KR102135220B1 - COMPOSITION FOR SKIN WHITENING COMPRISING Polyopes affinis AS AN ACTIVE INGREDIENT - Google Patents

Abstract

The present invention relates to a composition for whitening comprising an extract of gamjamaksal as an active ingredient. Thereby, the composition for whitening comprising the extract of the tuna kelp of the present invention exhibits almost no side effects to the human body by using the tuna, which is a kind of sea horse that grows in the ocean by replacing natural products previously obtained on land. , The skin whitening efficacy according to melanin biosynthesis inhibition is very excellent effect.

Description

Composition for skin whitening that includes tuna skin as an active ingredient {COMPOSITION FOR SKIN WHITENING COMPRISING Polyopes affinis AS AN ACTIVE INGREDIENT}

The present invention is polyopes affinis ) relates to a composition for whitening comprising an extract as an active ingredient, and more specifically, a cosmetic composition for skin whitening comprising an extract of blackcurrant that inhibits melanin production as an active ingredient and a pharmaceutical for preventing or treating melanin pigmentation disease It relates to a composition.

The skin is an externally exposed organ that acts as a protective barrier between the inside and outside of the human body and protects the living body from the surrounding environment by defending against the invasion of external environmental pollutants and microorganisms, including chemicals and ultraviolet rays.

Melanin is a dark brown grain pigment and is present on the skin, hair, and eyes. The cells that make melanin are called melanocytes, and melanin is made from small, bag-shaped organelles called melanosomes in melanocytes. Melanocytes extend melanocytes and transmit melanin to other cells, including keratinocytes. Depending on the type of melanocyte, the melanosome may be maintained as it is. The skin color is determined by the amount of melanin. The higher the amount of melanin, the darker the skin color. Skin color differences among races are not due to different numbers of melanocytes, but because of the different size of melanocytes and the amount of melanin produced.

Melanin absorbs more than a certain amount of ultraviolet rays and blocks harmful ultraviolet rays from entering the human body, thereby protecting the human body. When the skin turns brown by sunlight, the melanin cells present in the lower layer of the skin are stimulated by UV rays to create melanin to protect the body, and send the melanin made to the top of the skin to prevent UV rays from penetrating deep into the skin Because. With the same logic, the more people exposed to the sun, the more melanin is produced to protect the skin from UV rays, and the darker the skin.

When melanin is excessively generated due to external environmental changes such as excessive exposure to ultraviolet rays, air pollution, and stress, pigmentation occurs in the skin, causing melanin, blemishes, and freckles. Darkening of the skin is caused by the reaction of skin cells to internal and external factors, and a representative factor is due to exposure to ultraviolet rays. Specifically, when the skin is exposed to ultraviolet light, tyrosinase is activated. By tyrosinase acts on tyrosine present in the skin tissue, it is caused by an oxidation process that produces dopa and dopaquinone. In a melanosome in the skin pigment cell melanocyte, a polymer called melanin is synthesized, and this melanin is delivered to the keratinocytes, the keratinocytes of the skin, and reaches the skin surface by the keratinization process. It protects the skin from UV rays. Therefore, melanin is an indispensable UV protection agent for our human body, and also plays a role of an effective free radical scavenger that removes various radicals that modify biological components such as proteins, lipids, and nucleic acids.

However, when melanin is excessively synthesized topically, or skin physiological functions are reduced due to skin lesions and aging, melanin is deposited on the surface of the skin, causing spots, freckles, and various pigmentation. As the cause and mechanism of skin darkening are revealed as described above, substances having an inhibitory effect on the activity of tyrosinase, an enzyme involved in the skin darkening process in the production of whitening cosmetics, are blended into cosmetics, or some of the process of melanin production A method of reducing the production of melanin by inhibiting the reaction has been used.

Representative materials used as skin whitening materials include chemical substances such as ascorbic acid, kojic acid, hydroquinone, and plant extracts such as epithelium extract and licorice extract. However, ascorbic acid is not suitable as a melanin production inhibitor due to its lack of tyrosinase activity inhibitory effect and low stability of the molecule itself, and kojic acid has excellent inhibitory activity of tyrosinase, but discolors and changes over time when blended into cosmetics. Therefore, not only is there a problem in stability such as a decrease in titer, but also has a limitation in use due to large skin irritation, and hydroquinone is limited to use as a cosmetic due to skin irritation and safety problems. Therefore, arbutin, a glycoside obtained by reacting a glyceryl group with hydroquinone, is used as a whitening component, and it is a principle that hydroquinone is not detected, but there are cases in which hydroquinone is detected in a very small amount due to process problems.

