Composition for controlling skin commensal flora

Abstract

To provide a novel composition for control of skin resident flora that can increase or improve the diversity of bacterial strains in skin resident flora compared to a state before treatment.SOLUTION: A composition contains an effective amount of earthworm lipids extracted from Lumbricus rubellus. The composition is, for example, ointment or liquid chemical. The composition is useful as, for example, raw materials for skin care products and cosmetic preparations.SELECTED DRAWING: Figure 2

Description

[Technical field]

[0001] The present invention relates to a composition for controlling normal skin flora, which mainly belongs to the technical field of skin care. More specifically, the present invention relates to a composition for controlling normal skin flora, which contains so-called earthworm lipids extracted from earthworms. [Background technology]

[0002] In addition to the epidermis and stratum corneum functioning as a barrier between the inside and outside of the body, the formation of the sebum film plays an important role in the human skin barrier mechanism. The sebum film is formed by fatty acid sodium salts that are generated when sebum secreted from the sebaceous glands is decomposed into free fatty acids by the enzyme lipase derived from skin microorganisms and then combined with sodium salts contained in sweat secreted from the sweat glands. The sebum film acts as a physical barrier to prevent the intrusion of foreign substances, while also suppressing water evaporation from the epidermis to prevent the skin from drying out, and stabilizing the environment on the skin surface. When the amount of sebum secreted is low, or when the amount of sweat produced is low even if the amount of sebum secreted is sufficient, it is thought that fatty acid sodium salts are difficult to produce, and the formation of the sebum film is insufficient, resulting in a reduced barrier effect.

[0003] Normally, more than 200 kinds of microorganisms that are suited to the skin environment maintain a balance and form a flora that prevents the proliferation of pathogenic bacteria. Some free fatty acid isomers produced when sebum is decomposed by lipase of skin microorganisms have the effect of suppressing the proliferation of certain types of bacteria, and it is thought that the ratio of constituent fatty acids affects the skin microflora. It is thought that the resistance to pathogenic microorganisms differs depending on the composition of the skin microflora. In this way, controlling the composition or diversity of the skin microbiota or resident skin flora to a state similar to that of healthy skin is important for maintaining healthy skin, preventing the invasion of pathogenic microorganisms, and suppressing inflammation such as atopic dermatitis. Non-Patent Document 1 introduces that the diversity of resident skin flora is low in sensitive skin.

[0004] As a technique for controlling the skin's resident flora, for example, a topical agent for controlling skin's resident bacteria, which contains magnesium lactate and / or calcium lactate as an active ingredient, is known (Patent Document 1). According to Patent Document 1, this topical agent for controlling skin's resident bacteria can reduce the number of harmful bacteria while suppressing the decrease in the number of beneficial bacteria in the skin's resident bacteria.

[0005] Meanwhile, earthworms, especially farmed red earthworms (Lumbricus rubellus), are dried and powdered and widely distributed as dietary supplements and medicines. These earthworms contain ingredients that have antipyretic and analgesic properties, as well as the enzyme lumbrokinase, which is believed to dissolve blood clots. [Prior art documents] [Patent documents]

[0006] [Patent Document 1] JP 2018-52891 A [Non-patent literature]

[0007] [Non-Patent Document 1] "Shiseido discovers that sensitive skin has low diversity of resident skin flora", August 19, 2020, Publisher: Shiseido Co., Ltd., Internet (URL: https: / / corp.shiseido.com / jp / news / detail.html?n=00000000002960) Summary of the Invention [Problem to be solved by the invention]

[0008] As mentioned above, controlling the composition and diversity of the resident skin flora to a state similar to that of healthy skin is important for maintaining healthy skin, preventing the invasion of pathogenic microorganisms, and suppressing inflammation such as atopic dermatitis. The main objective of the present invention is to provide a new composition for controlling the resident skin flora, which can increase or improve the diversity of bacterial species in the resident skin flora (the diversity of microorganisms on the skin) compared to the state before treatment. [Means for solving the problem]

[0009] As a result of extensive research, the present inventors discovered that so-called earthworm lipids extracted from earthworms have the effect of increasing the diversity of resident skin bacteria, and thus completed the present invention. The present invention can include, for example, the following aspects. [1] A composition for controlling the diversity of bacterial species in the normal skin flora, comprising an effective amount of earthworm lipids and a base. [2] The composition described in [1] above, wherein the earthworm lipids are obtained from the extraction residue remaining after extracting and removing proteins from raw earthworms. [3] The composition described in [1] or [2] above, wherein the earthworm is a red earthworm (Lumbricus rubellus). [4] The composition according to any one of the above [1] to [3], for application to a subject with low sebum secretion or a subject with atopic skin symptoms. [5] An agent for enhancing a monoenoic fatty acid having 16 carbon atoms on the skin, comprising the composition according to any one of [1] to [4] above. [6] The composition according to any one of [1] to [4] above, which is in the form of an ointment, cream, gel, liquid, cataplasm, tape, sheet, aerosol, external powder, spray, or bath additive, or the agent for enhancing a monoenoic fatty acid having a carbon number of 16 on the skin according to [5] above. [7] A cosmetic preparation comprising the composition according to any one of [1] to [6] above or an agent for enhancing the effect of a monoenoic fatty acid having 16 carbon atoms on the skin. [8] A composition for controlling the resident skin flora, obtained by extracting the residue (dehydrated earthworm cake) remaining after proteins have been extracted and removed from raw earthworms using a polar organic solvent, or by saponifying the dehydrated earthworm cake with an alkali and then extracting the saponified product using a nonpolar organic solvent. [9] Earthworm lipids for controlling the resident skin flora are obtained by extracting them with a polar organic solvent from the extraction residue (dehydrated earthworm cake) after the proteins have been extracted and removed from raw earthworms, or by saponifying the dehydrated earthworm cake with alkali and then extracting the saponified product with a nonpolar organic solvent.

