KR20150050995A - Composition for skin whitening comprising amaranthus spp. l. extract or fraction thereof - Google Patents

Abstract

The present invention relates to a composition for skin whitening containing an extract of amaranth (Amaranthus spp. L.) or a fraction thereof. More specifically, the invention relates to a cosmetic composition for skin whitening or for improving skin pigmentation, a quasi-drug composition, or to a food composition, which contain an extract of amaranth or a fraction thereof as an active ingredient. In addition, the present invention relates to a pharmaceutical composition for preventing or treating skin pigmentation diseases, which contains the extract of amaranth or the fraction thereof as an active ingredient. In addition, the present invention relates to a method for improving skin whitening and preventing or improving skin pigmentation by using the composition, or a method for preventing, improving, or treating skin pigmentation diseases.

Description

TECHNICAL FIELD [0001] The present invention relates to a skin whitening composition containing an amaranth extract or a fraction thereof. L. EXTRACT OR FRACTION THEREOF}

The invention amaranth (Amaranthus spp . L.) extract or a fraction thereof. More specifically, the present invention relates to a cosmetic composition, a quasi-drug composition or a food composition for improving skin whitening or coloring, which comprises an extract of Amaranth or a fraction thereof as an active ingredient.

The present invention also relates to a pharmaceutical composition for preventing or treating skin pigmentation diseases containing an extract of Amaranth or a fraction thereof as an active ingredient.

The present invention also relates to a method for improving skin whitening, preventing or improving pigmentation, or preventing, ameliorating or treating skin pigmentation diseases using the above compositions.

Melanin, which determines human skin color, is produced in melanocytes. Specifically, melanin is a black-brown pigment formed by a polymerization oxidation reaction with an enzyme such as tyrosinase existing in melanocytes, using tyrosine as an amino acid present in vivo as a substrate. The melanin thus formed is transferred to the epidermal cell called keratinocyte through the dendrites of melanocytes. Melanin, which has been transferred to keratinocytes, can be removed by keratinocytes falling off the skin when they fall off the epidermis. However, since there is no enzyme that degrades melanin in vivo, the once formed melanin is not degraded in vivo. Therefore, in order to lighten the skin, it is important to suppress the production of melanin.

Recently, the study of whitening agents as cosmetics has become increasingly necessary as skin blackness is recognized as skin aging due to ultraviolet rays, along with the improvement of living standards in Asian countries, which prefer emotionally white skin. Accordingly, substances exhibiting tyrosinase inhibitory activity such as ascorbic acid, hydroquinone, glutathione, and arbutin have been used in combination with cosmetics or medicines (Patent Documents 1 and 2) 2) Most of them are ineffective or unstable because of their shape. In particular, a compound such as hydroquinone exhibits a strong decolorizing action, has a skin sensitization itself, can induce skin allergies and the like, and exhibits adverse effects such as inducing vitiligo by changing normal skin functions. Thus, Its use is limited.

In addition, even if the substance inhibits the activity of tyrosinase as described above, if such a substance acts on keratinocytes, which are keratinocytes, to increase the production of factors promoting melanin biosynthesis, the whitening action is consequently weakened The active inhibitor of tyrosinase is not considered to be a good whitening agent because it can cause harmful effects, and many tyrosinase inhibitors have been developed, but most of them are known to show no excellent whitening effect.

Under these circumstances, the present inventors have conducted studies on substances which are not toxic and have excellent whitening action from natural natural resources, and Amaranthus spp . L.) extract inhibits the activity of tyrosinase and inhibits the production of melanin pigment, and has completed the composition for skin whitening utilizing the same.

Korean Patent Registration No. 10-0923141 Korean Patent Registration No. 10-0921947

The invention amaranth (Amaranthus spp . L.) extract of the present invention.

Specifically, one object of the present invention is to provide a cosmetic composition, a quasi-drug composition, a food composition, and a cosmetic composition for improving skin whitening or coloring, comprising the Amaranth extract or a fraction thereof as an active ingredient, Thereby providing a method of preventing or improving deposition.

Another object of the present invention is to provide a pharmaceutical composition for preventing or treating skin pigmentation diseases containing an amaranth extract or a fraction thereof as an active ingredient, and a method for preventing or treating skin pigmentation disease using the composition.

The present invention as an embodiment for solving the above-mentioned problem is amaranth (Amaranthus spp . L.) extract or a fraction thereof as an active ingredient for skin whitening or pigmentation improvement.

In the present invention, Amaranthus spp. L. is a dicotyledon belonging to Amaranthaceae. Although it is not classified as a cereal, it is known that it has similar properties to common grains and similar uses. Amaranth seeds contain about 48% to 69% of starch and contain high quality fatty acids with high fat content and high unsaturation compared to cereals. In addition, amaranth is rich in lysine and sulfur-containing amino acids, and contains a large amount of protein with superior amino acid composition, and has attracted attention as a new protein of vegetable protein. Amaranth is also used for industrial purposes as it is used as a lubricant for computer diskettes as well as for food because of high plant squalene content.

