wherein the non-polar organic solvent is a hydrocarbon organic solvent.

[0029]

[12] A method for producing a composition for controlling the diversity of bacterial species in the skin resident flora, comprising the steps of extracting earthworm lipids from earthworms and mixing the earthworm lipids with a base.

[0030] Effects of the Invention

[0031] According to the present invention, the diversity of microorganisms on the skin (diversity of skin-resident bacteria) can be increased or improved compared to the state before application, thereby improving the barrier function of the skin and suppressing the proliferation of pathogenic bacteria. BRIEF DESCRIPTION OF THE DRAWINGS

[0032] Figure 1The upper panel shows the results for healthy individuals, and the lower panel shows the results for individuals with atopic dermatitis.

[0033] Figure 2 The results of diversity analysis are shown. The left graph shows the results of Simpson's Index analysis, and the right graph shows the results of Shannon Entropy analysis. Each line represents the change in the control and the change in the case of earthworm lipid application, respectively.

[0034] Figure 3 This is a chromatogram (TIC) showing the results of GC / MS analysis. The left panel shows the results of the control, and the right panel shows the results of the application of earthworm lipid. The lines represent the chromatograms of sebum two weeks after application, sebum before application, and earthworm lipid, respectively. ▽ indicates the peak of 16-carbon monounsaturated fatty acids.

[0035] Figure 4 The results of diversity analysis are shown. The left graph shows the results of Simpson's index analysis, and the right graph shows the results of Shannon entropy analysis. Each line represents the change in the control and the change in the case of earthworm lipid application, respectively.

[0036] Figure 5 This is a chromatogram (TIC) showing the results of GC / MS analysis. The left panel shows the results of the control, and the right panel shows the results after applying earthworm lipid. The lines represent the chromatograms of sebum two weeks after application, sebum before application, and earthworm lipid, respectively. ▽ indicates the peak of 16-carbon monounsaturated fatty acids.

[0037] Figure 6 The results of diversity analysis are shown. The left graph shows the results of Simpson's index analysis, and the right graph shows the results of Shannon entropy analysis. Each line represents the change in the control and the change in the case of earthworm lipid application, respectively.

[0038] Figure 7 This is a chromatogram (TIC) showing the results of GC / MS analysis. The left panel shows the results of the control, and the right panel shows the results after applying earthworm lipid. The lines represent the chromatograms of sebum two weeks after application, sebum before application, and earthworm lipid, respectively. ▽ indicates the peak of 16-carbon monounsaturated fatty acids. DETAILED DESCRIPTION

[0039] Hereinafter, embodiments of the present invention will be described in detail.

[0040] 1 Composition of an embodiment of the present invention

[0041] A composition according to an embodiment of the present invention (hereinafter referred to as the "present composition") is characterized in that it is a composition for controlling the species diversity of the skin's resident bacterial flora (the diversity of microorganisms on the skin or the diversity of skin-resident bacteria), and contains an effective amount of earthworm lipid. The present composition can be preferably used to increase or improve the species diversity of the skin's resident bacterial flora compared to the state before application, or to maintain this diversity.

[0042] "Control" here refers to changing or maintaining the number of bacterial species in the resident skin flora or the occupancy rate of various bacterial species in the flora (skin resident flora control), including increasing or improving the diversity of the resident skin flora compared to the state before application of the composition of the present invention (skin resident flora improvement), and maintaining this diversity at a higher level due to skin resident flora improvement or other factors (skin resident flora maintenance). "Diversity" is a concept that encompasses both species richness (a large number of bacterial species) and species uniformity (low variation in the number of individuals between species). The greater the number of bacterial species, the more uniform the number of individuals of each species, or the smaller the variation in occupancy rate, the higher (better) the diversity is. Whether or not there is an increase or improvement, and the extent of the increase or improvement, can be determined by taking these two aspects into consideration, using indicators such as the Simpson's Diversity Index, Shannon Entropy, and the Nakamura RI Index (Nakamura, Setouchi Junior University Bulletin, No. 24: 37-41, 1994). The "effective amount" refers to an amount that allows a person skilled in the art to confirm the effect of the control (eg, improvement or maintenance of the resident bacterial flora of the skin).

[0043] "Earthworm lipids" refers to the diverse lipid groups contained in earthworms. In particular, it refers to the diverse lipid groups contained in all parts of earthworms. This lipid group is usually composed of more than 100 kinds of diverse lipids. This earthworm lipid can be obtained, for example, as follows: extracting and removing protein from living earthworms (earthworm dehydrate) and performing an extraction operation for obtaining lipids with a polar organic solvent to obtain an extract. Examples of such polar organic solvents include mixed organic solvents of chlorine-based organic solvents and alcohol-based organic solvents, specifically, mixed organic solvents of chloroform and methanol, or mixed organic solvents obtained by adding water to these. In addition, the earthworm dehydrate can be saponified with an alkali, and the alkaline solution of the saponified product can be made acidic, and then extracted with a non-polar organic solvent. Examples of such non-polar organic solvents include hydrocarbon-based organic solvents, specifically, hexane, cyclohexane, and heptane.

