WO2023113111A1 - Skin whitening composition for relieving melasma, age spots, etc. using water-soluble mastic gum - Google Patents

Abstract

Disclosed is a composition for improving skin whitening and skin astringency, the composition reducing the expression of tyrosinase genes without exhibiting cytotoxicity for CCD-986sk cells, which are human fibroblasts, and uses a water-soluble mastic gum having a high binding capacity to hemoglobin proteins.

Description

Composition for skin whitening using water-soluble mastic gum to improve melasma, age spots, etc.

The present invention relates to a skin whitening composition for improving melasma, age spots, etc. using water-soluble mastic gum.

As living standards improve in modern society, interest in beauty as well as health is gradually increasing. Among them, development of products for improvement of pigmentation-type skin diseases such as melasma and freckles caused by skin damage and aging caused by ultraviolet rays, air pollution, etc., and improvement of overall skin tone is active.

Melanin present in the basal layer of the epidermis is a major pigment that determines skin color and plays a role in protecting the skin by preventing damage caused by ultraviolet rays and removing active oxygen. However, when melanin is locally overproduced, it causes skin diseases such as melasma, freckles, dark spots, and skin cancer. In addition, when continuously overproduced melanin stays in the dermis, it turns the skin color black or dull, increasing the desire for whitening (Pigment Cell Res. 2007 Feb;20(1):2-13.; Curr Biol. 2011 Nov. 22;21(22):1906-11.; Ann Dermatol Venereol.

Melanin is a phenolic polymeric natural pigment widely present in living organisms. In the human body, it is synthesized in melanosomes in melanocytes in the basal layer of the epidermis, and is transferred to surrounding keratinocytes to show human skin color. When melanin is abnormally low, skin lesions such as vitiligo are induced, and when it is produced in excess, on the contrary, acquired hyperpigmentation such as melanoderma, postinflammatory melanoderma, and solar lentigo A variety of skin diseases, including

A series of processes in which melanin is synthesized are collectively referred to as melanogenesis. Melanin production uses tyrosine, one of the amino acids, as a substrate, and DOPA (3, After conversion to DOPA quinone through 4-dihydroxyphenylalanine), melanin is synthesized by polymerization with amino acids or proteins after undergoing a non-enzymatic reaction and spontaneous oxidation process (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 the 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 signal transduction pathway is the cAMP/PKA (cyclic monophosphate/protein kinase A) pathway. cAMP promotes the expression of MITF via PKA and CREB1 (cAMP responsive element binding protein 1), and MITF plays an important role in melanin synthesis. As a transcriptional regulator, it promotes the transcription of tyrosinase, TRP-1 and TRP-2 (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 in the first step of melanin production, studies on whitening agents are mainly focused on developing tyrosinase activity inhibitors or antagonists for tyrosinase substrates to control tyrosinase activity. Research has been conducted in this direction (Kor. J. Pharmacogn., 44(3):220, 2013).

Currently, the substances most commonly used as whitening agents include arbutin and kojic acid. Arbutin is a tyrosine antagonist, and kojic acid chelates the active site of tyrosinase to inhibit the process of tyrosine to dopa and dopaquinone. (Korean J. Plant Res., 26(5):526, 2013; J. Korean Soc. Food Sci. Nutr., 41(2):156, 2012). Although arbutin and kojic acid are known to have strong whitening effects, due to some side effects such as skin irritation, studies are being actively conducted to find substances having whitening activity from natural substances with relatively few side effects.

On the other hand, one of the functions that prevent skin aging and maintain healthy skin is an astringent action. Astringent action is a phenomenon in which skin proteins form cross-links with high-molecular flavonoids, resulting in skin contraction. The contraction action contracts skin and mucosal blood vessels, suppresses the secretion of mucus in the cell and lymphatic gaps, forms a sparingly soluble film on the surface of the skin and mucous membranes to protect local areas, and maintains elasticity by making tissues dense. It also functions to reduce cell membrane permeability. As such, the convergence effect can be said to give a sense of elasticity and flexibility to the skin by contracting excessively open pores or pores. Since substances with astringent activity have the property of binding to proteins, the degree of astringent effect can be judged by measuring the degree of binding to hemoglobin in general (Journal of the Korean Society of Fashion & Beauty 2007, 5( 3):99-104).

