KR20210047588A - Skin-lightening Composition Using an Extract of Xanthium canadense - Google Patents

Abstract

The present invention relates to a composition for skin whitening using a Xanthium canadense extract. The composition inhibits melanin synthesis in B16F10, a mouse-derived melanoma cell line, and has activity of inhibit the expression to tyrosinase and TRP-1 involved in melanin synthesis.

Description

Skin-lightening Composition Using an Extract of Xanthium canadense}

The present invention relates to a composition for skin whitening using the Xanthium canadense extract.

As the standard of living in the modern world improves, interest in beauty as well as health is gradually increasing. Among them, product development for improving skin tone and pigmentation-type skin diseases such as melanoma and freckle caused by skin damage and aging caused by ultraviolet rays and air pollution is actively developed.

Melanin, which is present in the base layer of the epidermis, is a major pigment that determines the color of the skin. It protects the skin by preventing damage caused by ultraviolet rays and removing free radicals. However, when melanin is locally overproduced, it causes skin diseases such as spots, freckles, black spots, and skin cancer. In addition, if the melanin that is continuously overproduced stays in the dermis, it induces the desire for whitening because it turns the skin color black or the complexion dull (Pigment Cell Res. 2007 Feb;20(1):2-13.; Curr Biol. 2011 Nov 22;21(22):1906-11.;Ann Dermatol Venereol.2012 Nov;139 Suppl 3:S73-7).

Melanin is a phenolic polymer natural pigment that is widely present in living organisms. In the human body, it is synthesized from melanocytes in melanocytes in the basement layer of the epidermis, and is transferred to surrounding keratinocytes to represent human skin color. Abnormally low levels of melanin cause skin lesions such as vitiligo. Conversely, when excessively produced, acquired hyperpigmentation such as melanoderma, postinflammatory melanoderma, solar lentigo, etc. Various skin diseases may be caused, including.

The synthesis of melanin is made by making melanosomes from melanocytes in the base layer of the skin, and a series of processes by which melanin is synthesized is collectively referred to as melanogenesis. Melanin production is based on tyrosine, one of the amino acids, and DOPA (3, tyrosinase related protein-1), TRP-2 (tyrosinase related protein-2) 4-dihydroxyphenylalanine) to dopaquinone (DOPA quinone), a non-enzymatic reaction, spontaneous oxidation, and then a polymerization reaction with amino acids or proteins to synthesize melanin (Korean J. FOOD SCI. TECHNOL., 2000. 32(3):736; Journal of Life Science, 2013. 23(12):1445; Pigment Cell Res. 1999. 12(1):4-12.). Recently, studies on inhibition of melanin synthesis have been conducted using proteins such as tyrosinase, TRP-1, and TRP-2 as well as microphtalmia associated transcription factor (MITF). The major intracellular signaling pathway is the cAMP/PKA (cyclic monophosphate/protein kinase A) pathway, where cAMP promotes the expression of MITF via PKA and CREB1 (cAMP responsive element binding protein 1), and MITF is important in the melanin synthesis process. It promotes transcription of tyrosinase, TRP-1, and TRP-2 as transcriptional regulators (Pharmazie. 2015 Oct;70(10):646-9.; Int J Mol Sci. 2015 Oct 13;16(10): 24219-42; Curr Pharm Biotechnol. 2015;16(12):1111-9.).

Since tyrosinase plays the most important role as the first step to produce melanin, research on whitening agents is mainly a tyrosinase activity inhibitor or an antagonist against the substrate of tyrosinase to regulate tyrosinase activity. Research has been conducted in the direction of developing (Kor. J. Pharmacogn., 44(3):220, 2013).

Arbutin, which is currently most commonly used as a whitening agent, is an antagonist of tyrosine, and kojic acid inhibits the process from tyrosine to dopa and dopaquinone by chelating the active site of tyrosinase (Korean J. Plant Res., 26(5):526, 2013; J. Korean Soc. Food Sci. Nutr., 41(2):156, 2012). Arbutin and kojic acid are known to have a strong whitening effect, but due to some side effects such as skin irritation, research has been actively conducted to find a material having whitening activity from natural materials with relatively few side effects.

The present invention discloses the whitening activity of the extract of the Big Dogwood.

It is an object of the present invention to provide a composition for skin whitening using an extract of a large dorado.

Other or specific objects of the present invention will be presented below.

