KR101832169B1 - Composition for skin whitening comprising extract of Picrasma quassioides as effective component - Google Patents

Abstract

The present invention relates to a whitening composition comprising an extract of an old-fashioned room as an active ingredient. Since the extract of the present invention has an antioxidative activity and is excellent in the effect of inhibiting melanin production and tyrosinase activity, Functional cosmetics and skin pigmentation diseases which can prevent or ameliorate skin pigmentation diseases.

Description

[0001] The present invention relates to a composition for skin whitening,

The present invention relates to a skin whitening composition containing an extract of a high-grade wood as an active ingredient.

Melanin is a key determinant of hair and skin color, and it protects skin cells from ultraviolet light. However, excessive accumulation of melanin causes hyperpigmentation such as freckles and spots. The melanin-forming cells in the basal layer of the epidermis have a specific structure called melanosomes, in which melanin is produced by the action of a melanogenic enzyme. Melanin has black and brown series of eumelanin and yellow and red series of pheomelanin. The synthesis of melanin is caused by tyrosinase, tyrosine is doped (DOPA) and dopa It begins with conversion to quinone (DOPA quinone). In addition, TRP-2 (tyrosinase-related protein 2) can be produced by converting DOPA chrome into 5,6-dihydroxyindole-2-carboxylic acid, TRP-1 tyrosinase-related protein 1 catalyzes the oxidation reaction to indole-5,6-quinone-2-carboxylic acid and inhibits tyrosinase (eu) production in eumelanin synthesis tyrosinase) and TRP-1 and 2 are involved. In addition, the microphthalmia-associated transcription factor (MITF), well known as a regulator of melanin formation, is a transcription factor with a helix-loop-helix-leucine zipper structure. Is known to regulate the process of blackening and the proliferation and survival of cells. Melanocyte stimulating hormone (α-MSH), adrenocorticotropic hormone (PGE2), and prostaglandin E2 (melanocyte stimulating hormone), which are involved in melanin biosynthesis, have been reported to regulate melanin synthesis through cAMP.

Currently, kojic acid, vitamin derivatives, arbutin and dibutylated hydroxytoluene (BHT), which are used as synthetic antioxidants, are inexpensive and exhibit excellent whitening effect due to their antioxidative activity. However, It is reported and regulates usage. Currently, studies are being actively carried out to find functional materials from natural products both domestically and externally.

Picasma quassioides is a small arboreous tree in the dicotyledonous plant family, derived from the bitter taste of the leaves and stems, as in cattle. It has the effect of dryness, humidification and sterilization. It is used as a remedy for symptoms such as indigestion, gastroenteritis, pulmonary tuberculosis, eczema, obesity, improvement, etc. in one room, and the whole tree is put into a pot, .

Korean Patent No. 1312025 discloses a method for producing ganoderma lucidum extract and use of the extract. Korean Patent Registration No. 0905386 discloses a herbal cosmetic composition for atopic skin containing herbal extracts. However, the whitening composition containing the extract of the present invention as an active ingredient has not yet been disclosed.

The present invention has been made in view of the above-mentioned needs, and it is an object of the present invention to provide an antioxidative activity of the extract of the old tree, which, when treated with melanoma cells (B16-F10 mouse melanoma cell), inhibits the production of melanin and tyrosinase And confirming that the effect of inhibiting the activity is excellent, thereby completing the present invention.

In order to solve the above-mentioned problems, the present invention provides a cosmetic composition for whitening skin, which comprises an extract of an old-fashioned room as an active ingredient.

In addition, the present invention provides a pharmaceutical composition for preventing or treating melanin pigment hyperproliferative disorder containing an extract of a high-grade wood as an active ingredient.

The present invention also provides a health functional food composition for preventing or ameliorating hyperpigmentation of melanin pigment, which comprises an extract of an old tree as an active ingredient.

Since the extract of the present invention has antioxidant activity and is excellent in the effect of inhibiting the activity of melanin production and tyrosinase, functional cosmetics for skin whitening and health functional foods capable of preventing or improving skin pigmentation diseases Or as a therapeutic agent for skin pigmentation diseases.