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

[11] The composition for controlling resident skin bacteria described in [8] above or the earthworm lipid for controlling resident skin flora described in [9] above, wherein the non-polar organic solvent is a hydrocarbon organic solvent.

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

[0010] According to the present invention, the diversity of microorganisms on the skin (the diversity of resident skin bacteria) can be increased or improved compared to the state before application, thereby enhancing the barrier function of the skin and suppressing the growth of pathogenic bacteria. [Brief description of the drawings]

[0011] [Figure 1] This shows the normal skin flora. The upper diagram shows the results for healthy subjects, and the lower diagram shows the results for subjects with atopic dermatitis. [Diagram 2] The diagram shows the results of diversity analysis. The left diagram shows the results of Simpson's Index analysis, and the right diagram shows the results of Shannon Entropy analysis. Each line type shows the progress of the control and the progress when earthworm lipid was applied, respectively. [Diagram 3]These are chromatograms (TIC) showing the results of GC / MS analysis. The left figure shows the control results, and the right figure shows the results when earthworm lipids were applied. Each line shows the chromatograms of sebum after two weeks of application, sebum before earthworm lipid application, and earthworm lipid, respectively, and the ▽ indicates the peak of monoenoic fatty acid with 16 carbon atoms. [Figure 4] The diagram shows the results of diversity analysis. The left diagram shows the results of Simpson's Index analysis, and the right diagram shows the results of Shannon Entropy analysis. Each line type shows the progress of the control and the progress when earthworm lipid was applied, respectively. [Diagram 5] These are chromatograms (TIC) showing the results of GC / MS analysis. The left figure shows the control results, and the right figure shows the results when earthworm lipids were applied. Each line shows the chromatograms of sebum after two weeks of application, sebum before earthworm lipid application, and earthworm lipid, respectively, and the ▽ indicates the peak of monoenoic fatty acid with 16 carbon atoms. [Figure 6] The diagram shows the results of diversity analysis. The left diagram shows the results of Simpson's Index analysis, and the right diagram shows the results of Shannon Entropy analysis. Each line type shows the progress of the control and the progress when earthworm lipid was applied, respectively. [Figure 7] These are chromatograms (TIC) showing the results of GC / MS analysis. The left figure shows the control results, and the right figure shows the results when earthworm lipids were applied. Each line shows the chromatograms of sebum after two weeks of application, sebum before earthworm lipid application, and earthworm lipid, respectively, and the ▽ indicates the peak of monoenoic fatty acid with 16 carbon atoms. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0012] Hereinafter, an embodiment of the present invention will be described in detail. 1. Composition according to an embodiment of the present invention The composition according to the embodiment of the present invention (hereinafter referred to as "the composition of the present invention") is a composition for controlling the diversity of bacterial species in the resident skin flora (the diversity of microorganisms on the skin or the diversity of resident skin bacteria), and is characterized by containing an effective amount of earthworm lipid. The composition of the present invention can be preferably used to increase or improve the diversity of bacterial species in the resident skin flora compared to the state before application, or to maintain said diversity.

[0013] Here, "control" refers to changing or maintaining the number of species in the resident skin flora or the occupancy rate of multiple species in the flora (control of resident skin flora), including increasing or improving the diversity of species in the resident skin flora from the state before application of the composition of the present invention (improvement of resident skin flora), and maintaining the diversity that is already high due to improvement of resident skin flora or other factors (maintenance of resident skin flora). "Diversity" is a concept that combines the aspects of abundance of species (large number of species of bacteria) and uniformity of species (small variation in the number of individuals between species). The more species there are, and further the more similar the number of individuals of each species is, or the smaller the bias in the occupancy rate is, the higher (better) the diversity is. The presence or absence and the degree of the improvement or enhancement can be determined taking into consideration the above two aspects, for example, using as indicators Simpson's Index, Shannon Entropy, and Nakamura's RI index (Nakamura, Journal of Setouchi Junior College, Vol. 24: 37-41, 1994). The term "effective amount" refers to an amount at which the effect of the control (e.g., improvement of resident skin flora, maintenance of resident skin flora) can be confirmed by a person skilled in the art.