The present invention discloses the skin whitening activity and skin astringent activity of water-soluble mastic gum.

An object of the present invention is to provide a composition for skin whitening using water-soluble mastic gum.

Another object of the present invention is to provide a composition for improving skin convergence using water-soluble mastic gum.

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

As confirmed in the following Examples and Experimental Examples, the present invention does not show any particular cytotoxicity to CCD-986sk cells, which is a human fibroblast cell, while water-soluble mastic gum does not show tyrosina It was completed by suppressing the expression of the enzyme gene and confirming that it has a high binding force with hemoglobin protein.

Considering the foregoing, in one aspect, the present invention can be identified as a skin peeling composition containing water-soluble mastic gum as an active ingredient, and in another aspect, skin convergence improvement containing water-soluble mastic gum as an active ingredient. composition can be identified.

In the present specification, "water-soluble mastic gum" is a suspension obtained by suspending mastic gum in water as a solvent by converting mastic gum into fine particles (average particle diameter in the range of 1 to 999 μm) or nanoparticles (average particle diameter in the range of 1 to 999 nm). ) (that is, to maintain a dispersed state without aggregation or precipitation in water as a solvent). Such water-soluble mastic gum can be obtained by pulverizing mastic gum into microparticles or nanoparticles, adding water thereto, stirring, and mixing. Particularly, it may be preferable to obtain by pulverizing mastic gum into microparticles, adding and mixing water thereto, and pulverizing the mixture (or mixed solution) again to nanoparticle the particles of mastic gum in the mixture. At this time, in order to stabilize the dispersed particles and prevent aggregation and precipitation, suspending agents may be added in appropriate amounts before and after mixing the fine or nanoparticle mastic gum with water and before and after grinding. Any suspending agent known in the art may be used, such as gum arabic, tragacanth gum, agar, karaya gum, and locust bean gum. ), guar gum, xanthan gum, ghatti gum, water-soluble polymers such as pectin, cyclodextrin, glycerin or its fatty acid ester derivatives, methylcellulose or hydrogel Modified cellulose such as oxymethyl cellulose, various surfactants, and the like can be used. These suspending agents may be used alone or in combination of two or more, and may be used in an appropriate amount for suspending, specifically, in the range of 20 to 200 parts by weight based on 100 parts by weight of mastic gum. For more specific details regarding the preparation of water-soluble mastic gum, refer to the following examples, or Korean Registered Patent No. 10 entitled "Method for Manufacturing Mastic Gum Aqueous Solution with Higher Dispersion and Water-soluble Mastic Gum Through Nanoparticles" -1865712, etc. may be referred to. Here, mastic gum refers to a resin obtained from a mastic tree ( Pistacia lentiscus ).

In addition, in the present specification, "active ingredient" means a component that exhibits the desired activity alone or can exhibit activity in combination with a carrier having no activity itself.

The composition of the present invention may contain the active ingredient in any amount (effective amount) as long as it can exhibit skin whitening activity, skin astringent activity, etc. depending on the product, formulation, etc., and a typical effective amount is based on the total weight of the composition. When it will be determined within the range of 0.001% by weight to 15% by weight. Here, "effective amount" means that when the composition of the present invention is administered to a mammal, preferably a human, to which it is applied, during the administration period according to the advice of medical experts, skin experts, etc., the intended medical / It refers to the amount of the active ingredient included in the composition of the present invention that can exhibit a dermatological effect. Such an effective amount can be determined empirically within the ordinary ability of one skilled in the art.

In addition to the active ingredients, the composition of the present invention is used for skin whitening effect, skin convergence improvement effect, skin wrinkle improvement activity, skin hypersensitivity reaction inhibition activity, skin protection activity (inhibition of skin damage by ultraviolet rays, skin moisturizing) Etc.), etc.), etc., in order to enhance the convenience of intake, intake, and use, any compound or natural extract known to have a corresponding activity and whose safety has already been verified in the art may be additionally included. .

