JP2018043946A - Melanogenesis inhibitor - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a skin-whitening agent having excellent skin-whitening effect, and a melanogenesis inhibitor having significant melanogenesis inhibitory effect.SOLUTION: The present invention provides a melanogenesis inhibitor containing sulfoquinovosylglycerol (SQG) or a salt thereof, a skin-whitening agent containing SQG or a salt thereof, and the like.SELECTED DRAWING: None

Description

  The present invention relates to a melanin production inhibitor and a whitening cosmetic. More specifically, the present invention relates to a melanin production inhibitor or a whitening cosmetic composition containing sulfoquinovosylglycerol or a salt thereof (also referred to herein as SQG or a salt thereof) as an active ingredient, and SQG or a salt thereof. The present invention relates to a method for whitening the skin used.

  Pigmentation due to sunburn, stains, freckles, liver spots, senile pigment spots, etc. are caused by increased production of black melanin by melanocytes and production of excess black melanin. As a whitening agent for suppressing the production of black melanin, a whitening agent containing a whitening component such as kojic acid, L-ascorbic acid, α-glucosylglycerol (α-D-glucopyranosylglycerol) has been proposed. Patent Document 1 describes a whitening agent containing α-D-glucopyranosylglycerol, which is a sugar glycerol, as an active ingredient. However, existing whitening agents may have problems in safety and the like, and development of whitening agents with high safety and whitening effect is required.

Chloroplast thylakoid membranes and membrane components of microalgae are known to contain a large amount of glycolipids such as galactosyl diacylglycerol, sulfoquinovosyl diacylglycerol, and digalactosyl diacylglycerol. Describes a method for producing sugar glycerol, comprising subjecting microalgae to medium temperature treatment and / or organic solvent treatment.
However, it has not been known so far that the microalgal membrane extract obtained by these methods and sulfoquinovosyl diacylglycerol have a melanin production inhibitory action.

International Publication No. 2004/331581 International Publication No. 2014/192940 International Publication No.2015 / 064648

  An object of the present invention is to provide a melanin production inhibitor having a significant melanin production inhibitory effect, and to provide a whitening cosmetic material having an excellent whitening effect.

  As a result of intensive studies to achieve the above object, the present inventors have found that an extract obtained by subjecting microalgae to intermediate temperature treatment has an excellent melanin production inhibitory effect without exhibiting cytotoxicity. I found out. As a result of analyzing the fraction showing the melanin production inhibitory effect, the present inventors found the melanin production inhibitory effect superior to sulfoquinovosylglycerol (SQG) and completed the present invention.

That is, the present invention is as follows.
[1] A melanin production inhibitor comprising sulfoquinovosylglycerol or a salt thereof.
[2] A melanin production inhibitor comprising a microalgal membrane extract.
[3] The agent according to [2], wherein the microalgal membrane extract contains sulfoquinovosylglycerol or a salt thereof.
[4] The microalgae membrane extract is a supernatant extract after intermediate temperature treatment obtained by subjecting microalgae to intermediate temperature treatment, followed by separation operation of a water-soluble fraction and a water-insoluble fraction. [3] The agent according to [3].
[5] The agent according to any one of [2] to [4], wherein the microalga is a microalga belonging to the green alga class.
[6] The agent according to [5], wherein the microalgae belong to the genus Desmodemus.
[7] The agent according to any one of [1] to [6], which is for skin whitening.
[8] A whitening cosmetic comprising sulfoquinovosylglycerol or a salt thereof.
[9] A whitening cosmetic comprising a microalgal membrane extract.
[10] The cosmetic according to [9], wherein the microalgal membrane extract contains sulfoquinovosylglycerol or a salt thereof.
[11] The microalgae membrane extract is a supernatant extract after intermediate temperature treatment obtained by subjecting microalgae to intermediate temperature treatment, followed by separation operation of a water-soluble fraction and a water-insoluble fraction. [9] The cosmetic according to [9].
[12] The cosmetic according to any one of [9] to [11], wherein the microalga is a microalga belonging to the green alga class.
[13] The cosmetic according to [12], wherein the microalgae belong to the genus Desmodemus.
[14] The cosmetic according to any one of [8] to [13], which is an external preparation for skin.

  Since SQG or a salt thereof can significantly suppress melanin production, exhibits high melanin production inhibitory activity such as melanin production inhibitory activity even at low concentrations, and exhibits little cytotoxicity, the present invention Thus, it is possible to provide a highly safe and excellent melanin production inhibitor. A melanin production inhibitor containing SQG or a salt thereof as an active ingredient can significantly inhibit excessive melanin production in the skin, thus preventing pigmentation due to sunburn, spots, freckles, dull skin, liver spots, etc. And / or can be used as an effective skin whitening agent, and is useful in the production of skin whitening cosmetics. Furthermore, since the agent of the present invention has high safety, it can be blended into a food / beverage product composition or a pharmaceutical composition and blended into various compositions having a whitening effect / melanin production inhibitory effect. In the present invention, SQG or a salt thereof can be provided by a microalgal membrane extract.

FIG. 1 is a diagram showing the results of high performance liquid chromatography (HPLC) analysis of a medium temperature treated supernatant (2). FIG. 2 is a graph showing the effect of kojic acid, medium temperature treated supernatant (1) or medium temperature treated supernatant (2) on the melanin production rate and survival rate of melanocytes. FIG. 3 is a graph showing the effect of kojic acid, sulfoquinovosylglycerol (SQG) or glucosylglycerol (GG) on the melanin production rate and survival rate of melanocytes.

  The present invention will be described below. The terms used in this specification have meanings commonly used in the art unless otherwise specified.

1. Melanin production inhibitor comprising sulfoquinovosylglycerol (SQG) or a salt thereof The present invention comprises a melanin production inhibitor comprising SQG or a salt thereof as an active ingredient (also referred to herein as the agent of the present invention). I will provide a.
Since SQG or a salt thereof has an action of suppressing the production of excessive melanin by skin melanocytes, the agent of the present invention can prevent pigmentation due to sunburn or the like, blotches, freckles, dull skin, liver spots, etc. Alternatively, it can be used as a skin lightening agent effective for improvement. Therefore, in one embodiment of the present invention, the agent of the present invention is a whitening agent for whitening the skin, and can be used as a whitening cosmetic by mixing the agent of the present invention and various components for cosmetics. .
The skin whitening effect achieved by the present invention includes not only an active whitening effect that whitens the skin color, but also a passive whitening effect that suppresses blackening of the skin color. This includes not only a whitening effect that improves pigmentation such as buckwheat but also a whitening effect that suppresses pigmentation.

The agent of the present invention can prevent and ameliorate local pigmentation such as stains, buckwheat, sunburn, etc. In addition to these uses, for example, systemic pigmentation such as Addison's disease and drugs It can be used for all purposes that are meaningful in exhibiting a melanin production inhibitory action or a whitening effect, such as prevention, improvement and treatment of induced pigment increase.
The agent of the present invention can also be used as a reagent for research on melanin, melanocytes, and diseases related to melanin (for example, melanoma).

  In this specification, melanin refers to black melanin (also called true melanin or eumelanin).

  In the present specification, “whitening” refers to suppressing the increase of melanocytes, suppressing the production of melanocytes by melanocytes, preventing excessive deposition of melanin on the skin, thinning already deposited melanin pigments Say one or more functions such as

  The agent of the present invention can be applied to any animal as long as the desired effect is exhibited, but is usually for human use.

  The agent of the present invention contains SQG or a salt thereof as an active ingredient. SQG or a salt thereof is present in the agent of the present invention in an effective amount for achieving desired effects such as a melanin production inhibitory effect and a whitening effect.

(1) Sulfoquinobosylglycerol Sulfoquinobosylglycerol (SQG) refers to a compound in which sulfoquinoboses are bonded to the hydroxyl group of glycerol. The SQG used in the present invention may be one in which sulfoquinose is bonded to the hydroxyl group of the 1st carbon of glycerol, or may be one in which sulfoquinovoose is bonded to the hydroxyl group of the 2nd carbon, Or a mixture thereof may be used. Preferably, the SQG used in the present invention is one in which sulfoquinose is bonded to the hydroxyl group of the 1-position carbon of glycerol. The structure of SQG in which sulfoquinose is bonded to the hydroxyl group of the 1-position carbon of glycerol is illustrated in the following formula (I).

The structure of SQG in which sulfoquinobose is bonded to the hydroxyl group of the 2-position carbon of glycerol is exemplified in the following formula (II).

  The SQG used in the present invention may be an isomer of SQG exemplified above, or may be a hydrate or a solvate.