[0014] "Earthworm lipid" refers to a wide variety of lipid groups contained in earthworms. In particular, it refers to a wide variety of lipid groups contained in all parts of earthworms. The lipid group is usually composed of more than 100 types of diverse lipids. The earthworm lipid can be obtained as an extract by, for example, performing an extraction operation to obtain lipids from the extraction residue (earthworm dehydrated cake) in which proteins are extracted and removed from raw earthworms using a polar organic solvent. Examples of the polar organic solvent include a mixed organic solvent of a chlorine-based organic solvent and an alcohol-based organic solvent, and specifically, for example, chloroform and methanol or a mixed organic solvent obtained by adding water to these can be mentioned. In addition, the earthworm dehydrated cake can be saponified with an alkali, and the alkaline solution of the saponified product is acidified, and then extracted with a nonpolar organic solvent. Examples of the nonpolar organic solvent include a hydrocarbon-based organic solvent, and specifically, for example, hexane, cyclohexane, and heptane can be mentioned. The obtained earthworm lipids (total lipids) can be in oily or solid form. The obtained extract may be mixed with additives and appropriately processed to form a composition containing the extract (solid, semi-solid, liquid, etc.). The earthworm according to the present invention is not particularly limited, but is preferably of the genus Lumbricus, and more preferably Lumbricus rubellus, so-called red earthworm.

[0015] The dosage form of the composition of the present invention is not particularly limited as long as it is a dosage form that can be applied directly to the surface of the skin, and examples thereof include ointments, creams, gels, liquids (suspensions, emulsions, lotions, etc.), poultices, tapes, sheets, aerosols, external powders, sprays, and bath additives.

[0016] In preparing these preparations, in addition to earthworm lipids, various ingredients can be appropriately selected and used as inactive bases or carriers used in preparing ordinary topical preparations. In the case of ointments, creams, gels, or lotions, such bases or ingredients include white petrolatum, yellow petrolatum, lanolin, white beeswax, cetanol, stearyl alcohol, stearic acid, hardened oils, gelling hydrocarbons, polyethylene glycol (macrogol), 1,3-butylene glycol, ethanol, isopropanol, liquid paraffin, squalane, and other bases; solvents and dissolving agents such as oleic acid, isopropyl myristate, glycerin triisooctanoate, crotamiton, diethyl sebacate, diisopropyl adipate, hexyl laurate, fatty acids, fatty acid esters, fatty alcohols, and vegetable oils; antioxidants such as thymol, tocopherol derivatives, ascorbic acid, dibutylhydroxytoluene, and butylhydroxyanisole; and phenoxyethanol. Examples of suitable active ingredients include preservatives such as glycerin, glycerin monostearate, propylene glycol, sodium hyaluronate, etc.; moisturizing agents such as glycerin, glycerin monostearate, propylene glycol, sodium hyaluronate, etc.; surfactants such as polyoxyethylene derivatives, glycerin fatty acid esters, sucrose fatty acid esters, sorbitan fatty acid esters, propylene glycol fatty acid esters, lecithin, etc.; thickeners such as carboxyvinyl polymers, xanthan gum, carboxymethylcellulose, carboxymethylcellulose sodium salts, hydroxypropylcellulose, hydroxypropylmethylcellulose, etc.; stabilizers (including chelating agents and antioxidants) such as sodium edetate hydrate, citric acid hydrate, sodium citrate hydrate, etc.; silicones such as dimethicone, cyclic silicone, modified silicone, etc.; and keratin softening ingredients such as salicylic acid. Furthermore, if desired, preservatives, absorption enhancers, pH adjusters, colorants (including pigments such as iron oxide), flavorings, UV absorbers, UV scattering agents (titanium oxide, zinc oxide, etc.), excipients, dispersants, emulsifiers, isotonicity agents, buffers, fillers, crosslinking agents, cooling agents, coating agents, and other suitable additives may be added.

[0017] Examples of the cataplasm include tackifiers such as polyacrylic acid and polyacrylic acid copolymers; crosslinkers such as aluminum sulfate, potassium aluminum sulfate, aluminum chloride, magnesium aluminometasilicate, and dihydroxyaluminum acetate; thickeners such as sodium polyacrylate, polyvinyl alcohol, polyvinylpyrrolidone, gelatin, sodium alginate, carboxymethylcellulose, sodium carboxymethylcellulose salts, hydroxypropylcellulose, and hydroxypropylmethylcellulose; polyhydric alcohols such as glycerin, polyethylene glycol (macrogol), propylene glycol, and 1,3-butanediol; surfactants such as polyoxyethylene derivatives; fragrances such as L-menthol; preservatives such as parahydroxybenzoic acid esters; purified water; and supports such as plastic films, nonwoven fabrics, and cotton. Furthermore, stabilizers, preservatives, absorption promoters, pH adjusters, and other suitable additives may be added as desired.

[0018] The tape preparation may contain 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, (meth)acrylic acid alkyl ester (co)polymer, polyacrylic acid ester, methacrylic acid ester and other acrylic resins; alicyclic saturated hydrocarbon resin, rosin resin, terpene resin and other tackifiers; liquid rubber, liquid paraffin and other softeners; dibutylhydroxytoluene and other antioxidants; polyhydric alcohols such as propylene glycol; oleic acid and other absorption promoters; polyoxyethylene derivatives and other surfactants, and other suitable additives. In addition, a water-absorbent polymer such as sodium polyacrylate or polyvinyl alcohol and a small amount of purified water may be added to prepare a water-absorbent tape preparation. Even in this case, stabilizers, preservatives, absorption promoters, pH adjusters and other suitable additives may be added as desired.

