Classifications

• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K31/00—Medicinal preparations containing organic active ingredients
  • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
  • A61K31/7016—Disaccharides, e.g. lactose, lactulose
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K8/00—Cosmetics or similar toiletry preparations
  • A61K8/02—Cosmetics or similar toiletry preparations characterised by special physical form
  • A61K8/04—Dispersions; Emulsions
  • A61K8/042—Gels
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K8/00—Cosmetics or similar toiletry preparations
  • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
  • A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
  • A61K8/33—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing oxygen
  • A61K8/34—Alcohols
  • A61K8/342—Alcohols having more than seven atoms in an unbroken chain
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K8/00—Cosmetics or similar toiletry preparations
  • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
  • A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
  • A61K8/60—Sugars; Derivatives thereof
  • A61K8/604—Alkylpolyglycosides; Derivatives thereof, e.g. esters
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P17/00—Drugs for dermatological disorders
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
  • A61Q19/00—Preparations for care of the skin
  • A61Q19/08—Anti-ageing preparations

Definitions

• the present invention relates to an activator containing an anionic biosurfactant and a skin condition improving agent containing an anionic biosurfactant. It also relates to a cosmetic or quasi-drug that uses the activator.
• cosmetics and quasi-drugs for topical skin preparations generally contain bioactive ingredients, such as moisturizing ingredients, emollient ingredients, astringent ingredients, keratin softening ingredients, whitening ingredients, anti-inflammatory (anti-inflammatory) ingredients, UV protection ingredients, antioxidant ingredients, cell activating ingredients, and blood circulation promoting ingredients, to achieve effects such as moisturizing, preventing rough skin, preventing blemishes and freckles, tightening the skin, anti-inflammation, promoting blood circulation, and preventing aging skin.
• bioactive ingredients such as moisturizing ingredients, emollient ingredients, astringent ingredients, keratin softening ingredients, whitening ingredients, anti-inflammatory (anti-inflammatory) ingredients, UV protection ingredients, antioxidant ingredients, cell activating ingredients, and blood circulation promoting ingredients, to achieve effects such as moisturizing, preventing rough skin, preventing blemishes and freckles, tightening the skin, anti-inflammation, promoting blood circulation, and preventing aging skin.
• Patent Document 1 describes a skin topical composition that contains Monk fruit extract and sophorolipid, which has fibroblast activation and collagen synthesis effects, as well as skin roughness improvement effects and UV protection effects.
• Patent Document 2 also describes that mannosylerythritol lipid (MEL) inhibits the formation of skin melanocytes, improves the overall skin tone, making the skin clearer, and provides whitening effects by improving and brightening the dark skin tone.
• MEL mannosylerythritol lipid
• compositions containing anionic surfactants are used as cleansing compositions that have cleansing power and an excellent feel on the skin.
• Patent Document 3 describes that a skin cleansing composition containing anionic surfactants such as acyl isethionate and its salts, alkyl sulfates or polyoxyethylene alkyl sulfates, and 2-ethylhexyl glyceryl ether, and a specific amount of water, has excellent foam volume and cleansing power when diluted, exfoliates a large amount of cells, leaves the skin smooth after cleansing, and provides excellent adhesion of makeup cosmetics applied to the skin after cleansing.
• anionic surfactants such as acyl isethionate and its salts, alkyl sulfates or polyoxyethylene alkyl sulfates, and 2-ethylhexyl glyceryl ether, and a specific amount of water
• one embodiment of the present invention aims to provide a novel activator and skin condition improving agent.
• anionic biosurfactants have good activating ability and are useful as cell activators, thus completing the present invention.
• surfactants in detergents are an exogenous stress
• many of the conventional anionic surfactants described in Patent Document 3 are compounds that are skin irritating.
• the inventors discovered that the above anionic biosurfactants are low in skin irritation and also have good foaming durability, and can be suitably used as products to be applied to human skin, such as cosmetics and quasi-drugs.
• the anionic biosurfactants can be suitably used as various skin condition improving agents. That is, one aspect of the present invention relates to an activator containing an anionic biosurfactant, a skin condition improving agent containing an anionic biosurfactant, and cosmetics and quasi-drugs containing the activator.
• a novel activator and skin condition improver can be provided.
• an activator that is useful as a cell activator, has low skin irritation, and has good foaming durability, various skin condition improvers, and cosmetics and quasi-drugs that contain the activator.
• the activating agent according to one embodiment of the present invention is characterized by including an anionic biosurfactant.
• activation refers to maintaining or enhancing cell function or cell activity.
• Activator refers to a substance that has activation ability and, as a result, can suppress the decline in cell function or cell activity. Therefore, “activator” is synonymous with “cell activator.”
• anionic biosurfactant refers to an anionic biosurfactant.
• anionic refers to a property in which hydrophilic groups ionize in water and become negatively charged.
• an acidic functional group such as a carboxyl group, a sulfonic acid group, or a sulfate ester group. Therefore, the anionic biosurfactant according to one embodiment of the present invention can be said to be a biosurfactant having an acidic functional group.
• biosurfactant is a general term for a substance (compound) that has surface activity and/or emulsification ability and is produced (biosynthesized) by a living organism (e.g., a microorganism).
• the anionic biosurfactant contained in the activator according to one embodiment of the present invention is not particularly limited as long as it is an anionic substance produced by an organism and has surface activity and/or emulsification ability, but examples include biosurfactants having a glycolipid structure, biosurfactants having an amino acid structure, biosurfactants having an organic acid structure, and biosurfactants having a polymer structure.
• the activator according to one embodiment of the present invention may contain only one type of the various anionic biosurfactants listed above, or may contain two or more types.
• biosurfactants having a glycolipid structure that may be contained in the activator according to one embodiment of the present invention include succinoyl trehalose lipid (hereinafter sometimes referred to as STL), sophorolipid, etc.
• STL succinoyl trehalose lipid
• sophorolipid etc.
• the anionic biosurfactant contained in the activator according to one embodiment of the present invention is preferably a biosurfactant produced by a microorganism, more preferably a biosurfactant having a glycolipid structure, and even more preferably a succinoyl trehalose lipid, from the viewpoints of foaming durability and low skin irritation.
• the activator according to one embodiment of the present invention preferably contains an anionic biosurfactant containing a biosurfactant produced by a microorganism, more preferably contains an anionic biosurfactant containing a biosurfactant having a glycolipid structure, and even more preferably contains an anionic biosurfactant containing a succinoyl trehalose lipid.
• the succinoyl trehalose lipid that may be contained in the activator of one embodiment of the present invention can be obtained by aerobically culturing Rhodococcus erythropolis SD-74 strain in a medium containing a carbon source such as a fatty acid or vegetable oil, or by aerobically culturing Rhodococcus sp. TB-42 strain in a medium containing a carbon source such as an unsaturated hydrocarbon or a halogenated hydrocarbon.
• Rhodococcus erythropolis SD-74 strain has been deposited at the National Institute of Technology and Evaluation, Patent Microorganism Depositary Center (Address: Room 122, 2-5-8 Kazusa Kamatari, Kisarazu City, Chiba Prefecture, Japan, Postal Code 292-0818) under "Accession Number: NITE BP-03788 (Deposit Date: December 1, 2022)".
• Succinoyltrehalose lipid obtained by culturing microorganisms in a medium containing a carbon source, is a glycolipid in which the sugar portion is trehalose and 1-2 moles each of succinic acid and fatty acid are ester-linked per mole of trehalose.
• the fatty acid portion of this glycolipid is derived from the carbon source, which is the culture substrate, and different fatty acids can be bound by changing the composition of the carbon source.
• succinoyl trehalose lipid that may be contained in the activator according to one embodiment of the present invention may be, for example, a compound having the structure of the following general formula 1.
• the substituents R1 and R2 are aliphatic hydrocarbon groups.
• the aliphatic hydrocarbon groups which are the substituents R1 and R2 in general formula 1 may each independently be linear or branched, and are preferably linear from the viewpoint of assimilation of the raw material carbon source.
• the aliphatic hydrocarbon groups which are the substituents R1 and R2 in the general formula 1 may each independently be an alkyl group having 5 to 30 carbon atoms.
• the alkyl group preferably has 8 or more carbon atoms, more preferably has 10 or more carbon atoms, and even more preferably has 12 or more carbon atoms.
• the alkyl group preferably has 25 or less carbon atoms, more preferably has 22 or less carbon atoms, and even more preferably has 20 or less carbon atoms.
