CN101316574A - Skin care cosmetic and skin roughness improving agent containing biosurfactant - Google Patents

Abstract

The present invention relates to a cosmetic for rough skin improvement/skin care comprising a biosurfactant, particularly MEL-A, MEL-B or MEL-C.

Description

Skin care cosmetic and skin roughness improving agent containing biosurfactant

technical field

The present invention relates to the use of a biosurfactant or a premix thereof for skin care/skin roughness improvement, in particular to the use of a biosurfactant as a cosmetic, and further to a skin care/skin roughness improvement cosmetic comprising a biosurface Active agents or premixes thereof. More specifically, the present invention relates to a cosmetic, characterized in that: the biosurfactant is mannose erythritol lipid (hereinafter referred to as "MEL"), such as mannose erythritol lipid A (hereinafter referred to as "MEL") MEL-A"), mannose erythritol lipid B (hereinafter referred to as "MEL-B") or mannose erythritol lipid C (hereinafter referred to as "MEL-C"); or mannose mannitol lipid ( Hereinafter referred to as "MML"). In addition, the present invention relates to an agent for improving rough skin.

Background technique

Rough skin disease means that the skin is in a dry state, and scaly exfoliation of keratinocytes is observed on the skin. The formation of such rough skin disease is attributed to the leaching of lipids such as cholesterol, ceramide, and fatty acid between keratinocytes, and degeneration of keratinocytes and disturbance of proliferation/keratinization balance in epidermal cells attributable to ultraviolet rays and detergents resulting in keratinization Layer permeability barrier is not fully formed. For the purpose of preventing or treating the aforementioned rough skin diseases, studies have been conducted on supplementing the skin with interkeratinocyte lipid components or synthetic interkeratinocyte lipid analogues.

Lamella granules biosynthesized in the spinous layer and granular layer of the cells are released in the intercellular space just below the stratum corneum, extend to form a lamellar structure, and expand in the intercellular space to produce the aforementioned intercellular lipids of the corneocytes . Lamellar granules are composed of glycosylceramides, cholesterol, ceramides, phospholipids, etc.; however, glycosylceramides are rarely included in interkeratinocyte lipids. That is, it is believed that glycosylceramide in lamellar granules is hydrolyzed by β-glucocerebrosidase and converted into ceramide, and that this ceramide forms a lamellar structure, resulting in The formation of the stratum corneum permeability barrier improves, acting as a barrier against pruritus. It has been reported that ceramide supplements are effective in overcoming skin roughness due to detergents and are highly effective in improving skin roughness (Non-Patent Document 1).

Plant extracts mainly consist of glycosylceramides, but are still not satisfactory substitutes for ceramides. There are many reaction steps during its synthesis, and its large-scale production is costly.

MEL is a natural surfactant produced by yeast, and various physiological actions thereof have been previously reported (Non-Patent Document 2). Recently, mannose-mannitol lipid (MML) obtained by substituting mannitol for erythritol has been found (Patent Document 1). Their use in external preparations and cosmetics as anti-inflammatory agents and antiallergic agents (Patent Document 2), as drugs for treating baldness and hair growth (Patent Document 3), and their antibacterial effects (Patent Document 4) and Surface tension lowering effect (Patent Document 5); however, the use of MEL as an effective substitute for improving skin roughness is unknown.

Patent Document 1: JP 2005-104837-A

Patent Document 2: JP 2005-68015-A

Patent Document 3: JP 2003-261424-A

Patent Document 4: JP Sho-57-145896-A

Patent Document 5: JP Sho-61-205450

Non-Patent Document 1: Skin and Beauty, 36: 210, 2004

Non-Patent Document 2: Journal of Bioscience and Bioengineering 94:187, 2002

Contents of the invention

Problems solved by the present invention

Ceramides are useful as ingredients in cosmetics for improving rough skin, but because synthetic products or extracts from plants are expensive, ceramides are currently only used in small amounts. The main object of the present invention is to provide topical preparations for skin which use biosurfactants produced by microorganisms as a substitute for ceramides. That is, the present invention provides a biosurfactant by which the formation of the stratum corneum permeability barrier is improved, thereby achieving improvement in pruritus, and which is a readily available lipid component as an external preparation for skin.

way of solving the problem

As a result of thorough research to solve the above problems, it was found that after adding the biosurfactant to a rough skin model made by degreasing a three-dimensional skin model with sodium dodecyl sulfate (SDS), the biosurfactant The role of alternatives to ceramides; the use of biosurfactants was further demonstrated by application on the site of pruritus (produced by treating human skin with SDS). In short, it was found that biosurfactants can be used not only as emulsifiers but also in place of ceramides; thereby completing the present invention.

The present invention relates to the following skin care cosmetics and skin roughness improving agents.

(1) Skin care cosmetics comprising at least one biosurfactant.

(2) The skin care cosmetic according to (1), which is used for improving rough skin.

(3) The skin care cosmetic according to (1), which is used for improving rough skin by the action of a surfactant.

(4) The skin care cosmetic according to (1), wherein the biosurfactant is mannose erythritol lipid (MEL) and/or mannose mannitol lipid (MML).

(5) The skin care cosmetic according to (1), wherein the biosurfactant is selected from the group consisting of mannose erythritol lipid A (MEL-A), mannose erythritol lipid B (MEL-B) and mannose At least one of sugar erythritol lipid C (MEL-C).

(6) The skin care cosmetic according to (1), wherein the biosurfactant is mannose erythritol lipid B (MEL-B).