SQG salts include salts with inorganic acids (eg, hydrochloride, hydrobromide, sulfate, nitrate, phosphate, etc.); salts with organic acids (eg, formate, acetate, trifluoro) Acetate, maleate, tartrate, citrate, fumarate, methanesulfonate, benzenesulfonate, p-toluenesulfonate, etc .; acidic or basic amino acids (eg aspartic acid, glutamic acid, Salts with arginine, lysine, ornithine, etc .; salts with inorganic bases [for example, salts with metals (alkali metals such as potassium and sodium; alkaline earth metals such as calcium and magnesium; aluminum), ammonium salts, etc.]; Organic bases (eg trimethylamine, triethylamine, pyridine, picoline, ethanolamine, diethanolamine, triethanolamine) , Dicyclohexylamine, N, N′-dibenzylethylenediamine, methylamine, diethylamine, ethylenediamine, tris (hydroxymethyl) methylamine, guanidine, choline, cinchonine, meglumine, etc.), and the like. As long as is obtained, it is not particularly limited. The SQG salt is preferably suitable for the desired application. For example, when intended for use in cosmetics, the SQG salt is preferably a cosmetically acceptable salt, and is suitable for the skin. For the purpose of application, the salt of SQG is preferably a dermatologically acceptable salt, and when used as a pharmaceutical, it is preferably a pharmaceutically acceptable salt.
Each salt of SQG can be obtained by reacting SQG with an inorganic base or an organic base according to a method known per se.

SQG or a salt thereof can be produced by a method known per se. For example, SQG can be produced by enzymatically or chemically linking sulfoquinose to a hydroxyl group of glycerol. Alternatively, the acyl group can be produced by enzymatically or chemically cleaving an acyl group from, but not limited to, an acylated sulfoquinovosylglycerol. For example, SQG can be produced using a method according to the method described in Reference Example 1 of International Publication No. 2014/192940, or the method for producing sugar glycerol described in International Publication No. 2014/192940 is used. However, the manufacturing method is not limited to these. SQG or a salt thereof can be purified using techniques such as extraction, distillation, crystallization, column chromatography and the like, which are usually used in organic synthesis.
SQG or a salt thereof contained in the agent of the present invention can be provided by a microalgal membrane extract described later. Therefore, as one aspect, the present invention provides a melanin production inhibitor containing a microalgal membrane extract.

(2) Formulation of a melanin production inhibitor containing SQG or a salt thereof as an active ingredient The agent of the present invention comprises a carrier suitable for a desired use (for example, a pharmaceutically acceptable carrier, a cosmetically acceptable carrier, It can be formulated and provided in an arbitrary dosage form according to a conventional method using a dermatologically acceptable carrier, etc., and other compositions (for example, cosmetics, food and beverage compositions or pharmaceutical compositions) Etc.) and can be used. As long as the desired effect is not impaired, the agent of the present invention can contain various components that can be usually blended in various compositions in addition to SQG, which is an active ingredient, or a salt thereof, depending on the application. Alternatively, the salt itself (or the microalgal membrane extract of the present invention alone) may be used as the agent of the present invention.

  The content of SQG or a salt thereof in the agent of the present invention is, for example, 0.00001 to 100% by weight, preferably 0.0001 to 100% by weight, more preferably 0.001 to 100% by weight, still more preferably 0.01 to 100% by weight, and even more. Preferably it is 0.1 to 100% by weight, even more preferably 1.0 to 100% by weight.

  The agent of the present invention may be in any dosage form as long as it is suitable for the desired application, and takes any form such as liquid, paste, gel, solid, powder, foam, jelly, etc. be able to. When the dosage form of the agent of the present invention is solid or powder, it may be dissolved and used at a desired concentration using an appropriate solvent or the like. When the dosage form of the agent of the present invention is a solution, it may be used by diluting in an appropriate solvent so as to obtain a desired concentration. When the dosage form of the agent of the present invention is a solution, the pH of the solution is preferably about 4 to about 10, more preferably about 5 to about 9, but is not limited thereto.

The agent of the present invention can be mixed with various components that can be used in ordinary whitening cosmetics to obtain the whitening cosmetics of the present invention.
In addition, the agent of the present invention includes excipients, moisture-proofing agents, preservatives, reinforcing agents, thickeners, emulsifiers, antioxidants, sweeteners, acidulants, seasonings, used in ordinary foods and beverages and pharmaceuticals. Various components suitable for a desired use such as a coloring agent and a fragrance can be appropriately blended and provided as a food or drink or a medicine.

2. A whitening cosmetic comprising sulfoquinovosylglycerol (SQG) or a salt thereof The present invention further comprises a whitening cosmetic comprising SQG or a salt thereof as an active ingredient (herein also referred to as the whitening cosmetic of the present invention). Provide). In one aspect, the agent of the present invention can be mixed with various components that can be used in normal cosmetics (including quasi-drugs) to make the whitening cosmetic of the present invention.
SQG or a salt thereof contained in the whitening cosmetic composition of the present invention can be provided by a microalgal membrane extract described later. Therefore, as one aspect, the present invention provides a whitening cosmetic containing a microalgal membrane extract.

  The amount of SQG or a salt thereof in the whitening cosmetic composition of the present invention is usually 0.000001% by weight or more, preferably 0.00001% by weight or more, more preferably 0.0001% by weight, although it is not limited as long as the desired effect is obtained. % Or more, more preferably 0.001% by weight or more, preferably 0.01% by weight or more, more preferably 0.1% by weight or more, still more preferably 1.0% by weight or more, and even more preferably 2.0% by weight from the viewpoint that an effective effect can be exhibited. The amount of SQG or a salt thereof in the whitening cosmetic composition of the present invention is usually 99 from the viewpoint that the effect reaches its peak even if it is too much, and there is no room for adding other ingredients. % By weight or less, preferably 50.0% by weight or less, more preferably 10.0% by weight or less. In the whitening cosmetic composition of the present invention, SQG or a salt thereof is usually 0.000001-30.0 wt%, preferably 0.00001-30.0 wt%, more preferably 0.0001-30.0 wt%, still more preferably 0.001-30.0 wt%, and even more. Preferably 0.01 to 20.0% by weight, and still more preferably 0.1 to 10.0% by weight.

  In the present invention, cosmetics are a concept including quasi-drugs. There is no restriction | limiting in particular in the form, Arbitrary forms, such as liquid form, paste form, gel form, solid form, powder form, foam form, and jelly form, can be taken. Specifically, for example, solution, suspension, lotion, cream, gel, stick, pencil, spray, ointment, mousse, patch, lotion, emulsion, serum, shampoo, hair rinse, hair conditioner, hair mousse, hair gel, Hair wax, foundation, eyeliner, eyebrow pencil, mascara, eye shadow, teak, lipstick, lipstick, lustrous, powder, pack, pack mask, perfume, eau de cologne, facial cleanser, facial cleanser, cleansing foam, cleansing oil, cleansing gel , Cleansing milk, gel, soap, aerosol, bath preparation, hair nourishing agent, sunscreen agent (sunscreen), wound dressing, adhesive bandage and the like.

  In addition to the above SQG or a salt thereof, the cosmetic for whitening of the present invention usually contains various cosmetically or dermatologically acceptable components that can be used in cosmetics (including quasi drugs). Depending on the desired application, it can be blended within a range that does not impair the effects of the present invention. For example, oil components, surfactants, amino acids, amino acid derivatives, lower alcohols, higher alcohols, polyhydric alcohols, sugar alcohols and their alkylene oxide adducts, water-soluble polymers, gelling agents, moisturizers, bactericides, and Antibacterial agent, anti-inflammatory agent, analgesic agent, antifungal agent, keratin softening release agent, skin colorant, hormone agent, UV absorber, antiperspirant and astringent active ingredient, sweat deodorant, vitamin agent, blood flow promoter ( Vasodilator, blood circulation promoter), crude drug, pH adjuster, sequestering agent, viscosity modifier, pearlizing agent, natural fragrance, synthetic fragrance, dye, pigment, antioxidant, preservative, emulsifier, fat and wax , Silicone compounds, perfume oils and the like.

