Antibacterial agent for skin resident bacteria

Abstract

To provide a novel improver of a skin resident bacteria flora.SOLUTION: There are provided: 1) a Staphylococcus aureus antibacterial agent including as an active ingredient, at least one kind of S-petasin, S-isopetasin, and fukinone; 2) a skin resident bacteria flora improver including as an active ingredient, at least one kind of S-petasin and S-isopetasin; 3) the skin resident bacteria flora improver described in the 2) which exhibits strong antibacterial activity to Staphylococcus aureus, and exhibits weak antibacterial activity to Staphylococcus epidermidis, thereby exhibiting an improvement action of the skin resident bacteria flora; 4) an anti-dandruff bacteria agent including as an active ingredient, at least one of condensed tannin and kaempferol 3-O-(6"-acetyl)-glucoside.SELECTED DRAWING: None

Description

Technical Field

[0001] The present invention relates to an antibacterial agent against skin commensal bacteria derived from perilla, and is applied as a material for, for example, food and drink products, pharmaceuticals, cosmetics, etc.

Background Art

[0002] On the surface of healthy human skin, a skin commensal flora, an aggregate of microorganisms called skin commensal bacteria, is formed. Although there are individual differences, site differences, seasonal variations, etc., Propionibacterium acnes (P. acnes) is the most dominant anaerobic bacterium in the skin commensal flora, and Staphylococcus aureus (Staphylococcus aureus: S. aureus) and Staphylococcus epidermidis (Staphylococcus epidermidis: S. epidermidis) are aerobic bacteria that inhabit the skin as skin commensal bacteria. It has been reported that the skin commensal flora plays two conflicting roles: biological defense against other pathogenic microorganisms and disease manifestation (endogenous infection). As a biological defense, there is a barrier function to prevent the entry of pathogenic bacteria from the outside. However, when the balance of the skin commensal flora is disrupted by trauma, poor general condition due to illness, mental stress, cold, dryness, inappropriate washing, etc., skin commensal bacteria such as S. aureus and P. acnes overgrow, inducing various skin symptoms and causing disease manifestation (Patent Document 1). Thus, in the skin commensal bacteria, it is preferable that the state is such that S. aureus is less and S. epidermidis is more.

[0003] Moreover, dandruff is composed of secretions of sebaceous glands, secretions of sweat glands, exfoliated or desquamated substances of the epidermal layer, and is usually caused by enhanced secretion of sebaceous glands or keratinization of cells. Furthermore, it is known that the growth of Malassezia, a type of scalp resident fungus and a lipophilic fungus, is involved in the occurrence of dandruff. Conventionally, Malassezia furfur has been known as the causative fungus of dandruff. Furthermore, in recent years, it has been reported that other Malassezia species such as Malassezia restricta and Malassezia globosa are also involved in dandruff and other scalp diseases (Patent Document 3).

[0004] In addition, the odor that occurs under the armpits (also called axillary osmidrosis) can be roughly classified into an acidic and stuffy odor (referred to as sweat odor, acid odor, etc.) and an apocrine odor (also called "waki ga"). The apocrine odor is caused by secretions derived from apocrine sweat glands distributed under the armpits, and due to its complex and strong odor, it is particularly likely to be perceived by the person himself or those around him. Therefore, for people who are concerned about axillary osmidrosis, the strength of the apocrine odor in particular is a matter of concern. Furthermore, the development of materials for suppressing the occurrence of axillary osmidrosis is also demanded.

[0005] The apocrine odor is mainly composed of (1) a sulfur-like and putrid odor and (2) an animal-like and spicy odor, and their main causative components are, respectively, alcohol compounds having a thiol group at the 3-position represented by 3-mercapto-3-methylhexan-1-ol (hereinafter, these compounds are also referred to as 3-mercapto alcohol compounds), and β-hydroxy acid compounds represented by 3-hydroxy-3-methylhexanoic acid.

[0006] To date, S. hominis is known as the producing bacterium of 3-mercapto alcohol compounds such as 3-mercapto-3-methylhexan-1-ol, which is a causative substance of the sulfur-like odor among apocrine odors.

[0007] Furthermore, sebum secreted from the skin is decomposed by lipase derived from skin microorganisms to generate fatty acids, which further undergo oxidative decomposition to produce malodorous substances. In addition, these fatty acid decomposition products stimulate skin cells, causing inflammations such as pimples, acne, and rough skin, as well as bad odors.

[0008] In particular, Propionibacterium acnes, along with Staphylococcus, is the most prevalent on human skin and forms a bacterial flora on healthy skin. It plays a major role in the development of acne and has also been reported as a cause of chronic infections. On the skin, Propionibacterium acnes secretes lipase in hair follicles to decompose sebum and produce fatty acids. Fatty acids accumulated in hair follicles are oxidized by ultraviolet rays, oxygen, etc., which forms plugs, causes suppuration, and the resulting inflammatory state is acne. Also, when sebum is decomposed by lipase derived from Propionibacterium acnes, various substances that cause body odor are formed (Patent Document 4). In particular, seborrheic areas such as around the nose, forehead, and behind the ears have densely packed sebaceous glands and are prone to sebum secretion, so Propionibacterium acnes is likely to proliferate.

[0009]

Patent Document 1

Patent Document 2

Patent Document 3

Patent Document 4

Disclosure of the Invention

Problems to be Solved by the Invention

[0010] Under such circumstances, the inventors of the present invention have found that among the extracts of Petasites japonicus and its constituents, S-petasin and S-isopetasin have strong antibacterial activity against Staphylococcus aureus among the skin commensal bacteria, and have weak antibacterial activity against Staphylococcus epidermidis, thus having an improving effect on the skin commensal flora. They have also found that fukinone has antibacterial activity against Staphylococcus aureus, and have completed the present invention. An object of the present invention is to provide a novel agent for improving the skin commensal flora.

Means for Solving the Problems

[0011] To solve the above problems, the technical features of the present invention are as follows. 1. An antibacterial agent against Staphylococcus aureus containing at least one of S-petasin, S-isopetasin, and fukinone as an active ingredient. 2. An agent for improving the skin commensal flora containing at least one of S-petasin and S-isopetasin as an active ingredient. 3. The agent for improving the skin commensal flora according to 2. above, characterized in that it has a strong antibacterial activity against Staphylococcus aureus and a weak antibacterial activity against Staphylococcus epidermidis, thereby having an improving effect on the skin commensal flora. 4. An antifungal agent containing at least one of condensed tannin and kaempferol 3-O-(6"-acetyl)-glucoside as an active ingredient. 5. An agent for improving the skin commensal flora containing the fat-soluble extract of Petasites japonicus as an active ingredient. 6. An antibacterial agent against axillary osmidrosis containing the fat-soluble extract of Petasites japonicus as an active ingredient. 7. An antibacterial agent against Malassezia containing the fat-soluble extract of Petasites japonicus as an active ingredient. 8. An antibacterial agent against acne bacteria at seborrheic sites containing the fat-soluble extract of Petasites japonicus as an active ingredient.