The amaranth in the present invention is more preferably amaranth Tuscan crew enteoseu (Amaranthus cruentus), amaranth Tuscan hive Reader (Amaranthus hybridus), or amaranth Tuscan hypo mitochondrial carcass (Amaranthus hypocondriacus ) and amaranthus hive leader .

The term "extract" used in the present invention refers to an extract obtained by extracting amaranth, a diluted solution or concentrate of the extract, a dried product obtained by drying the extract, a preparation or a purified product of the extract, Extracts themselves and extracts of all formulations which can be formed using extracts.

The Amaranth extract of the present invention can be extracted from various organs of the natural, hybrid or variant plants of Amaranth and can be used as a seed of Amaranth such as seeds, roots, ground parts, stems, leaves, flowers, It can be extracted not only from the bark of the fruit but also from the plant tissue culture.

The amaranth extract of the present invention may preferably be an extract of seeds of amaranth.

In the amaranth extract of the present invention, the method for extracting the amaranth is not particularly limited and may be carried out according to a method commonly used in the art. Non-limiting examples of the extraction method include hydrothermal extraction, ultrasonic extraction, filtration, and reflux extraction. These may be performed alone or in combination with two or more methods.

In the present invention, the kind of the extraction solvent used for extracting the amaranth is not particularly limited, and any solvent known in the art can be used. Examples of the extraction solvent include water, distilled water, alcohol, or a mixed solvent thereof. When the alcohol is used as a solvent, it is more preferable to use a C ₁ to C ₄ alcohol such as methanol, ethanol, propanol , Butanol) can be used. In the present invention, water or distilled water may be more preferably used as the extraction solvent of amaranth.

The term "fraction " as used herein means a product obtained by performing fractionation to separate a specific component or a specific component group from a mixture containing various components.

The fractionation method for obtaining the fraction in the present invention is not particularly limited and may be carried out according to a method commonly used in the art. As a non-limiting example of the above-mentioned fractionation method, there can be mentioned a method in which a fraction obtained from the extract is treated with a predetermined solvent by treating the extract obtained by extracting amaranth.

The kind of the fraction solvent used for obtaining the fraction in the present invention is not particularly limited, and any solvent known in the art can be used. Non-limiting examples of the fraction solvent include polar solvents such as water, distilled water and alcohol; And non-polar solvents such as hexane, ethyl acetate, chloroform, and dichloromethane. These may be used alone or in combination of two or more. When alcohols are used in the fraction solvent, C ₁ to C ₄ alcohols can be preferably used. The fraction solvent used in the present invention may more preferably be water, distilled water, hexane or ethyl acetate.

The amaranth extract or its fractions of the present invention have an effect of effectively inhibiting the production of melanin, specifically showing the effect of inhibiting the activity of tyrosinase which catalyzes the step of determining the rate of melanin production, and as an enzyme associated with melanin production TRP-2, and MITF, which are enzymes associated with tyrosinase.

Tyrosinase is a metal-containing protein containing a pair of copper ions in its active site. L-tyrosine is hydroxylated by 3,4-dihydroxyphenylalanine (L-DOPA) in the production pathway of melanin Which is oxidized to phenylalanine-3,4-quinone (dopaquinone) and then involved in the step of synthesizing melanin through various intermediates such as dopachrome, indole-5,6-quinone .

The extract of Amaranth or the fraction thereof of the present invention has an effect of inhibiting the activity of tyrosinase, and particularly, it has an excellent effect of inhibiting autoxidation of DOPA of tyrosinase.

According to one embodiment of the present invention, melanocyte-derived cells derived from embryo of C57BL as a melanocyte cell line of the mouse are treated with the extract of Amaranth or a fraction thereof in a concentration-dependent manner, It has been found that the melanin content is reduced dependent on the treatment concentration of the amaranth extract or its fraction (Examples 2 and 6).

According to another embodiment, it has been confirmed that, when melan-A cells are treated with the extract of Amaranth or a fraction thereof, the autoxidation of DOPA of tyrosinase is inhibited depending on the treatment concentration of the amaranth extract or its fraction ( (Example 3), and the expression of tyrosinase-related enzymes TRP-1, TRP-2 and MITF was also found to be reduced depending on the concentration of the treatment of the extract of Amaranth or its fraction (Example 4).

As used herein, the term "skin whitening" means any action that inhibits the synthesis of melanin and inhibits or prevents skin deposition of melanin.

As used herein, the term "skin pigmentation" refers to any action or condition that causes skin color to darken due to increased synthesis of melanin.

As used herein, the term "improvement" means any action that at least reduces the degree of symptomatology, such as a parameter associated with relief or treatment of a condition.