(7) The skin care cosmetic according to (1), wherein the biosurfactant is mannose erythritol lipid C (MEL-C).

(8) The skin care cosmetic according to (1), wherein the biosurfactant is mannose erythritol lipid A (MEL-A).

(9) A rough skin improving agent consisting of at least one biosurfactant.

Effect of the present invention

In the present invention, it has been found that MELs such as MEL-A, MEL-B and MEL-C as well as MML, which are biosurfactants produced by microorganisms, can be used instead of ceramides for skin care and skin roughness improvement . The biosurfactant of the present invention can be produced on a large scale by culturing microorganisms. By using this ceramide substitute, rough skin improvement/skin care effect and emulsification effect can be expected. Thus, an external preparation for skin that is effective in improving rough skin can be obtained. In particular, MEL-B and MEL-C are highly hydrophilic and can be prepared as stable emulsifiers. Biosurfactants can be used in the form of premixes.

MELs are preferred because MELs can be incorporated in cosmetics and external preparations for skin by being dissolved in oil-based or oil-soluble ingredients, and can be used in aqueous solutions (such as skin) by introducing MEL into liposomes. lotion, moisturizing lotion) form, which can be introduced into the skin excellently. Liposomes can be used in the form of non-aqueous solutions. It is not necessary to form all MELs into liposomes, MELs in liposome form can be mixed with MELs in sheet form or monomeric MELs.

The biosurfactant of the present invention is particularly useful as a skin care cosmetic and as a cosmetic for improving skin roughness; it is also useful as a quasi-drug and a drug such as a therapeutic agent for skin diseases such as moderate to severe skin roughness , acne, eczema, sebum deficiency, senile dry skin and itchy skin.

Biosurfactants can be used as ingredients combined with cleansing cosmetics because biosurfactants have cleansing properties in addition to skin roughness improvement/skin care effects.

Brief description of the drawings

Fig. 1 is the effect diagram of MEL represented by the survival rate (the absorbance of nail hairpin) in the rough skin model (made using a three-dimensional skin model and processed with SDS);

Fig. 2 is a graph showing the water recovery effect in the stratum corneum when a cream containing MEL-A is applied to the skin of the upper part of the human arm roughened by SDS treatment;

Figure 3 is the effect diagram of MEL-A, MEL(OL) and MEL(MY) expressed by the survival rate (absorbance of nail hairpin) in the rough skin model (made with a three-dimensional skin model and processed with SDS); MEL-A = MEL produced by incubation with soybean oil; MEL (MY) = MEL produced by incubation with methyl myristate; MEL-A (OL) = MEL produced by incubation with olive oil;

Figure 4 is the effect of MEL-B and MEL-C in the rough skin model (made using a three-dimensional skin model and processed with SDS) expressed by the survival rate (absorbance of nail hairpin); MEL-B is more able to Improvement of rough skin; in the figure, MEL-A(OL) is MEL-A prepared using olive oil as a raw material, MEL-B(OL) is MEL-B prepared using olive oil as a raw material, and MEL-A( SB) is MEL-A prepared using soybean oil as a raw material, MEL-B (SB) is MEL-B prepared using soybean oil as a raw material, and MEL-C (SB) is MEL-C prepared using soybean oil as a raw material;

Figure 5 is a graph showing the effect of moisture recovery in the stratum corneum when a cream containing MEL-B is applied to the upper skin of a human arm roughened by SDS treatment;

Fig. 6 is a graph showing the dispersion stability of MEL-B and MEL-C. In terms of water dispersion stability, MEL-B and MEL-C are superior to MEL-A; hardly any change in turbidity is observed, because a stable suspension state is maintained after 6 hours; and

Fig. 7 is a graph showing the results when the skin roughness test was performed on the MEL-B liposome aqueous solution and the MEL-B suspension using a three-dimensional skin model, and the effect was examined.

Best Mode for Carrying Out the Invention

"Biosurfactant" herein is a general name for substances produced by living organisms that have surface activity and emulsification, which not only exhibit excellent surface activity and high biodegradability, but also have various physiological effects , thus showing the possibility of behavior and function different from those of synthetic surfactants.

The "premix" refers to a substance in which a dispersant is added in addition to a functional substance, or a substance that is diluted with a solvent for ease of use in the production of cosmetics. The biosurfactant of the present invention can be provided in the form of a premix in which a dispersant and a solvent are mixed as a cosmetic or a cosmetic additive for rough skin improvement/skin care.

Ceramides are sphingolipids that occupy approximately 50% of the intercellular lipids in the stratum corneum. Ceramides obtained from bovine brains were often used previously, but due to the spread of mad cow disease, ceramides obtained from plants for cosmetics are required.

The ceramide-like effect is the effect of improving skin color reduction and cosmetic adaptability reduction in the main intercellular component ceramide in the stratum corneum of the skin. A biosurfactant alone has a ceramide-like skin roughness improvement/skin care effect; by combining with a ceramide, the effect of a biosurfactant can be enhanced.

One advantage of biosurfactants with ceramide-like action is that they can be easily produced on a large scale and used in the form of emulsifiers. Therefore, the biosurfactant of the present invention has more applications than existing ceramides.

For biosurfactants, trehalolipids, rhamnolipids, sophorolipids, surfactants, californic acid, latex, MEL, MML, etc. can be used, but it is preferred to use biosurfaces that form a lamellar structure active agent. Among these biosurfactants, MEL and MML are preferred, MEL-A, MEL-B or MEL-C is more preferred, and MEL-B or MEL-C is particularly preferred.

Sheet-forming biosurfactants include MEL and MML, especially MEL.