  Examples of the oil component include saturated or unsaturated such as heptanoic acid, octanoic acid, nonanoic acid, decanoic acid, pentadecanoic acid, heptadecanoic acid, nonadecanoic acid, lauric acid, myristic acid, palmitic acid, stearic acid, arachidonic acid, and behenic acid. Saturated fatty acids; higher alcohols obtained from the saturated or unsaturated fatty acids; octyldodecyl lactate, octyldodecyl ricinoleate, myristyl myristate, myristyl palmitate, myristyl stearate, myristyl isostearate, myristyl oleate, myristyl behenate, Elca Myristyl acid, cetyl ethylhexanoate, cetyl caprate, cetyl myristate, cetyl palmitate, cetyl stearate, cetyl isostearate, cetyl oleate, cetyl behenate, cetyl erucate, Stearyl listinate, stearyl palmitate, stearyl stearate, stearyl hydroxystearate, stearyl isostearate, stearyl oleate, stearyl behenate, stearyl erucate, isostearyl myristate, isostearyl palmitate, isostearyl stearate, hydroxystearic acid Isostearyl acid, isostearyl isostearate, isostearyl oleate, isostearyl behenate, isostearyl myrate, behenyl myristate, behenyl palmitate, behenyl stearate, behenyl isostearate, behenyl oleate, behenyl behenate, erucic acid Behenyl, erucyl myristate, erucyl palmitate, erucyl stearate, erucyl isostearate, o Esters of linear or branched fatty alcohols such as erucyl inmate, erucyl behenate, erucyl erucate, cholesteryl nonanoate, cetearyl nonanoate; lauroyl glutamate di (cholesteryl / behenyl / octyldodecyl), lauroyl glutamate di (cholesteryl / Esters of amino acids such as octyldodecyl), dilauroyl glutamate (phytosteryl / octyldodecyl), dilauroyl glutamate (octyldodecyl / phytosteryl / behenyl), lauroyl sarcosine isopropyl, myristoylmethyl β-alanine (phytosteryl / decyltetradecyl) Manufactured by Ajinomoto Co., Inc., “El Dew” PS-203, PS-304, PS-306, APS-307, SL-205); dimer linoleic acid (phyto) Teryl / isostearyl / cetyl / stearyl / behenyl), dimer dilinoleate dimer dilinoleyl bis (behenyl / isostearyl / phytosteryl), dimer dilinoleyl hydrogenated rosin condensate, diisostearate dimer dilinoleyl, bis (lauroyl) Esters of dimer acids such as glutamic acid / lauroyl sarcosine) dimer linoleyl; squalane, squalene, castor oil and / or hydrogenated castor oil and derivatives thereof; hydroxystearic acid monoglyceride, hydroxystearic acid diglyceride, isostearic acid monoglyceride, isostearic acid diglyceride Oleic acid monoglyceride, oleic acid diglyceride, ricinoleic acid monoglyceride, ricinoleic acid diglyceride, linoleic acid monoglyceride, reno Glycerides such as diacid glyceride, linolenic acid monoglyceride, linolenic acid diglyceride, tartaric acid monoglyceride, tartaric acid diglyceride, citric acid monoglyceride, citric acid diglyceride, malic acid diglyceride, malic acid diglyceride and ethylene oxide 1-30 mol adducts of these glycerides; Beeswax, lanolins and their derivatives including liquid and refined lanolin; shea, linseed, camellia, almond, avocado, olive, rapeseed, safflower, soy, peanut, coconut, macadamia nut, jojoba Oil, cinnamon oil, grape oil, corn oil, sunflower oil, sasanqua oil, rice bran oil, rice germ oil, wheat germ oil, kyounin oil, tea seed oil, carnauba wax, sesame oil, cottonseed oil, cacao oil, rapeseed oil, evening primrose Oily raw materials derived from animals and plants such as oil, palm oil, palm oil, palm kernel oil, hydrogenated palm oil, mink oil, beef tallow, lard, wood wax, candelilla wax, whale wax; paraffin, isoparaffin, octane, decane, dodecane , Isododecane, hexadecane, isohexadecane, microcrystalline wax, liquid paraffin, petroleum jelly, ceresin and other oil-based raw materials; methylpolysiloxane, dimethylpolysiloxane, cyclomethicone, polyoxyethylene / methylpolysiloxane, polyoxypropylene・ Methyl polyoxysiloxane, poly (oxyethylene, oxypropylene), methyl polysiloxane, methylphenyl polysiloxane, fatty acid-modified polysiloxane, aliphatic alcohol-modified polysiloxane, amino acid-modified polysiloxane Silicones such silicone polymers such as; resin acids, fatty esters, and ketones.

Examples of the surfactant include an anionic surfactant, a nonionic surfactant, a cationic surfactant, and an amphoteric surfactant. Examples of the anionic surfactant include N-long chain acyl acidic amino acid salts (eg, N-long chain acyl glutamate, N-long chain acyl aspartate), N-long chain acyl neutral amino acid salts (eg, N-long chain acyl glycine salts, N-long chain acyl alanine salts, N-long chain acyl threonine salts, N-long chain acyl sarcosine salts, etc.) (for example, manufactured by Ajinomoto Co., Inc.) “Amisoft” CT-12, LT-12, CK-22, CS-22, CS-11, CK-11, LS-11, LK-11, MS-11, MK-11, GS-11, HS-11 CA, LA-D, “amino soap” AR-12, “amylite” GCK-12, GCK-11, GCS-11, ACT-12), N-long chain fatty acid acyl-N-methyltaurine salt, alkylsa Fate and its alkylene oxide adduct, fatty acid amide ether sulfate, fatty acid metal salt, weak base and amino acid salt, sulfosuccinic acid type surfactant, alkyl phosphate and its alkylene oxide adduct, higher alkyl sulfate ester salt, alkyl ether sulfate Examples of the nonionic surfactant include ether type surfactants (eg, glycerol ether and its alkylene oxide adducts), esters, and the like. Type surfactant (eg, glycerin ester and its alkylene oxide adduct), ether ester type surfactant (eg, sorbitan ester and its alkylene oxide adduct), fatty acid alkylo Luamide (eg, fatty acid monoethanolamide, fatty acid diethanolamide, etc.), ester type surfactant (eg, polyoxyalkylene fatty acid ester, polyoxyalkylene polyhydric alcohol fatty acid ester, polyoxyalkylene sorbitan fatty acid ester, polyoxyalkylene hydrogenated) Castor oil, glyceryl monostearate, glycerin ester, fatty acid polyglycerin ester, acylamino acid polyglycerin ester, sorbitan ester, sucrose fatty acid ester, etc.), nitrogen-containing nonionic surfactants (eg, alkyl glucosides, cured) Castor oil pyroglutamic acid diester and its ethylene oxide adduct, and fatty acid alkanolamide, etc.); cationic surfactants include, for example, aliphatic amine salts ( Examples, alkylammonium chloride, dialkylammonium chloride, alkyltrimethylammonium chloride (C ₁₆ -C ₂₂ ), dialkyldimethylammonium methosulfate salt, etc., aromatic quaternary ammonium salts (eg, quaternary ammonium salts of the above aliphatic amine salts) , Benzalkonium salts, etc.), fatty acid acyl arginine esters, N-long chain acyl arginine ethyl pyrrolidone carboxylates, amidoamines, stearamide propyl dimethylamine glutamate, stearamide propyl dimethylamine lactate, stearamide propyl dimethylamine pyrrolidone Carboxylate, behenamidopropyldimethylamine glutamate behenamidopropyldimethylamine lactate, behenamidopropyldimethylamine pyrrolidone carboxylate, etc. Examples of amphoteric surfactants include betaine type surfactants (eg, alkylbetaines, alkylamidobetaines, sulfobetaines, imidazolinium betaines, aminopropionates, carboxybetaines, etc.), N-long chain acyls Examples thereof include arginine, N- (3-alkyl (12,14) oxy-2-hydroxypropyl) arginine hydrochloride, aminocarboxylic acid type surfactant, imidazoline type surfactant and the like.

  Examples of amino acids include arginine, lysine, glutamic acid, aspartic acid, valine, leucine, isoleucine, serine, glycine, alanine, proline, hydroxyproline, threonine, histidine, phenylalanine, tryptophan, tyrosine, glutamine, asparagine, cysteine, cystine. , Methionine and their salts.

  Examples of amino acid derivatives include pyrrolidone carboxylic acid and its salt, trimethylglycine, lauroyl lysine and the like.

Examples of the lower alcohol include ethanol, propanol, isopropanol, butanol and the like.
Examples of higher alcohols include cetyl alcohol, lauryl alcohol, isostearyl alcohol, stearyl alcohol, oleyl alcohol, myristyl alcohol, octyldodecanol, behenyl alcohol, lanolin alcohol, cetostearyl alcohol, decyltetradecanol, hexyldecanol, cholesterol, phytosterol, etc. Is mentioned.

  Examples of the polyhydric alcohol include glycerin, diglycerin, ethylene glycol, 1,3-butylene glycol, propylene glycol, dipropylene glycol, and isoprene glycol.

  Examples of sugar alcohols and alkylene oxide adducts thereof include mannitol and erythritol.

  Examples of water-soluble polymers include polyglutamic acid, polyamino acids including polyaspartic acid and salts thereof, polyethylene glycol, high-polymerization polyethylene glycol, gum arabic, alginate, xanthan gum, hyaluronic acid, hyaluronic acid salt, chitin, Chitosan, water-soluble chitin, pectin, mannan, carrageenan, gellan gum, quince seed, guar gum, carboxyvinyl polymer, carboxymethylcellulose, hydroxymethylcellulose, hydroxyethylcellulose, hydroxypropylcellulose, hydroxypropylmethylcellulose, hydroxypropyl starch phosphate, polyacrylamide, polyvinyl Alcohol, polyvinylpyrrolidone, hydroxypropyltrimethylammonium chloride, poly Dimethylmethylene piperidinium hydride, polyvinylpyrrolidone derivative quaternary ammonium, acrylic acid copolymer, cationized protein, collagen degradation product and derivatives thereof, acylated protein, dextrin palmitate ester, aluminum stearate, polyglycerin, dibenzylidene sorbitol, 12- Examples thereof include hydroxystearic acid.

  Examples of the gelling agent include N-2-ethylhexanoyl-L-glutamic acid dibutylamide, N-lauroyl-L-glutamic acid dibutylamide (for example, GP-1, EB-21 manufactured by Ajinomoto Co., Inc.) and the like.