The cosmetic composition of the present invention can be used as a cosmetic composition in the form of a solution, a topical ointment, a cream, a foam, a nutritional lotion, a softening lotion, a pack, a soft water, a latex, a makeup base, But are not limited to, those selected from the group consisting of solutions, pastes, gels, lotions, powders, soaps, surfactant-containing cleansing, oils, powdered foundations, emulsion foundations, wax foundations, patches and sprays no.

The cosmetic composition of the present invention may further comprise at least one cosmetically acceptable carrier to be incorporated in a cosmetic composition for general skin. Examples of the cosmetic composition include ordinary ingredients such as oil, water, a surfactant, a moisturizer, a lower alcohol, , A chelating agent, a coloring agent, a preservative, a perfume, and the like may be appropriately compounded, but the present invention is not limited thereto.

The cosmetically acceptable carrier contained in the cosmetic composition of the present invention varies depending on the formulation of the cosmetic composition.

When the formulations of the present invention are ointments, pastes, creams or gels, the carrier component may be an animal oil, a vegetable oil, a wax, a paraffin, a starch, a tracer, a cellulose derivative, polyethylene glycol, silicon, bentonite, silica, talc, zinc oxide May be used, but is not limited thereto. These may be used alone or in combination of two or more.

When the formulation of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate, polyamide powder and the like may be used as a carrier component. In particular, But are not limited to, propellants such as rocaborn, propane / butane or dimethyl ether. These may be used alone or in combination of two or more.

When the formulation of the present invention is a solution or an emulsion, a solvent, a dissolving agent or an emulsifying agent may be used as a carrier component, and examples thereof include water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, Propylene glycol, 1,3-butyl glycol oil and the like can be used, and particularly fatty acid esters of cottonseed oil, peanut oil, corn oil, olive oil, castor oil and sesame oil, glycerol aliphatic ester, polyethylene glycol or sorbitan May be used, but is not limited thereto. These may be used alone or in combination of two or more.

When the formulation 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, Crystalline cellulose, aluminum metahydroxide, bentonite, agar or tracant, but are not limited thereto. These may be used alone or in combination of two or more.

When the formulation of the present invention is a soap, use is made of an alkali metal salt of a fatty acid, a fatty acid hemiester salt, a fatty acid protein hydrolizate, isethionate, a lanolin derivative, an aliphatic alcohol, a vegetable oil, glycerol, But is not limited thereto. These may be used alone or in combination of two or more.

According to another embodiment of the present invention, there is provided a quasi-skin composition for skin whitening or improvement of pigmentation containing Amaranthus spp. L. extract or a fraction thereof as an active ingredient.

The term "quasi-drug product" used in the present invention means products that are less active than drugs, among the products used for diagnosing, treating, improving, alleviating, treating or preventing diseases of human or animal. For example, Quasi-drugs are products that are used for the purpose of treating or preventing diseases of humans or animals, products that are mild or have no direct action on the human body.

The quasi-drug composition of the present invention can be manufactured by, but not limited to, a body cleanser, a foam, a soap, a mask, an ointment, a cream, a lotion, an essence and a spray.

In the quasi-drug composition of the present invention, the description of the amaranth extract or its fractions, skin whitening, skin pigmentation, and improvement is the same as described above in connection with the cosmetic composition.

According to another embodiment of the present invention, Amaranthus spp . L.) extract or a fraction thereof as an active ingredient for skin whitening or pigmentation improvement.

The food composition of the present invention is not particularly limited and includes a health functional food composition.

When the health functional food composition of the present invention is used as a food additive, the composition may be added as it is or may be used together with other food or food ingredients, and may be appropriately used according to a conventional method.

The kind of the food is not particularly limited, and includes food in a conventional sense. Non-limiting examples of the food to which the substance can be added include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gums, dairy products including ice creams, , A drink, an alcoholic beverage, and a vitamin complex.

When the health functional food composition of the present invention is a beverage composition, various flavoring agents, natural carbohydrates, and the like may be contained as additional components such as ordinary beverages. Non-limiting examples of such natural carbohydrates include monosaccharides such as glucose and fructose; Disaccharides such as maltose and sucrose; Natural sweetening agents such as dextrin, cyclodextrin; Synthetic sweetening agents such as saccharin and aspartame, and the like. The proportion of the additional component added may be appropriately determined by a person skilled in the art.

In addition to the above, the health functional food composition of the present invention may contain various nutrients, vitamins, electrolytes, flavors, colorants, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloid thickening agents, pH adjusting agents, stabilizers, , Alcohols, carbonating agents used in carbonated drinks, and the like. In addition, the health functional food composition of the present invention may contain pulp for the production of natural fruit juice, fruit drink or vegetable drink. These components may be used independently or in combination of two or more. The ratios of these additives can also be suitably selected by those skilled in the art.