[0044] The obtained earthworm lipid (total lipid) can be in the form of oil or solid.

[0045] The obtained extract can be added with additives and processed appropriately to prepare a composition (solid, semi-solid, liquid, etc.) containing the extract.

[0046] The earthworms of the present invention are not particularly limited, but are preferably of the genus Lumbricus, and more preferably Lumbricus rubellus, so-called red earthworms.

[0047] The dosage form of the composition of the present invention is not particularly limited as long as it can be directly applied to the skin surface. Examples thereof include ointments, creams, gels, liquids (suspensions, emulsions, lotions, etc.), papules, patches, tablets, aerosols, external powders, sprays, and bath preparations.

[0048] When preparing these preparations, in addition to earthworm lipids, various compounding ingredients can be appropriately selected and used in inert bases or carriers commonly used for preparing external preparations. As such bases or ingredients, in the case of ointments, creams, gels or lotions, there can be mentioned bases such as white petrolatum, yellow petrolatum, lanolin, white beeswax, cetyl alcohol, stearyl alcohol, stearic acid, hydrogenated oil, gelled hydrocarbons, polyethylene glycol (macrogol), 1,3-butylene glycol, ethanol, isopropyl alcohol, liquid paraffin, squalane; solvents and dissolution aids such as oleic acid, isopropyl myristate, triethylhexanoin, crotamiton, diethyl sebacate, diisopropyl adipate, hexyl laurate, fatty acids, fatty acid esters, aliphatic alcohols, and vegetable oils; thymol, tocopherol derivatives, ascorbic acid, butylated hydroxytoluene, butylated hydroxyanisole, etc. Antioxidants; preservatives such as phenoxyethanol and parabens; humectants such as glycerin, glyceryl monostearate, propylene glycol, sodium hyaluronate; surfactants such as polyoxyethylene derivatives, glyceryl fatty acid esters, sucrose fatty acid esters, sorbitan fatty acid esters, propylene glycol fatty acid esters, lecithin; thickeners such as carboxyvinyl polymers, xanthan gum, carboxymethyl cellulose, sodium carboxymethyl cellulose, hydroxypropyl cellulose, hydroxypropyl methylcellulose; stabilizers such as sodium edetate hydrate, citric acid hydrate, sodium citrate hydrate (including chelating agents, antioxidants); silicones such as dimethicone, cyclic silicones, modified silicones; keratin-softening ingredients such as salicylic acid. Furthermore, as desired, preservatives, absorption accelerators, pH adjusters, colorants (including pigments such as iron oxides), fragrances, ultraviolet absorbers, ultraviolet scatterers (such as titanium oxide and zinc oxide), excipients, dispersants, emulsifiers, isotonicity agents, buffers, fillers, crosslinking agents, cooling agents, film-forming agents, and other appropriate additives may be added.

[0049] Examples of patch preparations include: thickeners such as polyacrylic acid and polyacrylic acid copolymers; crosslinkers such as aluminum sulfate, potassium aluminum sulfate, aluminum chloride, magnesium aluminum metasilicate, and dihydroxyaluminum acetate; thickeners such as sodium polyacrylate, polyvinyl alcohol, polyvinyl pyrrolidone, gelatin, sodium alginate, carboxymethyl cellulose, sodium carboxymethyl cellulose, hydroxypropyl cellulose, and hydroxypropyl methyl cellulose; polyols such as glycerin, macrogol, propylene glycol, and 1,3-butylene glycol; surfactants such as polyoxyethylene derivatives; fragrances such as L-menthol; preservatives such as parabens; purified water; and supports such as plastic films, nonwoven fabrics, and cotton. Furthermore, stabilizers, preservatives, absorption enhancers, pH adjusters, and other appropriate additives may be added as desired.

[0050] About patch, can be mixed with adhesives such as natural rubber, isoprene rubber, polyisobutylene, polybutene, liquid polyisoprene, styrene-isoprene-styrene block copolymer (SIS block copolymer), styrene-butadiene-styrene block copolymer, styrene-ethylene / butylene-styrene block copolymer, (methyl) acrylic acid alkyl ester (to) polymer, polyacrylate, methacrylate etc. acrylic resins;Alicyclic saturated hydrocarbon resin, rosin resin, terpene resin etc. tackifying resin;Liquid rubber, liquid paraffin etc. softeners;BHT etc. antioxidants;Propylene glycol etc. polyols;Oleic acid etc. absorption promoter;Surfactant such as polyoxyethylene derivative, other suitable additives.In addition, can add sodium polyacrylate, polyvinyl alcohol etc. can be aqueous macromolecule and a small amount of purified water and make aqueous patch.In this case, and then can be mixed with stabilizer, preservative, absorption promoter, pH adjusting agent, other suitable additives according to expectation.