FIG. 1 shows cell viability of B16F10 mouse melanocytes treated at various concentrations according to one embodiment of the present invention. CON is a negative control group treated with DMEM medium.
FIG. 2 is a graph showing the relationship between the content of melanin in melanocytes of B16F10 mouse and the content of tyrosinase in melanocytes of B16F10 mouse, when α-MSH (α-melanocyte stimulating hormone) And the activity was measured. α-MSH is a melanogenesis inducer, and AR is a positive control for arbutin.

In order to accomplish the object of the present invention, the present invention provides a cosmetic composition for skin whitening comprising an extract of an old tree as an active ingredient.

In the cosmetic composition according to an embodiment of the present invention, the extract of the red-leafy cabbage is extracted with a solvent selected from the group consisting of water, a lower alcohol having 1 to 4 carbon atoms and a mixed solvent thereof, preferably ethanol will be.

For example, the oldwood extract

1) pulverizing the old wood chamber to produce a powder;

2) adding 5 ~ 15 times the amount of ethanol to the weight of the dead wood powder and extracting it by hot water bath at room temperature for 5 to 15 hours; And

3) filtering and concentrating the extract of the old wood obtained in the step 2) and freeze-drying the extract,

Preferably, 1) pulverizing the old wood chamber to produce a powder;

2) adding 70% ethanol to the dead skin of 10 times the weight of the dead wood powder and extracting it by hot water bath at room temperature for 10 hours; And

3) a step of filtering and concentrating the extract of the old wood obtained in the step 2), followed by lyophilization, but the present invention is not limited thereto.

In addition, the cosmetic composition may inhibit the production of melanin or the activity of tyrosinase, but is not limited thereto.

On the other hand, the whitening effect refers to an effect to improve or prevent glaring skin, spots, freckles, and dark circles caused by various factors such as exposure to ultraviolet rays, change in hormone balance, genetic program, Or an effect of maintaining a beautiful skin with a sense of transparency and an effect of reducing the dullness of the skin to increase gloss and tautness. In general, it is known that the skin, spots, freckles, and dark circles of the skin, stamens, freckles and dark circles are stimulated by melanocytes due to stimulation by ultraviolet rays or changes in hormone balance, and melanin pigment is deposited on the skin. Therefore, if the production of melanin can be inhibited, it is possible to prevent or improve gross skin, spots, freckles and dark circles.

The cosmetic composition for skin whitening may be in the form of a solution, suspension, emulsion, paste, gel, cream, lotion, powder, soap, surfactant-containing cleansing oil, powder foundation, emulsion foundation, wax foundation and spray , But the present invention is not limited thereto. More specifically, it is made up of formulations such as soft lotion, nutritional lotion, nutritional cream, massage cream, essence, eye cream, cleansing cream, cleansing foam, cleansing water, pack, spray, powder, fact, lip gloss, lipstick and shadow . The cosmetic composition comprising each of these formulations may contain various bases and additives necessary for formulation of the formulation, and the kinds and amounts of these ingredients can be easily selected by those skilled in the art.

The cosmetic composition of the present invention may contain, in addition to the active ingredient, at least one skin whitening active ingredient exhibiting the same or similar functions. Skin whitening active ingredients include kojic acid and its derivatives, arbutin, ascorbic acid and its derivatives, hydroquinone and its derivatives, resorcinol, cycloalkanone, methylenedioxyphenylalkanol, 2,7-dinitroindazole or Plant extracts such as mandarin orange extract, rice extract, licorice extract, and the like, but are not limited thereto.

When the formulation of the cosmetic composition of the present invention is a paste, a cream or a gel, an animal fiber, a plant fiber, a wax, a paraffin, a starch, a tragacanth, a cellulose derivative, a polyethylene glycol, a silicone, a bentonite, Etc. may be used.