In the food composition of the present invention, the description of the amaranth extract or its fractions, skin whitening, skin pigmentation, and improvement is the same as described above in connection with the cosmetic composition.

In the cosmetic composition, quasi-drug composition or food composition of the present invention, the Amaranth extract or its fraction may be contained preferably in an amount of 0.0001% by weight to 50% by weight, more preferably 0.01% By weight to 40% by weight, and more preferably 0.1% by weight to 30% by weight. There is an advantage of exhibiting excellent whitening effect within the above range.

According to another embodiment of the present invention, Amaranthus spp . L.) extract or a fraction thereof as an active ingredient. The present invention also provides a pharmaceutical composition for preventing or treating skin pigmentation diseases.

In the pharmaceutical composition of the present invention, the description of the amaranth extract or its fractions, and skin pigmentation is the same as described above in connection with the cosmetic composition.

The skin pigmentation disease of the present invention means all diseases caused by skin pigmentation due to an increase in the production of melanin. Non-limiting examples of the skin pigmentation diseases include stain, freckles, spots, daylight pigment, pigmentation after drug use, inflammation pigmentation, and pregnancy pigmentation.

In the pharmaceutical composition of the present invention, the effective amount of the amaranth extract or its fraction is not particularly limited and may be included in an amount of 0.00001 to 99.99% by weight based on the total weight of the pharmaceutical composition.

The pharmaceutical composition of the present invention may further comprise a pharmaceutically acceptable carrier in addition to containing an amaranth extract or a fraction thereof as an active ingredient.

In the present invention, the expression "pharmaceutically acceptable " means that it can be routinely used in the pharmaceutical field, without disturbing the biological activity and properties of the compound to be administered, without irritating the organism upon administration thereof.

The pharmaceutical composition of the present invention may be formulated together with the carrier to be used as food, medicines, feed additives, drinking water additives, and the like.

In the present invention, the type of the carrier is not particularly limited and any carrier conventionally used in the art can be used. Examples of the carrier include, but are not limited to, saline, sterilized water, Ringer's solution, buffered saline, albumin injection solution, lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, maltodextrin, glycerol, . These may be used alone or in combination of two or more.

In addition, the pharmaceutical composition of the present invention may be supplemented with other pharmaceutically acceptable additives such as excipients, diluents, antioxidants, buffers or bacteriostats, and may be used as fillers, extenders, wetting agents, disintegrants, , A binder, a lubricant, and the like may be additionally used.

 The pharmaceutical composition of the present invention may be formulated into various formulations suitable for oral administration or parenteral administration.

Non-limiting examples of such oral dosage forms include troches, lozenges, tablets, aqueous suspensions, oily suspensions, prepared powders, granules, emulsions, hard capsules, soft capsules, syrups or elixirs .

In order to formulate the pharmaceutical composition of the present invention for oral administration, a binder such as lactose, saccharose, sorbitol, mannitol, starch, amylopectin, cellulose, or gelatin; Excipients such as dicalcium phosphate and the like; Disintegrating agents such as corn starch or sweet potato starch; Lubricants such as magnesium stearate, calcium stearate, sodium stearyl fumarate or polyethylene glycol wax may be used, and sweeteners, fragrances, syrups and the like may also be used. .

Furthermore, in the case of capsules, in addition to the above-mentioned substances, liquid carriers such as fatty oils can be further used.

Non-limiting examples of the parenteral preparation include injectable solutions, suppositories, respiratory inhalation powders, aerosol formulations for spray, mouth spray, mouthwash, toothpaste, ointment, powder for application, oil, have.

In order to formulate the pharmaceutical composition of the present invention for parenteral administration, sterilized aqueous solutions, non-aqueous solvents, suspensions, emulsions, freeze-dried preparations and external preparations may be used. Examples of the non-aqueous solutions and suspensions include propylene glycol, polyethylene Glycerol, vegetable oils such as olive oil, injectable esters such as ethyl oleate, and the like.

More specifically, when the pharmaceutical composition of the present invention is formulated into an injection, the composition of the present invention is mixed with water or a stabilizer or buffer in water to prepare a solution or suspension, which is then mixed with an ampoule or a vial Unit dosage form. When the pharmaceutical composition of the present invention is formulated into an aerosol formulation, a propellant or the like may be added together with the additive such that the water-dispersed concentrate or the wet powder is dispersed.

When the pharmaceutical composition of the present invention is formulated into an ointment, cream, or the like, it is possible to use an animal oil, a vegetable oil, a wax, a paraffin, a starch, a tracer, a cellulose derivative, a polyethylene glycol, a silicone, a bentonite, Or the like can be used as a carrier.