In one embodiment of the present invention, preferred MELs include three types of MEL-A represented by formula (I), MEL-B represented by formula (II) and MEL-C represented by formula (III); or use a combination of these. MEL-A represented by formula (I) and MEL-B represented by formula (II) are more preferable, and MEL-B represented by formula (II) is most preferable.

MML is represented by formula (IV) (in the formula, any one or both of the acetyl groups at the 4-position and 6-position in mannose may be substituted with a hydroxyl group).

R ¹ and R ² each have 1 to 19 carbon atoms, preferably 1 to 17, and more preferably 7 to 15 carbon atoms.

R ¹ and R ² may be the same or different, and include C ₁ to C ₁₉ alkyl, C ₂ to C ₁₉ alkenyl, C ₅ to C ₁₉ dienyl, and C ₈ to C ₁₉ trienyl.

In formulas (I), (II), (III) and (IV), preferred groups for R ¹ and R ² include C ₁ to C ₁₉ alkyl groups such as CH ₃ (CH ₂ ) ₆ , CH ₃ (CH ₂ ) ₈ , CH ₃ (CH ₂ ) ₁₀ CH ₃ (CH ₂ ) ₁₂ , CH ₃ (CH ₂ ) ₁₄ , CH ₃ (CH ₂ ) ₁₆ and CH ₃ (CH ₂ ) ₁₈ . MEL in formula (I), (II) or (III) and MML in formula (IV), wherein R ^{and R} are C _to C _alkyl , can be obtained as follows: starting materials such as saturated carboxylic acids or Unsaturated monocarboxylic acids such as formic acid, acetic acid, propionic acid, butyric acid, valeric acid, lauric acid, myristic acid, palmitic acid, stearic acid, oleic acid and palmitoleic acid or their esters (monoalkyl esters, mono , di-, triglycerides, or oils including these saturated fatty acids or unsaturated mono-fatty acids) were added to the culture medium. When an unsaturated carboxylic acid such as oleic acid or palmitoleic acid is used as a raw material, the unsaturated carboxylic acid or the product during its β-oxidation is introduced, whereby for R ^and R ² the alkyl group becomes the main part, Alkenyl groups become minor moieties. Likewise, MEL in formula (I), (II) or (III) and MML in formula (IV), wherein R ¹ and R ² are C ₂ to C ₁₉ alkenyl, C ₅ to C ₁₉ dienyl or C ₈ to C ₁₉ trienyl can be obtained as follows: raw materials such as linoleic acid, linolenic acid, arachidonic acid, EPA, DHA or their esters (monoalkyl esters, mono , di-, triglycerides, or oils including their highly unsaturated fatty acids) were added to the medium. For example, when linoleic acid is used as a raw material, for R ¹ and R ² , an alkenyl group becomes a major part and a dienyl group becomes a minor part. When linolenic acid is used as a raw material, for R ¹ and R ² , a dienyl group becomes a major part, and an alkenyl or trienyl group becomes a minor part.

These biosurfactants may be used alone, or two or more biosurfactants may be used in combination.

In formulas (I) to (IV), R ¹ and R ² may be the same or different, and represent a hydrogen atom; straight or branched C ₁ to C ₁₉ , preferably C ₁ to C ₁₇ , and more preferably C ₇ to C ₁₅ alkyl; straight or branched C ₂ to C ₁₉ , preferably C ₂ to C ₁₇ , and more preferably C ₇ to C ₁₅ alkenyl; straight or branched C ₅ to C ₁₉ , preferably C ₅ to C ₁₇ , and more preferably C ₇ to C ₁₅ dienyl; or linear or branched C ₈ to C ₁₉ , preferably C ₈ to C ₁₇ , and more preferably C ₈ to C ₁₅ trienyl.

There is no particular limitation on the method of preparing the biosurfactant, and a fermentation method using microorganisms may optionally be selected. For example, MELs (MEL-A, MEL-B and MEL-C) can be prepared by culturing Pseudozymaantarctica NBRC 10736 according to standard methods. As the microorganism, Pseudozyma antarctica, Pseudozyma sp. and the like can be used. It is well known that MEL mixtures can be readily obtained from any microorganism. The MEL mixture can be purified using silica gel chromatography to isolate MEL-A, MEL-B and MEL-C. Pseudozyma antarctica and Pseudozyma tsukubaensis are considered as MEL-B-producing microorganisms, and these microorganisms can be used. Pseudozymahubeiensis is considered to be a MEL-C producing microorganism, and this microorganism can be used. There are no particular limitations on the microorganisms having the ability to produce MEL, and they can be appropriately selected according to the purpose.

For the medium for fermentative production of biosurfactants, N sources (such as yeast extract and peptone), C sources (such as glucose and fructose), inorganic salts (such as sodium nitrate, dipotassium hydrogen phosphate and magnesium sulfate) can be used. A common medium consisting of a mixture of seven waters), and to which one or two more non-aqueous substrates such as oils such as olive oil, soybean oil, sunflower oil, corn oil, canola oil and coconut oil and hydrocarbons such as Those of liquid paraffin and tetradecane.

Fermentation conditions such as pH, temperature and time period can be optionally set, and the medium after fermentation can be directly used as the biosurfactant of the present invention. If necessary, after fermentation, optional operations such as filtration, centrifugation, extraction, purification, and sterilization may be applied to the medium, and the obtained product may also be diluted, concentrated, and dried to the extent that the essence of the present invention is not impaired. obtained extract.