Effects of the Invention

[0012] According to the present invention, since S-petasin, S-isopetasin, and fukinone have antibacterial effects against Staphylococcus aureus, by containing at least one of these as an active ingredient, it is useful as a Staphylococcus aureus antibacterial agent. Further, according to the present invention, by having strong antibacterial activity against Staphylococcus aureus and weak antibacterial activity against Staphylococcus epidermidis, the ratio of Staphylococcus aureus to Staphylococcus epidermidis on the skin can be adjusted, thereby having an improving effect on the skin resident flora, and thus being useful as an agent for improving the skin resident flora. Furthermore, according to the present invention, since condensed tannin and kaempferol 3-O-(6"-acetyl)-glucoside have an inhibitory effect on pityrosporum, they are useful as anti-pityrosporum agents. Also, according to the present invention, the fuki liposoluble extract shows concentration-dependent antibacterial activity against the bad bacterium Staphylococcus aureus S. Aureus, but does not show antibacterial activity against the good bacterium Staphylococcus epidermidis S. Epidermidis etc. Thus, since it does not affect the growth of good bacteria but shows antibacterial activity against bad bacteria, it has an improving effect on the skin resident flora, and thus is useful as an agent for improving the skin resident flora. Furthermore, according to the present invention, by applying the fuki liposoluble extract, S. hominis that produces 3-mercaptoalcohol compounds, which cause axillary osmidrosis, is suppressed in the armpit. Thus, it is useful as an anti-axillary osmidrosis bacterium agent. Also, according to the present invention, by applying the fuki liposoluble extract, it has an effect of reducing the presence ratio of Malassezia bacteria. Thus, it is useful as an anti-Malassezia bacterium agent and is also effective for scalp care and health care of other hair growth sites. Furthermore, it has been confirmed that in seborrheic areas, it has an effect of suppressing the growth of excessive acne bacteria (P. Acnes), and thus is useful as an anti-acne bacterium agent in seborrheic areas. In the present application, the "seborrheic area" refers to an area where sebum secretion is active, for example, around the nose, forehead, behind the ears, etc., but is not limited to these as long as it is an area where sebum secretion is active.

Brief Description of the Drawings

[0013]

Figure 1

Figure 2

Figure 3

Figure 4

Figure 5

Best Mode for Carrying Out the Invention

[0014] Hereinafter, the present invention will be described in detail. The Staphylococcus aureus antibacterial agent of the present invention is characterized by containing at least one of S-petasin, S-isopetasin, and fukinone as an active ingredient. Further, the skin commensal flora improver of the present invention is characterized by containing at least one of S-petasin and S-isopetasin as an active ingredient. The method for obtaining the above compounds S-petasin, S-isopetasin, and fukinone is not particularly limited, but it is preferably extracted from Petasites japonicus. "Petasites japonicus" which is the raw material of the Staphylococcus aureus antibacterial agent and the skin commensal flora improver of the present invention (hereinafter referred to as "skin commensal flora improver, etc.") is a perennial herb of the genus Petasites of the Asteraceae family, and is called "Houttuynia cordata Thunb." in the Chinese crude drug name. It is dioecious and has a rhizome that runs horizontally underground, and broad leaf blades are attached to the petioles emerging from this rhizome. The flowers appear prior to the unfolding of the leaves in early spring, and bear a corymbose capitulum (flower stem: Petasites japonicus) composed of tubular flowers at the top of a short upright stem. It has been cultivated since ancient times as a vegetable native to Japan, and the inflorescence, flower stalk, or leaf stalk is used for food. Wild-growing ones are also collected and utilized. Cultivated varieties include Aichi Wase Buky, Mizu Fuki (Kyoto Buky), Akita Buky, Rawang Buky, etc.

[0015] As solvents for extracting active ingredients such as skin resident flora improvers from raw materials, water, methanol, ethanol, isopropyl alcohol, 1,3-butylene glycol, ethylene glycol, propylene glycol, glycerin, ethyl acetate, acetone, etc. can be used. Two or more of these solvents may be mixed. Desirably, hydrous ethanol may be used as the extraction solvent. In particular, hydrous ethanol is an extraction solvent that is less likely to reduce the activity of the active ingredient during extraction and is also preferable in terms of the safety of the extract for food use. The type of water for extraction is not particularly limited, and tap water, distilled water, mineral water, alkaline ion water, deep water, etc. can be used.

[0016] As the extraction temperature, for example, when using hydrous ethanol, the extraction may be carried out at an extraction temperature of 20 to 80°C, desirably about 60 to 70°C. This is because if the extraction temperature is too low, it becomes difficult to extract the active ingredient, and if the extraction temperature is too high, the activity of the active ingredient is likely to decrease.

[0017] The hydrous ethanol as the extraction solvent preferably has an ethanol concentration of 40 to 90% (wt / wt). The ethanol concentration is set at 40% (wt / wt) or more because if the ethanol content is too low, the extraction amount will be small, and the ethanol concentration is set at 90% (wt / wt) or less because if the ethanol concentration is too high, it becomes difficult to extract the active ingredient. Desirably, the ethanol concentration is 60 to 80% (wt / wt), and more desirably about 70% (wt / wt). Note that for ethanol extraction, in order to improve the content rate of the active ingredient, it is preferably repeated while changing the ethanol concentration step by step.

[0018] As an extraction method for the skin resident flora improver of the present invention, any method such as continuous extraction, immersion extraction, countercurrent extraction, supercritical extraction, etc. can be adopted, and any device can be used at room temperature or under reflux heating.

[0019] Specifically, the extraction raw material (Petasites japonicus) is put into a treatment tank filled with an extraction solvent, and the active ingredient is eluted while stirring. For example, when using hydrous ethanol as the extraction solvent, an extraction solvent about 5 to 100 times the amount (weight ratio) of the extraction raw material is used, and extraction is carried out for about 30 minutes to 2 hours. After eluting the active ingredient in the solvent, filtration is performed to remove the extraction residue to obtain an extract. Then, the extract is treated by dilution, concentration, drying, purification, etc. according to conventional methods, and the compounds S-petasin, S-isopetasin, and fukinone are isolated and purified to obtain the skin resident flora improver of the present invention. In the present invention, those obtained by isolating these compounds from Petasites japonicus extract may be used, but Petasites japonicus extract that does not require isolation and purification as long as it contains these compounds may also be used. Examples of the isolation and purification methods include methods such as activated carbon treatment, resin adsorption treatment, ion exchange resin, liquid-liquid countercurrent distribution, etc. Specifically, for example, isolation and purification can be carried out by the method of the examples in the present specification.

[0020] The skin resident flora improver of the present invention can be used as a raw material for various foods and beverages. Examples of foods and beverages include general foods (nutritional drinks, etc.) such as confectionery (gum, candy, caramel, chocolate, cookies, snacks, jelly, gummy, tablet confectionery, etc.), noodles (buckwheat noodles, udon noodles, ramen noodles, etc.), dairy products (milk, ice cream, yogurt, etc.), seasonings (miso, soy sauce, etc.), soups, and beverages (juice, coffee, black tea, tea, carbonated beverages, sports drinks, etc.). The skin resident flora improver of the present invention may be appropriately formulated in these foods and beverages.

[0021] These food and drink products can be formulated with various ingredients according to their types. For example, glucose, fructose, sucrose, maltose, sorbitol, stevioside, corn syrup, lactose, citric acid, tartaric acid, malic acid, succinic acid, lactic acid, L - ascorbic acid, dl-α - tocopherol, sodium erythorbate, glycerin, propylene glycol, glycerin fatty acid ester, polyglycerin fatty acid ester, sucrose fatty acid ester, sorbitan fatty acid ester, propylene glycol fatty acid ester, gum arabic, carrageenan, casein, gelatin, pectin, agar, vitamin Bs, nicotinamide, calcium pantothenate, amino acids, calcium salts, pigments, flavors, preservatives and other food materials can be used.

[0022] As a specific manufacturing method, the solvent extract of butterbur can be spray - dried or freeze - dried together with powdered cellulose, and then it can be easily incorporated into food and drink products (such as instant foods) by making it into powder, granules, tablets or solutions. Also, the solvent extract of butterbur can be dissolved in, for example, oils and fats, ethanol, glycerin or a mixture thereof to make it liquid, and then added to beverages or solid foods. If necessary, it can be mixed with binders such as gum arabic and dextrin to make it powdery or granular, and then added to beverages or solid foods.

[0023] When applying the skin resident flora improver of the present invention etc. to food and drink products, the addition amount is preferably such that the total content of the active ingredient is 1 - 20 wt% or less with respect to the food and drink products, since the main purposes are disease prevention and health maintenance.