The pharmaceutical composition of the present invention may preferably be formulated into a parenteral dosage form, and more preferably, it may be a gel, a patch, a spray, an ointment, a warning agent, a lotion agent, a liniment agent, a pasta agent and a cataplasma agent Lt; RTI ID = 0.0 > formulations < / RTI >

The pharmaceutically effective amount and the effective dose of the pharmaceutical composition of the present invention may be varied depending on the formulation method, administration method, administration time and / or administration route of the pharmaceutical composition, and the reaction to be achieved by the administration of the pharmaceutical composition The type and severity of the subject to be treated, the type of subject to be treated, age, weight, general health condition, symptom or degree of disease, sex, diet, excretion, drugs used simultaneously or simultaneously with the subject, And other factors well known in the medical arts, and those skilled in the art will readily determine and prescribe dosages that are effective for the desired treatment. The pharmaceutical composition of the present invention may be administered once a day or divided into several doses. Accordingly, the dosage is not limited in any way to the scope of the present invention.

A preferred dosage of the pharmaceutical composition of the present invention may be from 1 mg / kg to 1, OOO mg / kg per day.

The route of administration and the mode of administration of the pharmaceutical composition of the present invention may be independent of each other, and the method is not particularly limited, and any route of administration and administration method may be used as long as the pharmaceutical composition can reach the desired site You can follow. The pharmaceutical composition may be administered orally or parenterally.

The parenteral administration may be, for example, intravenous administration, intraperitoneal administration, intramuscular administration, transdermal administration or subcutaneous administration, and a method of applying, spraying or inhalation the composition to a diseased site may also be used But are not limited to.

The pharmaceutical composition of the present invention may preferably be administered parenterally, more preferably, topically to the skin of the site where the pigment is deposited.

As used herein, the term "prevention" means any action that inhibits or delays the onset of skin pigmentation disease by administering the pharmaceutical composition of the present invention to a subject.

As used herein, the term "treatment" refers to any action that causes the symptom of skin pigmentation disease to be improved or benefited by administering the pharmaceutical composition of the present invention to an individual.

According to another embodiment of the present invention, Amaranthus spp . Which comprises administering to a subject a cosmetic composition, a quasi-drug composition or a food composition containing the extract or fraction thereof as an active ingredient.

The term "individual" as used herein refers to all animals, including mammals including rats, livestock, humans, and the like.

In the present invention, the cosmetic composition, the quasi-drug composition or the food composition containing the extract of Amaranth or the fraction thereof as an active ingredient are the same as those described above, and the meaning of skin whitening, pigmentation, prevention and improvement is also the same as described above.

In the present invention, the description of the dose, route of administration, mode of administration, and the like of the composition is the same as that described in connection with the pharmaceutical composition of the present invention.

According to another embodiment of the present invention, Amaranthus spp . L.) extract or a fraction thereof as an active ingredient to a subject in need thereof.

In the present invention, the pharmaceutical composition containing the amaranth extract or a fraction thereof as an active ingredient is the same as that described above, and the meaning of individual, skin pigmentation disease, prevention and treatment is also the same as described above.

In the present invention, the dosage, route of administration, mode of administration and the like of the pharmaceutical composition are the same as those described in connection with the pharmaceutical composition of the present invention.

The composition containing the extract of Amaranth or the fraction thereof of the present invention has excellent skin whitening effect and is excellent in the effect of preventing, improving or treating pigment deposition. Specifically, the composition of the present invention has an excellent effect of inhibiting the activity of tyrosinase and inhibiting the production of melanin.

In addition, since the composition of the present invention is a substance derived from a natural material, it has no toxicity and does not cause side effects even when it is applied to a human body, and thus its application to cosmetics, quasi-drugs, foods and / or pharmaceutical compositions is high.

Figure 1 shows the cytotoxicity of Amaranth extract according to concentration.
Figure 2 shows changes in the melanin content of melan-A cells following treatment with amaranth extract.
Fig. 3 shows changes in tyrosinase activity of melan-A cells following treatment with amaranth extract.
Fig. 4 shows changes in the expression level of tyrosinase-related enzymes of Melan-A cells following treatment with amaranth extract.
FIG. 5 shows changes in TRP-2 expression level of Melan-A cells following treatment with amaranth extract.
Figure 6 shows the cytotoxicity of the extracted fraction of amaranth according to the concentration.
Figure 7 shows changes in the melanin content of melan-A cells following treatment with the extract of amaranth.

Hereinafter, the present invention will be described in more detail by way of examples. It should be understood, however, that these examples are for illustrative purposes only and are not to be construed as limiting the scope of the invention.

Manufacturing example  One: amaranth  Preparation of extract

The seeds of Amaranthus spp. L. were washed with running water and then hot-air dried at a temperature of 40 ° C for 3 days immediately to maintain germination, and then pulverized by a pulverizer. The pulverized product was immersed in tertiary distilled water, followed by stirring at room temperature for 2 to 3 days. The leached product was filtered with a filter, and the leaching and filtration process was repeated two to three times. The filtrate was concentrated under reduced pressure and lyophilized to prepare a powder. At this time, the yield was 5%, and the obtained extract powder was used in the following experiment.