Accordingly, there is a need to develop an effective whitening component that is excellent in whitening effect and is harmless to the human body as a natural-derived component.

Korean Registered Patent No. 1448351 Korean Patent Publication No. 2011-0068923

The object of the present invention is to replace the existing synthetic whitening substances or natural products obtained on land, but it has little side effects by using tuna, a kind of sea horse that grows in the ocean, and has very little skin whitening effect due to melanin biosynthesis inhibition. It is to provide an excellent skin whitening cosmetic composition, food composition and pharmaceutical composition for preventing or treating melanin pigmentation disease.

According to one aspect of the invention,

Provided is a cosmetic composition for skin whitening comprising an extract of kammaksalsal as an active ingredient.

The kammaksal extract may be extracted with water, alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof.

The cosmetic composition for whitening may be to inhibit melanin synthesis.

The cosmetic composition for whitening may be to inhibit tyrosinase activity.

The cosmetic composition for skin whitening may be any one selected from cleansing, lotion, cream, massage cream, eye cream, essence, gel, pack, spray, powder and sun cream for face wash.

According to another aspect of the invention,

Provided is a pharmaceutical composition for preventing or treating melanin pigmentation disease, which includes the tuna extract as an active ingredient.

The kammaksal extract may be extracted with water, alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof.

The melanin pigmentation disease may be any one selected from spots, freckles, blotch, liver, otamoban, sunburn, and pigmentation after trauma or inflammation.

The pharmaceutical composition may be any one selected from granules, powders, tablets, coatings, capsules, liquids, syrups, juices, suspensions, emulsions, drops and injection solutions.

According to another aspect of the present invention,

Provided is a food composition for skin whitening, comprising the extract of kammaksalsal as an active ingredient.

The skin whitening food composition may be added to any one selected from beverages, alcoholic beverages, confectionery, diet bars, dairy products, meat, chocolate, pizza, ramen, noodles, gums, ice cream, and vitamin complexes.

According to another aspect of the invention,

A method for producing a cosmetic composition for whitening is provided, which comprises extracting tuna skin with water, alcohol having 1 to 4 carbons, or a mixed solvent thereof to prepare tuna skin extract.

The kammaksal extract may be a fraction obtained by adding water and alcohol having 1 to 4 carbon atoms.

After the above step, the step of concentration under reduced pressure may be further performed.

According to another aspect of the invention,

Provided is a method for preparing a pharmaceutical composition for preventing or treating pigmentation disease, comprising extracting kamaksal with water, alcohol having 1 to 4 carbons, or a mixed solvent thereof to prepare kamaksal extract.

The kammaksal extract may be a fraction obtained by adding water and alcohol having 1 to 4 carbon atoms.

After the above step, the step of concentration under reduced pressure may be further performed.

The cosmetic composition for skin whitening, the food composition of the present invention, and the pharmaceutical composition for preventing or treating melanin pigmentation diseases replace the existing synthetic whitening substances or natural products obtained on the ground to replace tuna, a kind of sea horse that grows in the ocean. It shows little side effects to the human body, and has a very good effect of skin whitening effect due to melanin biosynthesis inhibition.

1 is a result of measuring the cell viability according to Experimental Example 1.
2 is a result of measuring the amount of melanin synthesis according to Experimental Example 2.
3 is a result of measuring the degree of tyrosinase activity according to Experimental Example 3.

The present invention can be applied to various transformations and can have various embodiments, and specific embodiments will be illustrated in the drawings and described in detail in the detailed description. However, this is not intended to limit the present invention to specific embodiments, and should be understood to include all conversions, equivalents, and substitutes included in the spirit and scope of the present invention. In the description of the present invention, when it is determined that a detailed description of known technologies related to the present invention may obscure the subject matter of the present invention, the detailed description will be omitted.

In the present invention, the kamaksal extract may be extracted using water, an organic solvent, or a mixture thereof as an extraction solvent. In this case, the type of the organic solvent used or the mixing ratio of water and the organic solvent is not particularly limited.