When the formulation of the cosmetic composition of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used as a carrier component. Especially, in the case of a spray, Propellants such as carbon, propane-butane or dimethyl ether.

When the formulation of the cosmetic composition of the present invention is a solution or an emulsion, a solvent, a solvent or an emulsifier is used as a carrier component, and examples thereof include water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, Glycol, 1,3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol or fatty acid esters of sorbitan.

When the formulation of the cosmetic composition of the present invention is a suspension, a carrier such as water, a liquid diluent such as ethanol or propylene glycol, a suspension such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, Microcrystalline cellulose, aluminum metahydroxide, bentonite, agar or tracant, etc. may be used.

When the formulation of the cosmetic composition of the present invention is an interfacial-active agent-containing cleansing, the carrier component may include aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, acethionate, imidazolinium derivative, methyltaurate, sarcosinate , Fatty acid amide ether sulfate, alkylamidobetaine, aliphatic alcohol, fatty acid glyceride, fatty acid diethanolamide, vegetable oil, linolenic derivative or ethoxylated glycerol fatty acid ester.

The cosmetic composition of the present invention may further contain an excipient including a fluorescent substance, a fungicide, a hygroscopic substance, a moisturizer, a fragrance, a fragrance carrier, a protein, a solubilizer, a sugar derivative, a sunscreen, a vitamin, .

In addition, the present invention provides a pharmaceutical composition for preventing or treating melanin pigment hyperproliferative disorder containing an extract of a high-grade wood as an active ingredient.

The term " melanin pigment hyperpigmentation " as used herein means darker or darker than other areas due to excessive increase of melanin in specific areas of skin or hands and toenails. The melanin pigment hyperpigmentation disorder can be selected from the group consisting of freckles, senile spots, liver, spots, brown or black spots, daylight pigmentation, cyanic melasma, hyperpigmentation after drug use, gravidic chloasma, Post-inflammatory hyperpigmentation due to burns or dermatitis including burns, and the like.

The pharmaceutical composition of the present invention may contain a pharmaceutically acceptable carrier in addition to the active ingredient. Such a carrier is usually used at the time of formulation, and includes lactose, dextrose, sucrose, sorbitol, mannitol, starch, Calcium carbonate, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methylcellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and minerals Oils, and the like, but are not limited thereto. The pharmaceutical composition of the present invention may further contain a lubricant, a wetting agent, a sweetening agent, a flavoring agent, an emulsifying agent, a suspending agent, a preservative, etc., in addition to the above components. Suitable pharmaceutically acceptable carriers and formulations are described in detail in Remington's Pharmaceutical Sciences, 19th ed., 1995.

The appropriate dosage of the pharmaceutical composition according to the present invention may vary depending on such factors as the formulation method, the administration method, the patient's age, body weight, sex, pathological condition, food, administration time, administration route, excretion rate, .

The pharmaceutical composition of the present invention may be administered orally or parenterally. In the case of parenteral administration, the composition may be administered topically to the skin, intravenously, subcutaneously, intramuscularly, intraperitoneally, or transdermally. In view of the fact that the pharmaceutical composition of the present invention is applied for the treatment or prevention of a melanin pigment hyperpigmentation disease, it is preferable that the composition is topically applied to the skin.

The concentration of the active ingredient contained in the composition of the present invention can be determined in consideration of the purpose of treatment, the condition of the patient, the period of time required, and the like, and is not limited to a specific range of concentration.

The pharmaceutical composition of the present invention may be manufactured in a unit dosage form by formulating it using a pharmaceutically acceptable carrier or excipient according to a method which can be easily carried out by those having ordinary skill in the art to which the present invention belongs Into a capacity container. The formulations may be formulated into any one of the formulations selected from injectables, creams, patches, sprays, ointments, alerts, lotions, liniments, pastes and cataplasms, and may additionally contain dispersing or stabilizing agents have.

The present invention also provides a health functional food composition for preventing or ameliorating hyperpigmentation of melanin pigment, which comprises an extract of an old tree as an active ingredient.