  Examples of the humectant include glycerin, diglycerin, dipropylene glycol, 1,3-butylene glycol, polyethylene glycol, hyaluronic acid, hyaluronic acid salt, chondroitin sulfate, chondroitin sulfate, honey, collagen, urea, DL-pyrrolidone carboxyl. Examples include sodium acid / L-pyrrolidone carboxylic acid mixed solution (AJIDEW NL-50), N-lauroyl-L-glutamate di (phytosteryl / 2-octyldodecyl), 1,3-pentanediol, 1,3-propanediol, and the like. It is done.

  Examples of the disinfectant and antibacterial agent include 4-hydroxybenzoic acid and its salts and esters, triclosan, chlorhexidine, phenoxyethanol, menthol, mint oil, glyceryl caprate, glyceryl caprylate, salicylic acid-N-alkylamide, N-coconut. Examples include oil fatty acid acyl-L-arginine ethyl DL-pyrrolidone carboxylate, benzalkonium chloride, and isopropylmethylphenol.

  Examples of the anti-inflammatory agent, analgesic agent, antifungal agent, keratin softener, skin colorant, and hormone agent include aloe extract, lily extract, hinokitiol, hydrocortisone (V), ε-aminocarboxylic acid, azulene, Allantoin, glycyrrhizic acids, glycyrrhizic acid derivatives, β-glycyrrhetinic acid and the like can be mentioned.

  An ultraviolet absorber is, for example, an organic substance (photoprotective filter) that is liquid or crystalline at room temperature, absorbs ultraviolet rays, and can release the absorbed energy as longer wavelength radiation (for example, heat). UV-B filter and UV-A filter. The UV-B filter can be oil-soluble or water-soluble. Examples of the oil-soluble substance include 3-benzylidene camphor or 3-benzylidene norcamphor and derivatives thereof (eg, 3- (4-methylbenzylidene) -camphor); 4-aminobenzoic acid derivatives (preferably 4- ( Dimethylamino) -benzoic acid-2-ethylhexyl ester, 4- (dimethylamino) -benzoic acid-2-octyl ester, 4- (dimethylamino) -benzoic acid pentyl ester); cinnamic acid ester (preferably 4-methoxycinnamic acid) Acid-2-ethylhexyl ester, 4-methoxycinnamic acid propyl ester, 4-methoxycinnamic acid isopropyl ester, 4-methoxycinnamic acid isopentyl ester, 2-cyano-3,3-phenylcinnamic acid-2-ethylhexyl ester [octocrylene (Octocrene Salicylic acid ester (preferably, salicylic acid-2-ethylhexyl ester, salicylic acid-4-isopropylbenzyl ester, salicylic acid homomenthyl ester); benzophenone derivatives (preferably 2-hydroxy-4-methoxybenzophenone, 2-hydroxy-4) -Methoxy-4'-methylbenzophenone, 2,2'-dihydroxy-4-methoxybenzophenone); benzalmalonic acid ester (preferably 4-methoxybenzalmalonic acid di-2-ethylhexyl ester); triazine derivatives (eg 2 , 4,6-trianilino- (p-carbo-2′-ethyl-1′-hexyloxy) -1,3,5-triazine, octyl triazone, dioctyl butamide triazone [Uvasorb® HEB ); Propane-1,3-dione (eg, 1- (4-t-butylphenyl) -3- (4′-methoxyphenyl) -propane-1,3-dione); ketotricyclo (5.2.1.); 0) Decane derivatives and the like. Examples of the water-soluble substance include 2-phenylbenzimidazole-5-sulfonic acid and its alkali metal salt, alkaline earth metal salt, ammonium salt, alkylammonium salt, alkanolammonium salt and glucamonium salt; benzophenone sulfonic acid Derivatives (preferably 2-hydroxy-4-methoxybenzophenone-5-sulfonic acid and its salts); sulfonic acid derivatives of 3-benzylidene camphor (eg 4- (2-oxo-3-bornylidenemethyl) -benzene Examples thereof include sulfonic acid and 2-methyl-5- (2-oxo-3-bornylidene) -sulfonic acid and salts thereof As the UV-A filter, benzoylmethane derivatives are used, for example, 1- (4′-t-butylphenyl) -3 (4′-methoxyphenyl) -propane-1,3-dione, 4-t-butyl-4′-methoxydibenzoylmethane (Parsol® 1789), 1-phenyl-3- (4′-isopropylphenyl) ) -Propane-1,3-dione, and enamine compounds, 4-methoxydibenzoylmethane, 4-t-butyl-4'-methoxydibenzoylmethane, titanium oxide, zinc oxide, octylmethoxycinnamate And 2-methoxyhexyl dimethoxybenzylidene dioxoimidazolidine propionate.

  Examples of antiperspirants and astringent active ingredients and antiperspirants include aluminum chloride, aluminum or zirconium salts such as aluminum chloride, aluminum chlorohydrate, aluminum zirconium trichlorohydrate, aluminum zirconium tetrachlorohydrate, pyrrolidone carboxylic acid Zinc etc. are mentioned.

  Examples of the vitamin agent include vitamin A, B1, B2, B6, E, and derivatives thereof.

  Examples of the blood flow promoting agent include ictamol, carpronium chloride, vitamin E or a derivative thereof, caffeine, capsaicinoid (eg, capsaicin, etc.), capsinoid (eg, dihydrocapsiate, etc.), cantalistin tincture, zingerone, tannic acid, minoxidil , Γ-oryzanol, zinc L-pyrrolidonecarboxylate, DL-pyrrolidonecarboxylic acid, DL-pyrrolidonecarboxylic acid sodium solution, DL-pyrrolidonecarboxylate sodium / L-pyrrolidonecarboxylic acid mixed solution (AJIDEW NL-50), and the like. .

  Examples of crude drugs include assembly extract, cephalanthin, garlic extract, carrot extract, aloe extract, gentian extract, yuzu extract, and altea extract.

  Examples of the pH adjuster include citric acid, adipic acid, phosphoric acid, glutamic acid, lactic acid, sulfuric acid, hydrochloric acid, ammonium, sodium hydroxide, potassium hydroxide, arginine, and γ-oryzanol.

  Examples of the sequestering agent include edetic acid and its salt, diethylenetriaminepentaacetic acid and its salt, ethylenediaminehydroxyethyl triacetic acid and its salt, hydroxyethanediphosphonic acid and its salt, phosphoric acid, citric acid, succinic acid, glucone Examples include acid, sodium polyphosphate, sodium metaphosphate, sodium hexametaphosphate, phytic acid, and sodium pentetate.

  Examples of the viscosity modifier include agar and organically modified clay mineral.

  Examples of the pearling agent include alkylene glycol esters, fatty acid alkanolamides, fatty acid monoglycerides, fatty ethers and the like.

  Examples of synthetic fragrances include esters, ethers, aldehydes, ketones, alcohols, and hydrocarbon-type fragrances.

  Examples of natural fragrances include flowers (eg, lily, lavender, rose, jasmine, neroli, Iran-Iran, etc.), stems and leaves (eg, geranium, patchouli, petitgren, etc.), fruits (eg, anise, coriander, cara). Way, juniper, etc.), pericarp (eg, bergamot, lemon, orange, etc.), root (eg, nutmeg, angelica, celery, cardamom, costas, iris, shobu, etc.), tree (eg, pine, sandalwood, guayak, cedar, Rosewood, herbs and grass (eg, tarragon, lemongrass, sage, thyme, etc.), needles and branches (eg, spruce, fir, pine, shrub pine, etc.), resins and balsam (eg, galvanum, elemi, benzoin) , Myrrh, frankincense, opopanax, etc.) and animal materials such as civet and beaver. It is. Furthermore, as a relatively low volatile essential oil often used as an aromatic component, for example, sage oil, chamomile oil, clove oil, Melissa oil, mint oil, cinnamon leaf oil, lime flower oil, juniper berry oil, vetiver oil , Balm oil, galvanum oil, labdanum oil and lavandin oil, bergamot oil, dihydromyrsenol, lyial, ryal, citronellol, phenylethyl alcohol, α-hexylcinnamaldehyde, geraniol, benzylacetone, cyclamenaldehyde, linalool, Boisambrene Forte, Ambroxan, indole, Hedione, Sandelice, citrus oil, mandarin oil, orange oil, allylpentyl glycolate, cyclovertal (Cyclo) vertal), lavandin oil, clary oil, β-damascone, geranium oil bourbon, cyclohexyl salicylate, Vertofix Coeur, Iso-E-Super, Fixolide NP, evanyl, Iraldein gamma, phenylacetic acid, benzyl acetate , Rose oxide, lomirat, irotil and floramat, peppermint oil, spearmint oil, anise oil, daikyo oil, caraway oil, eucalyptus oil, fennel oil, citrus oil, wintergreen oil, menthol Etc.

  Examples of the dye include cochineal red A (C.I. 16255), patent blue (C.I. 42051), chlorophyllin (C.I.75810), and the like.

  Examples of the pigment include titanium oxide, mica, sericite, and Nε-lauroyl-L-lysine.

  Examples of the antioxidant include vitamin E, vitamin E derivatives and salts thereof, and sodium sulfite.