• the succinoyl trehalose lipid having the structure of general formula 1 can be obtained, for example, by aerobically culturing the Rhodococcus erythropolis SD-74 strain in a medium containing a carbon source such as fatty acid or vegetable oil, or by aerobically culturing the Rhodococcus sp. TB-42 strain in a medium containing a carbon source such as unsaturated hydrocarbons or halogenated hydrocarbons.
• succinoyl trehalose lipid When a succinoyl trehalose lipid is produced by culturing a microorganism in a medium containing a carbon source containing an alkylene oxide, the resulting succinoyl trehalose lipid may contain a structural unit derived from an alkylene oxide adduct in the substituents R 1 and/or R 2.
• the succinoyl trehalose lipid preferably has a structure with a low content of structural units derived from alkylene oxide adducts, specifically, the content of structural units derived from alkylene oxide adducts in the substituents R 1 and R 2 in the general formula 1 is preferably 20% by mass or less, more preferably 10% by mass or less, even more preferably 5% by mass or less, and may be 0 parts by mass.
• the content of the anionic biosurfactant in 100 parts by mass of the activator according to one embodiment of the present invention is preferably 95 parts by mass or more, more preferably 98 parts by mass or more, and even more preferably 99 parts by mass or more, and may be 100 parts by mass, from the viewpoint of maintaining high purity and exerting stable performance (e.g., activation effect).
• the activator according to one embodiment of the present invention is characterized not only by its activating effect, but also by its low skin irritation and good foaming durability.
• the skin irritation and foaming durability of the activator can be measured and evaluated by the method described in the Examples.
• the critical micelle concentration (CMC) of the activator according to one embodiment of the present invention is preferably 1000 ⁇ 10 ⁇ 6 M or less, more preferably 500 ⁇ 10 ⁇ 6 M or less, and even more preferably 200 ⁇ 10 ⁇ 6 M or less, from the viewpoint of surface activity when used as a cleansing agent.
• the critical micelle concentration of the activator can be measured by the method described in the Examples.
• the activator according to one embodiment of the present invention preferably has a 50% cell proliferation inhibitory concentration (IC50) of 100 ppm or more, and more preferably 300 ppm or more.
• IC50 cell proliferation inhibitory concentration
• the IC50 of the activator can be measured by the method described in the Examples.
• the activator according to one embodiment of the present invention may also contain a solvent in addition to the anionic biosurfactant.
• the activator according to one embodiment of the present invention may be provided in the form of a solution in which the anionic biosurfactant is dissolved in a solvent.
• the solvent that may be contained in the activator according to one embodiment of the present invention is not particularly limited as long as it is capable of dissolving the anionic biosurfactant according to one embodiment of the present invention, and examples of the solvent include water, 1,3-butylene glycol, ethanol, acetone, and methanol. Of these, water or 1,3-butylene glycol is preferred from the viewpoint of use as a cosmetic raw material.
• the content (concentration) of the solvent is not particularly limited, but may be, for example, 0.1 to 50% by weight relative to 100% by mass of the activator according to one embodiment of the present invention.
• the activator according to one embodiment of the present invention may contain other additives in addition to the anionic biosurfactant and the solvent.
• additives that may be contained in the activator according to one embodiment of the present invention include, for example, fats and oils, waxes, mineral oils, fatty acids, alcohols, esters, metal soaps, gums and water-soluble polymeric compounds, vitamins, amino acids, skin whitening agents, moisturizing agents, hair growth agents, ⁇ -hydroxy acids, inorganic pigments, UV absorbers, astringents, antioxidants, anti-inflammatory agents, germicides and disinfectants, fragrances, pigments and colorants, hormones, sequestering agents, pH adjusters, chelating agents, preservatives and antifungal agents, cooling agents, stabilizers, animal and vegetable proteins and their decomposition products, animal and vegetable polysaccharides and their decomposition products, animal and vegetable glycoproteins and their decomposition products, blood flow promoters, anti-inflammatory and anti-allergic agents, cell activators, keratolytic agents, wound treatment agents, foaming agents, thickeners, oral agents, and deodorants and deodorants.
• a method for producing an activator according to one embodiment of the present invention includes, for example, a culture step of culturing a microorganism in a medium containing a carbon source, a precipitation step of precipitating a product obtained by the culture step, an extraction step of extracting an extract containing an anionic biosurfactant (e.g., an STL composition) from the precipitate obtained by the precipitation step, and a fat-soluble substance removal step of removing a fat-soluble substance from the extract obtained by the extraction step.
• an anionic biosurfactant e.g., an STL composition
• the step of culturing a microorganism in a medium containing a carbon source is carried out according to a conventional method. If necessary, nutrients such as a nitrogen source and inorganic salts may be added to the medium containing the carbon source.
• the "microorganism” in the culture process is not particularly limited as long as it can produce an anionic biosurfactant, but is preferably a microorganism belonging to the genus Rhodococcus, and more preferably Rhodococcus erythropolis SD-74 strain, Rhodococcus sp. TB-42 strain, or Rhodococcus baiconulensis NBRC 100611 strain, since they are capable of producing succinoyl trehalose lipids.
• the "carbon source” in the culture step is preferably a carbon compound that the microorganism assimilates during culture, particularly a natural oil, a hydrocarbon, a fatty acid, a fatty acid ester, or a higher alcohol.
• the carbon compound is intended to be a compound of carbon with hydrogen, nitrogen, or the like.
• the carbon source used in the culture step may be, for example, a natural oil, and may be either an animal oil or a vegetable oil, but is preferably a vegetable oil since it is easier to obtain.
• the vegetable oil used in the culture step is preferably, for example, palm oil, coconut oil, soybean oil, olive oil, safflower oil, rapeseed oil, corn oil, cottonseed oil, tall oil, etc., but is not limited thereto.
• hydrocarbons can be used as a carbon source in the culture process.
• Suitable hydrocarbons for use as carbon sources include normal alkanes such as n-decane, n-undecane, n-tridecane, n-tetradecane, n-pentadecane, n-hexadecane, n-heptadecane, n-octadecane, and n-nonadecane; normal alkenes such as 1-decene, 1-undecene, 1-dodecene, 1-tridecene, 1-tetradecene, 1-pentadecene, 1-hexadecene, 1-heptadecene, and 1-octadecene; and the like.
• the hydrocarbons used as the carbon source are preferably n-tridecane, n-tetradecane, n-pentadecane, n-hexadecane, or n-heptadecane, and more preferably n-tetradecane, n-pentadecane, or n-hexadecane.
• n-tetradecane is particularly preferred because it can improve various physiological functions of the resulting anionic biosurfactant (e.g., skin barrier properties, moisturizing properties, improvement of skin elasticity, inhibition of melanin production, etc.).
• Fatty acids can also be used as a carbon source in the culture process.
• Fatty acids that can be suitably used as carbon sources include decanoic acid, undecanoic acid, dodecanoic acid (lauric acid), tridecanoic acid, tetradecanoic acid (myristic acid), pentadecanoic acid, hexadecanoic acid (palmitic acid), heptadecanoic acid, octadecanoic acid (stearic acid), and oleic acid.
• fatty acid esters, and higher alcohols such as undecyl alcohol and dodecyl alcohol (lauryl alcohol) can also be used as carbon sources.
• tridecanoic acid tridecanoic acid
• tetradecanoic acid pentadecanoic acid, hexadecanoic acid (palmitic acid), and heptadecanoic acid
• tetradecanoic acid pentadecanoic acid
• hexadecanoic acid palmitic acid
• heptadecanoic acid tridecanoic acid
• tetradecanoic acid myristic acid
• pentadecanoic acid hexadecanoic acid
• heptadecanoic acid heptadecanoic acid
• tetradecanoic acid is particularly preferred, since it can improve various physiological functions of the resulting anionic biosurfactant (e.g., skin barrier properties, moisturizing properties, improvement of skin elasticity, inhibition of melanin production, etc.).
• the carbon source to be added to the medium the above-mentioned carbon sources are preferably used.
• the concentration of the carbon source added in the medium is preferably 5 to 20 mass%, more preferably 10 mass%.
• nitrogen-containing organic or inorganic substances that are usually used in culturing microorganisms are used, and for example, sodium nitrate, potassium nitrate, potassium hydrogen phosphate, potassium dihydrogen phosphate, etc. can be used.
• nutrients such as yeast extract and peptone may be added to the medium if necessary for the growth of the microorganism.
• the cultivation of the microorganisms in the cultivation step is preferably carried out under aerobic conditions with shaking and stirring.