Of the extracts obtained, four samples of Amaranth extract were selected and named Amaranth A, Amaranth B, Amaranth C, and Amaranth D. Amaranth A and B are extracts of seeds of Amaranthus cruentus and Amaranth C is an extract of hybrid seeds between Amaranthus hybridus and Amaranthus hypocondriacus , Amaranth D is an extract of Seed of Amaranthus hive leader.

Manufacturing example  2: amaranth  extraction Fraction  Produce

10 g of the extract powder prepared in Preparation Example 1 was dissolved in 0.1 L of distilled water, 0.1 L of hexane was mixed with a separating funnel, and the mixture was repeated three times with a hexane layer and an aqueous layer to obtain a hexane layer. Ethyl acetate was then added to the aqueous layer, and the ethyl acetate layer and the aqueous layer were repeated three times to obtain an ethyl acetate layer. Each of the fractions obtained above was then filtered and concentrated under reduced pressure and lyophilized to obtain 1.6 g of hexane fraction, 1.05 g of ethyl acetate fraction and 7.35 g of a water fraction.

Example  One: amaranth  Cytotoxicity of extracts

Melanocyte cells derived from the embryo of C57BL as a melanocyte cell line of non-tumorous mice were seeded at 1 × 10 ⁵ cells per well in a 24-well plate and cultured for 24 hours at 37 ° C. in a 10% CO ₂ cell incubator Then, each of the extracts of Amaranth A to D prepared in Preparation Example 1 was treated for 3 days at a concentration of 1 μg / ml, 10 μg / ml and 100 μg / ml, and cultured. After 72 hours of incubation, 200 μl of the MTT solution prepared at a concentration of 0.5 mg / ml of PBS was added and further incubated for 1 hour under the same culture conditions. The culture was then removed and 200 μl of DMSO was added per well and the absorbance was measured at 570 nm with an ELISA reader (Quant, USA). As a negative control, amaranth extract-untreated control group was used and arbutin was used as a positive control group. The results of the absorbance measurement are shown in FIG. 1, and the cell survival rate was expressed as a percentage of the cell survival ratio of the negative control group.

As a result of the above experiment, cell survival rate was 76% to 120% at the concentration of 1 ㎍ / ㎖, 10 ㎍ / ㎖ and 100 ㎍ / ㎖ of Amaranth extract. In particular, the extract of Amaranth C and D showed cell viability of 80% or more even at a concentration of 100 ㎍ / ㎖, and thus it could be used more safely without inducing side effects when applied to an individual. Overall, it was found that the amaranth extract preferably inhibited cell growth of melan-A without cytotoxicity when treated at a concentration of 100 μg / ml or less, more preferably at a concentration of 10 μg / ml or less .

Example  2: amaranth  Confirming the efficacy of extract to inhibit melanin formation

In order to confirm the melanin production inhibitory effect of the amaranth extract, the change of the melanin content was measured by an in vitro test on the melan-A cell line.

The melan-A cell line was dispensed into a 24-well plate at a cell number of 1 x 10 < ⁵ > per well. After confirming the attachment, the extracts of Amaranth A to D prepared in Preparation Example 1 were added at 1 / And 100 쨉 g / ml, and treated with PRMI 1640 (Sigma) containing 1% Antibiotic-Antimycotic (Gibco), 10% FBS, Hyclone, 100 nM TPA, L- glutamine and sodium bicarbonate (Gibco) for 72 hours at 37 ° C in a 10% CO ₂ cell incubator. After 72 hours of incubation, the cells of the wells were washed with PBS, lysed with 1 N NaOH, and the amount of melanin was measured at an absorbance of 405 nm. The amount of melanin was expressed as a percentage of the amount of melanin in the negative control group (no amaranth treatment group), and the measurement results are shown in Table 1 and FIG.

sample Concentration (/ / ml) Melanin content (%) Untreated control group - 100 Arbutin 100 86.00 200 79.03 Amaranth A One 75.65 10 65.60 100 68.61 Amaranth B One 93.76 10 74.54 100 65.97 Amaranth C One 71.01 10 67.37 100 71.66 Amaranth D One 73.42 10 65.35 100 70.37

As a result of the above experiment, it was confirmed that the melanin content of Melan-A cells was reduced in a concentration-dependent manner in the whole test group treated with Amaranth extract. In particular, when the amaranth extract was treated at a concentration of 10 μg / ml, the amount of melanin was reduced by approximately 25% to 35%. This result supports the fact that the amaranth extract has an excellent inhibitory effect on the production of melanin and thus can exert a skin whitening effect.

Example  3: amaranth  Identification of inhibitory activity of tyrosinase activity of extract

To confirm the inhibitory activity of tyrosinase activity of the amaranth extract, the cytotoxicity results of Example 1 and the results of the melanin production inhibitory effect of Example 2 among the Amaranth extracts prepared in Preparation Example 1 are summarized, and Amaranth C and The DOPA autoxidation inhibition test of tyrosinase was carried out using Amaranth D extract.