Vegetable oils and fats used as raw materials are not particularly limited and may be appropriately selected according to purpose; these may include soybean oil, rapeseed oil, corn oil, peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil and palm oil. Among them, soybean oil is particularly preferable in terms of improving production efficiency (production amount, production speed and yield). These may be used alone or in combination of two or more.

The source of inorganic nitrogen is not particularly limited and may be appropriately selected according to the purpose; examples include ammonium nitrate, urea, sodium nitrate, ammonium chloride and ammonium sulfate.

The method of collecting and purifying the biosurfactant is not particularly limited, and may be appropriately selected according to the purpose; for example, the biosurfactant is collected by centrifuging the medium to collect an oil fraction, and extracting with an organic solvent such as ethyl acetate, and concentrating.

As the extraction solvent, water and organic solvents such as alcohols (such as lower alcohols such as methanol, absolute ethanol and ethanol or polyalcohols such as propylene glycol and 1,3-butanediol), ketones such as acetone, diethyl ether, dioxane can be used alone. , acetonitrile, esters such as ethyl acetate, xylene, benzene, and chloroform, or two or more mixed liquids are used in combination, and those obtained by combining solvent extracts can also be used.

There is no particular limitation on the extraction method. Usually, the extraction can be performed at room temperature to the boiling point of the solvent under normal pressure. After extraction, filtration or ion exchange resins can be used to absorb, decolorize or purify the extract to prepare solutions, ointments, gels or powders. In many cases, the obtained product can be used as it is, but if necessary, it can be subjected to purification treatment such as deodorization treatment or decolorization within the range not affecting the effect of the product. For deodorizing means and decolorizing means, an activated carbon column may be used, and ordinary means generally suitable for extracting substances may optionally be selected and performed. If desired, silica gel column purification can be used to obtain high-purity biosurfactants.

As the surfactant, MEL-A, MEL-B and MEL-C are preferred, MEL-B and MEL-C are more preferred, and MEL-B is particularly preferred.

The biosurfactant of the present invention used as a cosmetic for skin care/skin roughness improvement can be produced by microbial fermentation as described above.

The amount of biosurfactant added to cosmetics, which can vary depending on the type of cosmetics to which it is applied, cannot be uniformly determined, but can be within a range in which roughness-improving/skin-care effects are not impaired. In general, the preferred amount is 0.001 to 50% by mass, more preferably 0.1 to 20% by mass, still more preferably 1 to 15% by mass, particularly preferably 3 to 10% by mass, relative to various cosmetics. Herein, the use form of the biosurfactant added to the cosmetic is optional. For example, a biosurfactant extracted from a culture medium may be used directly, or a highly purified biosurfactant may be used, or a biosurfactant may be used after being suspended in water or dissolved in an organic solvent such as ethanol.

Biosurfactants can be incorporated into cosmetics by dissolving in oil-soluble bases or ingredients, and are preferably incorporated as liposomes in water-based cosmetics such as facial toners and moisturizers liquid. Combining the biosurfactant in liposome form is preferred because it fuses with skin cells, thereby increasing its rate of absorption. There is no particular limitation on the method of preparing liposomes; any known preparation method such as ethanol injection method or Bangham method can be used.

There is no particular limitation on the method of using the biosurfactant to prepare the cosmetic of the present invention, and the biosurfactant can be dissolved in nonionic surfactants, lower alcohols, polyhydric alcohols, or natural oils such as olive oil, squalane, fatty acid or higher alcohols.

Examples of nonionic surfactants include sorbitan fatty acid esters (e.g., sorbitan monooleate, sorbitan monoisostearate, sorbitan monolaurate, sorbitan Sugar alcohol monopalmitate, sorbitan monostearate, sorbitan sesquioleate, sorbitan trioleate, diglyceryl sorbitan penta-2-ethylhexanoate esters, diglyceride sorbitan tetra-2-ethylhexanoate); glycerol/polyglycerol fatty acid esters (eg, monocottonseed fatty acid glycerides, glyceryl monoerucate, glyceryl sesquioleate, Glyceryl monostearate, glyceryl α,α'-oleic acid pyroglutamate, glyceryl monostearate malate); propylene glycol fatty acid esters (for example, propylene glycol monostearate); hardened castor oil derived substances; and glycerol alkyl ethers.

Examples of POE-based hydrophilic nonionic surfactants include POE-sorbitan fatty acid esters (for example, POE-sorbitan monooleate, POE-sorbitan monostearate, POE-Sorbitan Monooleate, POE-Sorbitan Tetraoleate); POE-Sorbitan Fatty Acid Ester (for example, POE-Sorbitan Monolaurate, POE-Sorbitan Monolaurate Oleate, POE-Sorbitan Pentaoleate, POE-Sorbitan Monostearate); POE-Glyceryl Fatty Acid Ester (for example, POE-Glyceryl Monostearate, POE-Glyceryl Monoisostearate fatty acid esters, POE-glyceryl triisostearate, POE-monooleate); POE-fatty acid esters (eg, POE-distearate, POE-mono-dioleate, distearic acid glycol esters); POE-alkyl ethers (e.g., POE-lauryl ether, POE-oleyl ether, POE-stearyl ether, POE-behenyl ether, POE-2-octyldodecane ethers, POE-cholestanol ethers); block polyether types (e.g., Pluronic); POE/POP-alkyl ethers (e.g., POE/POP-cetyl ether, POE/POP-2-decyl myristyl ether, POE/POP-monobutyl ether, POE/POP-hydrogenated lanolin, POE/POP-glyceryl ether); tetra-POE/tetra-POP-ethylenediamine condensates (e.g. tetronic acid (Tetronic)); POE-castor oil cured castor oil derivatives (for example, POE-castor oil, POE-cured castor oil, POE-cured castor oil monoisostearate, POE-cured castor oil triisostearate esters, POE-cured castor oil pyroglutamate monoisostearate diester, POE-cured castor oil maleate); POE-beeswax lanolin derivatives (for example, POE-sorbitol beeswax); alkanes Alcoholamides (e.g., palm oil fatty acid diethanolamide, lauric acid monoethanolamide, fatty acid isopropanolamide); POE-propylene glycol fatty acid esters; POE-alkylamines; POE-fatty acid amides; fatty acid sucrose esters; Oxydimethylamine Oxide; and Trioleyl Phosphate.