[0019] For aerosol preparations, bases such as white petrolatum, yellow petrolatum, lanolin, white beeswax, cetanol, stearyl alcohol, stearic acid, hardened oil, gelling hydrocarbon, polyethylene glycol (macrogol), liquid paraffin, and squalane, which are used in the preparation of ointments, creams, gels, and liquid preparations (suspensions, emulsions, and lotions), etc.; solvents and dissolving aids such as oleic acid, isopropyl myristate, diisopropyl adipate, isopropyl sebacate, glycerin triisooctanoate, crotamiton, diethyl sebacate, hexyl laurate, fatty acids, fatty acid esters, fatty alcohols, and vegetable oils; tocopherol derivatives, ascorbic acid, dibutylhydroxytoluene, butyric acid, and the like. preservatives such as parahydroxybenzoic acid esters; moisturizing agents such as glycerin, propylene glycol, sodium hyaluronate; surfactants such as polyoxyethylene derivatives, glycerin fatty acid esters, sucrose fatty acid esters, sorbitan fatty acid esters, propylene glycol fatty acid esters, lecithin; thickeners such as carboxyvinyl polymers, xanthan gum, carboxymethylcellulose, carboxymethylcellulose sodium salts, hydroxypropylcellulose, hydroxypropylmethylcellulose; propellants such as dimethyl ether, liquefied petroleum gas, and fluorohydrocarbons; compressed gases such as carbon dioxide, nitrogen, and nitrous oxide; and further, various stabilizers, buffers, flavorings, suspending agents, emulsifiers, fragrances, preservatives, solubilizing agents, and other appropriate additives may be blended.

[0020] Powders for external use may contain excipients such as potato starch, rice starch, corn starch, talc, zinc oxide, or other suitable additives. Even in this case, various stabilizers, preservatives, absorption promoters, sugars such as lactose, and other suitable additives may be further contained as desired.

[0021] The method for preparing the above-mentioned external preparation is not particularly limited, and it can be produced by a conventional method such as thoroughly kneading each component and the base component as required according to the desired dosage form. In addition, in the preparation of a cataplasm or tape, the kneaded mixture can be spread on a release paper, dried, and further laminated with a flexible support, and cut to the desired size.

[0022] The above-mentioned topical preparations, for example, ointments, creams, gels, liquids (suspensions, emulsions, lotions, etc.), aerosols, and topical powders, can be used in a conventional manner, such as by directly applying (spreading) them to the skin, or by applying or impregnating a support such as cloth, etc. Also, in the case of poultices and tapes, these preparations can be used by directly attaching them to the skin.

[0023] 2. Uses, manufacturing method, etc. of the composition of the present invention The composition of the present invention can be used to increase or improve the diversity of resident skin bacteria from the state before application. In addition, since the composition of the present invention can increase the ratio of monoenoic fatty acids having 16 carbon atoms, which are considered to have high antibacterial properties, from the state before application, it can also be used as an enhancer for increasing the ratio of monoenoic fatty acids having 16 carbon atoms on the skin (hereinafter referred to as the "monoene enhancer of the present invention").

[0024] As is clear from the experimental results described below, the composition of the present invention or the monoene enhancer of the present invention is highly effective for subjects with low sebum secretion or subjects with atopic skin symptoms, and is therefore preferably applied to these subjects. Here, "low sebum secretion" refers to, although it varies depending on gender, age, etc., approximately 1 / 2 or less, 1 / 3 or less, 1 / 5 or less, or 1 / 10 or less of the average sebum secretion of a healthy person.

[0025] The composition of the present invention can be produced, for example, by adding other ingredients (excipients, additives, etc.) to earthworm lipids in a range that does not impair the effects of the present invention, and mixing them in a conventional manner. The composition of the present invention can also be produced according to the production methods of each of the above-mentioned preparations.

[0026] Earthworm lipids can be prepared, for example, by extracting lipids from the residue (dehydrated earthworm cake) obtained after extracting and removing proteins from raw earthworms by known methods (e.g., enzymatic decomposition, insolubilization by addition of acid or organic solvent, heating or cooling, or physical removal by ultrafiltration, dialysis, or centrifugation) using an organic solvent. Alternatively, the dehydrated earthworm cake can be saponified with an alkali, the alkaline solution of the saponified product is acidified, and then extracted with an organic solvent. Dehydrated earthworm cake is known and can be obtained from a company that extracts proteins from raw earthworms. Specific methods for obtaining lipids from dehydrated earthworm cake include, for example, the Bligh-Dyer method, the Folch method, and direct saponification method. These methods can be used to obtain total lipids from earthworms.

[0027] In the case of the Bligh-Dyer method or the Folch method, a polar organic solvent is used as an extraction solvent, and earthworm lipids can be obtained, for example, by extraction from dehydrated earthworm cake. Examples of polar organic solvents include a mixed organic solvent of a chlorine-based organic solvent and an alcohol-based organic solvent, specifically, a mixed solvent of chloroform and methanol or a mixed solvent of chloroform, methanol and water. When a mixed solvent of chloroform and methanol is used as an extraction solvent, the mixing ratio of chloroform and methanol is appropriately selected, but a volume ratio within the range of 1:3 to 3:1 (chloroform:methanol) is appropriate, and a volume ratio within the range of 1:2 to 2:1 is preferable.