The amaranth extract was diluted with 80 mM sodium phosphate buffer (pH 6.8) to prepare a sample solution (1 μg / ml and 10 μg / ml), and 40 μl of the sample solution according to the concentration and a DOPA automatic 8.3 mM of L-DOPA (L-3,4-dihydroxyphenylalanine, Sigma) dissolved in 80 mM sodium phosphate buffer solution (pH 6.8) was added as an oxidation inhibiting substrate, and the tyrosinase buffer The solution was added to the test plate in order so that the volume of one column became 200 占 퐇. After reacting at 37 占 폚 for 30 minutes, the absorbance was measured at 475 nm. As a negative control, 80 mM sodium phosphate buffer without added Amaranth extract was used. The DOPA auto-oxidation inhibitory effect of the amaranth extract was expressed as a percentage of the negative control, and the results are shown in Table 2 and FIG. 3 below.

sample Concentration (/ / ml) Tyrosinase activity (%) 80 mM sodium phosphate
Control solution with buffer solution - 100 Amaranth C One 78.23 10 75.29 Amaranth D One 71.38 10 71.19

As a result of the above experiment, it was confirmed that about 30% of tyrosinase activity was inhibited when treated with Amaranth extract, which is statistically significant as compared with the negative control. As a result, it was found that Amaranth extract inhibited tyrosinase activity This is a result supporting that skin whitening effect can be exerted because of its excellent inhibitory effect.

Example  4: amaranth  Confirming the efficacy of the extract to inhibit the expression of melanin-related enzymes

To investigate the mechanism of melanin biosynthesis by amaranth extract treatment, the amounts of tyrosinase and tyrosinase-related enzymes TRP-1, TRP-2, and MITF were measured in cells treated with Amaranth extract .

Of the amaranth extracts prepared in Preparation Example 1, Amaranth C and D extracts were treated with melan-A cells for 3 days at concentrations of 1 μg / ml and 10 μg / ml, respectively. Then, each treatment group was harvested, and a cell lysis solution was used to obtain only proteins, and Western blotting was performed. Cells collected for each treatment group were dissolved in a dissolution buffer for 1 hour and centrifuged at 12,000 rpm for 20 minutes. Then, only the supernatant was collected, proteins were quantified using BSA, and developed to a total protein amount of 35 μg on a 10% SDS-polyacrylamide gel (100 V for 1 hour and 30 minutes). Then, the membrane was washed three times with TBST, and the protein developed on the gel was transferred to a membrane (80 V for 1 hour), and the antibody (tyrosinase, TRP-1, TRP-2, , And tubulin) were added and reacted at 4 캜 for one day. Then, it was washed with TBST three times for 10 minutes, then the secondary antibody was added, and reacted at room temperature for 1 hour. Then, the cells were repeatedly washed with TBST three times for 10 minutes, and then sensitized with ECL and confirmed in a fluorescence chemical image-analyzing system (Chemi-documentation system). As a negative control, amaranth extract-untreated control group was used and arbutin was used as a positive control group. The results of the experiment are shown in FIG.

As a result of the above experiment, it was confirmed that the expression of tyrosinase and related enzymes was markedly reduced when the amaranth extract was treated as compared with the negative control and the positive control. Among the enzymes with reduced expression, the expression level of TRP-2 showing the most remarkable decrease was calculated as a percentage of the negative control, and is shown in Table 3 and FIG. 5 below.

sample Concentration (/ / ml) TRP-2 expression level (%) Untreated control group - 100 Arbutin 100 90.59 200 90.27 Amaranth C One 51.24 10 34.43 Amaranth D One 55.63 10 46.54

As a result of the experiment, it was confirmed that the expression level of TRP-2 was decreased depending on the treatment concentration of Amaranth extract. In particular, when the extract of Amaranth was treated at a concentration of 10 μg / ml, it showed an inhibitory effect of about 50% to 60%, compared to that of arbutin, which is known as a conventional whitening ingredient, , Which is a significant result showing very good effects.

Example  5: amaranth  extraction Fraction  Cytotoxicity check

Cytotoxicity was measured in the same manner as in Example 1 using Amaranth C extract fractions (0.1 μg / ml, 1 μg / ml and 10 μg / ml concentration) of the amaranth extract fractions prepared in Preparation Example 2 . As a negative control, amaranth extract-untreated control group was used and arbutin was used as a positive control group. The results of the absorbance measurement are shown in FIG. 6, and the cell survival rate was expressed as a percentage of the cell survival ratio of the negative control group.

As a result of the above experiment, it was confirmed that all of the groups treated with Amaranth extract showed excellent cell viability, and when treated at a concentration of at least 10 μg / ml or less, it was possible to inhibit the growth of melan-A cells without cytotoxicity.