For example, the organic solvent may be one or more solvents selected from the group consisting of lower alcohol, hexane, acetone, ethyl acetate, chloroform, and diethyl ether. The lower alcohol may be an alcohol having 1 to 6 carbon atoms. For example, methanol, ethanol, propanol, butanol, normal-propanol, iso-propanol, normal-butanol, 1-pentanol, 2-butoxyethanol or ethylene glycol may be used as the lower alcohol. Other organic solvents include polar solvents such as acetic acid, dimethyl-formamide (DMFO), and dimethyl sulfoxide (DMSO), acetonitrile, ethyl acetate, methyl acetate, fluoroalkane, pentane, 2,2,4-trimethylpentane, and decane. , Cyclohexane, cyclopentane, diisobutylene, 1-pentene, 1-chlorobutane, 1-chloropentane, o-xylene, diisopropyl ether, 2-chloropropane, toluene, 1-chloropropane, chlorobenzene Non-polar solvents such as benzene, diethyl ether, diethyl sulfide, chloroform, dichloromethane, 1,2-dichloroethane, aniline, diethylamine, ether, carbon tetrachloride, and tetrahydrofuran (THF) can also be used.

As can be seen in the following examples, when extracting tuna with organic solvents, kamaksal extract is separated into supernatant and lower layer liquid, the supernatant being the oil fraction of tuna extract, and the lower layer liquid is common Corresponds to the solvent extract. When the lower layer liquid is separated again through centrifugation or filter paper, it is divided into liquid and solid residues. Therefore, the tuna extract of the present invention is interpreted as a concept including all of the oil fraction of the tuna extract, the liquid fraction of the tuna extract, and the wax fraction of the tuna extract. In one embodiment, the kammaksal extract may include any one or more selected from the group consisting of the oil fraction of kammaksal extract, the liquid fraction of kammaksal extract and the wax fraction of kammaksal extract.

In one embodiment, the chammaksal extract may be a lower alcohol extract of kammaksal, preferably ethanol extract of kammaksal.

As used herein, the term'extract' also includes a fraction in which an extract is additionally fractionated. That is, the extract of kammaksalsal is obtained not only by using the above-described extraction solvent, but also obtained by additionally applying a purification process. In addition, the extract or fraction obtained by passing through an ultrafiltration membrane having a constant molecular weight cut-off value, various chromatography (size, charge, hydrophobicity or separation by affinity), etc. Fractions obtained through various purification methods are also included in the extract of kammaksal of the present invention.

In one embodiment, the kammaksal extract may be a fraction of the organic solvent extract of kammaksal redistributed as a second organic solvent. Here, the term “organic solvent extract of tuna maksal” includes all of the oil fraction of kammaksal extract, the liquid fraction of gammakmaksal extract, and the wax fraction of kammaksal extract, in the broad sense. Refers to the liquid fraction of tuna black meat extract. Therefore, in one embodiment, the fraction obtained by re-fractionating the organic extract of tuna kamaksal with a second organic solvent may mean a fraction obtained by re-fractionating the liquid fraction of kammaksal extract with a second organic solvent. In another embodiment, the kammaksal extract may be a fraction obtained by re-fractionating the lower alcohol extract of kammaksal with a second organic solvent. In another embodiment, the kammaksal extract may be a fraction of ethanol extract of kammaksal re-fractionated with hexane. As can be seen in the following examples, fractions of tuna extract with a large amount of fat-soluble components of such tuna extract show a very good effect.

The term'extract', which is used in referring to tuna kamaksal in this specification, includes not only the crude extract obtained by treating the makkasal with an extraction solvent, but also a processed product of makkamaksal extract. For example, the black sea bream extract can be prepared in powder form by additional processes such as distillation under reduced pressure and freeze drying or spray drying.

In addition, the tuna extract of the present invention has a meaning in the broad sense also including chammaksal processed products, such as kammaksal powder, formulated to be able to administer the gammaksal itself to the human body. Although experiments were conducted with the extract of kamaksal in the present invention, it will be predictable to those skilled in the art that the desired effect can be achieved even in the same form as the kamaksal processed product.