[0051] Aerosols may be formulated with bases such as white petrolatum, yellow petrolatum, lanolin, white beeswax, cetyl alcohol, stearyl alcohol, stearic acid, hydrogenated oil, gelled hydrocarbon, polyethylene glycol (macrogol), liquid paraffin, and squalane for the preparation of ointments, creams, gels, and liquids (suspensions, emulsions, and lotions); solvents and dissolution aids such as oleic acid, isopropyl myristate, diisopropyl adipate, isopropyl sebacate, triethylhexanoin, crotamiton, diethyl sebacate, hexyl laurate, fatty acids, fatty acid esters, aliphatic alcohols, and vegetable oils; and tocopherol derivatives, ascorbic acid, butylated hydroxytoluene, and butylated hydroxyanisole. Antioxidants; preservatives such as parabens; humectants such as glycerin, propylene glycol, and sodium hyaluronate; surfactants such as polyoxyethylene derivatives, glycerol fatty acid esters, sucrose fatty acid esters, sorbitan fatty acid esters, propylene glycol fatty acid esters, and lecithin; thickeners such as carboxyvinyl polymers, xanthan gum, carboxymethyl cellulose, sodium carboxymethyl cellulose, hydroxypropyl cellulose, and hydroxypropyl methylcellulose; propellants such as methyl ether, liquefied petroleum gas, and fluorocarbons; compressed gases such as carbon dioxide, nitrogen, and nitrous oxide; and various stabilizers, buffers, flavoring agents, suspending agents, emulsifiers, fragrances, preservatives, dissolution aids, and other appropriate additives.

[0052] Powders for external use may be mixed with excipients such as potato starch, rice starch, corn starch, talc, zinc oxide, or other suitable additives. In this case, various stabilizers, preservatives, absorption enhancers, sugars such as lactose, or other suitable additives may be further mixed as desired.

[0053] There are no particular limitations on the method for preparing the external preparation. The preparation can be prepared by conventional methods such as thoroughly mixing the various components and the base components used as needed, depending on the desired dosage form. Furthermore, when preparing a poultice or patch, the mixed mixture can be spread on release paper, dried, and then attached to a flexible support, and cut into the desired size.

[0054] When the above-mentioned external preparations are, for example, ointments, creams, gels, liquids (suspensions, emulsions, lotions, etc.), aerosols, and external powders, they can be used by conventional methods, such as directly applying (applying) to the skin, or applying or impregnating a support such as cloth, etc. In addition, when they are patch preparations or patches, these preparations can be applied directly to the skin.

[0055] 2. Application and manufacturing method of the composition of the present invention

[0056] The composition of the present invention can be used to increase or improve the diversity of resident bacteria on the skin compared to the state before application. Furthermore, since the composition of the present invention can increase the ratio of C16 monounsaturated fatty acids, which are generally considered to have high antibacterial properties, compared to before application, it can also be used as an enhancer for increasing the ratio of C16 monounsaturated fatty acids on the skin (hereinafter referred to as the "monounsaturated enhancer of the present invention").

[0057] As shown in the experimental results described below, the composition or monounsaturated enhancer of the present invention is highly effective in subjects with low sebum secretion or subjects with atopic skin symptoms, and therefore is preferably used in these subjects. Here, "low sebum secretion" varies depending on gender, age, etc., but refers to, for example, less than 1 / 2, less than 1 / 3, less than 1 / 5, or less than 1 / 10 of the average sebum secretion of a healthy person.

[0058] The composition of the present invention can be produced, for example, by adding other components (excipients, additives, etc.) to earthworm lipids within a range that does not impair the effects of the present invention and mixing them according to conventional methods. Alternatively, the composition of the present invention can be produced according to the methods for producing the above-mentioned preparations.

[0059] Earthworm lipid can be prepared, for example, by performing an extraction procedure for obtaining lipid using an organic solvent on the residue (earthworm dehydrate) after protein has been extracted and removed from living earthworms by known methods (e.g., enzymatic decomposition, addition of acid or organic solvent, insolubilization by heating or cooling, or physical removal by ultrafiltration, dialysis, or centrifugation). Alternatively, the earthworm dehydrate can be saponified with an alkali, acidified by making the alkaline solution of the saponified product, and then extracted with an organic solvent. It should be noted that earthworm dehydrate is well known and can be obtained from manufacturers who extract protein from living earthworms.

[0060] Specific methods for obtaining lipids from earthworm dehydrates include, for example, the Bligh-Dyer method, the Folch method, and the direct saponification method. These methods can obtain total lipids from earthworms.