The present invention, as confirmed in the Examples and Experimental Examples below, when treated with the extract of chinensis B16F10, a mouse-derived melanoma cell line, inhibits melanin production in a concentration-dependent manner, and also expresses tyrosinase and TRP-1. It is completed by confirming that it is suppressed.

In consideration of the foregoing, the composition for skin whitening of the present invention is characterized in that it contains the extract of the Great Tokomari as an active ingredient.

In the present specification, the term "big dorado extract" refers to the extraction target of daikon daikon, leaves, stems, fruits, above-ground parts, roots, underground parts, or a mixture thereof with water, lower alcohols having 1 to 4 carbon atoms (methanol, ethanol, butanol, etc.), Methylene chloride, ethylene, acetone, hexane, ether, chloroform, ethyl acetate, butyl acetate, N,N-dimethylformamide (DMF), dimethyl sulfoxide (DMSO), 1,3-butylene glycol, propylene glycol, or their It refers to an extract obtained by leaching using a mixed solvent, an extract obtained using a supercritical extraction solvent such as carbon dioxide, pentane, or a fraction obtained by fractionating the extract, and the extraction method considers the polarity of the active substance, the degree of extraction, and the degree of preservation. Therefore, arbitrary methods such as cold settling, reflux, warming, ultrasonic radiation, and supercritical extraction can be applied. In the case of a fractionated extract, a fraction obtained by suspending the extract in a specific solvent and then mixing and policing it with a solvent having a different polarity, the crude extract is adsorbed on a column filled with silica gel, etc., and then a hydrophobic solvent, a hydrophilic solvent, or a mixed solvent thereof is added. It means including the fraction obtained as a mobile phase. In addition, the meaning of the extract includes a concentrated liquid extract or a solid extract from which the extraction solvent has been removed by a method such as freeze drying, vacuum drying, hot air drying, spray drying, or the like. Preferably, it means an extract obtained by using water, ethanol or a mixed solvent thereof as an extraction solvent, in particular, an extract using a mixed solvent of water and ethanol as an extraction solvent.

In addition, in the present specification, the term "active ingredient" refers to an ingredient that can exhibit a desired activity alone or exhibit activity with a carrier that is not itself active.

The composition of the present invention may contain the active ingredient in an arbitrary amount (effective amount) as long as it can exhibit skin whitening activity, etc., depending on the product, formulation, etc., and the usual effective amount is 0.001% by weight based on the total weight of the composition. To 15% by weight. Here, the term "effective amount" refers to intended medical and dermatological effects such as skin whitening effect when the composition of the present invention is administered to a mammal, preferably a human, during the administration period according to the recommendations of a medical expert, a skin expert, etc. It refers to the amount of the active ingredient contained in the composition of the present invention that can be represented. Such effective amounts can be determined empirically within the range of ordinary skill in the art.

In addition to the active ingredients, the composition of the present invention has similar activities such as skin whitening activity, skin wrinkle improvement activity, skin irritability suppression activity, skin protection activity (skin damage suppression by ultraviolet rays, skin moisturizing, etc.), etc. In order to enhance the convenience of administration, ingestion, and use through addition, any compounds or natural extracts that have already been tested for safety in the art and are known to have the corresponding activity may be additionally included.

These compounds or extracts include each country's Pharmacopoeia (“Korean Pharmacopoeia” in Korea), each country's health functional food code (in Korea, it is “health functional food standards and standards”), and each country's functional cosmetics code (in Korea, Compounds or extracts licensed in accordance with the laws of each country (“Pharmaceutical Affairs Act” in Korea) governing the manufacture and sale of compounds or extracts and pharmaceuticals listed in the fair documents such as "Functional Cosmetics Standards and Test Methods"), health In accordance with the laws of each country governing the manufacture and sale of functional foods (in Korea, it is the 「Act on Health Functional Foods」), national laws governing the manufacture and sale of compounds, extracts, and functional cosmetics with recognized functionality (in Korea, the 「Cosmetics Act」 ”), the functional compounds or extracts are included. For example, arbutin, niacinamide, ascorbyl glucoside, alpha-bisabolol, and oil-soluble licorice extract (Oil Soluble Licorice(Glycyrrhiza) Extract) are mentioned as skin whitening ingredients, and retinol, retinyl palmitate as skin wrinkle improvement ingredients. , Adenosine, polyethoxylated amide, and the like, and examples of the UV protection component include dromethrizol, dromethrizoltrisiloxane, digaloyltrioleate, dimethicodiethylbenzalmalonate, and diethylhexylbutamido. And complex extracts such as triazone and pine bark extract, phosphatidylserine, fingerroot extract powder, and complex extracts such as red ginseng and cornus milk. In addition, as a moisturizing ingredient, AP collagen enzyme decomposition peptide, collactive collagen peptide, N-acetylglucosamine, konjac potato extract, complex extract such as dandelion, rice bran extract, corn germ extract, low molecular weight collagen peptide, ground grass extract powder, phosphatidylserine, hyaluronic acid, etc. Examples of functional ingredients for'improving irritable skin condition' include L. sakei Probio 65, oils containing gam- marinolenic acid, L. plantarum CJLP133, a lactic acid bacteria derived from fruits and vegetables, and probiotics ATP. One or more of these compounds or natural extracts may be included in the composition of the present invention together with the active ingredient.