  Examples of the preservative include phenoxyethanol, paraben, sorbic acid, isopropylmethylphenol, benzalkonium chloride, benzoic acid, dehydroacetic acid, hexanediol, pentanediol, chlorhexidine sulfate, N-coconut oil fatty acid acyl-L-arginine ethyl DL-pyrrolidone carboxylate and the like can be mentioned.

  The emulsifier is, for example, a nonionic surfactant and contains a linear fatty alcohol having 8 to 22 carbon atoms, a fatty acid having 12 to 22 carbon atoms, and 8 to 15 carbon atoms in the alkyl group. Addition products of 2 to 30 moles of ethylene oxide and / or 0 to 5 moles of propylene oxide to alkylphenols and alkylamines containing 8 to 22 carbon atoms in the alkyl group; 8 to 22 in the alkyl (alkenyl) group Alkyl and / or alkenyl oligoglycosides containing 1 carbon atom, and their ethoxylates; castor oil and / or hydrogenated castor oil ethylene oxide 1-15 mol adduct; castor oil and / or hydrogenated castor oil ethylene oxide 15 ~ 60 mole adduct; unsaturated straight chain containing 12-22 carbon atoms or Partial esters of saturated branched fatty acids and / or hydroxycarboxylic acids containing 3 to 18 carbon atoms with glycerin and / or sorbitan, and adducts thereof with 1 to 30 mol of ethylene oxide; polyglycerin (2 -8 average self-condensation degree), polyethylene glycol (molecular weight 400-5000), trimethylolpropane, pentaerythritol, sugar alcohol (eg sorbitol), alkyl glucoside (eg methyl glucoside, butyl glucoside, lauryl glucoside and C16-C18 alkyl glucoside) ) And polyglucosides (eg cellulose), saturated and / or unsaturated linear or branched fatty acids containing 12 to 22 carbon atoms and / or hydroxycarboxylic acids containing 3 to 18 carbon atoms Partial esters and adducts thereof with 1 to 30 moles of ethylene oxide; pentaerythritol, fatty acids, mixed esters of citric acid and fatty alcohols, and / or fatty acids containing 6 to 22 carbon atoms, methyl glucose and polyols ( Preferably mixed esters with glycerol or polyglycerol); mono-, di-, trialkyl phosphates and mono-, di- and / or triPEG-alkyl phosphates and their salts; wool wax alcohols; polysiloxane / poly Alkyl / polyether copolymers and corresponding derivatives; block copolymers (eg polyethylene glycol-30 dipolyhydroxystearate); polymer emulsifiers (eg Goodrich's Pemulen type (TR-1, TR -2)); polyalkylene glycol, glycerol carbonate and the like. Ethylene oxide adducts include, for example, ethylene oxide of fatty alcohols, fatty acids, alkylphenols, or castor oil, and propylene oxide adducts include known commercial products. Examples of the partial glyceride include hydroxystearic acid monoglyceride, hydroxystearic acid diglyceride, isostearic acid monoglyceride, isostearic acid diglyceride, oleic acid monoglyceride, oleic acid diglyceride, ricinoleic acid monoglyceride, ricinoleic acid diglyceride, linoleic acid monoglyceride, linoleic acid diglyceride, linolenic acid Acid monoglyceride, linolenic acid diglyceride, erucic acid monoglyceride, erucic acid diglyceride, tartaric acid monoglyceride, tartaric acid diglyceride, citric acid monoglyceride, citric acid diglyceride, malic acid monoglyceride, malic acid diglyceride and the like. Furthermore, ethylene oxide 1-30 mol (preferably 5-10 mol) adducts of the above partial glycerides are also suitable. Examples of sorbitan esters include sorbitan monoisostearate, sorbitan sesquiisostearate, sorbitan diisostearate, sorbitan triisostearate, sorbitan monooleate, sorbitan sesquioleate, sorbitan dioleate, sorbitan trioleate, sorbitan monoelcate, sorbitan sesquito Elkato, sorbitan dielcate, sorbitan trielcate, sorbitan monoricinoleate, sorbitan sesquilicinolate, sorbitan diricinolate, sorbitan triricinolate, sorbitan monohydroxystearate, sorbitan sesquihydroxystearate, sorbitan dihydroxystearate, sorbitan trihydroxystearate Rate, sol Vitan monotartrate, sorbitan sesquitate, sorbitan ditartrate, sorbitan tritartrate, sorbitan monocitrate, sorbitan sesquitrate, sorbitan dicitrate, sorbitan tricitrate, sorbitan monomaleate, sorbitan sesquiterate, sorbitan dimate, mixed sorbitan dimaleate, sorbitan dimaleate Is mentioned. Furthermore, 1-30 mol (preferably 5-10 mol) addition product of ethylene oxide of the sorbitan ester is also suitable. Examples of the polyglycerol ester include polyglyceryl-2 dipolyhydroxystearate (Dehymuls (registered trademark) PGPH), polyglycerin-3-diisostearate (Lameform (registered trademark) TGI), polyglyceryl-4 isostearate ( Isolan® GI34), polyglyceryl-3 oleate, diisostearoyl polyglyceryl-3 diisostearate (Isolan® PDI), polyglyceryl-3 methylglucose distearate (Tego Care® 450), polyglyceryl- 3 Beeswax (Cera Bellina (registered trademark)), Polyglyceryl-4 Caprate (Polyglycerol Caprate T2010 / 90), Polyg Lyceryl-3 cetyl ether (Chimexane® NL), polyglyceryl-3 distearate (Cremophor® GS 32) and polyglyceryl polylysinolate (Admul® WOL 1403), polyglyceryl dimerate isostearate, and mixtures thereof Etc. Examples of polyol esters include mono-, di- and triesters of trimethylolpropane or pentaerythritol with lauric acid, coconut oil fatty acid, tallow fatty acid, palmitic acid, stearic acid, oleic acid, behenic acid, etc. Reacted with 1 to 30 mol of ethylene oxide, stearate, POE fatty acid glycerin ester, POE hydrogenated castor oil ester, POE sorbitan fatty acid ester, N-stearoyl-L-glutamate sodium, pyroglutamate isostearate PEG-40 hydrogenated Castor oil, pyroglutamic acid isostearic acid PEG-30 hydrogenated castor oil, pyroglutamic acid isostearic acid PEG-60 hydrogenated castor oil, pyroglutamic acid isostearic acid glycerase-25, and the like. That.

  Examples of the fat and wax include 12-hydroxystearic acid, lanolin, beeswax, candelilla wax, carnauba wax, petrolatum, solid paraffin, lanolin derivative, microcrystalline wax, ceresin, synthetic triglyceride, bees wax, and wood wax. .

  Examples of the silicone compound include chain silicones such as dimethylpolysiloxane, methylphenylpolysiloxane, and methylhydrogenpolysiloxane; cyclic silicones such as octamethylcyclotetrasiloxane, decamethylcyclopentasiloxane, and dodecamethylcyclohexasiloxane; and Amino-modified silicone compounds, fatty acid-modified silicone compounds, alcohol-modified silicone compounds, polyether-modified silicone compounds, epoxy-modified silicone compounds, fluorine-modified silicone compounds, glycoside-modified silicone compounds and / or alkyl-modified silicone compounds (Which can be liquid or resin-like at room temperature), dimethicone and hydrogenated silicate having an average chain length of 200 to 300 dimethylsiloxane units Such as a mixture of simethicone and the like.

  Perfume oils include mixtures of natural and synthetic fragrances.

In a preferred embodiment, the whitening cosmetic composition of the present invention is a skin external preparation.
When the whitening cosmetic composition of the present invention (or pharmaceutical composition containing SQG or a salt thereof as an active ingredient) is used as an external preparation for skin, auxiliary agents commonly used for external preparations for skin, such as disodium edetate and trisodium edetate , Sequestering agents such as sodium citrate, sodium polyphosphate, sodium metaphosphate, gluconic acid, caffeine, tannin, verapamil, tranexamic acid and its derivatives, grabrizine, hot water extract of karin fruit, various herbal medicines, tocopherol acetate , Drugs such as glycyrrhizic acid and its derivatives or salts, whitening agents such as vitamin C, magnesium ascorbate phosphate, glucoside ascorbate, arbutin, kojic acid, sugars such as glucose, fructose, mannose, sucrose, trehalose, retinoin Acid, retinol, retinol acetate Vitamin A such as retinol palmitate can also be appropriately blended.