On the other hand, in the present specification, the term'comprising as an active ingredient' means to include an amount sufficient to achieve the efficacy or activity of the tuna extract. In one embodiment of the present invention, the blackcurrant extract in the composition of the present invention is, for example, 0.001 mg/kg or more, preferably 0.1 mg/kg or more, more preferably 10 mg/kg or more, and more It is preferably 100 mg/kg or more, even more preferably 250 mg/kg or more, and most preferably 1 g/kg or more. As the natural tuna extract is not a side effect to the human body even when administered in an excessive amount, the upper limit of the quantity of the tuna extract contained in the composition of the present invention can be selected and carried out by a person skilled in the art within an appropriate range.

Ingredients included in the cosmetic composition for skin whitening comprising the extract of the kammaksal flesh of the present invention as an active ingredient include ingredients commonly used in cosmetic compositions in addition to the active ingredient as an active ingredient, for example, antioxidants, stabilizers Includes conventional adjuvants, such as solubilizers, vitamins, flavorings, and agents. The cosmetic composition of the present invention may be prepared in any formulation conventionally prepared in the art, for example, solutions, suspensions, emulsions, pastes, gels, creams, lotions, powders, soaps, cleansers containing surfactants, oils , Powder foundation, emulsion foundation, wax foundation, and the like, but is not limited thereto. More specifically, it can be prepared in the form of cleansing, lotion, cream, massage cream, eye cream, essence, gel, pack, spray, powder, sun cream for face wash.

When the formulation of the present invention is a paste, cream or gel, vegetable oil, wax, starch, cellulose, silicone, bentonite, silica, zinc oxide, etc. may be used as a carrier component, and when the formulation is a powder, lactose, silica as a carrier component , Powders such as aluminum hydroxide and calcium silicate may be used. When the formulation of the present invention is a solution or emulsion, a solvent, solubilizer or emulsifier is used as a carrier component, such as water, ethanol, glycerin, ethyl carbonate, ethyl acetate, propylene glycol, butylene glycol, 1 ,2-hexanediol, glycerol aliphatic ester, and sorbitan fatty acid ester.

When the formulation of the present invention is a suspension, as a carrier component, liquid diluents such as water, ethanol, and propylene glycol, suspensions such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, Microcrystalline cellulose, aluminum metahydroxide, bentonite, agar and the like can be used. In addition, when the formulation is a surfactant-containing cleansing, as the carrier component, aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, methyl taurate, sarcosinate, alkyl amidobetaine, aliphatic alcohol, fatty acid glycerol Rides, vegetable oils or ethoxylated glycerol fatty acid esters and the like can be used.

The pharmaceutical composition for prevention or treatment of pigmentation disease comprising the extract of the black sea bream of the present invention as an active ingredient may be prepared by using a pharmaceutically acceptable and physiologically acceptable adjuvant in addition to the active ingredient. Excipients, disintegrants, sweeteners, binders, coating agents, expanding agents, lubricants, lubricants or flavoring agents can be used.

The pharmaceutical composition may be preferably formulated into a pharmaceutical composition including one or more pharmaceutically acceptable carriers in addition to the active ingredients described above for administration.

Formulations of the pharmaceutical composition may be granules, powders, tablets, coatings, capsules, suppositories, liquids, syrups, juices, suspensions, emulsions, drops or injectables. For example, for formulation in the form of tablets or capsules, the active ingredient can be combined with an oral, non-toxic pharmaceutically acceptable inert carrier such as ethanol, glycerol, water, and the like. In addition, if desired or necessary, suitable binders, lubricants, disintegrants and colorants can also be included in the mixture. Suitable binders include, but are not limited to, natural sugars such as starch, gelatin, glucose or beta-lactose, corn sweeteners, natural and synthetic gums such as acacia, trachocanth or sodium oleate, sodium stearate, magnesium stearate, sodium Benzoate, sodium acetate, sodium chloride, and the like. Disintegrants include, but are not limited to, starch, methyl cellulose, agar, bentonite, xanthan gum, and the like.

Acceptable pharmaceutical carriers in compositions formulated as liquid solutions include, as sterile and biocompatible, saline, sterile water, Ringer's solution, buffered saline, albumin injection solution, dextrose solution, maltodextrin solution, glycerol, ethanol and One or more of these components may be mixed and used, and other conventional additives such as antioxidants, buffers and bacteriostatic agents may be added as necessary. In addition, diluents, dispersants, surfactants, binders, and lubricants may be additionally added to formulate into injectable formulations such as aqueous solutions, suspensions, emulsions, pills, capsules, granules or tablets.