[0061] When using the Bligh-Dyer method or the Folch method, a polar organic solvent can be used as an extraction solvent to extract earthworm lipid from, for example, earthworm dehydrate. Examples of polar organic solvents include mixed organic solvents of chlorine-based organic solvents and alcohol-based organic solvents, specifically, chloroform / methanol mixed solvents or chloroform / methanol / water mixed solvents. When using a chloroform / methanol mixed solvent as the extraction solvent, the mixing ratio of chloroform to methanol can be appropriately selected, and a volume ratio of 1:3 to 3:1 (chloroform:methanol) is suitable, and preferably 1:2 to 2:1.

[0062] In the direct saponification method, for example, fatty acid esters in earthworm dehydrate can be saponified with alkali, and the fatty acids can be extracted from the saponified product using a non-polar organic solvent to obtain earthworm lipids. Examples of the non-polar organic solvent include hydrocarbon organic solvents, specifically hexane, cyclohexane, and heptane.

[0063] The total lipids obtained as described above generally contain highly unsaturated fatty acids. Considering that these are easily oxidized and the oxides are irritating to the skin, they can be purified using conventional methods such as silica gel columns (to remove highly unsaturated fatty acids). Alternatively, instead of removing highly unsaturated fatty acids, antioxidants can be used to address skin irritation.

[0064] The earthworm lipid (total lipid) obtained by the above-mentioned extraction operation is usually an oily extract. Additives can be added to the extract and appropriately processed to prepare a composition containing the extract (solid, semi-solid, liquid, etc.), which can be used as earthworm lipid.

[0065] The composition or monounsaturated enhancer of the present invention can be used as a raw material for cosmetics. Cosmetics containing the composition or monounsaturated enhancer of the present invention (hereinafter referred to as "cosmetics of the present invention") can be functional cosmetics that increase or improve the diversity of resident bacteria on the skin compared to the state before application, or functional cosmetics that increase the ratio of 16-carbon monounsaturated fatty acids on the skin compared to before application. Furthermore, the cosmetics of the present invention are not limited to functional cosmetics and can also be, for example, skin care cosmetics, skin beautifying cosmetics, and body care cosmetics.

[0066] The cosmetic of the present invention may be in the form of, for example, a transparent liquid, a dispersion, an emulsion, a cream, a gel, a semi-solid, a solid, a foam, a powder / granular, an aerosol, or a sheet.

[0067] Here, "cosmetics" include not only "cosmetics" stipulated in laws and regulations on drugs and medical devices, but also "quasi-drugs" such as medicinal cosmetics.

[0068] When producing the cosmetics of the present invention, in addition to the composition of the present invention and the monounsaturated enhancer of the present invention, various other ingredients may be appropriately selected and used. Examples of such ingredients include water (purified water, distilled water, ion-exchanged water, etc.), monohydric alcohols (ethanol, isopropyl alcohol, etc.), moisturizers, oils, surfactants, pigments / colorants, fragrances, whitening active ingredients, anti-wrinkle ingredients, ultraviolet absorbers, anti-fading agents, antioxidants, cosmetic ingredients, preservatives, anti-inflammatory ingredients, antibacterial ingredients, and antipruritic ingredients.

[0069] Examples of the moisturizing agent include polyols such as glycerin, propylene glycol, dipropylene glycol, 1,3-butylene glycol, polyethylene glycol (300, 400, 1500, 4000), and polyglycerol; sugars such as glucose, sucrose, trehalose, pullulan, and maltitol; amino acids such as pyrrolidone carboxylic acid and citrulline; polysaccharides such as heparinoids and mucopolysaccharides; and biopolymers such as hyaluronic acid or its salts and collagen.

[0070] Examples of the oil include cocoa butter, shea butter, olive oil, camellia seed oil, macadamia nut oil, almond oil, coconut oil, sesame oil, corn oil, soybean oil, safflower oil, horse oil, egg yolk oil, mink oil, beef tallow, and horse fat; carnauba wax, candelilla wax, jojoba oil, beeswax, lanolin, Hoplostethus angustifolia, and safflower oil. atlanticus) oil; fatty acids such as lauric acid, myristic acid, palmitic acid, palmitoleic acid, stearic acid, oleic acid, linoleic acid, and linolenic acid; higher alcohols such as cetyl alcohol, stearyl alcohol, isostearyl alcohol, oleyl alcohol, lauryl alcohol, cholesterol, and sitosterol; hydrocarbons such as liquid paraffin, petrolatum, paraffin, ceresin, microcrystalline wax, squalane, and pristane; esters such as triisostearate, cetyl octanoate, isopropyl myristate, cetyl lactate, and diisostearyl malate; silicone oils such as siloxane, dimethicone, cyclodimethicone, methylpolysiloxane, and methylphenylpolysiloxane; and hydrocarbons, fats, waxes, hydrogenated oils, ester oils, fatty acids, higher alcohols, silicone oils, lanolin derivatives, oily gelling agents, and oil-soluble resins, of any origin, including animal oils, vegetable oils, and synthetic oils, and of any solid / liquid / volatile nature.