[0028] In the case of the direct saponification method, for example, fatty acid esters in the dehydrated earthworm cake are saponified with an alkali, and the fatty acids are extracted from the saponified product with a nonpolar organic solvent to obtain earthworm lipids. Examples of the nonpolar organic solvent include hydrocarbon organic solvents, specifically, for example, hexane, cyclohexane, and heptane.

[0029] The total lipids obtained as described above usually contain highly unsaturated fatty acids, which are easily oxidized. Taking into consideration the irritation of the oxidized acids to the skin, the lipids may be purified by a conventional method such as using a silica gel column (a procedure to remove highly unsaturated fatty acids). It is also possible to deal with the irritation to the skin by using an antioxidant without removing highly unsaturated fatty acids.

[0030] The earthworm lipids (total lipids) obtained by the above-mentioned extraction procedures are usually oily extracts, but additives may be added to such extracts and appropriately processed to form compositions containing the extracts (solid, semi-solid, liquid, etc.) that can be used as earthworm lipids.

[0031] The composition of the present invention or the monoene enhancer of the present invention can also be used as a raw material for cosmetics. A cosmetic containing the composition of the present invention or the monoene enhancer of the present invention (hereinafter referred to as the "cosmetic of the present invention") can be said to be a functional cosmetic that can increase or improve the diversity of normal skin bacteria compared to the state before application, or a functional cosmetic that increases the ratio of monoene fatty acids having 16 carbon atoms on the skin compared to the state before application. In addition, the cosmetic of the present invention is not limited to a functional cosmetic, and may be, for example, a skin care cosmetic, a skin-beautifying cosmetic, or a body care cosmetic. The cosmetic composition of the present invention may be in the form of, for example, a transparent liquid, dispersion, milk, cream, gel, semi-solid, solid, foam, powder or granules, aerosol, or sheet.

[0032] Here, "cosmetics" includes not only "cosmetics" as defined in the Pharmaceutical and Medical Device Act, but also "quasi-drugs" such as medicated cosmetics.

[0033] In producing the cosmetic of the present invention, various ingredients can be appropriately selected and used in addition to the composition of the present invention and the monoene enhancer of the present invention. Examples of such ingredients include water (purified water, distilled water, ion-exchanged water, etc.), monohydric alcohol (ethanol, isopropanol, etc.), moisturizers, oils, surfactants, pigments and coloring materials, fragrances, whitening active ingredients, anti-wrinkle ingredients, UV absorbers, anti-fading agents, antioxidants, cosmetic ingredients, preservatives, anti-inflammatory ingredients, bactericidal ingredients, antipruritic ingredients, etc.

[0034] Examples of the moisturizing agents include polyhydric alcohols such as glycerin, propylene glycol, dipropylene glycol, 1,3-butylene glycol, polyethylene glycol (300, 400, 1500, 4000), and polyglycerin; sugars such as glucose, sucrose, trehalose, pullulan, and maltitol; amino acids such as pyrrolidone carboxylic acid and citrulline; polysaccharides such as heparin-like substances and mucopolysaccharides; and biopolymers such as hyaluronic acid or salts thereof and collagen.

[0035] Examples of the oils include fats and oils such as cacao butter, shea butter, olive oil, camellia 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 tallow; waxes such as carnauba wax, candelilla wax, jojoba oil, beeswax, lanolin, and orange roughy 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 cetanol, stearyl alcohol, isostearyl alcohol, oleyl alcohol, lauryl alcohol, cholesterol, and sitosterol; and liquid paraffin. , petrolatum, paraffin, ceresin, microcrystalline wax, squalane, pristane, and other hydrocarbons; esters such as glyceryl triisostearate, cetyl octanoate, isopropyl myristate, cetyl lactate, diisostearyl malate, and other silicone oils such as siloxane, dimethicone, cyclic dimethyl silicone oil, methyl polysiloxane, methyl phenyl polysiloxane, and other silicone oils; and others, regardless of their origin (such as animal oils, vegetable oils, or synthetic oils) and their properties (such as solid, liquid, or volatile), including hydrocarbons, oils and fats, waxes, hardened oils, ester oils, fatty acids, higher alcohols, silicone oils, lanolin derivatives, oil-based gelling agents, and oil-soluble resins.

[0036] Examples of the surfactant include anionic surfactants such as higher fatty acid soaps, alkyl sulfate salts, alkyl ether sulfate salts, alkyl phosphate salts, alkyl ether phosphate salts, N-acylamino acid salts, and N-acyl-N-methyl taurine salts; cationic surfactants such as alkyltrimethylammonium chloride, dialkyldimethylammonium chloride, and benzalkonium chloride; amphoteric surfactants such as alkyldimethylaminoacetate betaine and alkylimidazolinium 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.), polyhydric alcohol ester type nonionic surfactants (glycerin fatty acid esters, polyglycerin fatty acid esters, sorbitan fatty acid esters, sucrose fatty acid esters, etc.), and block polymer type surfactants; polymeric surfactants such as acrylic acid / alkyl methacrylate (C10-30) copolymers, polyvinyl alcohol, sodium alginate, and cellulose derivatives; natural surfactants such as lecithin; and silicone type surfactants.

