JP2020045289A - Beauty composition - Google Patents

Abstract

To provide a novel beauty composition that is derived from natural plant component and highly safe.SOLUTION: A beauty composition contains hydrophilic organic solvent extract of Rosa×alba steam distillation residue as an active ingredient.SELECTED DRAWING: Figure 1

Description

  The present invention relates to a cosmetic composition, and more particularly, to a cosmetic composition containing rose extract as an active ingredient.

  Oxygen, which is indispensable for living organisms, is partially incorporated into the body and then converted into reactive oxygen species, which are more reactive than normal oxygen. Active oxygen having high reactivity is used for energy production and biological defense systems. However, excessive accumulation of active oxygen in the body causes oxidative stress that causes biological function damage and disease as described below.

  Collagen is a structural protein that forms connective tissues such as animal skin. Skin collagen is produced by fibroblasts present in the dermis of the skin and is important for maintaining the structure of the skin, keeping the skin young and not wrinkling. It is known that oxidative stress such as ultraviolet rays reduces the amount of collagen and degrades the extracellular matrix.

  Hyaluronic acid is mainly produced by fibroblasts of the skin dermis and cells of other connective tissues, and is a main component of the extracellular matrix. Hyaluronic acid is present at a high concentration in synovial fluid and is involved in various cell-cell interactions, such as maintaining water and lubricating joints. Hyaluronic acid is also decomposed by oxidative stress such as ultraviolet rays.

  The crosslinked structure of collagen and elastin in the extracellular matrix plays an important role in maintaining skin elasticity. Elastase, an elastin-degrading enzyme, is induced by stimulation with ultraviolet light. When elastin is degraded, the crosslinked structure becomes brittle and the elasticity of the skin is lost.

  The melanin pigment present in human skin is a general term for a substance in which a phenolic substance is polymerized into a pigment. It is made by the reaction of tyrosine → dopa → dopaquinone → melanin pigment in melanocytes. The most important role of melanin pigment is UV protection. When exposed to ultraviolet light, tyrosinase is activated and the above reactions are promoted. Cells containing melanin gradually move to the upper layer of the stratum corneum, and the adhesion between cells is degraded by the action of lipid metabolizing enzymes such as lipase and proteolytic enzymes such as elastase, and finally the stratum corneum detaches and falls off. Melanin pigment suppresses sunlight damage and malignant tumors caused by ultraviolet rays, and causes skin spots and freckles.

  Living organisms need an antioxidant functional substance or mechanism to prevent excessive generation of reactive oxygen species (ROS) due to exposure to ultraviolet rays or the like, and to capture the generated ROS. There are also antioxidants such as vitamin E and vitamin C in foods. Currently, the search for new antioxidants is also underway.

  For example, Patent Document 1 proposes an antioxidant consisting of one or more plant extracts of jasmine, rose, vetiver, and laurel. The external preparation for skin containing the perfume composition containing the antioxidant can impart a percutaneous antioxidant effect to the function as a perfume. Patent Document 2 proposes a cosmetic composition containing a hybrid tea rose or an extract thereof. This cosmetic composition has a high anti-aging effect, especially a high collagenase activity inhibitory effect. Patent Document 3 proposes a cosmetic containing a solvent extract of damask rose. This cosmetic is a cosmetic containing an extract having an antioxidant action, an anti-inflammatory action, and a whitening action as an active ingredient, and has an effect of preventing and improving skin aging (wrinkles, tarmi, etc.), skin firmness, and gloss. It has the effect of improving skin, protecting skin from oxidative damage and inflammation due to external factors (ultraviolet rays and chemical substances), and preventing and improving spots, freckles, spots and the like. Patent Document 4 proposes an external preparation for skin containing β-endorphin, a β-endorphin production promoter, and an anti-inflammatory agent. The β-endorphin production promoter is selected from Terminaria extract, St. John's wort extract, Calendula officinalis extract, Giant kelp extract, or rose water. In this skin external preparation, the antioxidant effect is synergistically enhanced, the effect of the antioxidant is sufficiently exerted by adding a small amount, and the production of lipid peroxide caused by active oxygen production in the skin is suppressed, It has the effect of preventing or improving skin irritation and rough skin.