[0071] Examples of the surfactant include anionic surfactants such as higher fatty acid soaps, alkyl sulfate ester salts, alkyl ether sulfate ester salts, alkyl phosphate ester salts, alkyl ether phosphate ester salts, N-acylamino acid salts, and N-acyl-N-methyl taurates; cationic surfactants such as alkyltrimethylammonium chloride, dialkyldimethylammonium chloride, and benzalkonium chloride; amphoteric surfactants such as alkyldimethylaminoacetic acid betaine and alkylimidazolium betaine; nonionic surfactants such as polyethylene glycol-type nonionic surfactants (polyethylene glycol alkyl ethers, polyethylene glycol fatty acid esters, polyethylene glycol sorbitan fatty acid esters, etc.), polyol ester-type nonionic surfactants (glycerol fatty acid esters, polyglycerol fatty acid esters, sorbitan fatty acid esters, sucrose fatty acid esters, etc.), and block polymer-type surfactants; polymeric surfactants such as acrylic acid / C10-30 alkyl methacrylate copolymers, polyvinyl alcohol, sodium alginate, and cellulose derivatives; natural surfactants such as lecithin; and silicone-based surfactants.

[0072] Examples of the pigments or colorants include ultraviolet absorbers such as para-aminobenzoic acid derivatives, benzophenone derivatives, methoxycinnamic acid derivatives, and salicylic acid derivatives; ultraviolet scatterers such as titanium oxide fine particles and zinc dioxide fine particles; white pigments such as titanium dioxide and zinc oxide; coloring pigments such as iron oxide, chromium oxide, carbon black, and ultramarine; extender pigments such as mica, sericite, talc, and kaolin; pearlescent pigments; organic synthetic pigments such as azo dyes, xanthene dyes, indigo dyes, lakes, azo pigments, and phthalocyanine pigments; natural pigments such as β-carotene, lycopene, crocin, radix rosin, perilla frutescens, carthamin, safflower yellow, chlorophyll, riboflavin, cochineal, alizarin, shikonin, and curcumin; and polymer powders such as polyethylene powder and nylon powder.

[0073] Examples of the fragrance include natural fragrances such as rose flower oil, jasmine flower oil, lavender oil, eucalyptus oil, patchouli oil, peppermint oil, citronella oil, lemon peel oil, lemon oil, bergamot oil, sandalwood, cinnamon essential oil, oakmoss, iris oil, vetiver oil, musk, civet, castoreum, and ambergris; and synthetic fragrances such as α-limonene, β-caryophyllene, cis-3-hexenol, linalool, farnesol, β-phenylethanol, 2,6-nonadienal, citral, α-hexylcinnamaldehyde, β-ionone, L-carvone, cyclopentadecanone, linalyl acetate, benzyl benzoate, γ-undecalactone, eugenol, rose oxide, and phenylacetaldehyde dimethyl acetal.

[0074] Examples of the whitening active ingredient include magnesium ascorbyl phosphate, ascorbic acid-2-glucoside, 3-O-ethyl ascorbic acid, ascorbyl tetraisopalmitate, kojic acid, hydroquinone β-D-glucoside, ellagic acid, 4-n-butylresorcinol, potassium 4-methoxysalicylate, 4-(4-hydroxyphenyl)-2-butanol, 5,5′-dipropylbiphenyl-2,2′-diol, linoleic acid, trans-4-aminomethylcyclohexanoic acid, cetyl tranexamate hydrochloride, chamomile extract, adenosine-1-phosphate-2Na, and niacinamide.

[0075] Examples of the anti-wrinkle ingredients include polyols such as glycerin, polyethylene glycol, propylene glycol, and 1,3-butylene glycol; ceramides; amino acids such as serine, glycine, and pyrrolidone carboxylic acid; collagen; hyaluronic acid; retinal (vitamin A); niacinamide; elastin; and proteoglycans.

[0076] The method for producing the cosmetics of the present invention is not particularly limited and can be produced using conventional methods depending on the desired dosage form. For example, the various ingredients described above, including the activator of the present invention, can be mixed using an emulsifier such as a batch vacuum emulsifier, a disperser such as a colloid mill, a mixer such as a Henschel mixer or a hammer mill, or a pulverizer. The desired dosage form can then be formed using a cooling machine, a molding machine, or the like, as needed.

[0077] Furthermore, the cosmetics of the present invention obtained by an appropriate production method can be packaged into tube containers, bottle containers (narrow mouth), cream containers (wide mouth), powder containers, compact containers, stick containers, pencil-type containers, containers with brushes, pump bottles, aerosol containers, etc. using, for example, a filling machine, a packaging machine, etc. to prepare cosmetic products.

[0078] Example

[0079] The present invention is further described in detail with reference to the following examples, but the present invention is not limited by the following examples.