Example  6: amaranth  extraction Fraction  Confirming the inhibitory effect on melanin formation

Using the extract fraction of Amaranth C (0.1 μg / ml, 1 μg / ml and 10 μg / ml concentration) of the amaranth extract fractions prepared in Preparation Example 2, the melanin production inhibitory effect Experiment was performed. The amount of melanin was expressed as a percentage of the amount of melanin in the negative control group (no amaranth treatment group), and the measurement results are shown in Table 4 and FIG.

sample Concentration (/ / ml) Melanin content (%) Untreated control group - 100 Arbutin 100 97.85 200 95.89 Amaranth C hexane fraction 0.1 102.60 One 85.68 10 83.35 Amaranth C
Ethyl acetate fraction 0.1 95.46 One 94.71 10 91.87 Amaranth C distilled water fraction 0.1 101.52 One 84.50 10 84.42

As a result of the above experiment, it was confirmed that melanin content of melan-A cells was significantly decreased in the group treated with 1 ㎍ / ㎖ and 10 ㎍ / ㎖ of the fraction extracted with amaranth. In particular, treatment with 10 ㎍ / ㎖ of Amaranthus hexane extract or distilled water extract showed a decrease of about 20% of melanin level compared to the negative control. This shows that the extract fraction of amaranth was excellent in inhibiting melanin production, .

It is to be understood that both the foregoing general description and the following detailed description of the present invention are exemplary and explanatory and are intended to provide further explanation of the invention as claimed. More variations are possible within a range that does not. Accordingly, the present invention may be embodied in other ways than those specifically described and illustrated herein, and it may be understood by those skilled in the art.

Claims (17)

Amaranthus spp . L.) extract or a fraction thereof as an active ingredient.
The method of claim 1, wherein the amaranth is selected from the group consisting of Amaranthus cruentus , Amaranthus hybridus ), and a hybrid between Amaranthus hypocondriacus and Amaranthus hive leaders. The cosmetic composition according to claim 1 , wherein the amaranthus hypocondriacus is amaranthus hypocondriacus .
The cosmetic composition according to claim 1, wherein the amaranth extract or a fraction thereof is obtained by extracting seeds of amaranth.
The cosmetic composition according to claim 1, wherein the improvement of skin whitening or pigmentation is carried out by inhibiting the formation of melanin pigment.
The cosmetic composition according to claim 1, wherein the skin whitening or pigmentation improvement is carried out by inhibiting expression of tyrosinase.
The cosmetic composition according to claim 1, wherein the cosmetic composition is at least one selected from the group consisting of a solution, an external ointment, a cream, a foam, a nutritional lotion, a flexible lotion, a pack, a flexible water, a latex, a makeup base, Wherein the composition is selected from the group consisting of suspensions, emulsions, pastes, gels, lotions, powders, soaps, surfactant-containing cleansers, oils, powder foundations, emulsion foundations, wax foundations, patches and sprays.
Amaranthus spp . L.) extract or a fraction thereof as an active ingredient.
Amaranthus spp . L.) extract or a fraction thereof as an active ingredient.
9. A method of improving skin whitening or preventing or ameliorating pigmentation, comprising administering the composition of any one of claims 1 to 8 to a subject.
Amaranthus spp . L.) extract or a fraction thereof as an active ingredient for preventing or treating skin pigmentation diseases.
The method of claim 10, wherein said amaranth is, Amaranth Tuscan crew enteoseu (Amaranthus cruentus), amaranth Tuscan hive Reader (Amaranthus hybridus ), and a hybrid between Amaranthus hypocondriacus and Amaranthus hive leaders. The pharmaceutical composition according to claim 1 , wherein the amaranthus hypocondriacus is amaranthus hypocondriacus .
11. The pharmaceutical composition according to claim 10, wherein the amaranth extract or a fraction thereof is obtained by extracting seeds of amaranth.
11. The pharmaceutical composition according to claim 10, wherein the prevention or treatment of the skin pigmentation disease is performed by inhibiting the production of melanin pigment.
11. The pharmaceutical composition according to claim 10, wherein the prevention or treatment of the skin pigmentation disease is performed by inhibiting expression of tyrosinase.
[Claim 11] The method of claim 10, wherein the skin pigmentation disease is one or more diseases selected from the group consisting of spots, freckles, spots, daylight pigmentation, pigmentation after drug use, pigmentation after inflammation and pregnancy pigmentation Composition.
[Claim 11] The pharmaceutical composition according to claim 10, wherein the pharmaceutical composition has a formulation selected from the group consisting of gel, patch, spray, ointment, warning agent, lotion, liniment, pasta and cataract.
16. A method for preventing or treating a skin pigmentation disorder, comprising administering the pharmaceutical composition of any one of claims 10 to 16 to a subject other than a human.

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