Examples of lower alcohols include ethanol, propanol, isopropanol, isobutanol and t-butanol.

Examples of polyhydric alcohols include dihydric alcohols (e.g., ethylene glycol, propylene glycol, trimethylene glycol, 1,2-butanediol, 1,3-butanediol, tetramethylene glycol, 2,3- butanediol, pentamethylene glycol, 2-butylene-1,4-diol, hexanediol, octanediol); triols (e.g., glycerol, trimethylolpropane); tetraols (e.g., pentaerythritol such as 1,2,6-hexanetriol); pentahydric alcohols (e.g., xylitol); hexahydric alcohols (e.g., sorbitol, mannitol); polyol polymers (e.g., Ethylene glycol, dipropylene glycol, triethylene glycol, polypropylene glycol, tetraethylene glycol, diglycerol, polyethylene glycol, triglycerol, tetraglycerol, polyglycerol); glycol alkyl ethers (e.g., ethylene glycol Monomethyl ether, ethylene glycol monoethyl ether, ethylene glycol monobutyl ether, ethylene glycol monophenyl ether, ethylene glycol monohexyl ether, ethylene glycol mono-2-methylhexyl ether, ethylene glycol isoamyl ether, ethylene glycol benzyl ether, ethylene glycol isopropyl ether, ethylene glycol dimethyl ether, ethylene glycol diethyl ether, ethylene glycol dibutyl ether); glycol alkyl ethers (for example, diethylene glycol monomethyl ether) ether, diethylene glycol monoethyl ether, diethylene glycol monobutyl ether, diethylene glycol dimethyl ether, diethylene glycol diethyl ether, diethylene glycol butyl ether, diethylene glycol methyl ethyl ether, Triethylene glycol monomethyl ether, triethylene glycol monoethyl ether, propylene glycol monomethyl ether, propylene glycol monoethyl ether, propylene glycol monobutyl ether, propylene glycol isopropyl ether, dipropylene glycol methyl ether, dipropylene glycol ethyl ether, dipropylene glycol butyl ether); two Alcohol ether esters (for example, ethylene glycol monomethyl ether acetate, ethylene glycol monoethyl ether acetate, ethylene glycol monobutyl ether acetate, ethylene glycol monophenyl ether acetate, ethylene glycol di Adipate, Ethylene Glycol Disuccinate, Diethylene Glycol Monoethyl Ether Acetate, Diethylene Glycol Monobutyl Ether Acetate, Propylene Glycol Monomethyl Ether Acetate, Propylene Glycol Monoethyl Ether Acetate, Propylene Glycol Monopropyl Ether Acetate, Propylene Glycol Monophenyl Ether Acetate); Glycerol Monoalkyl Ethers (e.g., Chimeryl Alcohol, Squalyl Alcohol, Bathyl Alcohol); Sugars and Sugar Alcohols (e.g., Sorbitose alcohol, maltitol, maltotriose, mannitol, sucrose, erythritol, glucose, fructose, starch-degrading sugar, maltose, xylose, starch-degrading sugar reduced alcohol); glysolid; tetrahydrofurfuryl alcohol; POE- Tetrahydrofurfuryl alcohol; POP-butyl ether; POP/POE-butyl ether; tripolyoxypropylene glyceryl ether; POP-glyceryl ether; POP-glyceryl ether phosphate; POP/POE-pentaerythritol ether, and polyglycerol.

Examples of oils include animal and vegetable oils such as avocado oil, olive oil, sesame oil, camellia oil, evening primrose oil, turtle oil, macadamia nut oil, corn oil, mink oil, canola oil, egg yolk oil, almond oil ( parsic oil), wheat germ oil, camellia oil, castor oil, linseed oil, safflower oil, cottonseed oil, perilla seed oil, soybean oil, peanut oil, tea oil, kaya seed oil, rice bran oil, tung oil, jojoba Oil, cocoa butter, rectified coconut oil, horse oil, palm oil, coconut kernel oil, tallow, suet, lard, lanolin, spermaceti, beeswax, carnauba wax, vegetable wax, candelilla wax and horn Squalane, its solidified oils, mineral oils such as liquid paraffin and petrolatum, and synthetic triglycerides such as tripalmitin.