These compounds or extracts include each country's Pharmacopoeia (Korean Pharmacopoeia), each country's Health Functional Food Codex (in Korea, the Ministry of Food and Drug Safety, "Health Functional Food Standards and Specifications"), and each country's Functional Cosmetics Codex (in Korea, the Ministry of Food and Drug Safety, Functional cosmetics standards and test methods”), compounds or extracts that have been approved as items under the laws of each country governing the manufacture and sale of pharmaceuticals (“Pharmaceutical Affairs Act” in Korea), and health functional foods In accordance with the laws of each country regulating the manufacture and sale of compounds or extracts whose functionality has been recognized (the 「Cosmetics Act」 in Korea), the laws of each country regulating the manufacture and sale of functional cosmetics (the 「Cosmetics Act」 in Korea) Accordingly, compounds or extracts whose functionality has been recognized are included.

These ingredients include, for example, retinol, retinyl palmitate, adenosine, and polyethoxylease recognized as skin wrinkle improvement ingredients in the Cosmetics Codex (Korean Ministry of Food and Drug Safety notification, 「Standards and Test Methods for Functional Cosmetics」) under the Korean 「Cosmetics Act」. tidamide, etc., and drometrizol, drometrizoletrisiloxane, digaloyltrioleate, dimethicodyethylbenzalmalonate, diethylhexylbutamidotriazone, etc. recognized as UV protection ingredients in the Korean Cosmetics Codex. , Arbutin, Niacinamide, Ascorbyl Glucoside, Alpha-Bisabolol, Oil Soluble Licorice (Glycyrrhiza) Extract, etc., recognized as skin whitening ingredients in the same Cosmetic Code, and Korea 「Health Functional Food Act」 Complexes such as pine bark extract, phosphatidylserine, finger root extract powder, complex extracts such as red ginseng and cornus officinalis, etc. recognized as UV protection ingredients according to the Korea Health Functional Food Act, green tea extract, bamboo leaf extract, melon extract, raspberry extract, beeswax alcohol, ubiquinol, coenzyme Q10, tomato extract, grape seed extract, French maritime pine bark extract, red ginseng concentrate, etc., and also improve sensitive skin conditions according to the Korean Health Functional Food Act. L. sakei Probio 65 recognized as a functional ingredient, gamma-linolenic acid-containing oil, L.plantarum CJLP133, a lactic acid bacterium 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.

In a specific aspect, the skin composition of the present invention can be identified as a cosmetic composition.

When the composition of the present invention is recognized as a cosmetic composition, the cosmetic composition may be classified as an arbitrary product in terms of its use and law, and specifically, may be a functional cosmetic having a purpose such as skin whitening, a non-functional general cosmetic, and the like. In product form, it may take any product form, specifically, solution, suspension, emulsion, paste, gel, cream, lotion, powder, soap, surfactant-containing cleansing, oil, powder foundation, emulsion foundation, wax It can be formulated as a foundation, spray, and the like. In a specific product form, it may be a softening lotion, nutrient lotion, nutrient cream, massage cream, essence, eye cream, cleansing cream, cleansing foam, cleansing water, pack, spray, or powder formulation.

The cosmetic composition of the present invention may include, in addition to the active ingredient, ingredients commonly used in cosmetic compositions, for example, stabilizers, solubilizers, surfactants, vitamins, conventional adjuvants such as pigments and fragrances, 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 a carrier component. 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, and in particular, in the case of a spray, additional chlorofluorohydrocarbon, propane / May contain a propellant such as butane or dimethyl ether.

When the formulation of the present invention is a solution or emulsion, a solvent, solubilizing agent or 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 esters, polyethylene glycol, fatty acid esters 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, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester, or polyoxyethylene sorbitan ester, microcrystals Acidic cellulose, aluminum metahydroxide, bentonite, agar, and the like may be used.

When the formulation of the present invention is surfactant-containing cleansing, as carrier components, aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyl taurate, 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 and the like can be used.