  When a state of high blood sugar concentration called glycation stress continues, a denatured protein called advanced glycation end products (AGE) is generated by a crosslinking reaction between the protein and saccharide. When AGE accumulates in the body, it promotes tissue aging. For example, AGEd collagen in the skin weakens skin elasticity and participates in wrinkle formation.

JP 2007-291031 (Antioxidant fragrance composition and a skin external preparation containing the same) JP 2011-236147 A (Cosmetic composition, food and drink composition and pharmaceutical composition containing hybrid tea rose or its extract) JP 2014-240375 A (cosmetics) JP-A-2006-8537 (External preparation for skin)

  An object of the present invention is to provide a novel and safe cosmetic composition derived from natural plant components.

  Rose, a plant belonging to the genus Rosaceae, is a plant that is historically familiar as a flower that symbolizes beauty. It is said that the number of registered rose varieties is 40,000 or more because rose has a large number of original varieties and has been crossed since ancient times. Above all, Alvarose, which is said to be the “father of white roses,” has a refreshing and elegant fragrance and has a relaxing effect. The present inventors have surprisingly discovered that the hydrophilic organic solvent extract of the steam distillation residue of Alvarose has various physiological activities useful for beauty and completed the present invention. That is, the present invention provides a cosmetic composition comprising, as an active ingredient, a hydrophilic organic solvent extract of the residue of steam distillation of Alvarose.

  The hydrophilic organic solvent is, for example, ethanol.

  The cosmetic composition of the present invention is preferably used to exhibit at least one of the following functions: antioxidant activity, skin collagen production, elastase inhibitory activity, tyrosinase inhibitory activity, and AGE formation inhibitory activity.

  The cosmetic composition of the present invention may further comprise cosmetic, dermatological and / or pharmaceutically acceptable additives.

The present invention also provides a step of subjecting Alvarose to steam distillation to obtain an Alvarose steam distillation residue and an Alvarose steam distillation residue, and subjecting the Alvarose steam distillation residue to solvent extraction using a hydrophilic organic solvent. Provided is a method for producing a cosmetic composition, comprising a step of obtaining a hydrophilic organic solvent extract of a distillation residue.

  The hydrophilic organic solvent is, for example, ethanol.

  The Alvarose distillation residue hydrophilic organic solvent extract, which is an active ingredient of the cosmetic composition of the present invention, has an antioxidant activity, a collagen production promotion, an elastase inhibitory activity, a tyrosinase inhibitory activity, as evidenced by the examples below. It shows an anti-photoaging effect such as suppression of melanin production and an anti-sugar aging effect such as an AGE formation inhibitory activity. Strong antioxidant activity is particularly useful for aging prevention and disease prevention. The collagen production promoting action is useful for preventing skin aging. Elastase activity inhibitory action and AGE (especially AGE-collagen) formation inhibitory action are useful for skin elasticity retention, tyrosinase activity inhibitory action and melanin production inhibitory action are whitening. Therefore, the cosmetic composition of the present invention containing the Alvarose distillation residue hydrophilic organic solvent extract is extremely useful for anti-aging cosmetics, UV cosmetics and the like.

  The distillation residue produced as a by-product after steam distillation of Alvarose is usually discarded, but the present invention makes it possible to effectively use the distillation residue.