When the health functional food composition of the present invention is used as a food additive, the health functional food composition may be added as it is, or may be used together with other food or food ingredients, and suitably used according to a conventional method. The amount of the active ingredient can be suitably used depending on its use purpose (prevention or improvement). Generally, the health functional food composition of the present invention is added in an amount of not more than 15 parts by weight, preferably not more than 10 parts by weight based on the raw material, when the food or beverage is produced. However, in case of long-term ingestion for health, the amount may be less than the above range, and since there is no problem in terms of safety, the active ingredient may be used in an amount exceeding the above range.

There is no particular limitation on the kind of the health functional food. Examples of the foods to which the health functional food composition can be added include dairy products including meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen and other noodles, gums, ice cream, soups, Drinks, alcoholic beverages, and vitamin complexes, all of which include health foods in a conventional sense.

In addition, the health functional food composition of the present invention can be produced as a food, particularly a functional food. The functional food of the present invention includes components that are ordinarily added in food production, and includes, for example, proteins, carbohydrates, fats, nutrients, and seasonings. For example, in the case of a drink, a natural carbohydrate or a flavoring agent may be included as an additional ingredient in addition to the active ingredient. The natural carbohydrate may be selected from the group consisting of monosaccharides (e.g., glucose, fructose, etc.), disaccharides (e.g., maltose, sucrose etc.), oligosaccharides, polysaccharides (e.g., dextrin, cyclodextrin, , Xylitol, sorbitol, erythritol, etc.). The flavoring agent may be a natural flavoring agent (e.g., tau Martin, stevia extract, etc.) and a synthetic flavoring agent (e.g., saccharin, aspartame, etc.).

In addition to the above health functional food composition, it is also possible to use various nutrients, vitamins, electrolytes, flavors, colorants, pectic acids and salts thereof, alginic acid and its salts, organic acids, protective colloid thickening agents, pH adjusting agents, stabilizers, preservatives, glycerin, A carbonating agent used in beverages, and the like.

Hereinafter, the present invention will be described in more detail with reference to Examples. It is to be understood by those skilled in the art that these embodiments are merely illustrative of the present invention and that the scope of the present invention is not limited thereto.

Materials and methods

1. Preparation of the extracts of old wood chambers and measurement of extraction yield

The extract of the present invention was obtained by adding 10 times 70% ethanol to the weight of the old wood powder of the present invention, and then extracting it with hot water at room temperature for 10 hours. To determine the extraction yield, the old wood extract was filtered and concentrated, Dried to determine the weight of the building, and then calculated as a percentage of the amount of raw material used for the extraction.

2. Total polyphenol content measurement

The total polyphenol content was determined by modifying the method of Gutfiger T., 1981, J Am Oil Chem Soc, Vol. 58, 966-968. 100 μl of Folin-Ciocalteau reaction solution was added to 100 μl of the diluted extract of the present invention, and the mixture was allowed to stand for 3 minutes, and then 1 ml of 0.7 M Na ₂ CO ₃ solution was added and mixed. After the mixture was reacted for 1 hour, the absorbance was measured at 750 nm using a spectrophotometer. The total polyphenol contents of the extracts of the old wood chambers were calculated by preparing a standard curve based on tannic acid, a standard substance.

3. DPPH  Measurement of radical scavenging activity

Measurement of DPPH (1,1-diphenyl-2-picryl-hydrazyl) radical scavenging activity was measured by modifying the method of Blois MS, Nature, 1958, 26, 1199-1200. 50 mu l of a 0.2 mM DPPH solution was added to 100 mu l of the extract of the old wood chamber, reacted for 10 minutes, and the absorbance was measured at 517 nm. The DPPH radical scavenging activity was calculated by the absorbance reduction rate of the treated and untreated extracts of the old woody extracts and the RC50 value, which is the concentration of the old woody extract required for the reduction of DPPH radical scavenging activity by 50%.