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

In a specific aspect, the composition of the present invention can be regarded as a food composition.

The food composition of the present invention can be prepared in any form, for example, beverages such as tea, juice, carbonated beverages, and ionic beverages, processed oils such as milk and yogurt, chewing gum, rice cakes, traditional Korean snacks, bread, snacks, noodles, etc. It can be manufactured into health functional food preparations such as foods, tablets, capsules, pills, granules, liquid, powder, flakes, pastes, syrups, gels, jellies, and bars.

In addition, the food composition of the present invention may take any product classification as long as it conforms to the enforcement regulations at the time of manufacture and distribution in legal and functional classification. For example, health functional food according to the Korean 「Health Functional Food Act」, or confectionery, legumes, teas, and beverages according to each food type according to the Food Code (MFDS notification 「Food Standards and Specifications」) of the Korea 「Food Sanitation Act」 , special purpose foods, etc.

The food composition of the present invention may include food additives in addition to the active ingredient. Food additives may generally be understood as substances that are added to, mixed with, or infiltrated into food in manufacturing, processing, or preserving food. Since food is consumed daily and for a long period of time, its safety must be guaranteed. In the Food Additives Code under the laws of each country regulating the manufacture and distribution of food (the 「Food Sanitation Act」 in Korea), food additives with guaranteed safety are limitedly regulated in terms of ingredients or functions. In the Korean Food Additives Codex (MFDS notification 「Standards and Specifications for Food Additives」), food additives are classified into chemical synthetic products, natural additives and mixed preparations in terms of ingredients and are regulated. It is divided into additives, preservatives, emulsifiers, acidulants, and thickeners.

Sweeteners are used to impart appropriate sweetness to foods, and both natural and synthetic sweeteners 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.

Flavors may be used to improve taste or aroma, and both natural and synthetic flavors may be used. Preferably, it is the case of using a natural one. In case of using natural ones, in addition to flavor, the purpose of enhancing nutrition can also be combined. As a natural flavoring agent, it may be obtained from apples, lemons, tangerines, grapes, strawberries, peaches, etc., or obtained from green tea leaves, roundworms, bamboo leaves, cinnamon, chrysanthemum leaves, jasmine, and the like. In addition, those obtained from ginseng (red ginseng), bamboo shoots, aloe vera, ginkgo, etc. can be used. Natural flavors can be liquid concentrates or solid extracts. In some cases, synthetic flavors may be used, and synthetic flavors may include esters, alcohols, aldehydes, terpenes, and the like.

Calcium sorbate, sodium sorbate, potassium sorbate, calcium sorbate, sodium benzoate, potassium benzoate, ethylenediaminetetraacetic acid (EDTA), etc. may be used as preservatives, and as emulsifiers, gum arabic, carboxymethylcellulose, xanthan gum, pectin and the like, and as the acidulant, acidulic acid, malic acid, fumaric acid, adipic acid, phosphoric acid, gluconic acid, tartaric acid, ascorbic acid, acetic acid, and the like may be used. Acidulants may be added to the food composition to have an appropriate acidity for the purpose of inhibiting the growth of microorganisms in addition to improving taste.

As the thickening agent, a suspending agent, a sedimentation 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 physiologically active substances or minerals known in the art and guaranteed to be stable as food additives for the purpose of supplementing or reinforcing functionality and nutrition.

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

The food composition of the present invention may include the above-mentioned food additives in an appropriate amount to achieve the purpose of addition according to the type of product.

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

In a specific aspect, the composition for skin whitening of the present invention can be understood as a pharmaceutical composition.

When the composition for skin whitening of the present invention is understood 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, melasma, brown or dark spots, solar pigment Melasma blue, hyperpigmentation after the use of drugs such as calcium antagonists, hyperpigmentation sequelae following tissue sclerotherapy, brown spots of pregnancy, melasma in women taking oral contraceptives, wounds or dermatitis including scratches and burns may include, but are not limited to, post-inflammatory hyperpigmentation, phototoxic reaction, or other similar small fixed pigmented lesions.