• the cultivation temperature is preferably 20 to 35°C, more preferably 30°C.
• the cultivation pH is preferably 5.5 to 9.5.
• the cultivation period is preferably until a mixture containing an anionic biosurfactant (e.g., succinoyl trehalose lipid) at a concentration of 15 to 150 g/L is produced.
• an anionic biosurfactant e.g., succinoyl trehalose lipid
• the succinoyl trehalose lipid concentration reaches 25 g/L after 332 hours of cultivation, so cultivation for 199 to 664 hours is preferable.
• the above microorganisms may be seed cultured before the main culture.
• succinoyl trehalose lipids can be produced efficiently.
• the precipitation step is a step of precipitating a product containing an anionic biosurfactant (e.g., a product containing STL) produced by the microorganism in the above-mentioned culture step. That is, precipitation is performed on a medium or culture solution containing an anionic biosurfactant produced in the medium by the microorganism.
• the medium or culture solution to be precipitated may be a medium or culture solution in which the microorganism is cultured, which is centrifuged to remove the bacterial cells and the remaining substrate from the culture solution.
• "precipitating” is intended to take out a substance dissolved in the medium as a solid. That is, in the precipitation step according to one embodiment of the present invention, an anionic biosurfactant such as a glycolipid produced in the medium by the microorganism in the culture step can be taken out as a solid from the medium.
• the method for precipitating the product containing the anionic biosurfactant is not particularly limited as long as it is a method that can separate the product containing the anionic biosurfactant as a solid from the culture solution, and a conventional method can be used.
• the medium in which the microorganism is cultured in the above-mentioned culture step can be acidified, and the acidic substance in the medium can be precipitated to precipitate the product containing the acidic anionic biosurfactant (acid precipitation).
• the product containing the anionic biosurfactant can be precipitated by lowering the pH in the culture solution (target). In order to lower the pH of the culture solution, an acidic substance, for example, HCl, can be added.
• the precipitated product can then be removed, for example, by performing a centrifugation process.
• the product obtained through the precipitation step will be referred to as the "precipitated product”.
• the extraction step is a step of extracting the STL composition from the precipitated product.
• a solvent that is incompatible with water and in which an anionic biosurfactant, particularly a glycolipid, is soluble is added to the precipitated product, thereby dissolving the anionic biosurfactant contained in the precipitated product in a solvent layer.
• the solvent layer in which the anionic biosurfactant is dissolved can be separated to obtain an anionic biosurfactant composition from which water-soluble substances have been removed.
• the "water-soluble substance” is a substance that is soluble in water, and can be said to be a water-soluble impurity such as a salt contained in the solid precipitate precipitated from the culture medium together with water.
• the "salt” is a compound in which the hydrogen atom of an acid is replaced with a metal or other metallic group.
• the above-mentioned precipitated product precipitated from the culture medium in which the microorganisms were cultured contains water and contains a large amount of water-soluble impurities.
• Such precipitated products also contain fat-soluble impurities, and because they are solid, it is difficult to sufficiently remove the water-soluble substances contained within the precipitate even when they are washed with water.
• by carrying out an extraction process using a solvent that is incompatible with water and in which the anionic biosurfactant is soluble it is possible to obtain an anionic biosurfactant composition from which the water-soluble substances have been sufficiently removed.
• the solvent to be added to the precipitated product can be one that is incompatible with other solvents (e.g., water) that can dissolve water-soluble substances and in which the anionic biosurfactant is soluble.
• solvents include ester-based solvents, alcohol-based solvents, and hydrocarbon-based solvents, and specific examples include ethyl acetate, 1-butanol, and xylene.
• the amount of solvent to be added to the precipitated product is preferably 0.1 to 10 times the amount of the precipitated product by mass, and more preferably is equal to the amount of the precipitated product.
• the extraction process will be described using an example in which ethyl acetate is used as the solvent.
• the culture product is precipitated from the culture liquid, and ethyl acetate, the solvent, is added to the precipitated product and thoroughly stirred to separate the solution containing the precipitated product into two layers, an ethyl acetate layer and an aqueous layer.
• the ethyl acetate layer formed in the upper layer is separated using a separating funnel or the like.
• the precipitation step and the extraction step are performed in sequence, but this is not necessarily limited to this.
• the medium in which the microorganism has been cultured may be centrifuged to remove the bacterial cells and residual substrates, and the resulting mixture may be used to remove water-soluble substances using a solvent.
• the material from which water-soluble substances are removed using a solvent in the extraction step may be a medium or culture solution containing an anionic biosurfactant produced in the medium by the microorganism during culture, or a mixture containing a solid anionic biosurfactant separated from the reaction system.
• the product containing the anionic biosurfactant obtained through this extraction process is called the "extraction product.”
• the fat-soluble substance removal step is a step of removing fat-soluble substances from the extraction product.
• fat-soluble substance refers to a substance that is soluble in fats and oils.
• the method for removing fat-soluble substances from the solid extract product from which the water-soluble substances have been removed is not particularly limited as long as it is a method capable of separating the anionic biosurfactant from the fat-soluble substances, and conventional methods can be used. For example, first, the solvent is distilled off from the extract product, and then a solvent capable of separating the anionic biosurfactant from the fat-soluble substances is added to the solid obtained by distilling off the solvent, and the solvent layer is removed to obtain an anionic biosurfactant from which the fat-soluble substances have been removed. This makes it possible to efficiently remove fat-soluble impurities from the extract product.
• a solvent capable of separating anionic biosurfactants from fat-soluble substances is a solvent in which anionic biosurfactants are poorly soluble or insoluble, and fat-soluble substances are soluble.
• hexane is used as such a solvent, the water-soluble substances are removed from the precipitated product precipitated from the culture liquid using the solvent, and then the solid (extraction product) obtained by distilling off the solvent is suspended in hexane and the hexane is removed by filtration or centrifugation. This allows fat-soluble impurities to be efficiently removed from the extraction product containing anionic biosurfactants.
• the activating agent according to one embodiment of the present invention may be used as an activating agent for various cells, but is preferably used as an activating agent for skin cells.
• skin cells to which the activating agent according to one embodiment of the present invention can be applied include epidermal keratinocytes and dermal fibroblasts, but it is preferable to use the activating agent for epidermal keratinocytes.
• the activating agent according to one embodiment of the present invention may be an epidermal keratinocyte activating agent used for epidermal keratinocytes.
• the activator according to one embodiment of the present invention is characterized not only by having an activating effect, but also by having low skin irritation and good foaming durability, and can be suitably used in cosmetics or quasi-drugs. That is, in one embodiment of the present invention, a cosmetic or quasi-drug containing the activator according to one embodiment of the present invention is provided.
• Cosmetics according to one embodiment of the present invention include skin cosmetics (lotion, beauty essence, milky lotion, cream, etc.), lipstick, sunscreen cosmetics, make-up cosmetics, etc., but are not limited to these. It is preferable to use it in aqueous cosmetics such as lotion, beauty essence, milky lotion, cream pack/mask, pack, hair washing cosmetics, fragrance cosmetics, liquid body cleanser, UV care cosmetics, deodorant cosmetics, oral care cosmetics, etc. (in the case of cosmetics).
• aqueous cosmetics such as lotion, beauty essence, milky lotion, cream pack/mask, pack, hair washing cosmetics, fragrance cosmetics, liquid body cleanser, UV care cosmetics, deodorant cosmetics, oral care cosmetics, etc. (in the case of cosmetics).
• quasi-drugs are quasi-drugs as defined in the Pharmaceutical Affairs Law and are designated by the Minister of Health, Labour and Welfare as provided for in Article 2, Paragraph 2 of the Pharmaceutical Affairs Law.
• quasi-drugs refer to items used for the purpose of diagnosing, treating, improving, mitigating, treating or preventing illnesses in humans or animals, and have a milder effect than pharmaceuticals.
• quasi-drugs exclude items used for pharmaceutical purposes, and include products used to treat or prevent illnesses in humans or animals, and products that have a mild effect on the human body or have no direct effect.
• quasi-drugs refer to items that have a mild effect on the human body and are used to prevent nausea, other discomfort, or bad breath or body odor; to prevent heat rash, sores, etc.; or to prevent hair loss, promote hair growth, or remove hair.
• Specific examples of quasi-drugs include medicated cosmetics, bath additives, antiperspirants, hair coloring agents, hair restorers, medicated soaps, and permanent wave agents.