It is the result of measuring the collagen-producing ability when adding Alvarose distillation residue ethanol extract (concentration: 10 to 400 μg / mL) to adult human dermal fibroblasts (NHDF-Ad). After culturing NHDF-Ad cells in EMEM medium supplemented with ethanol extract at a concentration of 10 μg / mL, 100 μg / mL or 400 μg / mL, the amount of collagen was measured by ELISA, and the cell viability was measured by MTT. . The collagen production ratio (%) and cell viability (%) of each test are shown with the collagen amount and cell viability of the control to which only DMSO was added as 100. In the figure, the polygonal line indicates the amount of collagen produced per cell. Addition of ethanol extract of Alvarose distillation residue at a concentration of 10 μg / mL or 100 μg / mL increased collagen production. Therefore, it was found that the ethanol extract of Alvarose distillation residue contains components that produce collagen and contribute to skin anti-aging. The results of measuring the amount of hyaluronic acid by ELISA in the above culture test are shown. No increase in the amount of hyaluronic acid was observed by addition of any concentration of the ethanol extract of Alvarose distillation residue. Therefore, it can be said that the ethanol extract of Alvarose distillation residue in these concentration ranges does not affect the hyaluronic acid-producing ability. The result of measuring the elastase inhibitory activity of the ethanol extract of Alvarose distillation residue (concentration: 10 to 400 μg / mL) is shown. About 40% of elastase inhibitory activity was observed in the ethanol extract of Alvarose distillation residue having a concentration of 100 μg / mL. From these results, it can be said that the ethanol extract of Alvarose distillation residue contains a component having a function of maintaining skin elasticity. The result of measuring the tyrosinase inhibitory activity of the ethanol extract of Alvarose distillation residue (concentration: 10 to 400 μg / mL) using L-DOPA as a substrate is shown. A concentration-dependent inhibitory effect on tyrosinase activity was observed in ethanol extracts of Alvarose distillation residue at concentrations of 100 μg / mL and 400 μg / mL. The result of measuring the tyrosinase inhibitory activity of the ethanol extract of Alvarose distillation residue (concentration: 10 to 400 μg / mL) using L-tyrosine as a substrate is shown. Inhibition of tyrosinase activity was observed by the addition of ethanol extracts of Alvarose distillation residue at concentrations of 100 μg / mL and 400 μg / mL. This effect is stronger when L-tyrosine is used as a substrate than when L-DOPA is used. The result of having measured the melanin production suppression effect of the ethanol extract of Alvarose distillation residue is shown. Addition of ethanol extracts of Alvarose distillation residue at concentrations of 10 μg / mL and 100 μg / mL reduced melanin production. The Alvarose distillation residue ethanol extract contains a component that inhibits the synthesis of melanin and promotes a whitening component. It is a figure which shows the AGE formation inhibitory activity by Alvarose distillation residue ethanol extract (400 microgram / mL-16 mg / mL density | concentration). In the ethanol extract of Alvarose distillation residue, a remarkable decrease in the amount of AGE production was observed in a concentration-dependent manner. Alvarose distilled ethanol extract had strong anti-AGE activity, suggesting the presence of substances that inhibit saccharification.

  Hereinafter, the present invention will be described in detail by showing one embodiment of the present invention. The cosmetic composition of the present invention contains a hydrophilic organic solvent extract of a steam distillation residue of Alvarose as an active ingredient.

  Alba Rose (scientific name Rosa Alba) is a strain of Old Rose and is said to be a natural cross between Rosa Damaskena and Rosa Kanina. The albarose raw material to be subjected to steam distillation may be flowers, petals, sepals, leaves, stems, seeds, roots, etc., preferably flowers, and particularly preferably petals. The steam distillation operation is based on a conventional method.

  The residue after distillation is solvent-extracted with a hydrophilic organic solvent. The distillation residue raw material subjected to the solvent extraction may be any of a water-containing residue itself, a frozen product, a dried product, a lyophilized product, and the like. Preferably, it is a freeze-dried product.

  Examples of the hydrophilic organic solvent include lower alcohols such as methanol, ethanol, propanol, butanol, glycerol, propylene glycol and 3-butylene glycol; ketones such as acetone and methyl ketone; ethers such as ethyl ether, isopropyl ether, diethyl ether and tetrahydrofuran. Carboxylic acid esters such as ethyl acetate, butyl acetate and methyl propionate. The hydrophilic organic solvents may be used alone or in combination of two or more. Preferably, it is ethanol.

  The temperature at the time of the solvent extraction may be usually from 20 to 28 ° C, and preferably 25 ° C. The extraction time may usually be 24 to 48 hours. It is preferable to use appropriate shaking means and stirring means at the time of extraction.