[0080] [Reference Example] Study on the Diversity of Skin Resident Bacteria

[0081] The skin resident bacteria of 50 healthy individuals (9 of whom had a history of atopic dermatitis) and 50 individuals with atopic dermatitis, a total of 100 individuals, were sampled and metagenomic analysis was used to investigate the species diversity of the skin resident bacteria (diversity of skin resident bacteria). This diversity was evaluated using the Simpson's Index (D) and Shannon Entropy (H). The ratio of the number of individuals of the i-th species to the total number of individuals in the skin resident bacteria was set to pi (|pi| < 1), and the values ​​of D and H were calculated using the following formulas.

[0082]

[0083]

[0084] Simpson's diversity index ranges from 0 to 1, and the closer it is to 1, the more evenly diverse the bacteria are. The larger the absolute value of Shannon's entropy, the higher the diversity. The analysis results are shown in Table 1 below.

[0085]

Table 1

[0086]

[0087] * :p<0.01

[0088] As shown in Table 1, it was found that the diversity of the group without atopic symptoms was significantly higher (more bacterial species) than that of the group with atopic symptoms.

[0089] Note that, although each is a single example, a comparison of the resident bacterial flora of the skin of a healthy person and the resident bacterial flora of the skin of a person with atopic dermatitis is shown in Figure 1 . Figure 1 The bacterial groups with a occupancy rate of 0.05% or more are shown in the table. Figure 1 As shown, 194 bacterial species were identified in healthy individuals, about one-third of which were non-pathogenic acne bacteria, whereas only 31 bacterial species were identified in individuals with atopic dermatitis, more than one-third of which were pathogenic Staphylococcus aureus.

[0090] Furthermore, according to the results of sebum sampling conducted simultaneously with the sampling of skin resident bacteria, 23 of the 50 people with atopic dermatitis had sebum secretion levels that were a fraction to less than one-tenth of those of healthy people.

[0091] As described above, people with atopic dermatitis tend to have low diversity of skin-resident bacteria. Furthermore, many people tend to have not only low diversity of skin-resident bacteria but also reduced sebum secretion.

[0092] [Example 1] Preparation of earthworm lipid

[0093] (1) Obtaining earthworm raw materials

[0094] The earthworm raw material for obtaining earthworm lipid used in the experiments described below was the residue (dehydrated earthworm) obtained by extracting protein from red earthworms (Lumiphidium edulis), and was provided by Miyazaki Blood Flow Research Institute Co., Ltd. (Miyazaki Prefecture).

[0095] (2) Preparation of earthworm lipids

[0096] Earthworm lipid (total lipid) was prepared using any of the following methods.

[0097] (2-1) Preparation by Bligh-Dyer method

[0098] Total lipids were extracted from 10 g of earthworm dehydrate using 30 mL of a chloroform / methanol mixed solvent (2:1), yielding approximately 2% earthworm lipids.

[0099] (2-2) Preparation by direct saponification method

[0100] Place 60 g of dehydrated earthworms in an eggplant-shaped flask, add 120 mL of 1 mol / KOH / ethanol, and heat at 80°C for 1 hour to saponify the fatty acid esters in the sample. After cooling, add purified water, filter the saponified ethanol solution into a liquid extraction apparatus, and reflux with isopropyl ether for 5 hours to remove unsaponifiable substances.

[0101] The alkaline solution in the liquid extraction device was taken out, sulfuric acid was added to make it strongly acidic, and then filtered. The solution was refluxed with hexane for 5 hours to extract fatty acids.

[0102] The hexane extract of the fatty acid was washed with purified water until neutral (methyl orange), filtered, concentrated to dryness, and then dissolved in acetonitrile and allowed to stand for 10 minutes to precipitate the acetonitrile-insoluble matter.

[0103] The supernatant of the acetonitrile solution was passed through a Florisil column (activated at 130°C for 3 hours), and the column was washed with acetonitrile. The eluted acetonitrile was concentrated to dryness under reduced pressure to obtain earthworm lipid.

[0104] (3) Purification of earthworm lipids - Removal of highly unsaturated fatty acids

[0105] Wakosil C-300 (activated at 110°C for 3 hours) was slurried in hexane and packed into a glass column (15 mm inner diameter, 30 cm length). Florisil PR (activated at 130°C for 3 hours) was slurried in hexane and layered on top of the Wakosil C-300. Anhydrous sodium sulfate was then layered to create a fatty acid purification column. The concentrated, dry fatty acids were dissolved in hexane and loaded onto the column, and eluted using ether / hexane.