  The whitening cosmetic composition of the present invention may contain other whitening ingredients in addition to the active ingredient SQG or a salt thereof. Other whitening components are not limited as long as they do not impair the desired effect. For example, hydroquinone such as hydroquinone, arbutin, hydroquinone glycoside and derivatives thereof; ascorbic acid, L-ascorbic acid-2-phosphate, and L -L-ascorbic acid esters such as ascorbic acid-2-sulfate, L-ascorbic acid glucosides such as L-ascorbic acid-2-glucoside and L-ascorbic acid-5-glucoside, ethyl 3-O-ascorbate Ascorbic acid and its derivatives such as L-ascorbic acid ethers such as ether and 3-O-ascorbic acid cetyl ether; Kojic acid and its derivatives such as kojic acid and kojic acid dipalmitate; Nicotinic acid derivatives such as nicotinamide ; Coumaroyl serotonin and Ferroy Serotonin derivatives such as serotonin; salicylic acid derivatives such as 3-methoxysalicylic acid, 4-methoxysalicylic acid and 5-methoxysalicylic acid; astaxanthin; α-lipoic acid; calcium pantethein sulfonate; apricot extract, wheat germ extract, acerola extract, gentian Extracts, cabbage extract, oil-soluble licorice extract, saicin extract, flaxseed extract, Dutch mustard extract, glaze extract, toki extract, bitumen extract, horse chestnut bark extract, Ibukitora noo extract, Clara extract, Ogon extract, Itadori extract, Extracts with whitening effects such as raspberry extract (raspberry ketone glucoside), birch extract, cypress root extract, chamomile extract, yeast extract and placenta extract; safflower extract, safflower seed extract, elderberry Extract, cypress extract, daisy extract, brown algae extract, ages extract, fire spine extract, persimmon tannin extract, ginkgo biloba extract, cassis extract, black tea extract, green tea extract, oolong tea extract, Plant extracts having a whitening effect, such as tamarind extract, rooibos extract, Sakuhakuhi extract, Koboku extract, Geoscoria compositor extract, rice bran extract, Tonin extract and Yukinoshita extract; ellagic acid, gallic acid , Pentagalloylglucose, resveratrol, grabrizine, glabrene, resorcin and derivatives thereof, etc .; polyphenols having a whitening effect; unsaturated fatty acids having a whitening effect such as linolenic acid and linoleic acid; glutathione (GSH) and collagen peptides, etc. Peptides having whitening effect of N-acetyl L-tyrosine etc. Rosin derivatives, cysteine derivatives such as N-acetyl L-cysteine, amino acids having a whitening effect such as ergothionein and tranexamic acid (t-AMCHA); nucleic acid derivatives having a whitening effect such as adenosine monophosphate; and lucinol (4- n-butylresorcinol), magnolignan, biphenyl compounds, rhododenol and derivatives thereof, such as one or more selected from alkylphenols having a whitening effect.

3. In one aspect of the microalgal membrane extract having a melanin production inhibitory effect, SQG or a salt thereof contained in the agent of the present invention or the whitening cosmetic of the present invention is obtained by subjecting the microalgae membrane component to a defatty acid group treatment. It can also be provided by the obtained microalgal membrane extract having an inhibitory effect on melanin production (also simply referred to as microalgal membrane extract in the present specification). The microalgal membrane extract contains SQG or a salt thereof. The membrane component of microalgae used in the present invention is a membrane component containing acylated SQG, such as a thylakoid membrane.
A method for producing a microalgal membrane extract is described in detail below.

(1) Microalgae In the present specification, “microalgae” are generally microscopic-sized algae that exist in water, and most of them have chlorophyll like plants, and carbon dioxide ( Fixes CO ₂ ) and produces oxygen (O ₂ ). Microalgae includes prokaryotes, cyanobacteria, and eukaryotes, Glaucophyta, red plant algae (Rhodophyta), green plant gate (Chlorophyta), crypto. Phytophyta (Cryptophyta), Haptophyta (Haptophyta), Heterokontophyta, Dinophyta (Dinophyta), Euglenophyta and Euglenophyta Various unicellular organisms classified as Chlorarachniophyta are included.

  Examples of algae belonging to the green plant family include algae belonging to classes such as Chlorophyceae, Trebouxiophyceae, Prasinophyceae, Ulvophyceae, and Charophyceae. Is mentioned. Examples of algae belonging to the green alga class include Neochloris oleoabundans (Tornabene, TG et al. 1983. Enzyme and Microb. Technol. 5: 435-440), Nanochloris sp. Nannochloris sp.) (Takagi, M. et al. 2000. Appl. Microbiol. Biotechnol. 54: 112-117), Chlamydomonas reinhardtii, Chlamydomonas genus algae, Seddesmus (Scenedesmus) algae and Desmodesmus (Desmodesmus) algae can be mentioned. Examples of the algae belonging to the Trevoxia algae include Chlorella algae such as Chlorella kessleri.

The microalgae used in the present invention is classified into such microalgae, and any microalgae may be used as long as a microalgae membrane extract having a desired effect can be obtained. As the microalgae used in the present invention, microalgae belonging to the green plant gate are preferable, microalgae belonging to the green alga net or Trevoxia algae are more preferable, and microalgae belonging to the genus Chlorella, Chlamydomonas, Desmodesmus or Senedesmus More preferred, more preferred are microalgae belonging to the genus Desmodemus, and most preferred is the FERM BP-22277 strain.
The microalgae used in the present invention is usually a living microalgae, but as long as a microalgae membrane extract exhibiting melanin production inhibitory activity is obtained, even a dead microalgae such as dried microalgae can be obtained. I do not care.
The microalgae used in the present invention may be microalgae cultured in vitro. In the case of using microalgae cultured in vitro, the culture method of microalgae is not particularly limited as long as microalgae can grow, and can be cultured by a method known per se, for example, in International Publication No. 2014/192940 It can be cultured by a method according to the described method for culturing microalgae.

(2) Defatty acid grouping of membrane components of microalgae Defatty acid grouping of membrane components of microalgae involves defatty acid grouping of the entire microalgae or one or more parts of microalgae containing membrane components. Is done. The defatty acid group formation of the membrane component of microalgae is not particularly limited as long as the fatty acid group (preferably acyl group) can be eliminated from the lipid contained in the membrane component of microalgae. (I) Medium temperature of microalgae The treatment can be carried out by (ii) organic solvent treatment using an organic solvent such as diethyl ether, ethanol or methanol. Preferably, the defatty acid grouping of the membrane component of the microalgae comprises (iii) subjecting the microalgae to an intermediate temperature treatment, and selecting a water-soluble fraction from the obtained intermediate temperature treatment product, (iv) diethyl ether, ethanol Alternatively, an organic solvent treatment using an organic solvent such as methanol is performed to select a soluble fraction. The “medium-temperature treated product” of microalgae means a product obtained by subjecting an aqueous solvent containing microalgae to intermediate temperature treatment. From the viewpoint of reducing the possibility that an organic solvent that can exhibit toxicity remains, the above-described (i) or (iii) is preferable as the defatty acid grouping of the membrane component of microalgae.
The cultured microalgae may be used as the microalgae subjected to the defatty acid group treatment. In the case of using cultured microalgae, the microalgae may be subjected to a defatty acid group treatment while contained in the medium, recovered from the medium, and put into an aqueous medium (aqueous solvent) such as water or an aqueous buffer. After resuspension, it may be subjected to a defatty acid group treatment.
The method for recovering microalgae from the medium is not particularly limited, and for example, a known method (Grima, EM et al. 2003. Biotechnol. Advances 20: 491-515) can be used. For example, natural sedimentation, centrifugation, filtration, etc. By this method, microalgae can be recovered from the medium. At that time, a flocculant may be used. The collected microalgae can be appropriately washed and / or resuspended using an appropriate medium. Examples of media that can be used for washing and suspension include aqueous media (aqueous solvents) such as water and aqueous buffers.
From the viewpoint of avoiding the introduction of medium components, it is preferable to use microalgae recovered from the medium and resuspended in an aqueous solvent. After collection from the medium, washing is performed, and microalgae resuspended in an aqueous solvent is used. preferable.

Further, the microalgae may be subjected to a defatty acid group treatment after being subjected to a pretreatment such as dilution, concentration, freezing, thawing, and drying, and these pretreatments may be appropriately combined.
Microalgae are diluted or concentrated so that the concentration of microalgae in the suspension is, for example, 25 g / L or more, or 250 g / L or more in terms of dry weight. You may use for a chemical conversion process.

  Prior to the defatty acid grouping treatment, the pH of the reaction system may be adjusted to be weakly acidic or weakly alkaline. The weak acidity may be, for example, pH 3.0 to 7.0, pH 4.0 to 6.0, or pH 4.5 to 6.0. The weak alkalinity may be, for example, pH 7.5 to 12.0, pH 9.0 to 11.0, or pH 9.0 to 10.5. The pH can be adjusted using, for example, an acidic substance such as hydrochloric acid, or an alkaline substance such as NaOH or KOH. By adjusting the pH, microalgae may or may not be hydrolyzed.

When performing a medium temperature treatment as a defatty acid group treatment, a medium temperature treatment of microalgae can be performed according to the method described in International Publication No. 2014/192940.
The microalgae can be subjected to an intermediate temperature treatment in a state suspended in an appropriate aqueous medium as described above. In the present specification, “medium temperature treatment” refers to treatment at an intermediate temperature. As the medium temperature treatment, for example, treatment at a medium temperature described in International Publication No. 2011/013707 and International Publication No. 2014/192940 can be referred to. The intermediate temperature can be appropriately set according to various conditions such as processing time. Although it is not particularly limited as long as a desired effect such as suppression of melanin production can be obtained, the intermediate temperature is usually 35 ° C. or higher, preferably 40 ° C. or higher, more preferably 45 ° C. or higher, more preferably 50 ° C. or higher. 70 ° C. or lower, preferably 65 ° C. or lower, more preferably 60 ° C. or lower. The time for the medium temperature treatment can be appropriately set according to various conditions such as the treatment temperature. The duration of the medium temperature treatment can be, for example, 1 hour or more, or 5 hours or more. Further, the intermediate temperature treatment time may be, for example, 48 hours or less, or 24 hours or less. The pH of the medium temperature treatment is not particularly limited as long as an extract having a desired effect is obtained, and is usually pH 3.0 to 11.0, may be weakly acidic, may be near neutral, and weakly alkaline. It may be. The weak acidity may be in the above weak acid range, for example, pH 4.5 to 6.0. The vicinity of neutrality may be, for example, pH 7.0 to 9.0. The weak alkalinity may be in the above weak alkaline range, for example, pH 9.0 to 10.5.
The medium temperature treatment may be performed by standing or may be performed while stirring or shaking.