Examples of fatty acids include lauric acid, myristic acid, palmitic acid, oleic acid, linoleic acid, linolenic acid, stearic acid, behenic acid, 12-hydroxystearic acid, isostearic acid, undecylenic acid, Tolic acid, eicosapentaenoic acid and docosahexaenoic acid. Examples of higher alcohols include lauryl alcohol, cetyl alcohol, stearyl alcohol, behenyl alcohol, myristyl alcohol, oleyl alcohol, cetostearyl alcohol, jojoba alcohol, lanolin alcohol, batyl alcohol, 2 - Decyltetradecyl Alcohol, Cholesterol, Phytosterols and Isostearyl Alcohol. Examples of synthetic esters include cetyl caprylate, octyl dodecyl myristate, isopropyl myristate, myristyl myristate, isopropyl palmitate, butyl stearate, Hexyl Laurate, Decyl Oleate, Dimethyl Caprylate, Cetyl Lactate, and Myristyl Lactate. Examples of silicones include chain polysiloxanes such as dimethylpolysiloxane and methylphenylpolysiloxane, cyclic polysiloxanes such as decamethylcyclopolysiloxane, and silicone resins. Three-dimensional network structure.

The skin care cosmetics of the present invention include emulsions, cosmetic lotions, creams, lotions, skin care oils, cleansing oils, bath oils, or face washes, makeup removers, shampoos, and body soaps.

In terms of ease of use, the MELs of the present invention, particularly MEL-A, MEL-B and MEL-C, are superior to ceramides because they are easy to prepare and have an emulsifying effect.

Example

The present invention will be described in more detail in the following examples; however, the present invention is not limited to these examples.

(Evaluation method of rough skin improvement effect)

Tissues were collected according to the operation manual attached to the test skin (Test Skin) LSE-002 or 003 kit (Toyobo Co., Ltd.). A ring securing the exposure site of the agent was attached to the surface of the LSE tissue, 0.1% sodium dodecyl sulfate (SDS) aqueous solution was added to the ring, and it was left to stand at room temperature for 5 minutes. Subsequently, the SDS was removed using an aspirator, and the tissue was washed by spraying 3 ml of assay medium using a pipette. Through these operations, the moisturizing ingredients in the stratum corneum are dissolved, thereby making dry skin.

Subsequently, 80 µL each of purified water or cosmetic solution (Fenatty oil-based cosmetic solution; FANCL Corporation) as a test object was added to the surface of the LSE (living skin equivalent) tissue, which was then left to stand at room temperature for 60 minutes . Then, the test substance was drawn out and removed using an aspirator. Subsequently, the LSE tissue was placed on a test plate without test medium, and cultured for 24 hours in a CO _incubator adjusted to a temperature of 37 °C and a relative humidity of 15 to 20% RH. Then, remove the LSE tissue from the CO _incubator , and follow the essentials of the operation manual attached to the LSE-003 kit, add 1.2 ml of the mixed solution of the test medium containing 0.333 g/ml tetrazolium salt (MTT) reagent to the to the test disk. Subsequently, the LSE tissue in the test dish was incubated for 3 h in a CO _incubator adjusted to a temperature of 37 °C and a relative humidity of 15 to 20% RH.

After processing with MTT, use an 8mmφ biopsy punch (biopsy punch) to drill out the center of the LSE tissue including the polycarbonate film to obtain a piece of LSE tissue, which is then transferred to a small test tube, and inserted into the Add 700 μL of 0.04N hydrochloric acid-isopropanol and extract in the dark for two hours. After the extraction is terminated, the extract solution is stirred and mixed thoroughly. Subsequently, at 562 nm, the absorbance of the extracted blue-violet formazan was measured. The absorbance obtained by this method is closely related to the effect of improving rough skin. Thus, this method can be effectively used to evaluate the improvement of rough skin quantitatively, simply and economically.

Embodiment 1: the preparation of MEL

One loop (loop) of Pseudozyma antarctica NBRC 10736 was inoculated in a seed medium (20 ml/500 ml Sakaguchi flask) for inoculum culture. Overnight incubation was performed at 30°C. The obtained medium was used as an inoculum. The seed medium consisted of the following: 4% glucose, 0.3% NaNO ₃ , 0.02% MgSO ₄ ·H ₂ O, 0.02% KH ₂ PO ₄ and 0.1% yeast extract. The above inoculum (75 ml) was inoculated in 1.5L (5L-tank) production medium and cultured at 30°C, 300rpm (stirring frequency) and 0.5L/min0 (air) (using 5L-tank). The production medium consisted of 3% soybean oil, 0.02% MgSO ₄ ·H ₂ O, 0.02% KH ₂ PO ₄ and 0.1% yeast extract. The medium (250 ml) was centrifuged (6,500 rpm, 30 minutes), the supernatant was removed, and the precipitate (cells) was collected. Ethyl acetate (50 mL) was added to the precipitate, which was then thoroughly stirred and centrifuged (8,500 rpm, 30 minutes) to separate the supernatant from the precipitate. Concentrate the supernatant using an evaporator. By using silica gel and eluting with hexane:acetone=5:1 and hexane:acetone=1:2, MEL fractions (MEL-A, MEL-B and MEL-C) were obtained.

Example 1A: Preparation of MEL-B

In a 500-ml Sakaguchi flask, frozen-stored P. tsukubaensis (0.2 ml) was inoculated in 20 ml of YM seed medium and cultured overnight at 26°C and 180 rpm to prepare seed inoculum. In a 500 ml Sakaguchi flask, the seed inoculum was inoculated again in 20 ml of YM seed medium and cultured overnight at 26°C, 180 rpm, as an inoculum. Inoculum (20 ml) was inoculated in 2 L of YM medium in a 5 L tank and cultured at 26° C. at 300 rpm (1/4 VVM, 0.5 L air/min) for 8 days. The medium was centrifuged at 7,900 rpm at 4° C. for 60 minutes to separate bacterial cells (including MEL-B) from the supernatant. Ethyl acetate (80 ml) was added to the cell fraction, which was then shaken until thoroughly suspended, followed by centrifugation at 7,900 rpm at 4°C for 30 minutes. An equal amount of brine was added to the obtained supernatant, and the mixture was stirred to yield an ethyl acetate layer. An appropriate amount of anhydrous sodium sulfate was added to the ethyl acetate layer, which was then left to stand for 30 minutes and evaporated to obtain crude purified MEL-B. The obtained crude purified MEL-B (20 g) was further purified using a silica gel column (200 g) and eluted with hexane/acetone to obtain the purified MEL-B product.