  The Alvarose distillation residue hydrophilic organic solvent extract is appropriately filtered and centrifuged to be purified. Further, as long as the effect of the cosmetic composition is not impaired, purification such as decolorization treatment and deodorization treatment may be performed.

  The purified Alvarose distillation residue hydrophilic organic solvent extract may be appropriately concentrated or dried by a concentration means such as evaporation or lyophilization. Further, the concentrated or dried extract may be dissolved again in a solvent such as water or alcohol.

  The Alvarose distillation residue hydrophilic organic solvent extract exhibits antioxidant activity, skin collagen production, elastase inhibitory activity, tyrosinase inhibitory activity, and AGE formation inhibitory activity, as described in Examples below. Therefore, the cosmetic composition of the present invention is preferably used to exhibit at least one of the above-mentioned effects.

  The cosmetic composition of the present invention may further comprise cosmetic, dermatological and / or pharmaceutically acceptable additives. Examples of additives include water; ethylene glycol, polyethylene glycol, propylene glycol, 1,3-butylene glycol, 1,4-butylene glycol, dipropylene glycol, glycerin, diglycerin, polyglycerin, pentylene glycol, isoprene glycol. Polyhydric alcohols such as glucose, maltose, fructose, xylitol, sorbitol, maltotriose, erythritol; lower alcohols such as methanol, ethanol, propyl alcohol, isopropyl alcohol, butyl alcohol and isobutyl alcohol; oleic acid, isostearic acid, lauric acid , Myristic, palmitic, stearic, behenic, undecylenic acids and other higher fatty acids; olive oil, corn oil, camellia oil, macaque Oils and fats such as annut oil, avocado oil, rapeseed oil, sesame oil, castor oil, safflower oil, cottonseed oil, jojoba oil, coconut oil, palm oil; waxes such as carnauba wax, candelilla wax, beeswax, lanolin; sorbitol, mannitol, Saccharides such as glucose, sucrose, lactose, trehalose and the like; methylpolysiloxane, methylphenylpolysiloxane, methylcyclopolysiloxane, octamethylcyclotetrasiloxane, octamethylcyclopentasiloxane, decamethylcyclopentasiloxane, methylhydrogenpolysiloxane, etc. Silicones; carrageenan, xanthan gum, gelatin, pectin, agarose, alginate, dextrin, methylcellulose, ethylcellulose, hydroxypropylcellulose, hydroxyethyl Thickeners such as cellulose, carboxymethylcellulose, carboxyvinyl polymer, polyvinyl alcohol, polyvinylpyrrolidone, gum arabic, karaya gum, tragacanth gum, tamarind gum; nonionic surfactants such as sodium lauroyl sulfate and polyoxyethylene sorbitan monooleate; alkyl Sulfate salts, anionic surfactants such as sodium normal dodecylbenzenesulfonate, and cationic surfactants such as polyoxyethylene dodecylmonomethylammonium chloride; phenoxyethanol, methylparaben, ethylparaben, propylparaben, butylparaben, paraoxybenzoate, Benzoic acid, salicylic acid and its salts, sorbic acid and its salts, dehydroacetic acid and its salts, chlorcresol, hexachloro Preservatives such as fen; minerals such as bentonite, smectite, beidellite, nontronite, saponite, hectorite, sauconite, and stevensite; bengala, iron oxide yellow, iron oxide black, cobalt oxide, ultramarine, navy blue, titanium oxide, oxide Inorganic pigments such as zinc; colorings such as red No. 202, yellow No. 4, blue No. 404; fragrances; balms; vitamins such as vitamin A, vitamin D, vitamin E, vitamin F, vitamin K, pyridoxine dicaprylate, Vitamin derivatives such as pyridoxine palmitate, ascorbyl dipalmitate, ascorbyl monopalmitate and ascorbyl monostearate; higher aliphatic hydrocarbons such as squalane, squalene and liquid paraffin; sphingolipids such as ceramide, cerebroside and sphingomyelin; Sterols such as phytosterols; antioxidants such as flavonoids, carotenoids, tannins, gallic acid, tocopherol, superoxide dismutase (SOD), thioredoxin, thioredoxin reductase, butylhydroxytoluene, and butylhydroxyanisole; Inflammatory agents; ultraviolet absorbers such as para-aminobenzoic acid, para-aminobenzoic acid monoglycerin ester, methyl anthranilate, homomenthyl-N-acetylanthranilate, octyl para-methoxycinnamate, ethyl-4-isopropylcinnamate; t-cyclo Amino acid derivatives, kojic acid, ascorbic acid, hydroquinone, ellagic acid, nicotinic acid, resorcinol, tranexamic acid, potassium 4-methoxysalicylate, mag Whitening agents such as lignan, 4-HPB, hydroxybenzoic acid, vitamin E, α-hydroxy acid, AMP; 1,3-butylene glycol (BG), dipropylene glycol (DPG), betaine, trehalose, sodium hyaluronate, water-soluble And moisturizing agents such as soluble collagen and urea.