[0037] Examples of the above-mentioned pigments or coloring materials include ultraviolet absorbers such as para-aminobenzoic acid derivatives, benzophenone derivatives, methoxycinnamic acid derivatives, and salicylic acid derivatives; ultraviolet scattering agents such as titanium oxide microparticles and zinc dioxide microparticles; 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, indigoid dyes, lakes, azo pigments, and phthalocyanine pigments; natural pigments such as beta-carotene, lilopine, crocin, raffin, shisonin, carthamine, safflower yellow, chlorophyll, riboflavin, cochineal, alizarin, shikonin, and curcumin; and polymer powders such as polyethylene powder and nylon powder.

[0038] Examples of the fragrances include natural fragrances such as rose oil, jasmine oil, lavender oil, eucalyptus oil, patchouli oil, peppermint oil, lemongrass oil, lemon oil, lime oil, bergamot oil, sandalwood, cinnamon, bark oil, oakmoss, iris oil, vetiver oil, musk, civet, cathurium, and ambergris; and synthetic fragrances such as α-limonene, β-caryophyllene, cis-3-hexenol, linalool, farnesol, β-phenylethyl alcohol, 2,6-nonadenal, citral, α-hexylcinnamic aldehyde, β-ionone, L-carvone, cyclopentadecanone, linalyl acetate, benzyl benzoate, γ-undecalactone, eugenol, rose oxide, and phenylacetaldehyde dimethyl acetal.

[0039] Examples of the whitening active ingredient include magnesium ascorbic acid phosphate, ascorbic acid-2-glucoside, 3-O-ethyl ascorbic acid, ascorbyl tetrahexyldecanoate, kojic acid, hydroquinone β-D-glucoside, ellagic acid, 4n-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 nicotinamide.

[0040] Examples of the anti-wrinkle components include polyhydric alcohols 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.

[0041] The method for producing the cosmetic composition of the present invention is not particularly limited, and it can be produced by a conventional method depending on the desired formulation. For example, the above-mentioned various blended components including the activator of the present invention can be mixed using an emulsifier such as a batch-type vacuum emulsifier, a dispersing machine such as a colloid mill, or a mixer / pulverizer such as a Henschel mixer or a hammer mixer, and the desired formulation can be obtained using a cooling machine, a molding machine, etc. as necessary.

[0042] In addition, the cosmetic of the present invention obtained by an appropriate manufacturing method can be packaged into a tube container, a bottle container (narrow mouth), a cream container (wide mouth), a powder container, a compact container, a stick container, a pencil container, a container with a brush, a pump bottle, an aerosol container, etc. using a filling machine, packaging machine, etc. to produce a cosmetic product. EXAMPLES

[0043] The present invention will be described in more detail below with reference to examples, but the present invention is not limited to the following examples in any way.

[0044] [Reference Example] Examination of the diversity of skin flora A total of 100 individuals, including 50 healthy individuals (9 of whom had a history of atopy) and 50 atopy carriers, were sampled from each individual's resident skin bacteria, and the diversity of bacterial species in the resident skin flora (diversity of resident skin bacteria) was investigated by metagenomic analysis. The diversity was evaluated using Simpson's Diversity Index (D) and Shannon Entropy (H). The values ​​of D and H are the proportion of the number of individuals of the i-th species in the total number of bacterial species in the resident skin flora to the total number of individuals in the flora, expressed as p i (|p i |<1) and calculated using the following formula:

[0045]

number

[0046]

number

[0047] Simpson's diversity index takes values ​​between 0 and 1, and the closer to 1 the value, the more uniformly diverse the bacteria are. In Shannon entropy, the larger the absolute value of the number, the higher the diversity. The analysis results are shown in Table 1 below.

[0048] [Table 1]

[0049] As shown in Table 1, it is clear that the group without atopic symptoms had significantly higher diversity (more bacterial species) than the group with atopic symptoms. A comparison of normal skin flora in healthy subjects and normal skin flora in atopic dermatitis holders is shown in Figure 1, with only one example for each. Figure 1 shows flora with an occupancy rate of 0.05% or more. As shown in Figure 1, 194 types of flora were identified in healthy subjects, of which about one-third were non-pathogenic acne bacteria, whereas only 31 types of flora were identified in atopic dermatitis holders, of which more than one-third were pathogenic Staphylococcus aureus.

[0050] Furthermore, the results of sebum sampling, which was conducted at the same time as the sampling of resident skin bacteria, showed that 23 of the 50 atopy carriers had sebum secretion levels that were one-tenth or less than that of healthy individuals. As mentioned above, atopic dermatitis carriers tend to have a low diversity of resident skin bacteria, and a certain number of individuals tend to have not only a low diversity of resident skin bacteria but also reduced sebum secretion.

[0051] [Example 1] Preparation of earthworm lipids (1) Obtaining earthworm materials The earthworm material used to obtain the earthworm lipids used in the experiments described below was the extraction residue (dehydrated earthworm cake) after protein extraction from Lumberg's rubellus (red earthworm), and was provided by Miyazaki Blood Flow Research Institute Co., Ltd. (Miyazaki Prefecture).

[0052] (2) Preparation of earthworm lipids Earthworm lipids (total lipids) were prepared by one of the following methods. (2-1) Preparation by Bligh-Dyer Method Total lipids were extracted from 10 g of dehydrated earthworm cake using 30 mL of a chloroform / methanol mixture (2:1). Approximately 2% of earthworm lipids were obtained.