4. ABTS  Measurement of radical scavenging activity

Measurement of ABTS (2,2'-azino-bis (3-ethylbenzthiazoline-6-sulfonic acid) radical scavenging activity was measured by modifying the method of Blois MS, Nature, 1958, 26, 1199-1200. ABTS and 2.45 mM K ₂ S ₂ O ₈ (potassium persulfate) were mixed and allowed to stand in a dark place for 1 day to form ABTS ^{· +} . Then, the mixed solution was stirred at 734 nm for 0.7 to 0.9 using molar extinction coefficient diluted with. by the ABTS ^{· +} solution was diluted in an extract of old trees room 50㎕ 100㎕ added to measure the change in the absorbance after five minutes. ABTS radical scavenging activity was calculated to be treated and the absorption rate of decrease of the untreated yarn of the old tree extract , And the RC50 value, which is the concentration of the extract of the old wood chamber, required to reduce the ABTS radical scavenging activity by 50%.

5. Measurement of cell viability

The cell viability of MTT was determined by MTT (3- (4,5-dimeth-ylthiazol-2-yl) -2,5-diphenyltetrazolium bromide) analysis. B16F10 mouse melanoma cells were cultured in a 96-well plate containing 100 占 퐇 of DMEM medium for 1 day and then treated with various concentrations (25, 50 and 100 占 퐂 / ml) of the extracts of the woody roots and cultured for 24 hours. Then, 10 μl of 5 mg / ml MTT solution was added to each well, followed by incubation for 3 hours to induce a reduction reaction. 100 μl of DMSO (dimethyl sulfoxide) solution was added to form purple formazan, The crystals were completely dissolved. The absorbance at 570 nm was measured using a spectrophotometer. The cell viability was calculated based on the survival rate of the untreated control cells cultured in the cells alone.

6. Biosynthesis of melanin and tyrosinase ( tyrosinase ) Activity measurement

B16F10 mouse melanoma cells were inoculated on a 12-well plate at a concentration of 1 x 10 ⁵ cells / ml and cultured for 24 hours. Melanocyte production was induced by treating α-MSH (melanocyte-stimulating hormone) Arbutin (100 μg / ml) and the extracts of old wood (25, 50 and 100 μg / ml) prepared for each concentration were treated for 48 hours. After washing with PBS (phosphate buffered saline), the cells were centrifuged to make a cell suspension. 200 μl of 1 N NaOH solution containing 10% DMSO (dimethyl sulfoxide) was added and dissolved at 60 ° C for 1 hour And absorbance was measured at 405 nm. The activity of tyrosinase was determined by adding 160 의 of substrate solution and 10 ㎕ L-DOPA (L-DOPA) dissolved in 1 ml of 0.1 M sodium phosphate buffer (pH 6.8) Was reacted at 37 캜 for 1 hour, and Dopamine chrome (DOPA chrome) generated in the reaction solution was measured at 475 nm. The inhibitory activity of tyrosinase was expressed by the absorbance reduction rate of the treated and untreated control of the extract of the woody thread.

Example  1. Extraction yields of the extracts of the old wood pellets, total polyphenol Content and Antioxidant  Active measurement

From the industrial point of view, the extraction yield of natural products is very important in terms of value added. Thus, in Example 1, the extraction yield of the extract of the old wood room was measured. As a result, the extraction yield was 15.95% as shown in Table 1 below.

Polyphenolic materials are aromatic compounds that give plants specific colors and act as substrates in redox reactions, and have two or more OH (phenolic hydroxy) groups in a molecule. Its principal components, flavonoids and tannins, have various physiological activities such as tooth decay prevention, hypertension inhibition, anti-AIDS, antioxidant, and anti-cancer. Flavonoids are one of the largest naturally occurring groups of phenolic compounds and can be divided into anthocyanin, chalcone, aurone, flavone, flavonol and their derivatives. The antioxidant activity of the extracts of old woody persimmon extracts was measured to be 55.90 mg / g. The antioxidative activity of the extracts of old woody persimmon extracts was measured to be 13.53% The radical scavenging activity was measured to be 6.85%.