The composition for skin whitening of the present invention can be understood as a cosmetic composition in a specific embodiment.

When the composition of the present invention is identified as a cosmetic composition, the cosmetic composition may be classified as a product for its use or legally, and specifically, may be a functional cosmetic having a use such as skin whitening, a non-functional general cosmetic, and the like. The product form can also take any product form, specifically solutions, suspensions, emulsions, pastes, gels, creams, lotions, powders, soaps, surfactant-containing cleansing, oils, powder foundations, emulsion foundations, waxes. It can be formulated as a foundation, spray, or the like. In a specific product form, it may be a flexible lotion, a nutritional lotion, a nutritional cream, a massage cream, an essence, an eye cream, a cleansing cream, a cleansing foam, a cleansing water, a pack, a spray, or a powder formulation.

In addition to the active ingredient, the cosmetic composition of the present invention may include components commonly used in the cosmetic composition, such as stabilizers, solubilizers, surfactants, vitamins, conventional adjuvants such as pigments and perfumes, and carriers.

When the formulation of the present invention is a paste, cream or gel, animal oil, vegetable oil, wax, paraffin, starch, tracanth, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc, or zinc oxide may be used as carrier components. I can.

When the formulation of the present invention is a powder or spray, lactose, talc, silica, aluminum hydroxide, calcium silicate, or polyamide powder may be used as a carrier component. In particular, in the case of a spray, additionally chlorofluorohydrocarbon, propane / May contain propellants such as butane or dimethyl ether.

When the formulation of the present invention is a solution or emulsion, a solvent, a solubilizing agent or an emulsifying agent is used as a carrier component, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1 ,3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol, fatty acid ester of sorbitan, and the like can be used.

When the formulation of the present invention is a suspension, as a carrier component, a liquid diluent such as water, ethanol or propylene glycol, an ethoxylated isostearyl alcohol, a suspending agent such as polyoxyethylene sorbitol ester, polyoxyethylene sorbitan ester, and crystallites Castle cellulose, aluminum metahydroxide, bentonite, agar, and the like can be used.

When the formulation of the present invention is a surfactant containing cleansing, as a carrier component, aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyltaurate, sarcosinate, fatty acid amide Ether sulfates, alkylamidobetaines, fatty alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, lanolin derivatives or ethoxylated glycerol fatty acid esters may be used.

The cosmetic composition of the present invention can be prepared according to a method for preparing a cosmetic composition conventionally performed in the art, except for including the active ingredient that exhibits skin whitening activity.

In a specific aspect, the composition of this invention can be grasped as a food composition.

The food composition of the present invention may be prepared in any form, such as beverages such as tea, juice, carbonated drinks, ion drinks, processed oils such as milk, yogurt, gums, rice cakes, Korean sweets, bread, snacks, noodles, etc. Foods, tablets, capsules, pills, granules, liquids, powders, flakes, pastes, syrup, gels, jelly, can be prepared in health functional food preparations such as bars.

In addition, the food composition of the present invention can be classified as a product as long as it conforms to the enforcement regulations at the time of manufacture and distribution in terms of legal and functional classification. For example, it is a health functional food according to the Korean 「Health Functional Food Act」, or confectionery, bean, tea, and beverages according to each food type according to the food code of the Korean 「Food Sanitation Act」 (``Food Standards and Standards'' notified by the Ministry of Food and Drug Safety). , Special use food, etc.