  Specific uses of the beauty composition of the present invention include skin care cosmetics such as suntan cosmetics, essences, moisturizing lotions, flexible lotions, and astringent lotions; body cosmetics such as firming cosmetics and anti-cellulite cosmetics; basic cosmetics (cosmetics) Water, emulsion, cream), foundation, base cream, face powder, waterproof, and other makeup cosmetics; baths such as soaps, shampoos, rinses, and bath additives; hair preparations;

  The form of the cosmetic composition of the present invention may be a solution, a solution, an aerosol, an emulsion, an emulsion, an emulsion, a lotion, a cream, an ointment, a powder, or a solid, depending on the use. From the viewpoint of immediate effect, it is processed into solutions, solutions, aerosols, lotions, emulsions, emulsions, emulsions and the like.

  The blending amount of the Alvarose distillation residue hydrophilic organic solvent extract in the beauty composition of the present invention is appropriately determined according to the purpose of use, and is usually 0.0001 to 0.5 mass as a solid content of the extract. %, Preferably 0.0001 to 0.3% by mass, more preferably 0.001 to 0.1% by mass.

1 shows a formulation example of the cosmetic composition of the present invention. However, the present invention is not limited to the following formulation examples.
[Formulation Example 1] Lotion

[Formulation Example 2] Emulsion

[Prescription Example 3] Cream

[Formulation Example 4] Bath agent

  The present invention also provides a method for producing the cosmetic composition. The production method comprises subjecting the Alvarose to steam distillation to obtain an Alvarose steam distillation residue and an Alvarose steam distillation residue, and subjecting the Alvarose steam distillation residue to solvent extraction using a hydrophilic organic solvent. Obtaining a hydrophilic organic solvent extract of the distillation residue. The description of each step is as described above.

  Hereinafter, the present invention will be described in more detail by showing examples of the cosmetic composition of the present invention. However, the present invention is not limited to the following examples.

(1) Preparation of Ethanol Extract of Alvarose Steam Distillation Residue The raw material comprising the flower part of Alvarose was subjected to steam distillation to obtain an Alvarose steam distillate and an Alvarose distillation residue. The moisture content of the Alvarose distillation residue was 86.7%.

  The residue of Alvarose steam distillation was dried with a freeze dryer, and the dried product was subjected to ethanol extraction. The ethanol extraction was performed at room temperature for 24 hours while shaking at a stirring speed of 200 rpm. The ethanol extract was filtered, concentrated at a temperature of 45 ° C. with a rotary evaporator, and then dried in a desiccator. This dried product is referred to as Alvarose distillation residue ethanol extract. The ethanol extraction ratio of the ethanol extract of Alvarose distillation residue (= weight of dried product / weight of raw material used for extraction) was 29.9%.