[0024] The skin resident flora improver and the like of the present invention may be used as a material for drugs (including pharmaceuticals and quasi-drugs). The raw material for drug preparations can be produced by appropriately blending an antibacterial agent against the skin resident bacteria of the present invention. Examples of the formulation raw materials that can be blended in the skin resident flora improver and the like of the present invention include excipients (glucose, lactose, sucrose, sodium chloride, starch, calcium carbonate, kaolin, crystalline cellulose, cacao butter, hardened vegetable oil, kaolin, talc, etc.), binders (distilled water, physiological saline, ethanol water, simple syrup, glucose solution, starch solution, gelatin solution, carboxymethyl cellulose, potassium phosphate, polyvinylpyrrolidone, etc.), disintegrants (sodium alginate, agar, sodium hydrogen carbonate, calcium carbonate, sodium lauryl sulfate, monoglyceride stearate, starch, lactose, powdered gum arabic, gelatin, ethanol, etc.), disintegration inhibitors (sucrose, stearin, cacao butter, hydrogenated oil, etc.), absorption promoters (quaternary ammonium base, sodium lauryl sulfate, etc.), adsorbents (glycerin, starch, lactose, kaolin, bentonite, silicic acid, etc.), lubricants (purified talc, stearate, polyethylene glycol, etc.), and the like.

[0025] The administration method of the skin resident flora improver and the like according to the present invention can generally be administered orally in the form of tablets, pills, soft and hard capsules, fine granules, powders, granules, liquids, etc., but parenteral administration may also be used. When administered as a parenteral preparation, it can be administered into local tissues, intradermally, subcutaneously, intramuscularly, and intravenously in a solution state or in a state where a dispersant, a suspending agent, a stabilizer, etc. are added. It may also be in the form of suppositories or the like.

[0026] The dosage can vary depending on the administration method, the condition of the disease, the age of the patient, etc. In adults, usually, 0.5 to 5000 mg of the active ingredient (perilla extract) per day can be administered, and in children, usually about 0.5 to 3000 mg can be administered. The blending ratio of the skin resident flora improving agent and the like can be appropriately changed depending on the dosage form. Usually, when administered orally or by mucosal absorption, it is about 0.3 to 15.0 wt%, and when administered parenterally, it is preferably about 0.01 to 10 wt%. Note that the dosage varies depending on various conditions, so there may be cases where an amount less than the above dosage is sufficient, and there may also be cases where it is necessary to administer beyond the range.

[0027] Examples of the forms of cosmetics that can contain the skin resident flora improving agent and the like of the present invention include, for example, emulsions, soaps, facial washes, bath agents, creams, emulsions, lotions, colognes, beard shaving creams, beard shaving lotions, makeup oils, sunburn / sunblock lotions, face powders, foundations, perfumes, packs, nail creams, enamels, enamel removers, eyebrow pencils, blushes, eye creams, eye shadows, mascaras, eyeliners, lipsticks, lip creams, shampoos, rinses, hair dyes, dispersions, cleaning agents, and the like. Examples of the forms of pharmaceuticals or quasi-drugs that can contain the skin resident flora improving agent and the like of the present invention include ointments, creams, external liquid preparations, and the like.