The pharmaceutical composition of the present invention may be formulated into an oral dosage form or a parenteral dosage form according to the route of administration by a conventional method known in the art, including a pharmaceutically acceptable carrier in addition to the active ingredient. The route of administration herein may be any appropriate route including topical route, oral route, intravenous route, 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, one drug is firstly administered intravenously and the other drug is secondly administered through a topical route.

Pharmaceutically acceptable carriers are well known in the art depending on the route of administration or dosage form, and specific reference may be made to the pharmacopoeia of each country including the "Korean Pharmacopoeia".

When the pharmaceutical composition of the present invention is prepared as an oral dosage form, powder, granule, tablet, pill, dragee, capsule, liquid, gel, syrup, suspension, wafer according to a method known in the art together with a suitable carrier. It can be prepared in formulations such as Examples of suitable carriers include sugars such as lactose, glucose, sucrose, dextrose, sorbitol, mannitol and xylitol, starches such as corn starch, potato starch and 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, glycerol etc. can be mentioned. In the case of formulation, appropriate binders, lubricants, disintegrants, coloring agents, diluents, etc. may be included as needed. Suitable binders include starch, magnesium aluminum silicate, starch ferrite, gelatin, methylcellulose, sodium carboxymethylcellulose, polyvinylpyrrolidone, glucose, corn sweetener, sodium alginate, polyethylene glycol, wax and the like, and oleic acid as a lubricant Sodium, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride, silica, talcum, stearic acid, magnesium salts and calcium salts thereof, polyethylene glycol, and the like, and disintegrants include starch, methyl cellulose , agar, bentonite, xanthan gum, starch, alginic acid or its sodium salt, and the like. Moreover, as a diluent, lactose, dextrose, sucrose, mannitol, sorbitol, cellulose, glycine, etc. are mentioned.

When the pharmaceutical composition of the present invention is prepared as a parenteral formulation, it may be formulated in the form of an injection, transdermal administration, nasal inhalation, and suppository along with a suitable carrier according to a method known in the art. When formulated as an injection, an aqueous isotonic solution or suspension may be used as a suitable carrier, and specifically, an isotonic solution such as phosphate buffered saline (PBS) containing triethanolamine, sterile water for injection, or 5% dextrose may be used. there is. When formulated as a transdermal formulation, it may be formulated in the form of ointments, creams, lotions, gels, external solutions, pastas, liniments, air rolls, and the like. In the case of nasal inhalation, it can be formulated in the form of an aerosol spray using a suitable propellant such as dichlorofluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide, etc., and when formulated as a suppository, the carrier is Witepsol ( witepsol), tween 61, polyethylene glycols, cacao fat, laurin fat, polyoxyethylene sorbitan fatty acid esters, polyoxyethylene stearates, sorbitan fatty acid esters, and the like can be used.

Specific formulations of pharmaceutical compositions are known in the art, and reference may be made to, for example, Remington's Pharmaceutical Sciences (19th ed., 1995). These documents are 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 10 g/kg, depending on the patient's condition, body weight, sex, age, severity of the patient, and route of administration. It may be in the range of 1 g/kg. Administration can be done once a day or divided into several times. These dosages should not be construed as limiting the scope of the present invention in any respect.

As described above, according to the present invention, it is possible to provide a composition for skin whitening and improving skin convergence using water-soluble mastic gum.

The composition of the present invention can be commercialized as cosmetics, food, medicine, and the like.

1 is a result of measuring the toxicity of water-soluble mastic gum in CCD-986sk.

2 is a result of tyrosinase gene expression of water-soluble mastic gum.

3 is a skin astringent activity measurement result through the degree of binding to hemoglobin.