Furthermore, it can be preferably formulated according to each disease or ingredient using methods disclosed in Remington's Pharmaceutical Science, Mack Publishing Company, Easton PA by appropriate methods in the art.

The pharmaceutical composition of the present invention may be administered orally or parenterally, and in the case of parenteral administration, intravenous injection, subcutaneous injection, intramuscular injection, intraperitoneal injection, transdermal administration, etc., preferably oral administration.

Suitable dosages of the pharmaceutical compositions of the invention vary by factors such as formulation method, mode of administration, patient's age, weight, sex, morbidity, food, time of administration, route of administration, rate of excretion, and response sensitivity, usually As a result, a skilled doctor can easily determine and prescribe a dose effective for the desired treatment or prevention. According to a preferred embodiment of the invention, the daily dosage of the pharmaceutical composition of the invention is 0.001-10 g/kg.

The pharmaceutical composition of the present invention is prepared in a unit dose form by formulating using a pharmaceutically acceptable carrier and/or excipient according to a method that can be easily carried out by a person skilled in the art to which the present invention pertains. Or it can be manufactured by incorporating it into a multi-dose container. At this time, the formulation may be in the form of a solution, suspension, or emulsion in an oil or aqueous medium, or may be in the form of ex, powder, granule, tablet, or capsule, and may further include a dispersant or stabilizer.

The present invention also provides a food composition for skin whitening comprising an extract of blackcurrant as an active ingredient.

The food composition according to the present invention may be formulated in the same manner as the pharmaceutical composition and used as a functional food, or added to various foods. Foods to which the composition of the present invention can be added include, for example, beverages, alcoholic beverages, confectionery, diet bars, dairy products, meat, chocolate, pizza, instant noodles, other noodles, gums, ice cream, vitamin complexes, and health supplements And so on.

The food composition of the present invention, as an active ingredient, may include not only kamaksal extract or kamaksal powder, but also ingredients that are commonly added during food production, for example, proteins, carbohydrates, fats, nutrients, seasonings And flavoring agents. Examples of the carbohydrates described above include monosaccharides, such as glucose, fructose, and the like; Disaccharides such as maltose, sucrose, oligosaccharides, etc.; And polysaccharides, for example, conventional sugars such as dextrin, cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As a flavoring agent, natural flavoring agents (tauumatin, stevia extract (for example, rebaudioside A, glycyrrhizine, etc.)) and synthetic flavoring agents (saccharin, aspartame, etc.) can be used. For example, when the food composition of the present invention is made of a drink and a beverage, citric acid, liquid fructose, sugar, glucose, acetic acid, malic acid, juice, and various plant extracts, etc. may be additionally included in addition to the kamakmak extract of the present invention. have.

The present invention provides a health functional food comprising a food composition for skin whitening that includes the tuna extract as an active ingredient. Health functional foods are foods made by adding kamaksal extract or kamaksal powder to food materials such as beverages, tea, spices, chewing gum, confectionery, etc., or foods prepared by encapsulation, powdering, suspension, etc. It means that it has a certain specific effect, but it has the advantage of not having side effects that can occur when taking the drug for a long time using food as a raw material, unlike general drugs. The health functional food of the present invention obtained in this way is very useful because it can be taken on a daily basis. The added amount of tuna black meat extract or black tuna powder in the health functional food cannot be uniformly defined depending on the type of the health functional food, but can be added in a range that does not impair the original taste of the food. , It is usually in the range of 0.01 to 50% by weight, preferably 0.1 to 20% by weight relative to the target food. In addition, in the case of a health functional food in the form of pills, granules, tablets or capsules, it is usually added in the range of 0.1 to 100% by weight, preferably 0.5 to 80% by weight. In one embodiment, the dietary supplement of the present invention may be in the form of pills, tablets, capsules or beverages.

The present invention also provides the use of a blackcurrant extract for the manufacture of cosmetic or pharmaceutical products for whitening. As described above, kamaksal extract or kamaksal powder can be used for whitening purposes.

In addition, the present invention provides a method for preventing or treating melanin pigmentation disease, including a method of whitening, comprising administering an effective amount of tuna extract to the human body.