The food composition of the present invention may contain food additives in addition to the active ingredients. Food additives can generally be understood as substances that are added to food and mixed or infiltrated in manufacturing, processing, or preserving food. Since they are consumed daily and for a long time with food, their safety must be ensured. Food additives with guaranteed safety are limited in terms of ingredients or functions in the Food Additives Code according to the laws of each country governing the manufacture and distribution of food (the “Food Sanitation Act” in Korea). In the Korean Food Additives Code (“Food Additive Standards and Standards” notified by the Ministry of Food and Drug Safety), food additives are classified into chemical synthetic products, natural additives, and mixed preparations in terms of ingredients.These food additives are sweeteners and flavors in terms of function. It is categorized into agents, preservatives, emulsifiers, acidulants, and thickeners.

Sweeteners are used to impart an appropriate sweet taste to foods, and both natural and synthetic can be used in the composition of the present invention. Preferably, a natural sweetener is used, and examples of the natural sweetener include sugar sweeteners such as corn syrup solids, honey, sucrose, fructose, lactose, and maltose.

Flavoring agents can be used to enhance taste or aroma, and both natural and synthetic can be used. Preferably, it is the case of using a natural one. In the case of using natural ones, the purpose of nutrient enhancement can be combined in addition to flavor. As a natural flavoring agent, it may be obtained from apples, lemons, tangerines, grapes, strawberries, peaches, or the like, or from green tea leaves, roundtails, bamboo leaves, cinnamon, chrysanthemum leaves, jasmine, and the like. In addition, you can use those obtained from ginseng (red ginseng), bamboo shoots, aloe vera, and ginkgo. The natural flavoring agent may be a liquid concentrate or a solid extract. In some cases, synthetic flavoring agents may be used, and synthetic flavoring agents may include esters, alcohols, aldehydes, terpenes, and the like.

As a preservative, sodium calcium sorbate, sodium sorbate, potassium sorbate, calcium benzoate, sodium benzoate, potassium benzoate, EDTA (ethylenediaminetetraacetic acid), etc. can be used, and as an emulsifier, acacia gum, carboxymethylcellulose, xanthan gum, Pectin, etc. may be mentioned, and as the acidulant, arithmetic, malic acid, fumaric acid, adipic acid, phosphoric acid, gluconic acid, tartaric acid, ascorbic acid, acetic acid, phosphoric acid, and the like can be used. In addition to the purpose of enhancing taste, the acidulant may be added so that the food composition has an appropriate acidity for the purpose of inhibiting the growth of microorganisms.

As the thickening agent, a suspending agent, a settling agent, a gel-forming agent, a swelling agent, and the like may be used.

In addition to the food additives described above, the food composition of the present invention may include a physiologically active substance or minerals known in the art for the purpose of supplementing and reinforcing functionality and nutritional properties and ensuring stability as a food additive.

Examples of such bioactive substances include catechins contained in green tea, vitamins such as vitamin B1, vitamin C, vitamin E, and vitamin B12, tocopherol, dibenzoyl thiamine, and the like, and minerals include calcium preparations such as calcium citrate, magnesium stearate. Magnesium preparations such as, iron preparations such as iron citrate, chromium chloride, potassium iodide, selenium, germanium, vanadium, and zinc.

In the food composition of the present invention, the food additive described above may be included in an appropriate amount to achieve the purpose of addition according to the product type.

Regarding other food additives that may be included in the food composition of the present invention, reference may be made to the food code of each country or the food additive code.

The composition for skin whitening of the present invention can be understood as a pharmaceutical composition in a specific embodiment.

When the composition for skin whitening of the present invention is identified as a pharmaceutical composition, the target disease can be understood as skin hyperpigmentation, and specific examples of such hyperpigmentation include freckles, senile spots, liver spots, spots, brown or black spots, sun pigments. Plaque, blue melanoma, hyperpigmentation after use of drugs such as calcium antagonists, sequelae of hyperpigmentation according to tissue sclerotherapy, gestational brown spots, black skin in women taking oral contraceptives, wounds or dermatitis including abrasions and burns Hyperpigmentation after inflammation caused by, phototoxic reactions, or other similar small fixed pigmented lesions may be included, but are not limited thereto.

The pharmaceutical composition of the present invention may be prepared in an oral dosage form or a parenteral dosage form according to an administration route by a conventional method known in the art, including a pharmaceutically acceptable carrier in addition to the active ingredient. Here, the route of administration may be any suitable route including a local route, an oral route, an intravenous route, an intramuscular route, and direct absorption through mucosal tissue, and two or more routes may be used in combination. An example of a combination of two or more routes is a case in which two or more formulations of drugs are combined according to the route of administration, for example, when one drug is first administered by an intravenous route and the second drug is administered by a local route.