[0028] In addition to the skin resident flora improving agent and the like according to the present invention, the above-mentioned skin external preparations can be formulated with components, oils, higher alcohols, fatty acids, ultraviolet absorbers, powders, pigments, surfactants, polyhydric alcohols / sugars, polymers, physiologically active components, solvents, antioxidants, fragrances, preservatives, etc. that are formulated in cosmetics, quasi-drugs, etc. within a range that does not impair the flora improving effect on the skin resident flora. Specific examples are listed below, but the present invention is not limited to these examples. (1) Examples of oils Ester-based oil phase components: glyceryl tri(2-ethylhexanoate), cetyl 2-ethylhexanoate, isopropyl myristate, butyl myristate, isopropyl palmitate, ethyl stearate, octyl palmitate, isocetyl isostearate, butyl stearate, butyl myristate, ethyl linoleate, isopropyl linoleate, ethyl oleate, isocetyl myristate, isostearyl myristate, isostearyl palmitate, octyldodecyl myristate, isocetyl isostearate, diethyl sebacate, diisopropyl adipate, isoaralkyl neopentanoate, glyceryl tri(caprylic / capric acid), trimethylolpropane tri(2-ethylhexanoate), trimethylolpropane triisostearate, pentaerythritol tetra(2-ethylhexanoate), cetyl caprylate, decyl laurate, hexyl laurate, decyl myristate, myristyl myristate, cetyl myristate, stearyl stearate, decyl oleate, cetyl ricinoleate, isostearyl laurate, isotridecyl myristate, isocetyl myristate, isostearyl myristate, isocetyl palmitate, isostearyl palmitate, octyl stearate, isocetyl stearate, isodecyl oleate, octyldodecyl oleate, octyldodecyl linoleate, isopropyl isostearate, cetostearyl 2-ethylhexanoate, stearyl 2-ethylhexanoate, hexyl isostearate, ethylene glycol dioctanoate, ethylene glycol dioleate, propylene glycol dicaprate, propylene glycol di(caprylic / capric acid), propylene glycol dicaprylate, neopentyl glycol dicaprylate, neopentyl glycol dioctanoate, glyceryl tricaprylate, glyceryl triundecanoate, glyceryl triisopalmitate, glyceryl triisostearate, octyldodecyl neopentanoate, isostearyl octanoate, octyl isononanoate, hexyldecyl neodecanoate, octyldodecyl neodecanoate, isocetyl isostearate, isostearyl isostearate, octyldodecyl isostearate, polyglyceryl oleate, polyglyceryl isostearate, dipropyl carbonate,Dialkyl (C12-18) carbonates, tricisocetyl citrate, triisoarachidyl citrate, triisooctyl citrate, lauryl lactate, myristyl lactate, cetyl lactate, octyldecyl lactate, triethyl citrate, acetyltriethyl citrate, acetyltributyl citrate, trioctyl citrate, diisostearyl malate, 2-ethylhexyl hydroxystearate, di-2-ethylhexyl succinate, diisobutyl adipate, diisopropyl sebacate, dioctyl sebacate, cholesteryl stearate, cholesteryl isostearate, cholesteryl hydroxystearate, cholesteryl oleate, dihydrocholesteryl oleate, phytosteryl isostearate, phytosteryl oleate, isocetyl 12-stearoylhydroxystearate, stearyl 12-stearoylhydroxystearate, isostearyl 12-stearoylhydroxystearate, etc. can be mentioned. Hydrocarbon-based oil phase components: Squalane, liquid paraffin, α-olefin oligomer, isoparaffin, ceresin, paraffin, liquid isoparaffin, polybutene, microcrystalline wax, petrolatum, etc. can be mentioned. Animal and vegetable oils and their hydrogenated oils, and waxes of natural origin: Animal oils such as beef tallow, hydrogenated beef tallow, lard, hydrogenated lard, horse oil, hydrogenated horse oil, mink oil, orange raffia oil, fish oil, hydrogenated fish oil, egg yolk oil and their hydrogenated oils, vegetable oils such as avocado oil, almond oil, olive oil, cocoa butter, apricot kernel oil, kukui nut oil, sesame oil, wheat germ oil, rice germ oil, rice bran oil, safflower oil, shea butter, soybean oil, evening primrose oil, perilla oil, tea seed oil, camellia oil, corn oil, rapeseed oil, hydrogenated rapeseed oil, palm kernel oil, hydrogenated palm kernel oil, palm oil, hydrogenated palm oil, peanut oil, hydrogenated peanut oil, castor oil, hydrogenated castor oil, sunflower oil, grape seed oil, jojoba oil, hydrogenated jojoba oil, macadamia nut oil, meadowfoam oil, cottonseed oil, hydrogenated cottonseed oil, coconut oil, hydrogenated coconut oil and their hydrogenated oils, waxes such as beeswax, high acid value beeswax, lanolin, reduced lanolin, hydrogenated lanolin, liquid lanolin, carnauba wax, montan wax, etc. can be mentioned. Silicone-based oil phase components: dimethylpolysiloxane, methylphenylpolysiloxane, methylcyclopolysiloxane, octamethylpolysiloxane, decamethylpolysiloxane, dodecamethylcyclosiloxane, methylhydrogenpolysiloxane, polyether-modified organopolysiloxane, dimethylsiloxane·methyl cetyloxysiloxane copolymer, dimethylsiloxane·methyl stearoxysiloxane copolymer, alkyl-modified organopolysiloxane, end-modified organopolysiloxane, amino-modified silicone oil, amino-modified organopolysiloxane, dimethiconol, silicone gel, acrylic silicone, trimethylsiloxysilicic acid, silicone RTV rubber, etc. Fluorine-based oil phase components: perfluoropolyether, fluorine-modified organopolysiloxane, fluorinated pitch, fluorocarbon, fluoroalcohol, fluoroalkyl·polyoxyalkylene copolymer-modified organopolysiloxane, etc. (2) Examples of higher alcohols Lauryl alcohol, myristyl alcohol, cetyl alcohol, stearyl alcohol, isostearyl alcohol, oleyl alcohol, behenyl alcohol, 2-ethylhexanol, hexadecyl alcohol, octyldodecanol, etc. (3) Examples of fatty acids Caprylic acid, capric acid, undecylenic acid, lauric acid, myristic acid, palmitic acid, palmitoleic acid, stearic acid, isostearic acid, oleic acid, linoleic acid, linolenic acid, arachidic acid, arachidonic acid, behenic acid, erucic acid, 2-ethylhexanoic acid, etc. (4) Examples of ultraviolet absorbers Para - aminobenzoic acid, amyl para - aminobenzoate, ethyl dihydroxypropyl para - aminobenzoate, glyceryl para - aminobenzoate, ethyl para - aminobenzoate, octyl para - aminobenzoate, octyldimethyl para - aminobenzoate, ethylene glycol salicylate, octyl salicylate, triethanolamine salicylate, phenyl salicylate, butylphenyl salicylate, benzyl salicylate, homomenthyl salicylate, benzyl cinnamate, octyl paramethoxycinnamate, 2 - ethylhexyl paramethoxycinnamate, glyceryl mono - 2 - ethylhexanoate diparamethoxycinnamate, isopropyl paramethoxycinnamate, diethanolamine salt of paramethoxyhydroxycinnamic acid, diisopropyl · diisopropyl cinnamate ester mixture, urocanic acid, ethyl urocanic acid, hydroxymethoxybenzophenone, hydroxymethoxybenzophenone sulfonic acid and its salts, dihydroxymethoxybenzophenone, sodium dihydroxymethoxybenzophenone disulfonate, dihydroxybenzophenone, dihydroxydimethoxybenzophenone, hydroxyoctoxybenzophenone, tetrahydroxybenzophenone, butyl methoxydibenzoylmethane, 2,4,6 - trianilino - p - (carbo - 2 - ethylhexyl - 1 - oxy) - 1,3,5 - triazine, 2 - (2 - hydroxy - 5 - methylphenyl) benzotriazole, methyl - O - aminobenzoate, 2 - ethylhexyl - 2 - cyano - 3,3 - diphenylacrylate, phenyl benzimidazole sulfate, 3 - (4 - methylbenzylidene) camphor, isopropyl dibenzoylmethane, 2 - ethylhexyl 4 - (3,4 - dimethoxyphenylmethylene) - 2,5 - dioxo - 1 - imidazolidinepropionate, and the like, and their polymer derivatives, silane derivatives, etc. (5) Examples of Powders and Pigments Dyes such as Red No. 104, Red No. 201, Yellow No. 4, Blue No. 1, Black No. 401, etc., Lake dyes such as Yellow No. 4 AL Lake, Yellow No. 203 BA Lake, etc., Nylon powder, Silk powder, Urethane powder, Teflon (registered trademark) powder, Silicon powder, Methyl polymethacrylate powder, Cellulose powder, Starch, Silicon elastomer spherical powder, Fine molecules such as polyethylene powder, Colored pigments such as Yellow iron oxide, Red iron oxide, Black iron oxide, Chromium oxide, Carbon black, Ultramarine, Blue, etc., White pigments such as Zinc oxide, Titanium oxide, Cerium oxide, etc., Extender pigments such as Talc, Mica, Sericite, Kaolin, Plate-like barium sulfate, etc., Pearl pigments such as Mica titanium, Metal salts such as Barium sulfate, Calcium carbonate, Magnesium carbonate, Aluminum silicate, Magnesium silicate, etc., Inorganic powders such as Silica, Alumina, etc., Metal soaps such as Aluminum stearate, Magnesium stearate, Zinc palmitate, Zinc myristate, Magnesium myristate, Zinc laurate, Zinc undecylenate, etc., Bentonite, Smectite, Boron nitride, etc. There are no particular restrictions on the shape (spherical, rod-shaped, needle-shaped, plate-shaped, irregular, flaky, spindle-shaped, etc.) and particle size of these powders. These powders may or may not have been previously surface-treated by conventionally known surface treatments, such as fluorine compound treatment, silicone treatment, silicone resin treatment, pendant treatment, silane coupling agent treatment, titanium coupling agent treatment, oil treatment, N-acylation lysine treatment, polyacrylic acid treatment, metal soap treatment, amino acid treatment, lecithin treatment, inorganic compound treatment, plasma treatment, mechanochemical treatment, etc. (6) Examples of surfactants Anionic surfactants: Fatty acid soaps, α-acyl sulfonates, Alkyl sulfonates, Alkyl allyl sulfonates, Alkyl naphthalene sulfonates, Alkyl sulfates, POE alkyl ether sulfates, Alkyl amide sulfates, Alkyl phosphates, POE alkyl phosphates, Alkyl amide phosphates, Alkyloyl alkyl taurine salts, N-acyl amino acid salts, POE alkyl ether carboxylates, Alkyl sulfosuccinates, Sodium alkyl sulfacetate, Acylated hydrolyzed collagen peptide salts, Perfluoroalkyl phosphate esters, etc. Cationic surfactants: alkyltrimethylammonium chloride, stearyltrimethylammonium chloride, stearyltrimethylammonium bromide, cetostearyltrimethylammonium chloride, distearyldimethylammonium chloride, stearyldimethylbenzylammonium chloride, behenyltrimethylammonium bromide, benzalkonium chloride, behenic acid amidopropyldimethylhydroxypropylammonium chloride, diethylaminoethylamide