  The intermediate temperature treatment may be performed continuously or intermittently. Reaction conditions such as treatment temperature may or may not be constant throughout the medium temperature treatment. That is, for example, the microalgae may be continuously treated at the same temperature (hereinafter, also referred to as “continuous intermediate temperature treatment”), or may be treated by changing the temperature in the middle. The continuous intermediate temperature treatment may be performed, for example, in the above-exemplified intermediate temperature range and the above-exemplified intermediate temperature treatment range. As an aspect which changes temperature on the way, the aspect which reduces temperature on the way is mentioned, for example. As an aspect of lowering the temperature in the middle, for example, an aspect of treating at a temperature lower than the temperature of the first stage intermediate temperature treatment as the second stage intermediate temperature treatment after once treating at an intermediate temperature as the first stage intermediate temperature treatment. It is done. The temperature of the first stage intermediate temperature treatment may be, for example, the intermediate temperature range exemplified above. The temperature of the second stage intermediate temperature treatment may be, for example, 30 ° C or higher, 35 ° C or higher, or 40 ° C or higher. The temperature of the second stage intermediate temperature treatment may be, for example, 55 ° C. or lower, 50 ° C. or lower, or 45 ° C. or lower. The time of the first stage intermediate temperature treatment may be, for example, 1 minute or more, 5 minutes or more, 10 minutes or more, or 20 minutes or more. The time of the first stage intermediate temperature treatment may be, for example, 120 minutes or less, or 60 minutes or less. The second stage intermediate temperature treatment time may be, for example, 1 hour or more, 2 hours or more, or 4 hours or more. The time of the second stage intermediate temperature treatment may be, for example, 20 hours or less, or 15 hours or less.

The intermediate-temperature treated product obtained by the above-mentioned intermediate-temperature treatment may be used in the present invention as it is, but further, a water-soluble fraction and a water-insoluble fraction of the intermediate-temperature treated product are separated to obtain a water-soluble fraction. Is preferably recovered and used in the present invention. Moreover, as long as it has a desired effect, this water-soluble fraction may be further subjected to drying, dilution, concentration, freezing and the like. In the present specification, the water-soluble fraction itself and the dried, diluted, concentrated and frozen products of the water-soluble fraction are collectively referred to as the supernatant extract after the intermediate temperature treatment. The supernatant extract after the intermediate temperature treatment can be used in the present invention.
Separation operation of the water-soluble fraction and the water-insoluble fraction can be performed by a method known per se. Examples of the separation operation are not particularly limited as long as a water-soluble fraction having a desired effect can be obtained, and include natural sedimentation, filtration operation, separation operation by centrifugation, and the like, and combinations thereof.

  “Organic solvent treatment” refers to treatment with an organic solvent. The conditions for the organic solvent treatment are not particularly limited as long as a microalgal membrane extract having a desired effect can be obtained. The organic solvent treatment can be performed by bringing microalgae into contact with an organic solvent. For example, the collected microalgae may be suspended in an organic solvent, or an organic solvent may be added to the microalgae suspension. Examples of the organic solvent include alcohols such as methanol, ethanol, 2-propanol, butanol, pentanol, hexanol, heptanol, and octanol, ketones such as acetone, ethers such as dimethyl ether and diethyl ether, methyl acetate, and ethyl acetate. Such as esters, alkanes such as n-hexane, benzene, phenol, and chloroform. As an organic solvent, a water-soluble thing is preferable. As an organic solvent, alcohol is preferable and methanol is more preferable. As the organic solvent, one kind of organic solvent may be used, or two or more kinds of organic solvents may be used in combination. The organic solvent may be a pure product or a mixture with other components. Examples of other components include an aqueous medium (aqueous solvent) such as water and an aqueous buffer solution. That is, for example, an aqueous solution of an organic solvent can be used for the organic solvent treatment. The organic solvent concentration in the mixture may be, for example, 10% (v / v) or more, or 20% (v / v) or more, 90% (v / v) or less, 70% (v / v) or less Or 50% (v / v) or less. Further, the concentration of the organic solvent in the reaction solution for performing the organic solvent treatment may be, for example, 10% (v / v) or more, or 20% (v / v) or more, 90% (v / v) or less, It may be 70% (v / v) or less, or 50% (v / v) or less. The organic solvent treatment time may be, for example, 10 minutes or longer, 30 minutes or longer, or 1 hour or longer. Further, the time of the organic solvent treatment may be, for example, 10 hours or less, 5 hours or less, or 3 hours or less. The temperature of the organic solvent treatment may or may not be controlled. The temperature of the organic solvent treatment may be, for example, 10 to 70 ° C. or room temperature. The organic solvent treatment may be performed by standing or may be performed while stirring or shaking.

  After subjecting the microalgae to the organic solvent treatment, an operation for separating the soluble fraction and the insoluble fraction is performed, and the soluble algal membrane extract can be obtained by recovering the soluble fraction. As long as it has a desired effect, the soluble fraction may be further subjected to drying, dilution, concentration, freezing and the like. In the present specification, the soluble fraction itself and the dried, diluted, concentrated, frozen and the like of the soluble fraction are collectively referred to as an organic solvent extract. This organic solvent extract can also be used in the present invention.

  The microalgal membrane extract (eg, supernatant extract after intermediate temperature treatment) obtained as described above is further fractionated by sterilization such as filtration sterilization, cation exchange chromatography, etc., depending on the intended use. It can also be subjected to desired operations such as concentration of active ingredients by operation, removal of toxic components such as pheophorbide and proteins.

  When applying the microalgae membrane extract to a living body, it is considered preferable to have a high safety and low allergenicity, so the microalgae membrane extract used in the present invention is preferably sterilized, It is preferable not to contain toxic substances such as pheophorbide and / or proteins.

4). Skin whitening method The present invention provides a skin whitening method by applying the whitening cosmetic of the present invention to a skin region where whitening is desired. Furthermore, the present invention provides a method for suppressing excessive melanin production in the skin, comprising applying the whitening cosmetic composition of the present invention to a subject in need of suppression of excessive melanin production in the skin. Provide cosmetic or therapeutic methods. In these methods, the whitening cosmetic of the present invention is preferably prepared as a skin external preparation.

  Hereinafter, the present invention will be described more specifically with reference to examples. However, the present invention is not limited to these examples.

Example 1
Preparation of medium temperature treated supernatant (1)
Culture of algae strain Green algae strain AJ7850 (FERM BP-22277 strain) was cultured in a 500 mL Erlenmeyer flask containing 100 mL of 0.2 x Gamborg B5 medium (Nippon Pharmaceutical), with a culture temperature of 30 ° C, light intensity of 7,000 lux, 3 The pre-cultured solution was obtained by culturing with shaking in an incubator with a% CO ₂ concentration air / CO ₂ mixed gas atmosphere (culture device CLE-303 manufactured by TOMY) for 7 days. Add 80 mL of pre-culture solution to a 2 L jar fermenter (Ishikawa Seisakusho) containing 1.5 L of 0.2 × Gunborg B5 medium at 500 mL / min at a culture temperature of 30 ° C and a light intensity of 20,000 lux. Culturing was performed for 15 days while blowing air / CO ₂ mixed gas of 3% CO ₂ concentration. Note that white light from a fluorescent lamp was used as the light source.

(0.2 x Gamborg B5 medium)
KNO ₃ 500 mg / L
MgSO ₄・ 7H ₂ O 50 mg / L
NaH ₂ PO ₄・ H ₂ O 30 mg / L
CaCl ₂・ 2H ₂ O 30 mg / L
(NH ₄ ) ₂ SO ₄ 26.8 mg / L
Na ₂ -EDTA 7.46 mg / L
FeSO ₄・ 7H ₂ O 5.56 mg / L
MnSO ₄・ H ₂ O 2 mg / L
H ₃ BO ₃ 0.6 mg / L
ZnSO ₄・ 7H ₂ O 0.4 mg / L
KI 0.15 mg / L
Na ₂ MoO ₂・ 2H ₂ O 0.05 mg / L
CuSO ₄・ 5H ₂ O 0.005 mg / L
CoCl ₂・ 6H ₂ O 0.005 mg / L
120 ℃ 15 minutes Autoclave sterilization

A culture solution of medium temperature treated green algae AJ7850 strain (FERM BP-22277 strain) 2.3 L was centrifuged at 6,000 rpm for 10 minutes to remove the supernatant, and AJ7850 strain alga was obtained. Suspend the obtained alga bodies in a small amount of sterile water, centrifuge and remove the supernatant 4 times, and concentrate the alga bodies after the fourth centrifugation to 100 mL with sterile water. A liquid was prepared. The pH was about 6.5. Medium temperature treatment was performed by stirring at 53 ° C. for 2 hours 30 minutes. The treated product after the intermediate temperature treatment was centrifuged to obtain a supernatant after removing the residue. The obtained supernatant was designated as an intermediate temperature treated supernatant (1).