• the content of the activator according to one embodiment of the present invention in 100% by mass of the cosmetic or quasi-drug according to one embodiment of the present invention is preferably 0.00001% by mass or more, more preferably 0.00005% by mass or more, and even more preferably 0.00001% by mass or more.
• the upper limit of the content of the activator is preferably 10% by mass or less, more preferably 5% by mass or less, and even more preferably 3% by mass or less.
• the cosmetic or quasi-drug according to one embodiment of the present invention may contain a solvent such as water and/or other additives in addition to the activator according to one embodiment of the present invention.
• additives that may be contained in the cosmetic or quasi-drug according to one embodiment of the present invention include, for example, fats and oils, waxes, mineral oils, fatty acids, alcohols, esters, metal soaps, gums and water-soluble polymeric compounds, vitamins, amino acids, skin whitening agents, moisturizing agents, hair growth agents, ⁇ -hydroxy acids, inorganic pigments, UV absorbers, astringents, antioxidants, anti-inflammatory agents, germicides and disinfectants, fragrances, pigments and colorants, hormones, sequestering agents, pH adjusters, chelating agents, preservatives and antifungal agents, cooling agents, stabilizers, animal and vegetable proteins and their decomposition products, animal and vegetable polysaccharides and their decomposition products, animal and vegetable glycoproteins and their decomposition products, blood flow promoters, anti-inflammatory and anti-allergic agents, cell activators, keratolytic agents, wound treatment agents, foam boosters, thickeners, oral agents, and deodorants and deodorants.
• the anionic biosurfactant according to one embodiment of the present invention may have an enhancing effect on genes involved in skin barrier properties (e.g., CLDN1 gene, CLDN7 gene, DSC1 gene, etc.), genes that are collagen decomposition enzymes (e.g., MMP1 gene), genes that are moisturizing or epidermal keratinocyte differentiation markers (e.g., LOR gene, TGM1 gene, CSTA gene, KRT1 gene, KRT4 gene, FLG gene, FLG2 gene, CASP14 gene, etc.), genes involved in hyaluronic acid production (e.g., HAS1 gene, etc.), genes involved in elastin fiber formation, which is an extracellular matrix component of the dermis (e.g., EMILIN1 gene, LOXL1 gene, etc.), genes involved in antioxidant ability (e.g., GCLC gene, etc.), genes involved in anti-stress (e.g., HSPB1 gene, etc.), and/or genes involved in skin barrier
• the anionic biosurfactant according to one embodiment of the present invention can have various effects of improving skin conditions, such as a skin barrier property improving effect, a moisturizing improving effect, an antioxidant ability promoting effect, an anti-stress ability improving effect, a whitening effect, a wrinkle improving effect, and a wrinkle formation inhibiting effect. Therefore, the anionic biosurfactant according to one embodiment of the present invention can also be suitably used as a skin condition improving agent. That is, in one embodiment of the present invention, a skin condition improving agent containing the anionic biosurfactant according to one embodiment of the present invention is provided.
• skin condition improving agent refers to a substance (composition) that improves the barrier properties or moisture retention of the skin (epidermis), suppresses changes in skin condition due to external stimuli such as oxidative stress, suppresses wrinkle formation by promoting the breakdown of collagen fibers in the skin (dermis), the formation of elastin fibers, or the production of hyaluronic acid, and improves wrinkles that have already formed, or that has a whitening effect or suppresses the formation of age spots by suppressing the production of melanin.
• skin condition improving agents include skin barrier improving agents, skin moisturizing improving agents, skin wrinkle formation inhibitors, skin wrinkle improving agents, whitening promoters, oxidative stress inhibitors, stress inhibitors, and skin flora improving agents. That is, the skin condition improving agent according to one embodiment of the present invention is preferably a skin barrier improving agent, skin moisturizing improving agent, skin wrinkle formation inhibitor, skin wrinkle improving agent, whitening promoter, (skin) oxidative stress inhibitor, (skin) stress inhibitor, or skin flora improving agent.
• the anionic biosurfactant contained in the skin condition improving agent according to one embodiment of the present invention is preferably an anionic biosurfactant obtained using a carbon source with 14 carbon atoms, or an anionic biosurfactant obtained using a carbon source with 16 carbon atoms.
• an ⁇ -gel-containing composition includes an anionic biosurfactant according to one embodiment of the present invention and a higher alcohol.
• the ⁇ -gel-containing composition according to one embodiment of the present invention contains the anionic biosurfactant according to one embodiment of the present invention as a substance having surface activity and/or emulsification ability, and thus has the effect of being excellent in stability and excellent in usability when used in cosmetics or quasi-drugs.
• Activator is used as appropriate for the specific aspects of the anionic biosurfactant according to one embodiment of the present invention contained in the ⁇ -gel-containing composition according to one embodiment of the present invention.
• ⁇ -gel refers to a gel containing ⁇ -type hydrated crystals in which amphiphilic substances having hydrophilic and lipophilic groups, such as surfactants, form an association of bilayer membranes stacked in a lamellar shape, and water is held between the hydrophilic groups.
• ⁇ -gels include gels that not only hold water between the hydrophilic groups of the bilayer membrane, but also hold a large amount of water in the hydrophilic parts of the association.
• the amphiphilic substances are arranged in a hexagonal shape in the bilayer membrane, and are expected to have high thickening, water retention, and barrier effects.
• the ⁇ -gel-containing composition according to one embodiment of the present invention preferably contains the anionic biosurfactant in an amount of 0.001% by mass or more relative to the total mass of the ⁇ -gel-containing composition. This makes it possible to obtain an ⁇ -gel-containing composition that is excellent in stability and has an excellent feel when used in cosmetics or pharmaceuticals (quasi-drugs).
• the content of the anionic biosurfactant relative to the total mass of the ⁇ -gel-containing composition is more preferably 0.005% by mass or more, even more preferably 0.010% by mass or more, even more preferably 0.020% by mass or more, and particularly preferably 0.050% by mass or more.
• the upper limit of the content of the anionic biosurfactant relative to the total mass of the ⁇ -gel-containing composition is not particularly limited, but is preferably 10.000% by mass or less, more preferably 5.000% by mass or less, and even more preferably 3.000% by mass or less.
• the ratio of the content of the anionic biosurfactant to the content of the higher alcohol, in mass ratio is preferably greater than 0 and less than 0.500, more preferably greater than 0 and less than 0.400, even more preferably greater than 0.001 and less than 0.300, and particularly preferably 0.002 to 0.200. If the ratio of the content of the anionic biosurfactant to the content of the higher alcohol is within the above range, an ⁇ -gel-containing composition can be obtained that is excellent in stability and has an excellent feel when used in cosmetics or pharmaceutical (quasi-drug) products.
• the ⁇ -gel-containing composition according to one embodiment of the present invention contains an anionic biosurfactant and a higher alcohol.
• higher alcohol refers to a monohydric chain saturated alcohol having 6 or more carbon atoms.
• the higher alcohol may be a linear monohydric chain saturated alcohol or a branched monohydric chain saturated alcohol.
• the higher alcohol may be a primary alcohol, a secondary alcohol, or a tertiary alcohol.
• Examples of the higher alcohols include n-hexyl alcohol, n-heptyl alcohol, n-octyl alcohol, n-nonyl alcohol, n-decyl alcohol, n-undecyl alcohol, n-heptyl alcohol, lauryl alcohol, n-tridecyl alcohol, myristyl alcohol, n-pentadecyl alcohol, cetyl alcohol (cetanol), margaryl alcohol, stearyl alcohol, behenyl alcohol, n-nonadecyl alcohol, arachidyl alcohol, ceryl alcohol, melissyl alcohol, palmitoleyl alcohol, oleyl alcohol, eicosenyl alcohol, 2-methylpentyl alcohol, 2-ethylbutyl alcohol, 2-ethylhexyl alcohol, methylamyl alcohol, capryl alcohol, diisobutyl carbinol, isostearyl alcohol, octyldode
• the higher alcohol more preferably has a carbon number of 10 or more, even more preferably 12 or more, particularly preferably 14 or more, and most preferably 16 or more. There is no particular upper limit on the carbon number, but it is, for example, 24 or less, more preferably 22 or less, and even more preferably 20 or less.
• the higher alcohol is more preferably a linear monohydric chain saturated alcohol, and from the viewpoint of ⁇ -gel formation, it is more preferably a primary alcohol.
• more preferred examples of the higher alcohol include myristyl alcohol, cetanol, stearyl alcohol, behenyl alcohol, and mixtures of two or more of these. These are more preferred from the viewpoint of ⁇ -gel formation.