(2) Antioxidant activity test The superoxide elimination activity of the ethanol extract of Alvarose distillation residue obtained above was measured by the ORAC (Oxygen Radical Absorbance Capacity) method. The ORAC method is a method for evaluating the ability of an antioxidant to delay the quenching speed of a fluorescent reagent by active oxygen as the antioxidant ability of the antioxidant. For example, the fluorescent reagent fluorescein in the sample is oxidized to the active oxygen peroxyl radical generated by the oxidizing substance AAPH (2,2′-azobis (2-aminopropane) dihydrochloride). If antioxidants are present in the sample, the quenching speed of fluorescein is slowed down because the antioxidants scavenge peroxyl radicals.

  1.8 mL of a 48 nM fluorescein solution was added to 0.3 mL of the above-mentioned ethanol extract of Alvarose distillation residue (pH 7.4), and then 0.9 mL of 43 mM AAPH (pH 7.4) was added. After vortexing this mixture for 10 seconds, the fluorescence intensity (excitation wavelength: 485 nm, measurement wavelength: 520 nm) was measured with a spectrofluorometer (product name: FlexStation 3, manufactured by Molecular Devices).

Superoxide scavenging activity was calculated from the obtained fluorescence intensity (n = 4). Table 5 shows the results. The superoxide scavenging activity is a value obtained by expressing the antioxidant activity per 1 mg of sample as Trolox (a vitamin E derivative; an antioxidant used as an index) equivalent. The larger the value, the higher the antioxidant activity. Show.

  As shown in Table 5, the superoxide scavenging activity of the ethanol extract of the Alvarose steam distillation residue was 1.67 mgTE / mg. That is, the above ethanol extract showed high antioxidant activity about 1.67 times that of the antioxidant Trolox.

(3) Collagen / hyaluronic acid production test The ability to produce collagen and hyaluronic acid when the above Alvarose distillation residue ethanol extract was added to a fibroblast medium was measured. Specifically, adult human dermal fibroblasts (NHDF-Ad, 1.0 × 10 ⁴ cells / well) as model cells were added with 2 μL of an ethanol extract having a concentration of 10 μg / mL, 100 μg / mL or 400 μg / mL. The cells were cultured at 37 ° C. for 72 hours in the prepared EMEM medium (998 μL) in the presence of 5% CO ₂ + 95% air. After the culture, the amounts of collagen and hyaluronic acid were measured by ELISA, and the cell viability was measured by MTT. Ascorbic acid (100 μg / mL) was used as a positive control for collagen production test, and N-acetylglucosamine (100 μg / mL) was added as a positive control for hyaluronic acid test to fibroblast medium. .

  Assuming that the amount of collagen, the amount of hyaluronic acid, and the cell viability of the control to which only DMSO was added were 100, the collagen production ratio (%) and the hyaluronic acid production ratio (%) of each test are shown in FIGS. n = 3). 1 and 2, the broken lines indicate the amount of collagen production and the amount of hyaluronic acid production per cell, respectively.

  Looking at the collagen production in FIG. 1, the addition of the ethanol extract of Alvarose distillation residue at a concentration of 10 μg / mL or 100 μg / mL significantly increased the collagen production. Therefore, it was found that the ethanol extract of Alvarose distillation residue contains components that produce collagen and contribute to skin anti-aging. From this, the use of the above-mentioned ethanol extract of Alvarose distillation residue is expected to exert effects such as skin moisture retention effect and joint pain improvement effect.

  On the other hand, looking at the amount of hyaluronic acid produced in FIG. 2, no increase in the amount of hyaluronic acid was recognized by addition of any concentration of the ethanol extract of Alvarose distillation residue. Therefore, it was found that the ethanol extract of Alvarose distillation residue in these concentration ranges did not affect the hyaluronic acid performance.