As used herein, the term “effective amount” refers to the amount of an active ingredient or pharmaceutical composition that induces a biological or medical response in a tissue system, human, as considered by a researcher, physician, or other clinician, which indicates that the disease or disorder Contains the amount that induces relief of symptoms. It is apparent to those skilled in the art that the effective amount and the number of administrations for the active ingredient of the present invention will vary depending on the desired effect. Therefore, the optimal dosage to be administered can be easily determined by those skilled in the art, the type of disease, the severity of the disease, the content of active ingredients and other ingredients in the composition, the type of formulation, and the patient's age, weight, general health It can be adjusted according to various factors, including condition, sex and diet, time of administration, route of administration and secretion rate of the composition, duration of treatment, and drugs used simultaneously.

In the method of treatment of the present invention, the composition comprising the extract of blackcurrant as an active ingredient is conventional through oral, rectal, intravenous, intraarterial, intraperitoneal, intramuscular, intrasternal, transdermal, topical, intraocular or intradermal routes. Can be administered in a manner.

Advantages and features of the present invention, and methods for achieving them will be clarified with reference to embodiments described below in detail. However, the present invention is not limited to the embodiments disclosed below, but will be implemented in various different forms, and only the present embodiments allow the disclosure of the present invention to be complete, and common knowledge in the technical field to which the present invention pertains. It is provided to fully inform the holder of the scope of the invention, and the invention is only defined by the scope of the claims.

Hereinafter, a preferred embodiment is provided to help the understanding of the present invention, but the following examples are only illustrative of the present invention, and it is apparent to those skilled in the art that various changes and modifications are possible within the scope and technical scope of the present invention. It is no wonder that such variations and modifications fall within the scope of the appended claims.

[Example]

Example One: Blackcurrant Preparation of extract

The dried blackcurrant (Polyopes affinis) was placed in an Erlenmeyer flask, and 80% methanol was added thereto, followed by immersion extraction for 24 hours. After repeating this process twice, the obtained extract was filtered with filter paper to remove debris from the blackcurrant, and the obtained filtrate was concentrated using a vacuum concentrator. After diluting the fraction by adding tertiary distilled water and butanol to the fully concentrated sample, the obtained fraction is concentrated under reduced pressure, and the obtained product is dried using a freeze dryer to obtain a blackjack extract, followed by DMSO (dimethyl sulfoxide, dimethyl sulfoxide, Sigma) (USA, USA) to a final concentration of 100 mg/mL, and then passed through a membrane filter paper (0.2 μm, Millipore, USA) using a disposable syringe to prepare aseptically.

[Experimental Example]

Cell model processing method

DMEM (Dulbecco's Modified Eagle's Medium) DMEM containing 1% penicillin-streptomycin and 10% fetal bovine serum, respectively, from the mouse-derived melanoma cells B16F10 cell line at 37°C and 5% CO ₂ conditions. Cultured in medium (Gibco BRL, USA). The cultured B16F10 cell line is dispensed into 2 x 10 ⁴ /mL pieces in a 24-well plate or 10 ml culture dish, and after 24 hours, the existing medium is removed and α-MSH, a melanin synthesis inducing hormone having a concentration of 200 nM ( Sigma aldrich (USA), arbutin (Sigma aldrich), a melanin synthesis inhibitor having a concentration of 1 mM, and tuna perilla extract prepared according to Example 1 above, at concentrations of 1, 2.5, and 5 μg/ Cells were cultured after treatment to be ml. After 72 hours, the cultured cells were used in experiments for measuring the cell viability of Experimental Example 1, the amount of melanin synthesis of Experimental Example 2, and the degree of tyrosinase enzyme activity of Experimental Example 3.

Experimental Example One: Blackcurrant Measurement of cell viability in cell model of extract

In order to measure cell viability, a solution of MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium) at a concentration of 0.05 mg/ml was added to the cultured cells, and in a cell incubator for 1 hour. While inducing a reduction reaction while incubating, the MTT solution was removed and DMSO was added to completely dissolve formazan crystals. The resulting solution was put into each well of a 96-well plate, 100 µl, and measured at an absorbance of 570 nm using a microplate reader, and the results are shown in FIG. 1.

According to this, when treated with α-MSH (α-melanocyte-stimulating hormone) alone, it exhibited some toxicity, and extracts of tuna with concentrations of 1, 2.5 and 5 μg/ml treated with α-MSH were α- It did not show cytotoxicity in the B16F10 cell line compared to the MSH alone treatment group. In addition, when treated with arbutin (arbutin) also showed no significant change.