Example 2
Preparation of medium temperature treated supernatant (2)
Culture of algae strain Green algae strain AJ7850 (FERM BP-22277 strain) was cultured in a 500 mL Erlenmeyer flask containing 100 mL of 0.2 x Gamborg B5 medium (Nippon Pharmaceutical), with a culture temperature of 30 ° C, light intensity of 7,000 lux, 3 The pre-cultured solution was obtained by culturing with shaking in an incubator with a% CO ₂ concentration air / CO ₂ mixed gas atmosphere (culture device CLE-303 manufactured by TOMY) for 7 days. Add 80 mL of pre-culture solution to a 2 L jar fermenter (Ishikawa Seisakusho) containing 1.5 L of 0.2 × Gunborg B5 medium at 500 mL / min at a culture temperature of 30 ° C and a light intensity of 20,000 lux. The culture was performed for 12 days while blowing air / CO ₂ mixed gas of 3% CO ₂ concentration. Note that white light from a fluorescent lamp was used as the light source.
The culture solution 3.7 L of the medium temperature treated green algae AJ7850 strain (FERM BP-22277 strain) was centrifuged at 6,000 rpm for 10 minutes, and the supernatant was removed to obtain algal bodies. Suspend the obtained alga bodies in a small amount of sterile water, centrifuge and remove the supernatant 4 times, and concentrate the alga bodies after the fourth centrifugation to 200 mL with sterile water. A liquid was prepared. 0.1NH ₂ SO ₄ was added to adjust the pH to 5.51. Medium temperature treatment was performed by stirring at 53 ° C. for 2 hours 30 minutes. The treated product after the intermediate temperature treatment was centrifuged to obtain a supernatant after removing the residue. The obtained supernatant was designated as an intermediate temperature treated supernatant (2).

Example 3
Analysis of mesophilic supernatant The mesophilic supernatant (2) prepared in Example 2 was subjected to HPLC analysis. HPLC is an HPLC analyzer equipped with two sugar analysis columns SH-1011 (Showa Denko) in series with an autosampler L-7200, pump L-7100, and differential refractive index detector L-2490 (both from Hitachi). Made using. Water was used as the mobile phase, the flow rate was 1.0 mL / min, and the column temperature was 60 ° C. The HPLC analysis results are shown in FIG. Mass spectrometry of the peak detected by HPLC confirmed that the peak with a retention time of 10.40 minutes was sulfoquinovosylglycerol. By subjecting the preparation of sulfoquinovosylglycerol (WO2014192940 Reference Example 1) to the same HPLC analysis, the concentration of sulfoquinovosylglycerol in the mesophilic supernatant (2) was calculated to be 94 mg / mL.
The same HPLC analysis was performed on the intermediate-temperature treated supernatant (1) prepared in Example 1, and a peak having the same retention time as that of sulfoquinovosylglycerol was detected.

Example 4
Melanin production inhibitory activity test The melanin production rate was measured in order to compare the whitening effect of the intermediate-temperature treated supernatant (1) prepared in Example 1 and the intermediate-temperature treated supernatant (2) prepared in Example 2. B16 melanoma was cultured in DMEM (Dulbecco's Modified Eagle Medium) (high glucose, containing 10% serum) at 37 ° C, 5% CO2, and saturated steam. The confluent cells were seeded in a 96 well plate. As an evaluation sample, 10 ml of the medium-temperature treated supernatant (1) (or (2)) was dried by lyophilization and redissolved in 4 ml of the above-mentioned DMEM (without phenol red) as a positive control. Used kojic acid (manufactured by Sigma-Aldrich). On the day after seeding, dilution series of each evaluation sample or positive control was prepared with the above DMEM (without phenol red), and the medium in each well was changed. After culturing for 3 days, the absorbance at 450 nm was measured with a microplate reader (manufactured by Thermo Fisher Scientific). When the measurement value of the control (without sample addition) is defined as 100%, the measurement value 3 days after the addition of each sample's specified concentration is set to relative%, so that the amount of melanin in the control is 100%. The melanin production rate of the sample was calculated. The result is shown in figure 2.

Cytotoxicity test (neutral red method)
After measuring the absorbance in the melanin production inhibition test, the evaluation sample solution was removed from the plate, and each well was rinsed with 200 μl of DMEM (containing high glucose and 10% serum). DMEM containing NR (neutral red) was added dropwise at 200 μl / well and allowed to stand for 2 hours at 37 ° C., 5% CO 2 and saturated steam. Discard DMEM (with neutral red), add 200 μl / well of washing fixative (equal mix of 2 wt% calcium chloride solution and 2 wt /% formalin solution), let stand for 1 minute, then remove wash fixative It was. NR extract (acetic acid ethyl alcohol) 200 μl / well was added and shaken for 15 minutes with a plate shaker. Incorporation of NR into living cells was examined by measuring absorbance at 540 nm of the NR extract with a microplate reader (manufactured by Thermo Fisher Scientific). When the measured value (absorbance) of the control (without sample addition) NR extract is defined as 100%, the absorbance of the NR extract of each sample-added cell at a given concentration is set to relative%, so that the cells of each sample Toxicity was calculated.
It was confirmed that the intermediate temperature-treated supernatants (1) and (2) had the same melanin production inhibitory effect at a concentration without cytotoxicity (FIG. 2).

Example 5
Sulfoquinovosylglycerol suppresses melanin production (including comparison with glycosylglycerol)
Sulfoquinovosylglycerol was synthesized according to the method described in Reference Example 1 of WO2014192940. To compare the whitening effect of sulfoquinovosylglycerol (SQG) and glycosylglycerol (GG) (purchased product), melanin production rate was measured. B16 melanoma was cultured in DMEM (Dulbecco's Modified Eagle Medium) (high glucose, containing 10% serum) at 37 ° C, 5% CO2, saturated steam. The confluent cells were seeded in a 96 well plate. On the next day, a dilution series of each evaluation sample or positive control (kojic acid (manufactured by Sigma Aldrich)) was prepared with the above DMEM (without phenol red), and the medium in each well was changed. After culturing for 3 days, the absorbance at 450 nm was measured with a microplate reader (manufactured by Thermo Fisher Scientific). When the measurement value of the control (without sample addition) is defined as 100%, the measurement value 3 days after the addition of each sample's specified concentration is set to relative%, so that the amount of melanin in the control is 100%. The melanin production rate of the sample was calculated. The cytotoxicity test was performed by the method described in Example 4. The results are shown in Figure 3.
SQG showed an inhibitory effect on melanin production at a non-cytotoxic concentration. On the other hand, GG could not confirm the melanin production inhibitory effect even when the concentration was twice or more that of SQG.

  According to the present invention, it is possible to provide a melanin production inhibitor and a whitening cosmetic composition which are excellent in safety and have a high melanin production inhibitory action. The agent of the present invention can be used for various purposes meaningful for exhibiting a melanin production inhibitory action or a whitening effect, such as pigmentation due to sunburn, stains, freckles, dull skin, liver spots, etc. It can be used as a whitening agent effective for prevention, improvement, treatment, etc., and is particularly useful for the production of cosmetics for skin whitening.

Claims (14)

  A melanin production inhibitor comprising sulfoquinovosylglycerol or a salt thereof.   A melanin production inhibitor comprising a microalgal membrane extract.   The agent according to claim 2, wherein the microalgal membrane extract contains sulfoquinovosylglycerol or a salt thereof.   The microalgae membrane extract is a supernatant extract after intermediate temperature treatment obtained by subjecting microalgae to intermediate temperature treatment, followed by separation operation of a water-soluble fraction and a water-insoluble fraction. 3. The agent according to 3.   The agent according to any one of claims 2 to 4, wherein the microalga is a microalga belonging to the green alga class.   The agent according to claim 5, wherein the microalgae belong to the genus Desmodemus.   The agent according to any one of claims 1 to 6, which is for skin whitening.   A whitening cosmetic comprising sulfoquinovosylglycerol or a salt thereof.   Whitening cosmetics containing a microalgal membrane extract.   The cosmetic according to claim 9, wherein the microalgal membrane extract contains sulfoquinovosylglycerol or a salt thereof.   The microalgae membrane extract is a supernatant extract after intermediate temperature treatment obtained by subjecting microalgae to intermediate temperature treatment, followed by separation operation of a water-soluble fraction and a water-insoluble fraction. 9. Cosmetics according to 9.   The cosmetic according to any one of claims 9 to 11, wherein the microalga is a microalga belonging to the green alga class.   The cosmetic according to claim 12, wherein the microalgae belong to the genus Desmodemus.   The cosmetic according to any one of claims 8 to 13, which is a skin external preparation.