Pharmaceutically acceptable carriers are well known in the art depending on the route of administration or formulation, and specifically, reference may be made to the pharmacopoeia of each country, including the “Korean Pharmacopoeia”.

When the pharmaceutical composition of the present invention is prepared in an oral dosage form, powders, granules, tablets, pills, dragees, capsules, solutions, gels, syrups, suspensions, wafers according to a method known in the art together with a suitable carrier It can be prepared in a formulation such as. Examples of suitable carriers at this time include sugars such as lactose, glucose, sucrose, dextrose, sorbitol, mannitol, and xylitol, starches such as corn starch, potato starch, wheat starch, cellulose, methylcellulose, ethylcellulose, sodium carboxymethylcellulose, Celluloses such as hydroxypropylmethylcellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, magnesium stearate, mineral oil, malt, gelatin, talc, polyol, vegetable oil, ethanol, grease Serol, etc. are mentioned. In the case of formulation, appropriate binders, lubricants, disintegrants, colorants, and diluents may be included as needed. Suitable binders include starch, magnesium aluminum silicate, starch ferryst, gelatin, methylcellulose, sodium carboxymethylcellulose, polyvinylpyrrolidone, glucose, corn sweetener, sodium alginate, polyethylene glycol, wax, etc., and lubricants include oleic acid. Sodium, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride, silica, talcum, stearic acid, magnesium salts and calcium salts thereof, polydecylene glycol, etc., and as disintegrating agents starch, methyl cellulose , Agar, bentonite, xanthan gum, starch, alginic acid, or a sodium salt thereof. Moreover, lactose, dextrose, sucrose, mannitol, sorbitol, cellulose, glycine, etc. are mentioned as a diluent.

When the pharmaceutical composition of the present invention is prepared in a parenteral formulation, it may be formulated in the form of an injection, a transdermal administration, a nasal inhalation and a suppository according to a method known in the art together with a suitable carrier. When formulated as an injection, an aqueous isotonic solution or suspension may be used as a suitable carrier, and specifically, triethanol amine-containing PBS (phosphate buffered saline), sterile water for injection, an isotonic solution such as 5% dextrose, etc. may be used. . When formulated as a transdermal administration, it can be formulated in the form of an ointment, cream, lotion, gel, external solution, pasta, liniment, air roll, and the like. In the case of nasal inhalants, suitable propellants such as dichlorofluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, and carbon dioxide can be used in the form of an aerosol spray.When formulated as a suppository, the carrier is Withepsol ( witepsol), tween 61, polyethylene glycols, cacao butter, laurin paper, polyoxyethylene sorbitan fatty acid esters, polyoxyethylene stearates, sorbitan fatty acid esters, and the like.

The specific formulation of pharmaceutical compositions is known in the art, and for example, Remington's Pharmaceutical Sciences (19th ed., 1995) may be referred to. This document is considered as part of this specification.

The preferred dosage of the pharmaceutical composition of the present invention is in the range of 0.001 mg/kg to 10 g/kg per day, preferably 0.001 mg/kg to 1 g, depending on the patient's condition, weight, sex, age, patient severity, and route of administration. May be in the /kg range. Administration can be made once a day or divided into several times. Such dosage should not be construed as limiting the scope of the invention in any aspect.

As described above, according to the present invention, it is possible to provide a composition for skin whitening using an extract of the Great Tokomari.

The composition for skin whitening of the present invention may be commercialized as food, cosmetics, pharmaceuticals, and the like.

Figures 1 and 2 are results showing that the extract of the snailfish extract inhibits melanin production in a concentration-dependent manner in B16F10, which is a melanoma cell line derived from a mouse.
FIG. 3 is a Western blot result showing that the extract of tyrosine chinensis suppresses the expression of tyrosinase and TRP-1 in the mouse-derived melanoma cell line B16F10.

Hereinafter, the present invention will be described with reference to Examples and Experimental Examples. However, the scope of the present invention is not limited to these Examples and Experimental Examples.

< Example > Big Cockerel Extract of Whitening activity

1. Materials and test methods

1.1 Sample extraction

After adding 10 times the weight of 70% ethanol to each dry powder of the aerial part and the subterranean part, extracting at room temperature for 24 hours, filtering (Whatman No.2), and then concentrating the filtrate with a vacuum concentrator. I did. Then, it was dried with a freeze dryer to obtain powdery above-ground part extract (XC-A) and underground part extract (XC-S).