stearate, dimethylaminopropylamide stearate, lanolin derivative quaternary ammonium salts, etc. Amphoteric surfactants: carboxybetaine type, amidobetaine type, sulfobetaine type, hydroxysulfobetaine type, amidosulfobetaine type, phosphobetaine type, aminocarboxylate type, imidazoline derivative type, amidoamine type, etc. Nonionic surfactants: propylene glycol fatty acid ester, glycerin fatty acid ester, polyglycerin fatty acid ester, sorbitan fatty acid ester, POE sorbitan fatty acid ester, POE sorbit fatty acid ester, POE glycerin fatty acid ester, POE alkyl ether, POE fatty acid ester, POE hydrogenated castor oil, POE castor oil, POE·POP copolymer, POE·POP alkyl ether, polyether-modified silicone lauric acid alkanolamide, alkylamine oxide, hydrogenated soy phospholipid, etc. Natural surfactants: lecithin, saponin, sugar-based surfactants, etc. (7) Examples of polyhydric alcohols and sugars Ethylene glycol, diethylene glycol, polyethylene glycol, propylene glycol, dipropylene glycol, polypropylene glycol, glycerin, diglycerin, polyglycerin, 3-methyl-1,3-butanediol, 1,3-butylene glycol, sorbitol, mannitol, raffinose, erythritol, glucose, sucrose, fructose, xylitol, lactose, maltose, maltitol, trehalose, alkylated trehalose, mixed isomerized sugar, sulfated trehalose, pullulan, etc. Also, chemical modifiers thereof, etc. can be used. (8) Examples of polymers Acrylic acid ester / methacrylic acid ester copolymer (plus size, manufactured by Gohsei Chemical Industry Co., Ltd.), vinyl acetate / crotonic acid copolymer (Resin 28-1310, manufactured by NSC), vinyl acetate / crotonic acid / vinyl neodecanoate copolymer (28-2930, manufactured by NSC), methyl vinyl ether maleic acid half ester (Gantrez ES, manufactured by ISP), t-butyl acrylate / ethyl acrylate / methacrylic acid copolymer (Lubimer, manufactured by BASF), vinyl pyrrolidone / vinyl acetate / vinyl propionate copolymer (Lubrisol VAP, manufactured by BASF), vinyl acetate / crotonic acid copolymer (Lubset CA, manufactured by BASF), vinyl acetate / crotonic acid / vinyl pyrrolidone copolymer (Lubset CAP, manufactured by BASF), vinyl pyrrolidone / acrylate copolymer (Lubiflex, manufactured by BASF), acrylate / acrylic amide copolymer (Ultrahold, manufactured by BASF), vinyl acetate / butyl maleate / isobornyl acrylate copolymer (Advantage, manufactured by ISP), anionic polymer compounds such as carboxyvinyl polymer (Carbopol, manufactured by BFGoodrich), acrylic acid / methacrylic acid alkyl copolymer (Pemulen, manufactured by BF Goodrich), etc., amphoteric polymer compounds such as amphoteric acetate of dialkylaminoethyl methacrylate polymer (Yukafoamer, manufactured by Mitsubishi Chemical), octyl acrylamide / acrylic acid hydroxypropyl / butylaminoethyl methacrylate copolymer (AMPHOMER, manufactured by NSC), etc., cationic polymer compounds such as quaternized product of vinyl pyrrolidone / dimethylaminoethyl methacrylate (GAFQUAT, manufactured by ISP), methyl vinyl imidazolium chloride / vinyl pyrrolidone copolymer (Lubricote, manufactured by BASF), etc., nonionic polymer compounds such as polyvinyl pyrrolidone (Lubrisol K, manufactured by BASF), vinyl pyrrolidone / vinyl acetate copolymer (Lubrisol VA, manufactured by BASF), vinyl pyrrolidone / dimethylaminoethyl methacrylate copolymer (Copolymer 937, manufactured by ISP), vinyl caprolactam / vinyl pyrrolidone / dimethylaminoethyl methacrylate copolymer (Copolymer VC713, manufactured by ISP), etc.In addition, natural polymer compounds such as cellulose or its derivatives, keratin and collagen or its derivatives, calcium alginate, pullulan, agar, gelatin, tamarind seed polysaccharide, xanthan gum, carrageenan, high-methoxyl pectin, low-methoxyl pectin, guar gum, gum arabic, crystalline cellulose, arabinogalactan, karaya gum, tragacanth gum, alginic acid, albumin, casein, curdlan, gellan gum, dextran, etc. can also be preferably used. (9) Examples of physiologically active ingredients Examples of physiologically active ingredients include substances that impart some physiological activity to the skin when applied thereto. For example, whitening ingredients, anti-inflammatory agents, anti-aging agents, UV protectants, slimming agents, tightening agents, antioxidants, hair growth stimulants, hair tonics, moisturizers, blood circulation promoters, antibacterial agents, fungicides, desiccants, cooling agents, warming agents, vitamins, amino acids, wound healing promoters, irritation relievers, analgesics, cell activators, enzyme components, and the like. Examples of these preferred compounding ingredients include, for example, ashitaba extract, avocado extract, amacha extract, hollyhock extract, arnica extract, aloe extract, apricot extract, apricot kernel extract, ginkgo extract, perilla extract, turmeric extract, oolong tea extract, schizandra extract, etinashi leaf extract, saffron extract, cinnamon extract, peony extract, ginger extract, barley extract, chickweed extract, anemone extract, nasturtium extract, hydrolyzed elastin, hydrolyzed wheat flour, hydrolyzed silk, chamomile extract, carrot extract, mugwort extract, licorice extract, calcarea extract, kakyo extract, kiwi extract, cinchona extract, cucurbit extract, guanosine, gardenia extract, kumasasa extract, clara extract, walnut extract, grapefruit extract, clematis extract, chlorella extract, mulberry extract, gentiana extract, black tea extract, yeast extract, burdock extract, rice bran ferment extract, rice germ oil, comfrey extract, collagen, loquat extract, saikosaponin extract, saiko extract, saiboku-to extract, sage extract, soapwort extract, sasame extract, Japanese quince extract, sansho extract, shiitake extract, rhubarb extract, shikon extract, perilla extract, cinnamon tree extract, burnet extract, European barberry extract, European quince extract, European chickweed extract, European mint extract, sage extract, Japanese butterbur extract, senkyu extract, centella extract, soybean extract, thallus extract, thyme extract, tea extract, clove extract, chigaya extract, chinpi extract, toki extract, toki-senka extract, tonin extract, thuja extract, houttuynia extract, tomato extract, natto extract, carrot extract, garlic extract, rosa extract, hibiscus extract,Examples include Bacopa monnieri extract, parsley extract, honey, witch hazel extract, Parietaria judaica extract, Coix lacryma-jobi extract, bisabolol, loquat extract, Flos farfarae extract, Buchloe dactyloides extract, Butcher's broom extract, grape extract, propolis, Luffa cylindrica extract, safflower extract, peppermint extract, Ficus carica extract, button extract, hop extract, pine extract, horse chestnut extract, Japanese skunk cabbage extract, Chinese photinia extract, Melissa officinalis extract, peach extract, Leontopodium alpinum extract, eucalyptus extract, Saxifraga stolonifera extract, yuzu extract, Fructus Gardeniae extract, mugwort extract, lavender extract, apple extract, lettuce extract, lemon extract, Aster ageratoides extract, rose extract, rosemary extract, Roman chamomile extract, royal jelly extract, etc. In addition, biopolymers such as deoxyribonucleic acid, mucopolysaccharides, sodium hyaluronate, sodium chondroitin sulfate, collagen, elastin, chitin, chitosan, hydrolyzed eggshell membrane, amino acids, hydrolyzed peptides, moisturizing components such as sodium lactate, urea, sodium pyrrolidone carboxylate, betaine, whey, trimethylglycine, oily components such as sphingolipids, ceramides, phytosphingosine, cholesterol, cholesterol derivatives, phospholipids, anti-inflammatory agents such as ε-aminocaproic acid, glycyrrhizic acid, β-glycyrrhetinic acid, lysozyme chloride, guaiazulene, hydrocortisone, vitamins such as vitamin A, vitamin B2, vitamin B6, vitamin C, vitamin D, vitamin E, calcium pantothenate, biotin, nicotinamide, vitamin C ester, active ingredients such as allantoin, diisopropylamine dichloroacetate, 4-aminomethylcyclohexanecarboxylic acid, antioxidants such as tocopherol, carotenoid, flavonoid, tannin, lignan, saponin, cell activators such as α-hydroxy acid, β-hydroxy acid, blood circulation promoters such as γ-oryzanol, vitamin E derivatives, wound healing agents such as retinol, retinol derivatives, skin lightening agents such as arbutin, kojic acid, placenta extract, sulfur, ellagic acid, linoleic acid, tranexamic acid, glutathione, cephalanthin, licorice extract, capsicum tincture, hinokitiol, garlic iodide extract, pyridoxine hydrochloride, DL-α-tocopherol, DL-α-tocopherol acetate, nicotinic acid, nicotine Hair growth agents such as acid derivatives, calcium pantothenate, D-panthenyl alcohol, acetyl panthenyl ethyl ether, biotin, allantoin, isopropyl methylphenol, estradiol, ethinyl estradiol, capronium chloride, benzalkonium chloride, diphenhydramine hydrochloride, takanal, camphor, salicylic acid, vanillyl nonanoate, vanillyl nonamide, piroctone olamine, glyceryl pentadecanoate, L-menthol, mononitroguaicol, resorcinol, γ-aminobutyric acid, benzethonium chloride, mexiletine hydrochloride, auxin, female hormones, cantharidin tincture, cyclosporine, zinc pyrithione, hydrocortisone, minoxidil, polyoxyethylene sorbitan monostearate, peppermint oil, sasani shiki extract, etc. can be mentioned. (10) Antioxidants Sodium bisulfite, sodium sulfite, erythorbic acid, sodium erythorbate, dilauryl thiodipropionate, tocopherol, tolbiguanide, nordihydroguaiaretic acid, para-hydroxyanisole, butylhydroxyanisole, dibutylhydroxytoluene, ascorbyl stearate, ascorbyl palmitate, octyl gallate, propyl gallate, carotenoids, flavonoids, tannins, lignans, saponins, plant extracts with antioxidant effects such as apple extract and clove extract, etc. can be mentioned. (11) Examples of solvents Purified water, ethanol, lower alcohols, ethers, LPG, fluorocarbons, N-methylpyrrolidone, fluoroalcohols, volatile linear silicones, next-generation chlorofluorocarbons, etc. can be mentioned.