[0053] (2-2) Preparation by direct saponification method 60g of dehydrated earthworm cake was collected in an eggplant-shaped flask, and 120mL of 1mol / KOH / ethanol was added, followed by heating at 80℃ for 1 hour to saponify the fatty acid esters in the sample. After cooling, purified water was added, and the ethanol solution after saponification was filtered in a liquid extraction apparatus and refluxed with diisopropyl ether for 5 hours to extract and remove unsaponifiable matter. The alkaline solution in the liquid extraction device was removed, sulfuric acid was added to make it strongly acidic, and then it was filtered and refluxed with hexane for 5 hours to extract fatty acids.

[0054] The hexane extract of fatty acids was washed with purified water until it became neutral (methyl orange), filtered, concentrated to dryness, and then acetonitrile was added to dissolve the fatty acids. The mixture was then allowed to stand for 10 minutes to precipitate the acetonitrile insolubles. 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 lipids.

[0055] (3) Purification of earthworm lipids: Removal of polyunsaturated fatty acids Wakosil C-300 (activated at 110°C for 3 hours) was slurried with hexane and packed into a glass column (inner diameter 15 mm, length 30 cm). Florisil PR (activated at 130°C for 3 hours) was slurried with hexane and layered on top of Wakosil C-300. Anhydrous sodium sulfate was then layered on top to create a column for purifying fatty acids. The concentrated and dried fatty acids were dissolved in hexane and loaded onto the column, and developed and eluted with ether / hexane.

[0056] [Example 2] Examination of the effect of earthworm lipids on improving the normal skin flora (1) Experimental method A total of 12 subjects, 9 atopy holders and 3 healthy subjects shown in Table 2, were used as subjects. The control / squalane and 1% earthworm lipid / squalane prepared in Example 1 (direct saponification method) were applied to the inside of the left and right elbows of each subject, with the subjects being blinded as to which one contained earthworm lipid to avoid a placebo effect. The same was applied twice a day, one drop each in the morning and evening (after bathing), on the same side. Three healthy subjects and one atopy holder applied the samples for 4 weeks, and the other eight atopy holders applied the samples for 2 weeks. Microorganisms on the skin were sampled by scraping the sample application area with a swab kit cotton swab for metagenomic analysis. The 16srRNA gene of bacteria was extracted from the swab, and its sequence was read. The sequence was compared with the reference total bacterial sequence database (approximately 15,000 species) to identify the bacterial species to the species level, and the number of individuals, Simpson's Index, and Shannon Entropy were calculated based on the number of DNA fragments of each bacterial species.

[0057] [Table 2]

[0058] After the two-week application, sebum was sampled from the application sites (both left and right) using oil blotting paper. Extraction, purification, and derivatization were then performed, followed by GC / MS analysis to examine the amount of sebum secreted and to analyze the ratio of monoenoic acid with 16 carbon atoms (C16:1), which is considered to have high antibacterial properties, to linear saturated acid with 16 carbon atoms (C16:0).

[0059] (2) Analysis results of subject A-109 Of the nine atopic dermatitis patients, four had extremely low sebum secretion, one of which was A-109. In A-109, in addition to a change in diversity, a change in the top bacterial species was observed when comparing before application and after 15 days. The results for the number of bacterial species on days 0, 8, and 15 are shown in Table 3. Note that bacterial species with an occupancy rate of 0.05% or more in the bacterial flora were counted. As shown in Table 3, application of earthworm lipids resulted in a significant increase in the number of bacterial species compared to the control.

[0060] [Table 3]

[0061] For A-109, we calculated Simpson's Index (the closer to 1, the higher the diversity) and Shannon Entropy (the higher the value, the higher the diversity) and performed diversity analysis. The results are shown in Figure 2. As shown in Figure 2, the diversity of the control A-109 tended to decrease, but when earthworm lipids were applied, the diversity tended to increase.

[0062] In addition, a GC / MS analysis of a separately sampled sebum gave the results shown in Figure 3. In Figure 3, the chromatographic scale of C16:0 (saturated fatty acid with 16 carbon atoms) was also used for comparison. As shown in Figure 3, the ratio of carbon number 16 monoenoic acids, which are considered to have high antibacterial properties, to carbon number 16 linear saturated acids did not change in the control compared to before application, but it increased significantly when earthworm lipid was applied. In addition, since the amount of isomers different from monoenoic acids contained in the applied earthworm lipid increased, it is speculated that carbon number 16 monoenoic acid isomers are produced from earthworm lipid by the action of resident skin bacteria.

[0063] (3) Analysis results of subject A-107 Subject A-107 was also one of those with low sebum secretion and had low bacterial flora diversity before the experiment. The application of earthworm lipids increased the diversity of the bacterial flora, and a change in the most dominant bacterial species was observed in both the control and earthworm lipid applications. The results for the number of bacterial species are shown in Table 4. As shown in Table 4, application of earthworm lipids resulted in a significant increase in the number of bacterial species compared to the control.

[0064] [Table 4]

[0065] For A-107, we calculated Simpson's Index and Shannon Entropy and performed a diversity analysis, obtaining the results shown in Figure 4. As shown in Figure 4, in A-107, although there was little change in the control, the diversity tended to decrease, and when earthworm lipid was applied, the diversity increased.