Example 2

Although soybean oil is used as a raw material in the preparation of MEL in Example 1, olive oil is used instead as a raw material for production in this example to separate and purify MEL-A, MEL-B and MEL-C, and use and Example 1 Cultivate in the same way. The MEL fractions obtained at this time were referred to as MEL-A(OL), MEL-B(OL) and MEL-C(OL) in order to distinguish them from the MEL obtained in Example 1.

Example 3

Although soybean oil was used as a production raw material in the preparation of MEL in Example 1, methyl myristate was used instead as a production raw material to separate and purify MEL-A, MEL-B and MEL-C in this example, and Cultivate in the same manner as in Example 1. The MEL fractions obtained at this time were referred to as MEL-A(MY), MEL-B(MY) and MEL-C(MY) in order to distinguish them from the MEL obtained in Example 1.

Example 4: Evaluation of MEL-A in the Rough Skin Model

A rough skin model using a three-dimensional skin model is produced as follows. A rough skin model was prepared by treating test skin (LSE-002 or 003; Toyobo Co., Ltd.) with 1% SDS to remove lipid components in the stratum corneum. The skin roughness preventive effect was examined as follows: MEL-A-dissolved olive oil was added to the cells, which were left overnight, and then the cell viability was calculated using a commercially available MTT kit. As shown in Figure 1, MEL-A increased cell viability in a concentration-dependent manner, demonstrating that MEL-A functions as a substitute for ceramide. Meanwhile, olive oil alone showed no such effect.

Example 5: Evaluation of MEL-B in a rough skin model

A rough skin model using a three-dimensional skin model is produced as follows. A rough skin model was prepared by treating test skin (LSE-002 or 003; Toyobo Co., Ltd.) with 1% SDS to remove lipid components in the stratum corneum. The skin roughness preventive effect was examined as follows: olive oil in which MEL-B obtained in Example 1A was dissolved was added to the cells, left overnight, and then cell viability was calculated using a commercially available MTT kit. As shown in Figure 4, MEL-B increased cell viability in a concentration-dependent manner, demonstrating that MEL-B functions as a substitute for ceramide. Meanwhile, olive oil alone showed no such effect.

Example 6: Evaluation of MEL-C in a rough skin model

A rough skin model using a three-dimensional skin model is produced as follows. A rough skin model was prepared by treating test skin (LSE-002 or 003, supplied by Toyobo Co., Ltd.) with 1% SDS to remove lipid components in the stratum corneum. The skin roughness preventive effect was examined as follows: olive oil in which MEL-C obtained in Example 1 was dissolved was added to the cells, left overnight, and then cell viability was calculated using a commercially available MTT kit. As shown in Figure 4, MEL-C increased cell viability in a concentration-dependent manner, demonstrating that MEL-C functions as a substitute for ceramide. Meanwhile, olive oil alone showed no such effect.

Example 7: Evaluation of MEL-A(OL) and MEL-A(MY) in rough skin model

Similar to Example 4, a skin roughness model using a three-dimensional skin model was produced as follows. A rough skin model was prepared by treating test skin (LSE-002 or 003, supplied by Toyobo Co., Ltd.) with 1% SDS to remove lipid components in the stratum corneum. The skin roughness preventive effect was examined as follows: olive oil in which MEL-A (Example 1), MEL-A(OL) (Example 2) or MEL-A(MY) (Example 3) was dissolved was added to the cells, This was left overnight and then cell viability was calculated using a commercially available MTT kit. As shown in Figure 3, MEL-A increased cell viability in a concentration-dependent manner, demonstrating that MEL-A functions as a substitute for ceramide. Meanwhile, olive oil alone showed no such effect.

Example 8: Effect of a cream containing MEL-A in the human skin roughness test

The skin was roughened by contacting the inner side of the human upper arm with a 1% SDS solution for 10 min. Immediately thereafter, the above-mentioned cream containing 5% MEL-A was applied, and after 3 hours, the skin was washed with warm water. Oil was wiped off with a Kim towel, and the moisture content of the stratum corneum of the skin was measured using a Skicon. As shown in Figure 2, recovery of moisture content was observed when the cream containing MEL-A was applied.

Example 9: Efficacy of MEL-B-containing creams in the human skin roughness test

The skin was roughened by contacting the inner side of the human upper arm with a 1% SDS solution for 10 min. Immediately thereafter the above cream containing 5% MEL was applied and after 3 hours the skin was washed with warm water. Oil was wiped off with a Kim towel, and the moisture content of the stratum corneum of the skin was measured using a Skicon. As shown in Figure 5, recovery of moisture content was observed when creams containing MEL-B or C were applied.

Embodiment 10: Dispersion stability test of MEL-B and MEL-C

Add MEL-B or MEL-C to water at a concentration of 10 mg/ml, stir and suspend. Absorbance was measured at 650 nm. The results are shown in FIG. 6 . As is apparent from the results in Fig. 6, MEL-B and MEL-C are particularly excellent in dispersion stability.