[0029] The anti-dandruff agent of the present invention is characterized by containing at least one of condensed tannin and kaempferol 3-O-(6"-acetyl)-glucoside as an active ingredient. The method for obtaining the above-mentioned condensed tannin and kaempferol 3-O-(6"-acetyl)-glucoside is not particularly limited, but it is preferably extracted from fuki. In addition, the extraction solvent, method, etc. can be carried out in the same manner as the skin resident flora improver described above, etc. Specifically, for example, it can be carried out by the method of the examples in the specification of the present application. Furthermore, the anti-dandruff agent of the present invention can be used as a raw material for various food and drink products, drugs (including pharmaceuticals and quasi-drugs), external skin preparations, and cosmetics, in the same manner as the skin resident flora improver described above, etc.

[0030] Furthermore, the skin resident flora improver, anti-Malassezia agent, and anti-acne agent of the present invention contain perilla fat-soluble extract as an active ingredient. The above-mentioned "perilla fat-soluble extract" refers to an extract obtained by extracting perilla with a fat-soluble solvent. As the fat-soluble solvent for extracting the perilla fat-soluble extract, hexane, methanol, ethanol, isopropyl alcohol, 1,3-butylene glycol, ethylene glycol, propylene glycol, glycerin, ethyl acetate, acetone, etc. can be used. These solvents may be mixed in two or more kinds. Desirably, hexane and ethanol may be used as the extraction solvent. In particular, the hexane-ethanol mixture is an extraction solvent that is less likely to reduce the activity of the active ingredient during extraction and is preferable in terms of safety in food use of the extract.

[0031] As the extraction temperature, for example, when using a hexane-ethanol mixture, the extraction temperature is preferably 20 to 60°C, more preferably about 40 to 50°C. This is because if the extraction temperature is too low, the active ingredient is difficult to extract, and if the extraction temperature is too high, the activity of the active ingredient is likely to decrease.

[0032] The hexane-ethanol mixture as the extraction solvent preferably has an ethanol concentration of 5 to 100%. The ethanol concentration is set at 5% or more because if the ethanol content is too low, the active ingredient is difficult to extract. Desirably, the ethanol concentration is 30 to 70%, and more desirably about 50%.

[0033] As the method for extracting the perilla fat-soluble extract of the present invention, any method such as continuous extraction, immersion extraction, countercurrent extraction, etc. can be adopted, and any device can be used under room temperature or reflux heating. Note that the specific extraction method is preferably carried out by the method of the examples of the present application. In addition, the skin resident flora improver, anti-Malassezia agent, and anti-acne agent containing the above perilla fat-soluble extract as an active ingredient can be used as materials for various foods and drugs (including pharmaceuticals and quasi-drugs), external skin preparations, and cosmetics, in the same manner as the above-described skin resident flora improver and the like.

Examples

[0034] Hereinafter, examples of the present invention will be described. Note that the examples shown below are for confirming the effect of improving the skin resident flora and the anti-dandruff effect of the composition obtained by the present invention, and the scope of the present invention is not limited to these products and manufacturing methods.

[0035] 1) Effect of the action of improving the skin resident flora in the perilla isolated component a) Isolation of perilla components The isolation of perilla components was carried out according to FIG. 1. That is, to the dried product (3.3 kg) of the aerial part (leaves and petioles) of perilla, 6 times the amount of 70% ethanol by weight was added, and extraction was carried out at 70 ° C for 2 hours. After filtering the extract, the solvent was distilled off under reduced pressure to obtain an extract (333 g) (yield: 10.1%). This was suspended in water (2 L), and partition was sequentially carried out with ethyl acetate (2 L, 3 times) and n-butanol (2 L, 3 times), and the partition liquids were concentrated respectively to obtain an ethyl acetate fraction (25.3 g) and a butanol fraction (17.2 g). The ethyl acetate fraction (24 g) was fractionated by normal phase silica gel column chromatography (500 g, n-hexane:ethyl acetate = 2:1 → 1:1 → ethyl acetate → methanol) to obtain fractions (Fr.) 1 to 9. Fr. 2 (0.55 g) was sequentially purified by normal-phase silica gel column chromatography (10 g, n-hexane:ethyl acetate = 10:1 → 1:1 → ethyl acetate), preparative thin-layer chromatography (TLC), and normal-phase HPLC (Chromatorex Si, 20 φmm × 250 mm, Fuji Silysia, hexane:ethyl acetate = 10:1) to obtain fukinone (36 mg). Fr. 3 was repeatedly purified by reverse-phase HPLC (Cosmosil 5C18ARII, 20 φmm × 250 mm, Nacalai Tesque, MeOH:THF = 9:1) to obtain isopetasine (32 mg). Fr. 4 was repeatedly purified by reverse-phase HPLC (Cosmosil 5C18ARII, MeOH:THF = 9:1) and Sunrise C30 (MeOH:THF = 9:1) to obtain myristic acid (46 mg), caprylic acid (3 mg), S-petasine (6 mg), and S-isopetasine (5 mg). Fr. 5 was repeatedly purified by reverse-phase HPLC (Cosmosil 5C18ARII, 70% MeOH) to obtain ethyl cafeate (4 mg) and 3,4-dihydroxyphenylpiruvic acid (5 mg). Fr. 6 was repeatedly purified by reverse-phase HPLC (Cosmosil 5C18ARII, 65% MeOH) to obtain ethyl cafeate (6 mg) and caffeic acid (62 mg). Fr. 7 (1.0 g) was fractionated into Fr. 7-1 to 10 by reverse-phase ODS column chromatography (30 g, 30% methanol → methanol). Then, Fr. 7-4 (95 mg) was purified by normal-phase HPLC (ethyl acetate) to obtain 2β-hydroxyfukinone (13 mg). Fr. 8 was repeatedly purified by reverse-phase HPLC (Cosmosil 5C18ARII, 65% MeOH) to obtain condensed tannin (31 mg) and kaempferol 3-O-(6"-acetyl)-glucoside (7 mg). Fr. 9 was repeatedly purified by reverse-phase HPLC (Cosmosil 5C18ARII, 55% MeOH) to obtain quercetin 3-O-glucoside (2 mg) and kaempferol 3-O-glucoside (6 mg). The obtained components were identified by comparing various NMR spectra with literature values. As a result, they were identified as the components shown in the following chemical formulae.

[0036] [Chemical formula]

[0037] b) Antibacterial test method Each strain was obtained from the National Institute of Technology and Evaluation (NBRC). Staphylococcus aureus Rosenbach 1884 (NBRC102135) Staphylococcus epidermidis Evans 1916 (NBRC12993) Cutibacterium acne subsp. acnes Nouioui et al. 2018 (NBRC107605) Corynebacterium tuberculostearicum Feurer et al. 2004 (NBRC113812) Malassezia furfur Baillon (NBRC0656) The following media were used. S. Aureus and S. epidermidis: NBRC medium No. 802 P. acne and C. tuberculostearicum: NBRC medium No. 312 M. furfur: NBRC medium No. 103

[0038] Antibacterial activity test for organisms other than M. furfur Add medium (90 μL), sample solution diluted with 10% DMSO (10 μL), and bacterial solution (10 μL) to a 96-well microplate. After stirring, culture at 37°C for 1 to 3 days, and then evaluate by visually observing the growth of the bacteria and measuring the turbidity (600 nm). Note that C. Acne was cultured under anaerobic conditions. The concentration of the inoculated bacterial solution was 5×10 3 ~2.56×10 4 cfu / 10 μL).