• the higher alcohol contains at least one of a higher alcohol having 16 carbon atoms and a higher alcohol having 18 carbon atoms. This is preferred because it provides a better feel when used in cosmetics or pharmaceuticals (quasi-drugs).
• higher alcohols include cetanol or a mixture of cetanol and other higher alcohols, stearyl alcohol or a mixture of stearyl alcohol and other higher alcohols, a mixture of cetanol and stearyl alcohol, or a mixture of the mixture and other higher alcohols.
• the ratio of the higher alcohol having 16 carbon atoms to the higher alcohol having 18 carbon atoms is preferably 90:10 to 10:90, more preferably 80:20 to 20:80, even more preferably 70:30 to 20:80, even more preferably 60:40 to 20:80, and particularly preferably 55:45 to 20:80.
• More specific examples of such higher alcohols include a combination of cetanol and stearyl alcohol in the above ratio, or cetearyl alcohol, which is a mixture of cetanol and stearyl alcohol.
• the ⁇ -gel-containing composition according to one embodiment of the present invention may further contain an oil in addition to the anionic biosurfactant and the higher alcohol.
• an oil By including an oil, the ⁇ -gel-containing composition according to one embodiment of the present invention can emulsify the oil in a gel composed of ⁇ -hydrate crystals and an aqueous phase to form a homogeneous gel consisting of three phases: ⁇ -hydrate crystals, an aqueous phase, and an oil phase.
• a homogeneous gel consisting of three phases: ⁇ -hydrate crystals, an aqueous phase, and an oil phase the stability of the composition can be further improved, which is preferable.
• the oil is preferably a liquid or paste-like oil at 1 atmosphere and 25°C, and a liquid oil is particularly preferred.
• liquid oil agent examples include waxes, hydrocarbons, higher alcohol esters, higher fatty acid esters, triglycerides, silicone oils, higher fatty acids, animal and vegetable oils, cholesterol fatty acid esters, sterols, sterol esters, polyphenols, etc.
• More preferred examples include waxes such as cownauba wax, candelilla wax, jojoba oil, beeswax, and lanolin; hydrocarbons such as mineral oil, isododecane, squalane, petrolatum, ceresin, and microcrystalline wax; esters such as isopropyl palmitate, isopropyl myristate, isooctyl myristate, isotridecyl myristate, octadecyl myristate, octyldodecyl myristate, cholesteryl isostearate, triethylhexanoin, cetyl ethylhexanoate, and tri(caprylic/capric)glyceryl; ethers; dimethyl silicone oils such as octamethyltrisiloxane and dimethicone; cyclic silicone oils such as cyclotetrasiloxane, cyclopentasilox
• the ratio of the content of the oil to the content of the higher alcohol, by mass is preferably greater than 0 and less than 800, more preferably greater than 0 and less than 150, even more preferably greater than 0 and less than 100, particularly preferably 0.05 to 80.0, and may be 0.1 to 80.0, or may be 0.5 to 80.0. If the ratio of the content of the oil to the content of the higher alcohol is within the above range, the stability of the ⁇ -gel-containing composition can be further improved, which is preferable.
• the ⁇ -gel-containing composition according to one embodiment of the present invention preferably contains, in addition to the anionic biosurfactant, higher alcohol, and oil, a polyhydric alcohol and/or a thickener, as described below, from the viewpoint of providing a superior feel when used in cosmetics or pharmaceuticals (quasi-drugs).
• the ⁇ -gel-containing composition according to one embodiment of the present invention may further contain a polyhydric alcohol in addition to the anionic biosurfactant and the higher alcohol.
• a polyhydric alcohol By containing a polyhydric alcohol, the ⁇ -gel-containing composition according to one embodiment of the present invention is preferable because it has a better feel when used in cosmetics or pharmaceuticals (quasi-drugs).
• the particle size of the ⁇ -gel-containing composition tends to be finer by containing a polyhydric alcohol.
• the finer particle size of the ⁇ -gel-containing composition further has the advantage of stabilizing the emulsion particles and improving skin compatibility.
• the polyhydric alcohol is not particularly limited, but examples thereof include propylene glycol, dipropylene glycol, 1,3-butylene glycol, pentylene glycol, glycerin, polypropylene glycol, diglycerin, polyglycerin, ethylene glycol, diethylene glycol, polyethylene glycol, 1,3-propanediol, sorbitol, etc. These may be used alone or in combination of two or more.
• the ratio of the content of the polyhydric alcohol to the content of the higher alcohol in terms of mass ratio, is preferably 0.1 or more, more preferably 1.0 or more, even more preferably 3.0 or more, and particularly preferably 5.0 or more.
• the upper limit of the mass ratio is not particularly limited, but is, for example, 100.0. If the ratio of the content of the polyhydric alcohol to the content of the higher alcohol is within the above range, the stability of the ⁇ -gel-containing composition can be further improved, and this is preferable.
• the ⁇ -gel-containing composition according to one embodiment of the present invention preferably contains the oil agent and/or the thickener described below in addition to the anionic biosurfactant, higher alcohol, and polyhydric alcohol, from the viewpoint of providing a superior feel when used in cosmetics or pharmaceuticals (quasi-drugs).
• the ⁇ -gel-containing composition according to one embodiment of the present invention may further contain a thickener in addition to the anionic biosurfactant and the higher alcohol.
• a thickener By containing a thickener, the ⁇ -gel-containing composition according to one embodiment of the present invention is preferably because separation of the aqueous phase is further suppressed and the stability is improved.
• the thickener is not particularly limited, but examples thereof include xanthan gum, carbomer, hydroxyethyl cellulose, methyl cellulose, ethyl cellulose, gum arabic, gellan gum, sclerotium gum, etc. These may be used alone or in combination of two or more kinds.
• the ratio of the content of the thickener to the content of the higher alcohol is preferably 0.01 or more, more preferably 0.03 or more, even more preferably 0.05 or more, and particularly preferably 0.10 or more, by mass.
• the upper limit of the mass ratio is not particularly limited, but is, for example, 10.00. If the ratio of the content of the thickener to the content of the higher alcohol is within the above range, the stability of the ⁇ -gel-containing composition can be further improved, and this is preferable.
• the ⁇ -gel-containing composition according to one embodiment of the present invention preferably contains the oil and/or polyhydric alcohol in addition to the anionic biosurfactant, higher alcohol, and thickener, from the viewpoint of providing a superior feel when used in cosmetics or pharmaceuticals (quasi-drugs).
• the ⁇ -gel-containing composition according to one embodiment of the present invention may contain, in addition to an anionic biosurfactant, a higher alcohol, and, if necessary, at least one selected from the group consisting of oils, polyhydric alcohols, and thickeners, other components may be included within a range that does not adversely affect the effects of the present invention.
• the other components include, but are not limited to, ultraviolet absorbers, antioxidants, emollients, emulsifiers, solubilizers, anti-inflammatory agents, moisturizers, preservatives, pH adjusters, dyes, fragrances, powders, etc.
• the emulsifier is intended to be an emulsifier other than the anionic biosurfactant, and for example, various anionic surfactants, cationic surfactants, amphoteric surfactants, and nonionic surfactants can be used.
• the content of the other components is not particularly limited as long as it does not adversely affect the effects of the present invention, but is, for example, 0 to 99% by mass relative to the total mass of the ⁇ -gel-containing composition.
• the ⁇ -gel-containing composition according to one embodiment of the present invention contains an anionic biosurfactant, a higher alcohol, and, optionally, at least one selected from the group consisting of an oil, a polyhydric alcohol, and a thickener, and, optionally, the other components described above, with the remainder being water.
• the amount of water contained in the ⁇ -gel-containing composition is not particularly limited, but is preferably 1% by mass to 99% by mass, more preferably 5% by mass to 95% by mass, and even more preferably 10% by mass to 90% by mass, relative to the total mass of the ⁇ -gel-containing composition.
• the water content of the ⁇ -gel-containing composition is 1% by mass or more relative to the total mass of the composition. Also, it is preferable that the water content is 99% by mass or less from the viewpoint of forming ⁇ -gel.
• the water used can be distilled water, ion-exchanged water, tap water, etc.
• the ⁇ -gel-containing composition preferably has a pH of 5.0 or more. If the pH is 5 or more, the stability of the ⁇ -gel-containing composition is excellent, which is preferable.
• the ⁇ -gel-containing composition more preferably has a pH of 6.0 or more.