(4) Elastase inhibitory activity test The elastase inhibitory activity of the ethanol extract of Alvarose distillation residue was measured. Specifically, an adult human dermal fibroblast (NHDF-Ad, 1.2 × 10 ⁶ cell / well) as a model cell was dissolved in a lysis buffer [0.2 M Tris-HCl (pH 8.0), 1 mM PMSF, 0 mM [1.5% Triton X-100] was added thereto, sonicated in ice water for 2 minutes, and left at room temperature for 10 minutes. The lysate was centrifuged at 400 g x 15 minutes at 4 ° C. To 25 µL of the supernatant after centrifugation, 23 µL of 0.2 M Tris-HCl (pH 8.0) and 2 µL of the ethanol extract of the above-described Alvarose distillation residue having a concentration of 10 µg / mL, 100 µg / mL or 400 µg / mL were added. Leave at temperature for 15 minutes. Next, 50 μL of a 5 mM substrate (Suc-Ala-Ala-Ala-pNA) was added and left at 37 ° C. for 2 hours. Thereafter, the absorbance at 406 nm was measured using an absorptiometer (product name ELx800, manufactured by Bio Teck). The measurement was also performed in a system to which 0.04 mM phosphoramidone was added as an elastase activity inhibitor (positive control).

  The absorbance ratio (elastase activity) of each test is shown in FIG. 3 with the absorbance of the control to which only DMSO was added as 100 (n = 3). In addition, in the ethanol extract of Alvarose distillation residue having a concentration of 400 μg / mL, elastase activity could not be measured due to coloring of the sample itself.

  As shown in FIG. 3, the elastase inhibitory activity of about 40% was recognized in the ethanol extract of Alvarose distillation residue having a concentration of 100 μg / mL. This result suggests that the ethanol extract of Alvarose distillation residue contains a component having a function of maintaining skin elasticity. Therefore, the above-mentioned ethanol extract of Alvarose distillation residue is expected to be used for exerting a function of maintaining skin elasticity.

(5) Tyrosinase inhibitory activity test The tyrosinase inhibitory activity of the above-mentioned ethanol extract of Alvarose distillation residue was evaluated. Specifically, L-DOPA (3- (3,4-dihydroxyphenyl) -L-alanine) or L-tyrosine was added in the presence of an ethanol extract at a concentration of 10 μg / mL, 100 μg / mL or 400 μg / mL. The absorbance at 475 nm, which indicates the amount of DOPA chromium produced by mushroom tyrosinase as a substrate, was measured with the absorptiometer. As a positive control, the absorbance of the system to which kojic acid (3.3 μg / mL) as an inhibitor was added was also measured.

  Tyrosinase activity (%) of each system is shown in FIG. 4 for L-DOPA, and FIG. 5 for L-tyrosine (n = 3), assuming that the absorbance of the control containing only DMSO is 100.

  When L-DOPA of FIG. 4 was used as a substrate, concentration-dependent tyrosinase activity inhibitory effects were observed in ethanol extracts of Alvarose distillation residue at concentrations of 100 μg / mL and 400 μg / mL.

  Even when L-tyrosine in FIG. 5 was used as a substrate, a concentration-dependent inhibitory effect on tyrosinase activity was observed by adding ethanol extracts of Alvarose distillation residue at concentrations of 100 μg / mL and 400 μg / mL. This effect was stronger when L-tyrosine was used as the substrate than L-DOPA.

  From the above results, it was suggested that the ethanol extract of Alvarose distillation residue contains a component that inhibits tyrosinase and prevents melanin fixation, and that a whitening effect can be expected.

(6) Melanin production inhibition test The melanin production inhibition effect of the ethanol extract of Alvarose distillation residue was evaluated. Specifically, human melanoma cells ^{(G361,10 5 cells / mL, 1mL} / well) was used as a model cell concentration 10μg / mL, 100μg / mL or 400 [mu] g / mL of Alba Rose vinasse ethanol extract 2μL additives The cells were cultured at 37 ° C. for 72 hours in the prepared EMEM medium (998 μL) in the presence of 5% CO ₂ + 95% air. After 1 mL of 1M NaOH was added to a part of the culture solution and stored at room temperature overnight (4 hours) under light shielding, the amount of melanin generated in the culture solution was determined from the absorbance of the plate reader at 405 nm. Further, 50 μL of the MTT staining solution was added to the remainder of the above culture solution, left at 37 ° C. for 4 hours in the presence of 5% CO ₂ + 95% air, and then 1 mL of HCL-isopropanol was added thereto. Time), and after preservation from light, the cell viability was determined from the absorbance of the plate reader at 570 nm. As a positive control, 100 μg / mL of arbutin exhibiting melanin production inhibitory activity was added in place of the above-mentioned ethanol extract of Alvarose distillation residue, and the amount of melanin produced in the system was also measured.