[0039] Antibacterial activity test of M. furfur Add medium (160 μL), sample solution diluted with 10% DMSO (20 μL), and bacterial solution (200 μL) to a 96-well microplate. After stirring, culture at 28°C for 3 days, and then evaluate by visually observing the growth of the bacteria and measuring the turbidity (600 nm). The concentration of the inoculated bacterial solution was 2.14×10 4 cfu / 20 μL.

[0040] c) Results and effects of the examples The results of the above tests are shown in Table 1 below. S-petasin, S-isopetasin, and fukinone, which are characteristic components of Petasites japonicus, showed antibacterial effects against Staphylococcus aureus (S. Aureus), and the MIC values were 25, 50, and 100 μg / mL, respectively, with the strongest activity of S-petasin. Thus, it was confirmed that S-petasin, S-isopetasin, and fukinone are useful as antibacterial agents against Staphylococcus aureus. S-petasine and S-isopetasine, which contain sulfur compounds, also showed antibacterial activity against Staphylococcus epidermidis (S. Epidermidis) and Propionibacterium acnes (P. Acne), which are so-called good bacteria, but the MIC value was 1 / 2 of that against S. Aureus. Thus, S-petasine and S-isopetasine have strong antibacterial activity against Staphylococcus aureus and weak antibacterial activity against Staphylococcus epidermidis, and thus have an improving effect on the skin resident flora of Staphylococcus aureus and Staphylococcus epidermidis. As a result, it was confirmed that these compounds are useful as skin resident flora improvers. The MIC values of myristic acid and caprylic acid, which are fatty acids, against S. Aureus were 100 μg / mL each, but their antibacterial effects against S. Epidermidis and P. Acne were weaker. Condensed tannin and kaempferol 3-O-(6"-acetyl)-glucoside showed antibacterial activity against Malassezia furfur, the causative agent of dandruff (MIC value: 100 μg / mL). In addition, since condensed tannin and kaempferol 3-O-(6"-acetyl)-glucoside, which are polyphenols, showed antibacterial activity against dandruff-causing bacteria, it was confirmed that these compounds are useful as anti-dandruff agents.

[0041]

Table 1

[0042] 2) Evaluation of the effect of the fat-soluble extract essential oil fraction of perilla on the skin resident flora of humans a) Preparation of the fat-soluble extract of perilla for evaluation The perilla fat-soluble extract for evaluation was prepared according to the following procedure. Hexane:ethanol mixture (1:1 by volume) was added to crushed perilla dry matter (1 kg), and stirred extraction was carried out at 40 °C for 2 hours. After the extract was concentrated to dryness, 70% v / v ethanol was added for dispersion, and citric acid was added until the pH reached around 4, followed by stirring. After filtration through celite (#100) (to remove chlorophyll), the filtrate was allowed to stand at 4 °C for 3 hours and then filtered through filter paper (No. 5) (to remove wax components). The filtrate was concentrated to dryness, resuspended by adding hexane:ethanol mixture (1:1 by volume) again, allowed to stand at 4 °C for 1 hour, and then filtered through filter paper (No. 5) (second wax removal). Water was added to the filtrate and stirred, and the upper hexane layer was separated. By concentrating the hexane layer to dryness, the perilla fat-soluble extract for evaluation was obtained.

[0043] b) Evaluation method, results and effects of the examples The antibacterial activity of the perilla fat-soluble extract was measured in the same manner as in the above test examples. The results are shown in Figure 2. As shown in Figure 2, the perilla fat-soluble extract showed concentration-dependent antibacterial activity against the pathogenic bacterium Staphylococcus aureus, but showed no antibacterial activity against the beneficial bacterium Staphylococcus epidermidis. From these results, it was found that the perilla fat-soluble extract is an ideal extract that does not affect the growth of beneficial bacteria and shows antibacterial activity against pathogenic bacteria.

[0044] 3) Evaluation of the skin resident flora in the axilla and behind the ear using sequence amplicon analysis a) Preparation of the perilla fat-soluble extract-containing emulsion Next, an emulsion containing the obtained perilla fat-soluble extract was prepared. The formulation is shown below. The placebo emulsion was one without the perilla fat-soluble extract. Hexyldecyl myristoyl methylaminopropionate 6.00% Dimethicone 1.50 Glyceryl stearate 0.50 Diglycerin distearate 1.50 Hydrogenated lecithin 0.50 Maltitol 5.00 Butylene glycol 10.00 Glycerin 5.00 Carbomer 1% aqueous solution 10.00 Water 59.89 Arginine 0.10 Perilla fat-soluble extract 0.01 Total amount 100.00%

[0045] b) Evaluation of the effect on the skin microbiota According to the following scheme, the influence on the skin microbiota was examined. An emulsion containing perilla fat-soluble extract (0.01%) and a placebo emulsion were each applied twice a day (after bathing and upon waking up) for 7 days to the right and left axillae and behind the ears of male subjects (5 people). Next-generation sequencing amplicon analysis was performed on the collected skin swabs at Technosugar Lab. The main results are shown below.

[0046] i) Staphylococcus epidermidis (a good bacterium, also known as the epidermal staphylococcus) and other Staphylococcus species The evaluation results of Staphylococcus epidermidis (a good bacterium, also known as the epidermal staphylococcus) and other Staphylococcus species are shown in Figure 3. As shown in Figure 3, Staphylococcus species were frequently found in relatively older age groups. S. epidermidis, the good bacterium, often decreased upon application of the perilla extract. It was found that the perilla extract has no effect of increasing S. epidermidis. Among other Staphylococcus species, S. hominis (which produces thioalcohol and causes body odor) is mainly mentioned in the axilla. Since other Staphylococcus in the axilla decreased in 4 out of 5 cases upon application of the perilla extract, it was confirmed that the perilla extract is useful as an axillary osmidrosis improver. Note that S. aureus, the bad bacterium, was not detected because the subjects were healthy.

[0047] ii) Propionibacterium acnes (acne bacterium) The evaluation results for Propionibacterium acnes (acne bacteria) are shown in Figure 4. P. acnes was not present in the armpits. In the seborrheic skin area behind the ears, P. acnes decreased in 4 out of 5 cases with the application of perilla extract. Thus, it was confirmed that it has an effect on improving seborrheic odor from behind the ears in middle-aged and elderly people. iii) Malassezia The evaluation results for Malassezia are shown in Figure 5. M. restricta and M. globosa are predominantly present. Except for some cases close to 100%, a decrease in the presence ratio of Malassezia with the application of perilla extract was observed.

[0048] Effects of the examples From the above results, it was confirmed that perilla fat-soluble extract suppresses the excessive growth of acne bacteria (P. Acnes) in seborrheic areas (around the nose, forehead, behind the ears), thereby suppressing inflammation and seborrheic odor. Also, it was confirmed that by suppressing the growth of S. Hominis predominantly present in the armpits, body odor from the armpits is suppressed. Moreover, since a decrease in the presence ratio was also observed for Malassezia, it was confirmed that it is also effective for scalp care and health care of other hair growth areas.