[0106] [Example 2] Study on the effect of improving the skin resident flora of earthworm lipids

[0107] (1) Experimental methods

[0108] 9 people with atopic dermatitis and 3 healthy people, a total of 12 subjects, were selected as shown in Table 2. The control / squalane and 1% earthworm lipid / squalane prepared in Example 1 (direct saponification method) were added to the same side of the inner side of the left and right elbows of each subject, 1 drop of each substance was applied twice a day, morning and evening (after bathing), without knowing which one was added with earthworm lipid, so as to avoid the placebo effect. 3 healthy people and 1 person with atopic dermatitis applied for 4 weeks, and the other 8 people with atopic dermatitis applied for 2 weeks. The sample application area was wiped with a cotton swab in a swab kit for metagenomic analysis to sample microorganisms on the skin. The 16srRNA gene of the bacteria was obtained from the wiping cotton swab, and its sequence was read. The sequence was compared with the reference total bacterial sequence database (about 15,000 species) to perform species identification at the species level. The number of individuals, Simpson index and Shannon entropy were calculated based on the number of DNA fragments of each species.

[0109]

Table 2

[0110] Personal ID History of atopic dermatitis age gender time A-101 have 51 male 4 weeks A-102 have 25 male 2 weeks A-103 have 34 male 2 weeks A-104 have 38 male 2 weeks A-105 have 27 male 2 weeks A-106 have 26 female 2 weeks A-107 have 30 female 2 weeks A-108 have 29 female 2 weeks A-109 have 45 female 2 weeks B-101 none 53 male 4 weeks B-102 none 30 male 4 weeks B-103 none 35 female 4 weeks

[0111] Sebum samples were collected from the applied area (left and right sides) using degreasing paper two weeks after application. Extraction, purification, and derivatization were then performed, followed by GC / MS analysis to investigate sebum secretion levels and analyze the ratio of monounsaturated acids (C16:1), generally considered to have high antibacterial properties, to linear saturated acids (C16:0).

[0112] (2) Analysis results of subject A-109

[0113] Among nine individuals with atopic dermatitis, four had extremely low sebum secretion, one of whom was A-109. Comparison of A-109 before application and on day 15 revealed not only changes in diversity but also shifts in the most abundant bacterial species. Table 3 shows the bacterial species counts on days 0, 8, and 15. Species with a population share of 0.05% or greater were counted.

[0114] As shown in Table 3, when earthworm lipid was applied, the number of bacterial species was significantly increased compared to the control.

[0115]

Table 3

[0116]

[0117] For A-109, the Simpson index (the closer to 1, the higher the diversity) and Shannon entropy (the larger the value, the higher the diversity) were calculated to perform diversity analysis. The results are shown in Figure 2 .

[0118] like Figure 2 As shown, the diversity of the A-109 control tended to decrease, while the diversity of the A-109 treated with earthworm lipids tended to increase.

[0119] In addition, GC / MS analysis of sebum samples obtained Figure 3 The results shown. It should be noted that Figure 3 In the comparison, the chromatograms were standardized using C16:0 (saturated fatty acid with 16 carbon atoms).

[0120] like Figure 3 As shown, the ratio of monounsaturated acids with 16 carbon atoms, which are generally considered to have high antibacterial properties, to linear saturated acids with 16 carbon atoms remained unchanged in the control compared to before application, but increased significantly after application of earthworm lipid. Furthermore, the amount of isomers other than monounsaturated acids contained in the applied earthworm lipid increased, suggesting that the production of monounsaturated acid isomers with 16 carbon atoms is due to the action of skin-resident bacteria on earthworm lipid.

[0121] (3) Analysis results of subject A-107

[0122] Subject A-107 was also one of the people with low sebum secretion and had low bacterial flora diversity before the experiment.

[0123] The diversity of the bacterial community increased with the application of earthworm lipid, and a shift in the most populous bacterial species was observed in both the control and the earthworm lipid-applied samples. The results of the bacterial species count are shown in Table 4.

[0124] As shown in Table 4, when earthworm lipid was applied, the number of bacterial species was significantly increased compared to the control.

[0125]

Table 4

[0126]

[0127] For A-107, the Simpson index and Shannon entropy were also calculated to perform diversity analysis, resulting in Figure 4 The results shown. Figure 4 As shown, in A-107, although the change in the control was small, there was a tendency for the diversity to decrease, while the diversity increased after applying earthworm lipid.

[0128] Sebum samples were analyzed by GC / MS in the same manner, and the Figure 5 The results shown. It should be noted that Figure 5 The chromatogram ratios were also standardized using C16:0 (saturated fatty acid with 16 carbon atoms) for comparison.

[0129] In A-107, perhaps due to the influence of the replacement of bacterial species, the ratio of monounsaturated acids with 16 carbon atoms, which are generally considered to have high antibacterial properties, to straight-chain saturated acids with 16 carbon atoms increased in both the control and earthworm lipid-coated cases compared with before coating, but the increase rate of monounsaturated acids was higher when earthworm lipid was coated.

[0130] (4) Analysis results of subject A-105

[0131] Subject A-105 was one of those with normal sebum secretion and high bacterial diversity before application. Application reduced the proportion of highly concentrated acne bacteria, accompanied by a further increase in diversity. A shift in the most concentrated bacterial species was observed in both the control and earthworm lipid application groups. The results of bacterial species counts are shown in Table 5.