Embodiment 11: Preparation of MEL-B liposome solution

Using the ethanol injection method, a MEL-B liposome solution was prepared as follows. Dissolve 10 mg of MEL-B in 0.5 ml of ethanol, and add 1 ml of distilled water prewarmed to about 70°C. The mixture was shaken and mixed briefly, and residual ethanol was distilled off using a rotary evaporator. Using a water bath type sonicator (W-220R; Honda Electronics Co., Ltd.), ultrasonication was applied thereto for about 5 minutes, and then distilled water was added to make the total volume 1 ml.

MEL-B liposome solutions were prepared using the Bangham method as follows. Dissolve 10 mg of MEL-B in 1 mL of chloroform, and distill off the solvent using a rotary evaporator to prepare a thin film. 1 ml of distilled water was added thereto, and ultrasonic treatment was applied thereto for about 5 minutes using a water bath type sonicator (W-220R; Honda Electronics Co., Ltd.).

A MEL-B suspension was prepared as follows. Add 1 mL of distilled water to 10 mg of MEL-B and stir using a vortex mixer to prepare a suspension.

Using the three-dimensional skin model shown in Example 4, the skin roughness test was carried out on the MEL-B liposome aqueous solution and the MEL-B suspension prepared by the ethanol injection method, the Bangham (Bangham) method, and their effects were checked . The results are shown in FIG. 7 . The results showed that the MEL-B liposome aqueous solution and suspension prepared by any one of the ethanol injection method and the Bangham (Bangham) method showed a similarity with the MEL-B olive oil solution (MEL-B concentration: 1%) ) same rough skin improvement effect.

Example 12: Preparation of cosmetic liquid

A beauty serum with the composition shown below was prepared using standard methods. As a control, a beauty serum without MEL was also prepared using standard methods.

(composition) (weight%)

Sorbitol 4.0

Dipropylene glycol 6.0

Polyethylene glycol 1500 5.0

POE(20) oleyl ether 0.5

Fatty acid sucrose ester 0.2

Methylcellulose 0.2

MEL-B 5.0

Purify water to 100% of the total

Example 13: Preparation of emulsion

Emulsions having the compositions shown below were prepared using standard methods. As a control, emulsions without MEL were also prepared using standard methods.

(composition) (weight%)

Glyceryl ether 1.5

Fatty acid sucrose esters 1.5

Sorbitan monostearate 1.0

Squalane 7.5

Dipropylene glycol 5.0

MEL-B 5.0

Purify water to 100% of the total

Example 14: Preparation of cream

A cream having the composition shown below was prepared using standard methods. As a control, a cream without MEL was also prepared using standard methods.

(composition) (weight%)

Propylene Glycol 6.0

Dibutyl phthalate 19.0

Stearic acid 5.0

Glyceryl monostearate 5.0

Sorbitan monostearate 12.0

Polyethylene sorbitan monostearate 38.0

Sodium edetate 0.03

MEL-B 5.0

Purify water to 100% of the total

Example 15: Preparation of Cosmetic Oil

(composition) (weight%)

olive oil 90

MEL-A 10

Example 16: Skin care oil

(composition) (weight%)

olive oil 50

MEL-C 30

Squalane 10

sesame oil 10

Example 17: Skin care oil

(composition) (weight%)

olive oil 39

MEL-C 59

sesame oil 1

Lavender Oil 0.4

Rosemary oil 0.4

Sage Oil 0.1

δ-tocopherol 0.1

Example 18: Cleansing oil

(composition) (weight%)

olive oil 40

MEL-B 28

Methylphenyl polysiloxane 2

Ethanol 0.3

Isostearic acid 0.1

Cetyl 2-ethylhexanoate 20

Polyethylene glycol diisostearate 2

Palm oil fatty acid diethanolamide 0.1

Polyethylene glycol monoisostearate 2

δ-tocopherol 0.1

Purified water 1

Aromatics Appropriate amount

Example 19: Bath Oil

(composition) (weight%)

olive oil 25

MEL-A 25

Liquid paraffin 25

Neopentyl glycol dicaprylate 10

Polyoxyethylene oleyl ether 10

Purified water 0.5

δ-tocopherol 0.1

Aromatics Appropriate amount

Industrial Applicability

According to the present invention, skin roughness can be prevented by supplementing the lost ceramide with MEL-A, MEL-B or MEL-C of the present invention, which is used not only as an emulsifier, but also They can also be used as a substitute for ceramides, and since they are easier to prepare than ceramides, they are expected to contribute greatly to the industry.

Claims (9)

Claims 1. A skin care cosmetic comprising at least one biosurfactant. 2. The skin care cosmetic according to claim 1, which is used for improving rough skin. 3. The skin care cosmetic according to claim 1, which is used for improving skin roughness by the action of a surfactant. 4. The skin care cosmetic according to claim 1, wherein the biosurfactant is mannose erythritol lipid (MEL) and/or mannose mannitol lipid (MML). 5. The skin care cosmetic according to claim 1, wherein said biosurfactant is selected from the group consisting of mannose erythritol lipid A (MEL-A), mannose erythritol lipid B (MEL-B) and At least one of mannose erythritol lipid C (MEL-C). 6. The skin care cosmetic according to claim 1, wherein said biosurfactant is Mannose Erythritol Lipid B (MEL-B). 7. The skin care cosmetic according to claim 1, wherein said biosurfactant is Mannose Erythritol Lipid C (MEL-C). 8. The skin care cosmetic according to claim 1, wherein said biosurfactant is mannose erythritol lipid A (MEL-A). 9. A rough skin improving agent consisting of at least one biosurfactant.

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