[0049] [Formulation examples] Formulation examples of the skin resident flora improving agent and anti-dandruff agent (perilla extract, perilla fat-soluble extract) according to the present invention are shown. Formulation example 1: Chewing gum Sugar 53.0 wt% Gum base 20.0 Glucose 10.0 Maltose 16.0 Fragrance 0.5 Perilla extract 0.5 100.0 wt%

[0050] Formulation example 2: Gummy candy Reduced maltose 40.0 wt% Granulated sugar 20.0 Glucose 20.0 Gelatin 4.7 Water 9.68 Ume Fruit Juice 4.0 Ume Flavor 0.6 Dye 0.02 Perilla extract 1.0 100.0wt%

[0051] Formulation Example 3: Candy Sugar 50.0wt% Maltose 33.0 Water 14.4 Organic Acid 2.0 Spice 0.2 Perilla extract 0.4 100.0wt%

[0052] Formulation Example 4: Yogurt (Hard - Soft) Milk 41.5wt% Skim Milk Powder 5.8 Sugar 8.0 Agar 0.15 Gelatin 0.1 Lactic Acid Bacteria 0.005 Fuki Extract 0.4 Spice Trace Water remainder 100.0wt%

[0053] Formulation Example 5: Tea Beverage (Liquid) Tea Leaves 6.0wt% Sodium Ascorbate 0.1 Fuki Extract 1.0 Water remainder 100.0wt%

[0054] Formulation Example 6: Tea Beverage (Powder) Tea Leaf Extract 90.0wt% Dextrin 7.0 Perilla extract 3.0 100.0wt%

[0055] Formulation Example 7: Soft Drink Fructose Glucose Syrup 30.0wt% Emulsifier 0.5 Perilla extract 0.05 Appropriate amount of fragrance Purified water remainder 100.0wt%

[0056] Formulation example 8: Chocolate Vitachocolate 18.0 Cocoa butter 16.5 Powdered sugar 42.0 Whole milk powder 22.0 Lentin 0.5 Perilla extract 1.0 Fragrance appropriate amount 100.0wt%

[0057] Formulation example 9: Caramel Sugar 39.0 Corn syrup 38.0 Condensed milk 15.5 Wheat flour 5.0 Butter 0.5 Shortening oil 1.0 Perilla extract 1.0 Fragrance appropriate amount 100.0wt%

[0058] Formulation example 10: Jelly Sugar 38.0 Corn syrup 42.5 Agar 1.5 Glucose 5.0 Water 12.0 Perilla extract 1.0 Colorant and fragrance appropriate amount 100.0wt%

[0059] Formulation example 11: Biscuit Wheat flour 67.5 Sugar 10.0 Shortening 15.0 Table salt 1.0 Leavening agent 1.0 Invert sugar 5.0 Perilla extract (antibacterial agent against skin resident bacteria) 0.5 100.0wt%

[0060] Formulation Example 12: Soft Capsule Brown rice germ oil 87.0 wt% Emulsifier 12.0 Perilla extract 1.0 100.0 wt%

[0061] Formulation Example 13: Tablet Lactose 54.0 wt% Crystalline cellulose 30.0 Starch hydrolyzate 10.0 Glycerin fatty acid ester 5.0 Perilla extract 1.0 100.0 wt%

[0062] Formulation Example 14: Chewable Tablet Sugar 76.4 wt% Glucose 19.0 Sucrose fatty acid ester 0.2 Butterbur extract 0.5 Purified water 3.9 100.0 wt%

[0063] Formulation Example 15: Cosmetic Cream Squalane 20.0 wt% Beeswax 5.0 Refined jojoba oil 5.0 Glycerin 5.0 Glycerin monostearate 2.0 Polyoxyethylene (20) sorbitan - Monostearate 2.0 Butterbur fat-soluble extract 0.1 Preservative appropriate amount Fragrance appropriate amount Purified water remainder 100.0 wt%

[0064] Formulation Example 16: Lotion Ethanol 5.0 wt% Glycerin 2.0 1,3 - Butylene glycol 2.0 Polyethylene oleyl ether 0.5 Sodium citrate 0.1 Citric acid 0.1 Perilla frutescens fat-soluble extract 0.1 Purified water remainder 100.0 wt%

[0065] Formulation Example 17: Emulsion Squalane 4.0 wt% Petrolatum 2.5 Cetanol 2.0 Glycerin 2.0 Hydrophobic glycerol monostearate 1.0 Stearic acid 1.0 L-Arginine 1.0 Perilla frutescens fat-soluble extract 0.1 Potassium hydroxide 0.1 Fragrance Trace amount Purified water remainder 100.0 wt%

[0066] Formulation Example 18: Body Gel Macadamia nut oil 2.0 wt% Octyldodecyl myristate 10.0 Methylphenylpolysiloxane 5.0 Behenyl alcohol 3.0 Stearic acid 3.0 Batyl alcohol 1.0 Glyceryl monostearate 1.0 Polyoxyethylene sorbitol tetraoleate 2.0 Hydrogenated soy phosphatide 1.0 Ceramide 0.1 Retinol palmitate 0.1 Preservative Appropriate amount Adenophora triphylla extract 1.0 Perilla frutescens extract 0.1 1,3-Butylene glycol 5.0 Purified water remainder 100.0 wt%

[0067] Formulation Example 19: Peel-off Pack Glycerin 5.0 Propylene Glycol 4.0 Polyvinyl Alcohol 15.0 Ethanol 8.0 Polyoxyethylene Glycol 1.0 Houttuynia cordata 5.0 Water 57.0 Angelica Extract 0.1 Fragrance and preservative appropriate amount 100.0 wt%

[0068] Formulation Example 20: Cold Cream White Beeswax 11.0 Liquid Paraffin 26.0 Lanolin 10.0 Almond Oil 15.0 Borax 0.5 Water 34.5 Angelica Extract 0.1 Fragrance Appropriate amount Preservative appropriate amount 100.0 wt%

[0069] Formulation Example 21: Bath Agent (Liquid) Propylene Glycol 50.0 wt% Ethanol 20.0 Sodium Sulfate 5.0 Angelica Extract 0.1 Lanolin 0.5 Avocado Oil 0.5 Dye 1.5 Fragrance 22.4 100.0 wt%

[0070] Formulation Example 22: Bath Agent (Granules) Sodium Hydrogen Carbonate 64.9 Anhydrous Sodium Sulfate 32.0 Borax 3.0 Perilla extract 0.1 100.0 wt%

[0071] Formulation Example 23: Shampoo Triethanolamine lauryl sulfate 5.0 Sodium polyoxyethylene lauryl ether sulfate 12.0 1,3-Butylene glycol 4.0 Diethanolamine laurate 2.0 Disodium edetate 0.1 Water 73.9 Perilla extract (antidandruff agent) 0.1 Fragrance Appropriate amount Preservative appropriate amount 100.0 wt%

[0072] Formulation Example 24: Conditioner Stearyl trimethyl ammonium chloride 2.0 Cetostearyl alcohol 2.0 Polyoxyethylene lanolin ether 3.0 Propylene glycol 5.0 Water 84.0 Perilla extract (antidandruff agent) 0.1 pH adjuster Appropriate amount Preservative appropriate amount 100.0 wt%

[0073] Formulation Example 25: Body soap Potassium laurate 15.0 Potassium myristate 5.0 Propylene glycol 5.0 Water 72.0 Perilla extract 3.0 pH adjuster Appropriate amount Preservative appropriate amount 100.0 wt%

[0074] Formulation Example 26: Hair liquid Ethanol 29.0 Polyoxypropylene butyl ether phosphate 10.0 Polyoxypropylene monobutyl ether 5.0 Triethanolamine 1.0 Water 50.0 Butcher's broom extract (anti-dandruff agent) 0.1 Preservative appropriate amount 100.0 wt%

[0075] Formulation Example 27: Hair Toner Ethanol 46.9 Ethyl oleate 1.0 Polyoxyethylene (40) hydrogenated castor oil 2.0 Water 52.0 Perilla extract (anti-dandruff agent) 0.1 100.0 wt%

Industrial Applicability

[0076] As described above, the present invention can provide an agent for improving the novel skin resident flora.

Claims

1. A skin resident flora improver containing perilla fat-soluble extract as an active ingredient.

2. An anti-axillary odor bacteria agent containing perilla fat-soluble extract as an active ingredient.

3. An anti-Malassezia bacteria agent containing perilla fat-soluble extract as an active ingredient.

4. An anti-acne bacteria agent for seborrheic areas containing perilla fat-soluble extract as an active ingredient.

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