• the pH of the ⁇ -gel-containing composition is preferably 12.0 or less, more preferably 11.5 or less, and even more preferably 11.0 or less.
• the pH of the ⁇ -gel-containing composition according to one embodiment of the present invention can be adjusted by adding a pH adjuster when preparing the ⁇ -gel-containing composition.
• pH adjusters that adjust the ⁇ -gel-containing composition to the alkaline side include sodium hydroxide, sodium bicarbonate, potassium hydroxide, magnesium hydroxide, triethanolamine, arginine, sodium citrate, sodium lactate, disodium succinate, etc.
• pH adjusters that adjust the ⁇ -gel-containing composition to the acid side include citric acid, lactic acid, malic acid, etc., or salts of these.
• the average particle size of the ⁇ -gel-containing composition according to one embodiment of the present invention is preferably 200.0 ⁇ m or less, more preferably 100.0 ⁇ m or less, and even more preferably 50.0 ⁇ m or less.
• the average particle size of the ⁇ -gel-containing composition refers to the average particle size of oil particles enclosed in an aggregate of lamellar multilayer films, and is a dimension that includes the multilayer films.
• the average particle size of the ⁇ -gel-containing composition is 200.0 ⁇ m or less, it is preferable from the viewpoint of emulsion stability.
• the average particle size of the ⁇ -gel-containing composition means the median size (volume basis), and is a value measured by the method described in the Examples below.
• the method for producing an ⁇ -gel-containing composition according to one embodiment of the present invention is not particularly limited as long as it is a method that can produce the desired ⁇ -gel-containing composition, and various methods can be adopted.
• the method for producing such an ⁇ -gel-containing composition may include an oil phase preparation step for preparing an oil phase containing an anionic biosurfactant, a higher alcohol, and an oil, an aqueous phase preparation step for separately preparing an aqueous phase containing water, and an emulsification step for mixing and emulsifying the aqueous phase and the oil phase.
• the method for preparing the oil phase containing the anionic biosurfactant, the higher alcohol, and the oil agent is not particularly limited as long as it is a method that allows these components to be mixed.
• a method can be suitably adopted in which the anionic biosurfactant and the higher alcohol are mixed and dissolved in each other, and then the oil agent is added to this mixture.
• the order in which the components are mixed is not limited to this, and any order is acceptable.
• the temperature at which the components are mixed in the oil phase preparation step is not particularly limited as long as it is a temperature at which the components are mutually dissolved.
• the aqueous phase preparation step is not particularly limited as long as it is a method that can mix water and, as necessary, water-soluble components such as polyhydric alcohols and pH adjusters.
• the order in which the components are mixed is not particularly limited, and any order is acceptable.
• the temperature at which the components are mixed in the aqueous phase preparation step is also not particularly limited, and any temperature at which the components dissolve in each other may be used.
• the pH of the aqueous phase obtained in the aqueous phase preparation step is preferably 5.0 or higher, and more preferably 6.0 or higher.
• the emulsification step is not particularly limited as long as it is a step that can emulsify the aqueous phase obtained in the aqueous phase preparation step and the oil phase obtained in the oil phase preparation step.
• the aqueous phase and the oil phase are mixed and stirred at 40°C to 90°C, more preferably 60°C to 90°C.
• the method of mixing the aqueous phase and the oil phase is not particularly limited, and the oil phase may be added to the aqueous phase, or the aqueous phase may be added to the oil phase.
• the addition method is also not particularly limited, and the mixture may be added gradually or all at once.
• the aqueous phase and the oil phase are heated to 40°C to 90°C, more preferably 60°C to 90°C, when mixed.
• the method of mixing and stirring the aqueous phase and the oil phase is not particularly limited, and the mixture may be stirred using a homomixer, homodisper, three-one motor, or the like. Among these, a homomixer is more preferably used.
• the stirring speed at which the aqueous phase and the oil phase are mixed and stirred is not particularly limited, but is, for example, 1000 rpm or more, and more preferably 2000 rpm or more.
• the stirring time is also not particularly limited, but is, for example, 10 seconds or more, and more preferably 1 minute or more.
• the aqueous phase and the oil phase are emulsified in the emulsification step, it is preferable to leave the resulting ⁇ -gel-containing composition to cool or cool it using a cooling device while stirring.
• the ⁇ -gel-containing composition according to one embodiment of the present invention has excellent stability and excellent usability when used in cosmetics or quasi-drugs, and therefore can be suitably used in cosmetics or medicinal (quasi-drug) products. That is, in one aspect of the present invention, a cosmetic or quasi-drug product containing the ⁇ -gel-containing composition according to one embodiment of the present invention is provided.
• Examples of cosmetics or quasi-drugs containing the ⁇ -gel-containing composition according to one embodiment of the present invention include, but are not limited to, skin cosmetics (lotion, serum, emulsion, cream, etc.), lipstick, sunscreen cosmetics, and makeup cosmetics.
• the ⁇ -gel-containing composition according to one embodiment of the present invention is preferably used in aqueous cosmetics such as lotion, serum, emulsion, cream pack/mask, pack, hair washing cosmetics, fragrance cosmetics, liquid body cleanser, UV care cosmetics, deodorant cosmetics, oral care cosmetics, etc. (in the case of cosmetics).
• a cosmetic or quasi-drug containing the ⁇ -gel-containing composition according to one embodiment of the present invention may contain a solvent such as water and/or other additives in addition to the ⁇ -gel-containing composition according to one embodiment of the present invention.
• One embodiment of the present invention may have the following configuration.
• a skin condition improving agent containing an anionic biosurfactant containing an anionic biosurfactant.
• the skin condition improving agent according to [7] which is a skin bacterial flora improving agent.
• An ⁇ -gel-containing composition comprising an anionic biosurfactant and a higher alcohol.
• Example 1 Acquisition of STL C14 Rhodococcus erythropolis SD-74 strain was seed cultured under the following conditions according to the method described in Y. Uchida et al., Agric. Biol. Chem., 53(3):765-769 (1989).
• the Rhodococcus erythropolis SD-74 strain used in this example was isolated as a vegetable oil-utilizing bacterium and deposited at the Patent Microorganisms Depositary Center of the National Institute of Technology and Evaluation under the accession number NITE AP-03788.
• Rhodococcus erythropolis SD-74 strain was inoculated into 100 ml of FPY medium (2% fructose (Fujifilm Wako Pure Chemical Industries, Ltd.), 0.5% polypeptone (Fujifilm Wako Pure Chemical Industries, Ltd.), 0.5% yeast extract powder (Oxoid Corporation), 0.1% NaNO 3 (Fujifilm Wako Pure Chemical Industries, Ltd.), 0.1% KH 2 PO 4 (Fujifilm Wako Pure Chemical Industries, Ltd.), 0.1% MgSO 4 -7H 2 O (Fujifilm Wako Pure Chemical Industries, Ltd.)) in a 500 ml Sakaguchi flask and cultured with shaking at 30°C for 72 hours to obtain a seed culture solution.
• FPY medium 2% fructose (Fujifilm Wako Pure Chemical Industries, Ltd.), 0.5% polypeptone (Fujifilm Wako Pure Chemical Industries, Ltd.), 0.5% yeast extract powder (Oxoid Corporation),
• a 10 L jar culture tank (Able) was used for culture at a culture temperature of 30° C. with stirring at 300 rpm.
• the pH of the medium was maintained at 7.0 by adding 50% KOH (Fujifilm Wako Pure Chemical Industries, Ltd.).
• the STL C14 obtained in Example 1 is a succinoyltrehalose lipid having a carboxyl group produced using tetradecane as a carbon source.
• Example 2 Obtaining STL C16 100 g of a white solid (STL C16) was obtained by the method described in Example 1, except that the carbon source was changed to hexadecane (Tokyo Chemical Industry Co., Ltd.).
• the STL C16 obtained in Example 2 is a succinoyltrehalose lipid having a carboxyl group produced using hexadecane as a carbon source.
• Example 3 Obtaining STLC18 100 g of a white solid (STL C18) was obtained by the method described in Example 1, except that the carbon source was changed to octadecane (Tokyo Chemical Industry Co., Ltd.).
• the STL C18 obtained in Example 3 is a succinoyltrehalose lipid having a carboxyl group produced using octadecane as a carbon source.
• a cell suspension was prepared using Humedia-KG2 medium (Kurashiki Boseki) so that normal human epidermal keratinocytes (Kurashiki Boseki) were at 8.5 x 10 4 cells/mL, and the human epidermal keratinocytes were seeded at 100 ⁇ L/well in a 96-well multi-well plate (Thermo Fisher).