  The melanin production (absorbance) and cell viability (absorbance) of the control to which 50% BG, 50% ethanol, jojoba oil and DMSO were added were taken as 100, and the melanin production ratio (%) and cell viability (%) of each test system were determined. ). FIG. 6 shows the results. In the figure, the polygonal line indicates the amount of melanin produced per cell.

  FIG. 6 shows that the addition of the ethanol extracts of Alvarose distillation residue at concentrations of 100 μg / mL and 400 μg / mL reduced melanin production comparable to that of the positive control arbutin. Therefore, it was suggested that the ethanol extract of Alvarose distillation residue contains components that inhibit the synthesis of melanin and promote whitening components.

(7) AGE formation inhibitory activity test Whether or not AGE was inhibited by coexistence of ethanol extract of Alvarose distillation residue in an in vitro test system for forming AGE was evaluated by the following procedure.

(Preparation of reaction solution)
BSA was dissolved in PBS to prepare a 20 mg / mL BSA solution. Glyceryl aldehyde was dissolved in PBS to prepare a 45 mg / mL glyceryl aldehyde solution. A DMSO solution of the ethanol extract was mixed at 5% (v / v) in ultrapure water. A 5% (v / v) DMSO solution was prepared as a negative control, and a 5% (v / v) DMSO solution mixed with 2.2 mg / mL aminoguanidine was prepared as a positive control.

  To each well of a 96-well plate, 50 μL of the BSA solution, 10 μL of the glyceryl aldehyde solution, and 40 μL of a negative control, a positive control, or an ethanol extract (concentration: 400 μg / mL, 4 mg / mL, or 16 mg / mL) were added and mixed. . The reaction time was the 18 hour period required for the AGE formation reaction to occur sufficiently in the test performed beforehand.

(Measurement of AGE formation amount)
The amount of AGE formed in the reaction solution was measured from the fluorescence intensity (Ex: 360 nm, Em: 465 nm) of the reaction solution using the spectrofluorometer (n = 3). The fluorescence intensities of the reaction solution at the 0th and 18th hours after the reaction were expressed as values where the fluorescence intensity at the 0th hour of the reaction of the negative control was set to 1. FIG. 7 shows the results.

  As shown in FIG. 7, after 18 hours, a remarkable decrease in the amount of AGE production was observed in the ethanol extract of Alvarose distillation residue in a concentration-dependent manner. Moreover, the AGE production was significantly lower than that of the positive control aminoguanidine. Therefore, the ethanol extract of Alvarose distilled has a strong AGE inhibitory activity and is expected to be applied to a cosmetic composition exhibiting an anti-glucose aging effect.

Claims (6)

  A cosmetic composition comprising, as an active ingredient, a hydrophilic organic solvent extract of Alvarose steam distillation residue.   The cosmetic composition according to claim 1, wherein the hydrophilic organic solvent is ethanol.   The cosmetic composition according to claim 1 or 2, wherein the cosmetic composition exhibits at least one function of antioxidant activity, skin collagen production, elastase inhibitory activity, tyrosinase inhibitory activity, and AGE formation inhibitory activity.   The cosmetic composition according to any one of claims 1 to 3, further comprising a cosmetically, dermatologically and / or pharmaceutically acceptable additive. Subjecting the Alvarose to steam distillation to obtain an Alvarose steam distillation residue and an Alvarose steam distillation residue, and subjecting the Alvarose steam distillation residue to solvent extraction using a hydrophilic organic solvent to remove the Alvarose steam distillation residue from the hydrophilic organic solvent. A method for producing a cosmetic composition, comprising a step of obtaining a solvent extract.   The method for producing a cosmetic composition according to claim 5, wherein the hydrophilic organic solvent is ethanol.