[0132] As shown in Table 5, when sebum secretion is normal, the number of bacterial species only slightly increases even when earthworm lipid is applied, while a significant decrease in the number of bacterial species is observed in the control.

[0133]

Table 5

[0134]

[0135] For A-105, the Simpson index and Shannon entropy were also calculated to perform diversity analysis, and the results were Figure 6 The results shown. Figure 6 As shown, there was little difference in the Simpson index between the control and the earthworm lipid coating, but the Shannon entropy of the earthworm lipid coating showed high diversity. This is presumably due to the high number of bacterial species with low occupancy.

[0136] Sebum samples were separately analyzed by GC / MS. Figure 7 The results shown. It should be noted that Figure 7 In the comparison, the chromatogram ratio was standardized using C16:0 (saturated fatty acid with 16 carbon atoms) for comparison.

[0137] In A-105, almost no change in the monounsaturated acid ratio was observed after application. This suggests that the effect of lipid application is minimal for individuals with naturally high diversification and high sebum secretion.

[0138] Industrial applicability

[0139] The composition of the present invention can increase or improve the diversity of resident skin bacteria compared to the state before application, thereby enhancing the skin's barrier function and inhibiting the proliferation of pathogenic bacteria. Therefore, it is useful as a raw material for skin care products and cosmetics, for example. Furthermore, since increasing or improving the diversity of resident skin bacteria is related to the amelioration and prevention of atopic symptoms and the maintenance of healthy skin, the composition of the present invention is also useful as a raw material for pharmaceuticals.

Claims

1. A composition for controlling the diversity of resident bacterial flora on the skin, comprising an effective amount of earthworm lipid and a base, wherein the earthworm lipid is prepared by the following method: The dehydrated earthworms are saponified with alkali, and the alkaline solution of the saponified product is made acidic, and then the saponified product is extracted with a non-polar organic solvent. The earthworm dehydrate is the residue after extracting and removing protein from earthworms. The non-polar organic solvent is at least one selected from the group consisting of hexane, cyclohexane and heptane, The earthworm is Lumbricus rubellus, The base is at least one selected from the group consisting of white petrolatum, yellow petrolatum, lanolin, white beeswax, cetyl alcohol, stearyl alcohol, stearic acid, hydrogenated oil, gelled hydrocarbon, macrogol, 1,3-butylene glycol, ethanol, isopropyl alcohol, liquid paraffin, and squalane.

2. The composition according to claim 1, which is used for application to a subject with low sebum secretion or a subject with atopic skin symptoms.

3. A C16 monounsaturated fatty acid enhancer for skin, comprising the composition according to claim 1.

4. The composition according to claim 1 or the C16 monounsaturated fatty acid enhancer on the skin according to claim 3, wherein The dosage forms are ointment, cream, gel, liquid, papule, patch, tablet, aerosol, external powder, spray or bath preparation.

5. A cosmetic comprising the composition according to claim 1 or 2 or the C16 monounsaturated fatty acid enhancer on the skin according to claim 3 or 4.

6. A composition for controlling resident bacterial flora on the skin, comprising an effective amount of earthworm lipid, wherein the earthworm lipid is obtained by saponifying earthworm dehydrate with an alkali, making the alkaline solution of the saponified product acidic, and then extracting the saponified product with a non-polar organic solvent. The earthworm dehydrate is the residue after extracting and removing protein from earthworms. The non-polar organic solvent is at least one selected from the group consisting of hexane, cyclohexane and heptane, The earthworm is Lumbricus rubellus.

7. An earthworm lipid for controlling resident bacterial flora on the skin, the earthworm lipid being obtained by saponifying earthworm dehydrate with an alkali, making the alkaline solution of the saponified product acidic, and then extracting the saponified product with a non-polar organic solvent. The earthworm dehydrate is the residue after extracting and removing protein from earthworms. The non-polar organic solvent is at least one selected from the group consisting of hexane, cyclohexane and heptane, The earthworm is Lumbricus rubellus.

8. A method for producing a composition for controlling the diversity of resident skin flora, comprising the steps of extracting earthworm lipid from earthworms and mixing the earthworm lipid with a base, wherein the earthworm lipid is prepared by the following method: The dehydrated earthworms are saponified with alkali, and the alkaline solution of the saponified product is made acidic, and then the saponified product is extracted with a non-polar organic solvent. The earthworm dehydrate is the residue after extracting and removing protein from earthworms. The non-polar organic solvent is at least one selected from the group consisting of hexane, cyclohexane and heptane, The earthworm is Lumbricus rubellus, The base is at least one selected from the group consisting of white petrolatum, yellow petrolatum, lanolin, white beeswax, cetyl alcohol, stearyl alcohol, stearic acid, hydrogenated oil, gelled hydrocarbon, macrogol, 1,3-butylene glycol, ethanol, isopropyl alcohol, liquid paraffin, and squalane.

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