• the experiment was performed with 5 technical replicates in the cell culture experiment. After culturing for 24 hours at 37°C in a 5% CO 2 atmosphere, the medium was discarded, and the STL sample-supplemented medium shown in Table 1 was added at 100 ⁇ L/well.
• the ATP amount of each well was measured using CellTiter-Glo (registered trademark) 2.0 Cell Viability Assay kit (Promega) according to the protocol attached to the kit, and the relative value was calculated when the ATP amount of the additive-free medium was set to 100.
• the progress of the increase in cell number in each well was measured using Provi CM20 (Evident), and the relative value was calculated when the number of cells in the additive-free medium was set to 100.
• the relative value of the ATP amount relative to the additive-free medium at 48 hours after addition and the relative value of the number of cells relative to the additive-free medium were used to calculate the relative value of the ATP amount per cell according to the following formula.
• Relative value of ATP amount per cell (relative value of each well obtained by ATP measurement relative to the supplement-free medium)/(relative value of each well obtained by cell count measurement relative to the supplement-free medium) The results are shown in Table 1.
• a cell suspension was prepared using DMEM medium (Fujifilm Wako Pure Chemical Industries, Ltd.) containing heat-inactivated fetal bovine serum (final concentration 10%) so that L929 mouse fibroblasts (KAC Corporation) were 2.0 x 10 5 cells/mL, and L929 mouse fibroblasts were seeded at 150 ⁇ L/well on a 96-well multi-well plate (Thermo Fisher). Note that the number of technical replicates in the cell culture experiment was 3. After 24 hours of culture at 37°C under a 5% CO 2 atmosphere, 150 ⁇ L/well of STL sample-supplemented medium was added.
• a 50% ethanol aqueous solution was prepared using ethanol (Fujifilm Wako Pure Chemical Industries, Ltd.), and glacial acetic acid (Fujifilm Wako Pure Chemical Industries, Ltd.) was added to the 50% ethanol aqueous solution to a final concentration of 1%.
• This 1% glacial acetic acid ethanol aqueous solution was added at 100 ⁇ L/well to fix the cells. After 5 minutes, the absorbance at 546 nm was measured using a Corona Multigrating Microplate Reader SH-9000Lab (Hitachi High-Tech Science Corporation), and the live cells that had incorporated Neutral Red were quantified.
• the 50% cell proliferation inhibition concentration (IC50) was calculated as an evaluation score of cytotoxicity. For STL C18, 50% cell proliferation inhibition was not observed even at 1000 ppm, which was the highest STL concentration tested this time.
• the IC50 values for sodium lauryl sulfate, sodium deoxycholate, and sodium caprylate are taken from L. Vian, J. Vinvent, J. Maurin, I. Fabre, J. Giroux, J. P. Cano, Toxicol in vitro, 9(2), p185-190,1995.
• STL C14, STL C16, and STL C18 all had IC50s of 300 ppm or higher, indicating low cytotoxicity.
• STL physiological function evaluation test Using STL C14, STL C16, and STL C18 obtained by the methods described in Examples 1 to 3 as activators for various physiological functions (i.e., using an activator containing 100 parts by mass of STL C14, STL C16, or STL C18 per 100 parts by mass of the activator), STL sample-added media were prepared so that the final concentration of the activator was 10 ppm, and physiological function evaluation tests were performed.
• a cell suspension of normal human epidermal keratinocytes was prepared using Humedia-KG2 medium (Kurashiki Boseki) at 1.53 x 10 cells/mL, and the human epidermal keratinocytes were seeded at 5 mL/well in a 4-Well Rectangular Dish, TC Surface (Thermo Fisher). Note that the number of technical replicates in the cell culture experiment was 3.
• the medium was discarded, and the medium (STL C14-added group, STL C16-added group, and STL C18-added group) containing each of the above activators (activators containing 100 parts by mass of STL C14, STL C16, or STL C18 per 100 parts by mass of activator) added to Humedia-KG2 medium (Kurashiki Boseki Co., Ltd.) to a final concentration of 10 ppm, or Humedia-KG2 medium (Kurashiki Boseki Co., Ltd.) as a non-added group was added at 5 mL/well to a 4-Well Rectangular Dish, TC Surface (Thermo Fisher Scientific).
• RNAprotect Cell Reagen Qiagen
• RNA purity and degradation were measured using NanoDrop One (Thermo Fisher) and TapeStation (Agilent), and it was confirmed that 260/280, an index of RNA purity, was 1.9 or more, 260/230 was 1.9 or more, and the RIN value, an index of RNA degradation, was 8 or more.
• RNA concentration After measuring the RNA concentration using Quant-iT (registered trademark) RNA Assay Kits (Thermo Fisher Scientific), a reverse transcription reaction was performed using ReverTra Ace qPCR RT master mix (Toyobo Co., Ltd.) by adding total RNA to a final RNA concentration of 50 ng/ ⁇ L to obtain cDNA.
• Quant-iT registered trademark
• ReverTra Ace qPCR RT master mix Toyobo Co., Ltd.
• Quantitative PCR experiments were carried out according to the protocol published by Qiagen using QIAcuty probe PCR kit (Qiagen), QIAcuty Nanoplate 8.5k 96-well (Qiagen), QIAcuty Four digital PCR system (Qiagen), and Tagman probe (Thermo Fisher) listed in Table 2 below, and the average value of the relative value of each gene was calculated when the untreated group was set to 1.
• the technical replicates of the quantitative PCR experiment were 2.
• the results of gene expression analysis by quantitative PCR are shown in Table 3.
• CMC critical micelle concentration
• STL protein denaturation test Using STL C14, STL C16, and STL C18 prepared in Examples 1 to 3, 10 mL (pH 8.0) of aqueous solution was prepared so that the final concentration of STL was 0.5%. The pH was adjusted using an aqueous sodium hydroxide solution (Fujifilm Wako Pure Chemicals). 0.03 g of zein powder (Fujifilm Wako Pure Chemicals), a protein derived from corn, was added to the aqueous solution of STL.
• the zein powder-added solution was transferred to a centrifuge tube and centrifuged at 6,200 rpm for 10 seconds to precipitate and remove undissolved zein powder.
• the protein concentration in the solution was measured using an ultramicrospectrophotometer NanoDrop (NanoDrop Technologies). The results are shown in Table 4.
• the average bubble area of the foam was then determined based on the images, and the creaminess (foaminess) of the foam was evaluated according to the following criteria. The results are shown in Table 4. Note that the creaminess of the foam is characterized by having a small final bubble diameter (measured as the average bubble area). The foam generated from the aqueous solutions containing STL C14, STL C16, and STL C18 had small bubble areas, suggesting that they could form creamy foam. ⁇ Evaluation criteria for creaminess of foam> ⁇ (Good): Bubble area is 20,000 ⁇ m2 or less. ⁇ (Pass): Bubble area is 20,001 ⁇ m2 or more and 30,000 ⁇ m2 or less. ⁇ (slightly poor): bubble area is 30,001 ⁇ m 2 or more and 40,000 ⁇ m 2 or less. ⁇ (bad): bubble area is 40001 ⁇ m2 or more.
• the foam change rate was also calculated using the following formula. The smaller the foam change rate, the better the foam retention, or in other words, the better the foam durability. As the foam change rate was small for STL C14, STL C16, and STL C18, it was believed that they were able to produce foam with good foam retention.
• Rate of change in foam (average bubble area 20 seconds after stirring has stopped) / (average bubble area 600 seconds after stirring has stopped) Similar foam characterization was performed using sodium lauryl sulfate, sodium deoxycholate, and sodium caprylate, and the results are shown in Table 4.
• the anionic biosurfactant is useful as a cell activator. It was also found that the anionic biosurfactant has low skin irritation and good foaming durability. It was also found that the anionic biosurfactant is useful as a skin condition improving agent.
• Example 4 0.100 parts by mass of the anionic biosurfactant obtained in Example 1 and 3.000 parts by mass of cetearyl alcohol as a higher alcohol were mixed at 80° C. to 85° C. to obtain a solution in which the anionic biosurfactant obtained in Example 1 was dissolved in a higher alcohol. 50.000 parts by mass of squalane as an oil agent was added to the obtained solution, and the solution was heated to 80° C. to 85° C. to obtain an oil phase.