1.2 Cell culture

B16F10 cells, a mouse-derived melanoma cell line, were purchased from the Korean Cell Line Bank, and 1% antibiotics (Gibco, USA) and 10% (100 mg/mL) fetal bovine serum (Gibco, USA). Grand Island, USA) containing DMEM medium (Dulbecco's modified Eagle's medium) _{was cultured in a 37°C, 5% CO 2} incubator, and passage culture was performed once every 4 days.

1.3 Measurement of melanin synthesis (Melanin contents)

B16F10 cells were cultured using DMEM medium containing 10% fetal calf serum and 100 nM α-MSH (melanocyte stimulating hormone) (Sigma-Aldrich Co.) in an environment of _{37°C and 5% CO 2.} Was used.

B16F10 cells cultured in DMEM medium were dispensed into a 24-well plate at 1 × 10 ⁵ cells/well, cultured for one day, and used in the experiment. Then, the samples were treated by concentration and incubated for 3 days. After incubation, the cells were washed with phosphate buffer (pH 7.4) and treated with 200 μl of 1N NaOH in each well to lyse the cells. The lysed cells were transferred to a 96-well plate and absorbance was measured at 405 nm using a microplate reader (Bioteck, cytation3). The inhibition rate of melanin production was calculated as a percentage compared to the control group, which is a sample untreated group (α-MSH-only treatment group).

1.4 Protein expression analysis by Western blot

In order to confirm the expression level of tyrosinase and TRP-1 among genes involved in melanogenesis, B16F10 cells were cultured and treated with XC-A and XC-S at a concentration of 5-25 μg/ml for 72 hours. RIPA buffer solution (10 mM Tris-HCl (pH 7.5), 1% NP-40, 0.1% sodium deoxycholate, 0.1 %) added with a protease inhibitor cocktail (Sigma, st. Louis, MO USA). SDS, 150 mM NaCl, 1 mM EDTA). Thereafter, the obtained cell lysate was centrifuged at 12,000×g for 5 minutes at 4° C., and then the protein concentration was measured using a BCA protein kit (Thermo Scientific). A sample buffer was added to the protein extracted from the cells, heated at 100° C. for 5 minutes, and then SDS-PAGE (sodium dodecyl sulfate polyacrylamide gel electrophoresis) was performed. After electrophoresis, proteins separated by molecular weight were transferred from a gel to a nitrocellulose membrane and quantified on a nitrocellulose membrane. At this time, in order to minimize the non-specific reaction, a buffer solution (phosphate buffered saline; PBS (pH 7.4), 0.1% Tween 20, 5% skimmed milk) was reacted at room temperature for 1 hour, and then the primary antibody was diluted as follows. Was used. The antibody was diluted by reacting for 1 hour at room temperature using a 5% non-fat dry milk solution, and the dilution ratio was 1:1000 for anti-tyrosinase antibody and 1:1000 for anti-TRP-1 antibody. After the reaction, wash 3 to 4 times for 5 minutes with a washing buffer (0.1% Tween-20, PBS), and then use a secondary antibody (HRP-conjugated anti-goat IgG antibody) solution diluted 1:5000 at room temperature for 1 hour. Reacted. After the secondary antibody reaction, the protein band to be analyzed was colored and quantified using an ECL (enhanced chemiluminescence) reagent.

2. Experiment result

2.1 Melanin contents measurement

The results are shown in FIGS. 1 and 2. Both the above-ground and subterranean extracts of Tokomari inhibited melanin production in a concentration-dependent manner compared to the α-MSH-treated group, which is a positive control group.

2.2 Western Blot Results

The results are shown in FIG. 3. As shown in FIG. 3, it was confirmed that as the sample treatment concentration increased, the expression levels of tyrosinase and TRP-1 protein decreased as compared with the α-MSH treatment group, which is a positive control. In the case of underground, tyrosinase increases in proportion to the treatment concentration, please check. Therefore, it is thought that the extract of chinensis is inhibited by melanin synthesis by regulating the expression of tyrosinase and TRP-1 protein involved in melanin synthesis.

Claims (5)

A composition for skin whitening, comprising the extract of the Big Dogwood as an active ingredient.
The method of claim 1,
The extract is obtained by extracting the above-ground part or the underground part of the large Dokomari with water, ethanol, or a mixed solvent thereof.
The method according to claim 1 or 2,
The composition is a composition for skin whitening, characterized in that the cosmetic composition.
The method according to claim 1 or 2,
The composition is a skin whitening composition, characterized in that the food composition.
The method according to claim 1 or 2,
The composition is a composition for skin whitening, characterized in that the pharmaceutical composition.

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