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

<Example> Evaluation of skin whitening activity and skin astringent activity of water-soluble mastic gum

<Example 1> Preparation of water-soluble mastic gum

Mastic gum powder was visually inspected to confirm color and inspect for foreign substances. Afterwards, 20% by weight of mastic gum powder, 75.7% by weight of purified water, 4% by weight of cyclodextrin, and 0.3% by weight of additives (ghatti gum) were added and heated at 30 to 50 ° C using a homomixer (KNS Co., Ltd.). was dispersed at 2,000 to 3,000 rpm for 30 to 60 minutes to a particle size of about 20 to 50 μm. The dispersion of the mastic gum was carried out using a bead mill (DNTech Co., Ltd.) at 20 to 40 ° C. for 3 to 6 hours so that the final dispersed particles had an average size of 600 to 1,000 nm. Thereafter, the mixture was filtered through a 40 to 60 μm housing filter to obtain an aqueous solution of mastic gum in a suspension state. In the following experiments, this water-soluble mastic gum was used as it is.

<Example 2> Cytotoxicity test

First, CCD-986sk cells, which are human fibroblasts, were subcultured at ATCC (Manassas, VA, USA) and frozen in cryovials. After dissolving this in a water bath, penicillin (100 U/ml), streptomycin (100 ug/ml), and 10% heat-inactivated serum were added to DMEM (Dulbecco's Modified Eagle's Medium) at 95% air, 5% CO ₂ and 37°C. cultured with. When the CCD-986sk cells adhered to the culture dish and the cells grew, the cells were collected using 0.25% trypsin/10 mM EDTA, divided at a ratio of 1:3, and maintained in 10% DMEM to culture the cells.

The reagent used in the experiment, CCK-8, was purchased from Dongin Biotech Co., Ltd. (Seoul, Korea). CCD-986sk cells were treated in 96-well plates at a concentration of 1 × 10 ³ cells/well by 100 ul and then cultured for 24 hours. When the cells attached to the bottom, the example samples were treated for 24 hours up to 2 mg/mL. After incubation for 4 hours under conditions of 10 ul of CCK-8 reagent and 100 ul of DMEM, absorbance was measured at 540 nm with a spectrophotometer.

As a result, as shown in FIG. 1, it was found that the sample of Example can be safely used without showing toxicity up to 0.5 mg/ml.

<Example 3> Measurement of tyrosinase gene expression

In the same manner as in Example 2, human fibroblasts were treated with 0.1 mg/ml of the sample and positive control, Ascorbic acid (AA), and then tyrosinase (TYR) known as a melanin production regulatory gene. The expression level of was confirmed through real-time PCR using Applied Biosystems' 'QuantStudio 3' according to the manufacturer's protocol. At this time, the data were expressed as mean ± standard deviation based on the results of three independent experiments for each condition. TYR primer sequences are shown in Table 1 below.

TYR-Forward AGGCAGAGGTTCCTGTCAGA TYR-Reverse ATCCCATGAAGTTGCCAGAG

As a result, as shown in FIG. 2, when CCD-986sk cells were treated with up to 0.3 mg/ml of the sample of Example, it was confirmed that the expression of TYR was significantly inhibited compared to the positive control group. <Experimental Example 2> Skin astringent activity test

500 ul each of the sample of Example was prepared for each concentration, mixed with the same amount of 0.1% hemoglobin solution mixed in PBS, mixed for 1 minute (Voltexing), and left at room temperature for 5 minutes. Thereafter, 100 ul each was dispensed into 96-well plates, and absorbance was measured at 405 nm with a spectrophotometer.

As a result, as shown in FIG. 3, it was found that the skin astringent activity of the example samples increased in a concentration-dependent manner.

Claims (6)

A composition for skin whitening comprising water-soluble mastic gum as an active ingredient. A composition for improving skin astringency comprising water-soluble mastic gum as an active ingredient. According to claim 2, Wherein the water-soluble mastic gum is a suspension obtained by suspending the mastic gum into fine particles (average particle diameter ranging from 1 to 999 μm) to nanoparticles (average particle diameter ranging from 1 to 999 nm) in water as a solvent. According to claim 2 or 3, The composition is a composition for skin whitening, characterized in that the cosmetic composition. According to claim 2 or 3, The composition is a composition for skin whitening, characterized in that the food composition. According to claim 2 or 3, The composition is a composition for skin whitening, characterized in that the pharmaceutical composition.

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