Experimental Example 2: Blackcurrant Measurement of melanin synthesis in cell model of extract

In order to measure the amount of melanin produced in the melanin synthesis-inducing cell model, 100 µl of the obtained product obtained by lysing the cells using 1N NaOH at a concentration of 10% DMSO in the cultured cells was added to each well of a 96-well plate. The result was measured in absorbance at 450 nm using a microplate reader, and the results are shown in FIG. 2.

According to this, in the case of treatment with α-MSH, melanin synthesis amount was decreased in a concentration-dependent manner when melanocyte synthesis-induced cells were treated with concentrations of 1, 2.5 and 5 µg/ml of blackcurrant extract. In addition, when arbutin was treated, the amount of melanin synthesis was also significantly decreased.

Experimental Example 3: Blackcurrant Measurement of tyrosinase activity in cell model of extract

In order to measure the degree of tyrosinase activity of the tuna extract, the amount of protein was quantified to 40 µg using the BCA protein quantification method of the supernatant of the cultured cells. In a 96-well plate, 100 µL of PBS and pH tyrosinase with 5 mM concentration of L-DOPA added, and 100 µl of the tyrosinase supernatant were added thereto and reacted for 1 hour. It was measured at an absorbance of 475 nm using a microplate reader, and the results are shown in FIG. 3.

According to this, in the case of treatment with α-MSH, in the cells in which melanin synthesis was induced, the concentration of 1, 2.5 and 5 µg/ml of black buckthorn extract was decreased in a concentration-dependent manner. In addition, when treated with arbutin, the degree of tyrosinase activity was also significantly reduced.

The embodiments of the present invention have been described above, but those skilled in the art can add, change, delete, or add elements within the scope of the present invention as set forth in the claims. It will be said that the present invention can be variously modified and changed by the like, and this is also included within the scope of the present invention.

Claims (17)

Contains the butanol fraction of methanol extract of polyopes affinis as an active ingredient,
Cosmetic composition for skin whitening having tyrosinase inhibitory activity. delete According to claim 1,
The cosmetic composition for whitening has a skin whitening cosmetic composition characterized in that to inhibit the synthesis of melanin. delete According to claim 1,
The cosmetic composition for skin whitening is a cosmetic composition for skin whitening, characterized in that it is any one selected from cleansing for face wash, lotion, cream, massage cream, eye cream, essence, gel, pack, spray, powder, and sun cream. Contains the butanol fraction of methanol extract of polyopes affinis as an active ingredient,
A pharmaceutical composition for preventing or treating melanin pigmentation disease having tyrosinase inhibitory activity. delete The method of claim 6,
The melanin pigmentation disease is a melanin pigmentation disease prevention or treatment pharmaceutical composition, characterized in that any one selected from pigmentation after freckles, freckles, blotch, liver, otamoban, sunburn, and trauma or inflammation. The method of claim 6,
The pharmaceutical composition is granules, powders, tablets, coatings, capsules, liquids, syrups, juices, suspensions, emulsions, drops and injection solutions, melanin pigmentation disease prevention or treatment pharmaceuticals characterized in that any one selected from Composition. Contains the butanol fraction of methanol extract of polyopes affinis as an active ingredient,
Food composition for skin whitening having tyrosinase inhibitory activity. The method of claim 10,
The food composition for skin whitening is added to any one selected from beverages, confectionery, dairy products, meat, noodles, gums, and ice creams. A method for producing a cosmetic composition for skin whitening having tyrosinase inhibitory activity, comprising extracting tuna kelp ( Polyopes affinis ) using methanol, followed by fractionation by adding distilled water and butanol to obtain a butanol fraction. delete The method of claim 12,
Method for producing a cosmetic composition for skin whitening having a tyrosinase inhibitory activity, characterized in that further performing a step of concentration under reduced pressure after the step. After extracting the tuna black pepper ( Polyopes affinis ) using methanol, fractionation by adding distilled water and butanol, and then obtaining a butanol fraction, comprising a pharmaceutical composition for preventing or treating melanin pigmentation disease having tyrosinase inhibitory activity Manufacturing method. delete The method of claim 15,
Method for producing a pharmaceutical composition for preventing or treating melanin pigmentation disease having tyrosinase inhibitory activity, characterized in that further performing a step of concentration under reduced pressure after the step.

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