• Example 5 A gel composition was obtained by the same procedure as in Example 4, except that 2.000 parts by mass of cetearyl alcohol and 1.000 parts by mass of behenyl alcohol were used instead of 3.000 parts by mass of cetearyl alcohol as the higher alcohol.
• Example 6 A gel composition was obtained by the same procedure as in Example 4, except that 2.100 parts by mass of cetanol and 0.900 parts by mass of stearyl alcohol were used instead of 3.000 parts by mass of cetearyl alcohol as the higher alcohol. As a result of checking the X-ray diffraction pattern, the obtained gel composition was an ⁇ -gel-containing composition.
• Example 7 A gel composition was obtained by the same procedure as in Example 4, except that 50,000 parts by mass of 2-ethylhexanoate was used instead of 50,000 parts by mass of squalane as the oil agent.
• Example 8 A gel composition was obtained by the same procedure as in Example 4, except that 50,000 parts by mass of dimethicone was used instead of 50,000 parts by mass of squalane as the oil agent. As a result of checking the X-ray diffraction pattern, the obtained gel composition was an ⁇ -gel-containing composition.
• Example 9 A gel composition was obtained by the same procedure as in Example 4, except that 50,000 parts by mass of squalane and 0.500 parts by mass of microcrystalline wax were used as the oil agent, and the amount of purified water was changed accordingly as shown in Table 5.
• Example 10 A gel composition was obtained by the same procedure as in Example 4, except that the amount of squalane used as the oil agent was changed from 50,000 parts by mass to 30,000 parts by mass, and the amount of purified water was accordingly changed as shown in Table 5. The X-ray diffraction pattern was confirmed to be an ⁇ -gel-containing composition.
• Example 11 In preparing the aqueous phase, 10,000 parts by mass of 1,3-butylene glycol (polyhydric alcohol) was added, and the amount of purified water was changed accordingly as shown in Table 5. The same procedure as in Example 4 was carried out to obtain a gel composition.
• Example 12 A gel composition was obtained by the same procedure as in Example 11, except that 10,000 parts by mass of 1,3-butylene glycol (polyhydric alcohol) was changed to 10,000 parts by mass of glycerin (polyhydric alcohol).
• Example 13 A gel composition was obtained by the same procedure as in Example 11, except that 10,000 parts by mass of 1,3-butylene glycol (polyhydric alcohol) was changed to 10,000 parts by mass of pentylene glycol (polyhydric alcohol).
• Example 14 In preparing the aqueous phase, 10,000 parts by mass of 1,3-butylene glycol (polyhydric alcohol) was added, and the amount of purified water was changed accordingly as shown in Table 5. The same procedure as in Example 8 was carried out to obtain a gel composition.
• 1,3-butylene glycol polyhydric alcohol
• Example 15 A gel composition was obtained by the same procedure as in Example 4, except that, as the surfactant, 0.100 parts by mass of glyceryl stearate (nonionic surfactant) was used in addition to 0.100 parts by mass of the anionic biosurfactant obtained in Example 1, and the amount of purified water was changed accordingly as shown in Table 6.
• the surfactant 0.100 parts by mass of glyceryl stearate (nonionic surfactant) was used in addition to 0.100 parts by mass of the anionic biosurfactant obtained in Example 1, and the amount of purified water was changed accordingly as shown in Table 6.
• Example 1 0.100 parts by mass of the anionic biosurfactant obtained in Example 1, 0.100 parts by mass of glyceryl stearate, and 3.000 parts by mass of cetearyl alcohol were mixed to obtain a solution in which the surfactant was dissolved in a higher alcohol, and 50.000 parts by mass of squalane was added to the obtained solution as an oil agent, followed by heating to 80°C to 85°C.
• Example 16 A gel composition was obtained by the same procedure as in Example 4, except that the amount of the anionic biosurfactant obtained in Example 1 was changed from 0.100 parts by mass to 0.300 parts by mass, and the amounts of purified water and sodium hydroxide were accordingly changed as shown in Table 6. The X-ray diffraction pattern was confirmed to be an ⁇ -gel-containing composition.
• Example 17 A gel composition was obtained by the same procedure as in Example 4, except that the amount of the anionic biosurfactant obtained in Example 1 was changed from 0.100 parts by mass to 0.030 parts by mass, and the amounts of purified water and sodium hydroxide were accordingly changed as shown in Table 6. The X-ray diffraction pattern was confirmed to be an ⁇ -gel-containing composition.
• Example 18 A gel composition was obtained by the same procedure as in Example 4, except that the amounts of the anionic biosurfactant and higher alcohol obtained in Example 1 were changed to 1.5 times the amounts, and the amounts of purified water and sodium hydroxide were accordingly changed as shown in Table 6.
• Example 1 the amount of the anionic biosurfactant obtained in Example 1 was changed from 0.100 parts by mass to 0.150 parts by mass, and the amount of cetearyl alcohol was changed from 3.000 parts by mass to 4.500 parts by mass.
• the amount of purified water was changed from 46.590 parts by mass to 45.030 parts by mass, and the amount of sodium hydroxide was changed from 0.013 parts by mass to 0.020 parts by mass.
• Example 19 A gel composition was obtained by the same procedure as in Example 4, except that no thickener (0.200 parts by mass of xanthan gum) was used and the amount of purified water was changed accordingly as shown in Table 6.
• Example 20 A gel composition was obtained by the same procedure as in Example 4, except that carbomer neutralized with sodium hydroxide (prepared by adding 0.100 parts by mass of sodium hydroxide to 10.000 parts by mass of a 2% aqueous solution of carbomer (0.200 parts by mass as carbomer)) was added instead of 0.200 parts by mass of xanthan gum as a thickener.
• the total amount of sodium hydroxide used in this case is shown in Table 6.
• Example 21 In preparing the aqueous phase, 0.029 parts by mass of citric acid was further added, and the amount of purified water was changed accordingly as shown in Table 6. The same procedure as in Example 4 was carried out to obtain a gel composition.
• Example 22 A gel composition was obtained by the same procedure as in Example 21, except that the amount of citric acid was changed from 0.029 parts by mass to 0.016 parts by mass.
• Example 23 A gel composition was obtained by the same procedure as in Example 21, except that the amount of citric acid was changed from 0.029 parts by mass to 0.009 parts by mass.
• Example 1 A gel composition was obtained by the same procedure as in Example 4, except that 0.100 parts by mass of the anionic biosurfactant obtained in Example 1 was not added and the amount of purified water was changed accordingly as shown in Table 6. The X-ray diffraction pattern was confirmed, and the obtained gel composition did not contain ⁇ -gel.
• Example 2 A gel composition was obtained by the same procedure as in Example 4, except that 0.100 parts by mass of the anionic biosurfactant obtained in Example 1 was changed to 0.100 parts by mass of glyceryl stearate (a nonionic surfactant).
• the ⁇ -gel-containing compositions obtained in Examples 11 to 14, which used a polyhydric alcohol in addition to an anionic biosurfactant and a higher alcohol had an extremely excellent feel when used.
• the ⁇ -gel-containing compositions obtained in Examples 11 to 14, which used a polyhydric alcohol in addition to an anionic biosurfactant and a higher alcohol had a small average particle size, indicating that the particle size of the ⁇ -gel-containing composition was finer.
• a comparison between Examples 19 and 20 showed that the ⁇ -gel-containing composition containing a thickener further suppressed separation of the aqueous phase, improving stability.
• Oil phase 1 and oil phase 2 were prepared by mixing and dissolving them at 80°C to 85°C in advance, and the two were mixed to prepare the oil phase. Separately, an aqueous phase was prepared by heating and mixing at 80°C to 85°C. Sodium hydroxide was added so that the aqueous phase had a pH of 8.
• the oil phase was gradually added to the water phase while stirring at 3000 rpm using a homomixer, and after the entire amount was added, the mixture was stirred at 5000 rpm for 5 minutes. After that, the mixture was cooled to 55°C while stirring at 1000 rpm, and additives 1 and 2 were added at that point, and the mixture was cooled to 35°C to obtain an emulsion composition.
• This emulsion composition had a rich feel when applied and was excellent in usability.
• the activator containing the anionic biosurfactant according to one embodiment of the present invention has excellent cell activation properties, low skin irritation, and good foaming durability. Therefore, the activator containing the anionic biosurfactant according to one embodiment of the present invention can be widely used not only as an activator, but also in cosmetics, quasi-drugs, etc. In addition, the anionic biosurfactant according to one embodiment of the present invention can be suitably used as various skin condition improving agents or ⁇ -gel-containing compositions.

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