The ABTS and DPPH scavenging activity of the extract of the present invention of the present invention sample Extraction yield (%) Total phenol content
(mg / Extract 1g) ABTS scavenging activity (%) DPPH scavenging activity (%) Vit.C (RC50) 15.95 Vit.C (RC50) 10.02 A wooden room 15.95 55.90 + - 1.74 13.53 + - 0.55 6.85 + - 0.33

Example  2. Analysis of cell viability according to the treatment of the extract of the tree of the present invention

In order to investigate the cytotoxicity of the extracts of old tree extracts from B16F10 mouse melanoma cells, 25, 50 and 100 μg / ml of the extracts of old tree extracts were treated for 24 hours before MTT analysis. As a result, as shown in Fig. 1, no inhibition of cell growth was observed at all concentrations of the extracts of the woody buds, compared with the control group treated with DMEM, and the cell survival rate was about 100% Were not found to be cytotoxic. Therefore, in Example 3, the whitening effect of 25, 50, and 100 μg / ml of the extract of the pine tree without cytotoxicity was tested on the cells.

Example  3. Production of melanin and inhibition of tyrosinase activity by the treatment of the extract of the present invention

The biosynthesis of melanin is carried out by using tyrosin, which is a kind of amino acid, as a precursor, through DOPA, DOPA quinone and indole-5,6-dihydroquinone, -5,6-dihydroquinone. ≪ / RTI > Melanin is produced in the melanocyte and is transferred to the epidermis and dermis boundary in the form of granules called melanosomes. Melanin is not only a direct cause of dark skin, but also causes serious problems in terms of skin beauty, such as promoting the formation of spots and freckles. In addition, over-producing melanin has been found to cause skin aging and skin cancer. Therefore, the content of melanin and the activity of tyrosinase according to the present invention were measured. As a result, as shown in FIG. 2, α-MSH was treated to induce melanin production When the extract of the present invention was treated, the amount of melanin was decreased in a concentration-dependent manner. In particular, when the extract of 100 μg / ml of the extract of the old tree was treated, the content of melanin was reduced by about 40% compared to the α-MSH treatment. In addition, the activity of tyrosinase inhibited the activity of tyrosinase in a dose-dependent manner as a result of treatment with the extract of the old tree. In particular, when treated with 100 μg / Decreased the activity of tyrosinase and showed an effect similar to the positive control group arbutin.

Claims (8)

A cosmetic composition for skin whitening comprising an extract of an old tree as an active ingredient. delete The method according to claim 1, wherein the high-molecular weight ethanol extract
1) pulverizing the old wood chamber to produce a powder;
2) adding 5 ~ 15 times the amount of ethanol to the weight of the dead wood powder and extracting it by hot water bath at room temperature for 5 to 15 hours; And
3) filtering and concentrating the ethanol extract of the old tree obtained in the step 2) and freeze-drying the cosmetic composition for skin whitening. The cosmetic composition for skin whitening according to claim 1, wherein the cosmetic composition inhibits the production of melanin or the activity of tyrosinase. The cosmetic composition as claimed in claim 1, wherein the cosmetic composition is a solution, suspension, emulsion, paste, gel, cream, lotion, powder, soap, surfactant-containing cleansing oil, powder foundation, emulsion, foundation, wax foundation and spray Wherein the cosmetic composition is one selected from the group consisting of a cosmetic composition and a cosmetic composition. A pharmaceutical composition for preventing or treating a melanin pigment hyperpigmentation disorder containing an extract of an old tree as an active ingredient. 7. The method of claim 6, wherein the melanin pigment hyperpigmentation disorder is selected from the group consisting of freckles, aging spots, liver, spots, brown or black spots, daylight pigmentation, cyanic melasma, hyperpigmentation after drug use, chloasma), or hyperpigmentation after inflammation due to scratches or dermatitis, including scratches and burns. A health functional food composition for prevention or amelioration of hyperpigmentation of melanin pigment comprising an extract of an old tree as an active ingredient.

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