[0066] Similarly, GC / MS analysis of separately sampled sebum was performed, and the results shown in Figure 5 were obtained. Note that in Figure 5, the chromatographic scale for C16:0 (saturated fatty acid with 16 carbon atoms) was also compared. In A-107, perhaps due to the change in bacterial species, the ratio of monoenoic acids with 16 carbon atoms, which are considered to have high antibacterial properties, to linear saturated acids with 16 carbon atoms, increased compared to before application in both the control and earthworm lipid application, but the increase rate of monoenoic acids was greater when earthworm lipid was applied.

[0067] (4) Analysis results of subject A-105 Subject A-105 had normal sebum secretion and a high bacterial flora diversity before application. The application reduced the proportion of acne bacteria, which had a high occupancy rate, and the diversity increased accordingly. A change in the most prevalent bacterial species was observed in both the control and earthworm lipid applications. The results of the bacterial species count are shown in Table 5. As shown in Table 5, when the amount of sebum secretion was normal, the application of earthworm lipids only slightly increased the number of bacterial species, whereas a clear decrease in the number of bacterial species was observed in the control.

[0068] [Table 5]

[0069] For A-105, we calculated Simpson's Index and Shannon Entropy and performed diversity analysis, and obtained the results shown in Figure 6. As shown in Figure 6, there was not much difference in Simpson's Index between the control and the earthworm lipid application, but the earthworm lipid application showed a higher diversity in Shannon Entropy. This was presumably due to the large number of bacterial species with low occupancy.

[0070] Similarly, GC / MS analysis of a separate sample of sebum was performed, and the results shown in Figure 7 were obtained. Note that in Figure 7, the chromatographic scale for C16:0 (saturated fatty acid with 16 carbon atoms) was also used for comparison. In A-105, almost no change in the monoenoic acid ratio was observed due to application. It was speculated that lipid application has little effect on people who have a high level of diversity and high sebum secretion. [Industrial Applicability]

[0071] The composition of the present invention can increase or improve the diversity of resident skin bacteria from the state before application, thereby enhancing the barrier function of the skin and suppressing the growth of pathogenic bacteria, and is therefore useful, for example, as an ingredient for skin care products and cosmetics. In addition, the increase or improvement of the diversity of resident skin bacteria leads to the improvement and prevention of atopic symptoms and the maintenance of healthy skin, so the composition of the present invention is also useful as an ingredient for pharmaceuticals.

Claims

1. A composition for controlling the diversity of bacterial species in the normal skin flora, comprising an effective amount of earthworm lipid derived from the genus Lumbricus and a base (excluding those containing ethyl esters or butyl esters of fatty acids constituting earthworm lipid that are not highly unsaturated fatty acids).

2. A method for producing the composition described in claim 1, comprising a step of obtaining earthworm lipids from the extraction residue remaining after extracting and removing proteins from raw earthworms.

3. The composition described in claim 1, wherein the earthworm lipid derived from the genus Lumbricus is red earthworm (Lumbricus rubellus) lipid.

4. The composition according to claim 1, for application to subjects with low sebum secretion or subjects with atopic skin conditions.

5. 5. An agent for regulating the amount of a monoenoic fatty acid having 16 carbon atoms on the skin, comprising the composition according to claim 1 or 3 to 4.

6. The composition according to any one of claims 1 and 3 to 4, or the agent for regulating the amount of a monoenoic fatty acid having 16 carbon atoms on the skin according to claim 5, wherein the dosage form is an ointment, a cream, a gel, a liquid, a cataplasm, a tape, a sheet, an aerosol, a powder for external use, a spray, or a bath additive.

7. A cosmetic preparation comprising the composition according to any one of claims 1 and 3 to 6 or an agent for adjusting the amount of a monoenoic fatty acid having 16 carbon atoms on the skin.

8. A method for producing a composition for controlling the resident skin flora, comprising the steps of extracting the composition using a polar organic solvent from the extraction residue (dehydrated earthworm cake) remaining after proteins have been extracted and removed from raw earthworms, or saponifying the dehydrated earthworm cake with an alkali and extracting the composition from the saponified product using a non-polar organic solvent.

9. A method for producing earthworm lipids for controlling the normal skin flora, comprising the steps of extracting from the extraction residue (dehydrated earthworm cake) remaining after proteins have been extracted and removed from raw earthworms using a polar organic solvent, or saponifying the dehydrated earthworm cake with an alkali and extracting from the saponified product using a non-polar organic solvent.

10. The method according to claim 8 or 9, wherein the polar organic solvent is a mixed solvent of a chlorine-based organic solvent and an alcohol-based organic solvent.

11. The method according to claim 8 or 9, wherein the non-polar organic solvent is a hydrocarbon organic solvent.

12. A composition for controlling the diversity of bacterial species in the normal skin flora (excluding those containing ethyl esters or butyl esters of fatty acids that are not highly unsaturated fatty acids and are constituents of earthworm lipids. ), comprising the steps of: Extracting earthworm lipids from earthworms of the genus Lumbricus; Mixing earthworm lipids from the genus Lumbricus with a base; The method includes:

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