KR100849678B1 - Extracts of soy hypocotyl with effects on skin whitening and composition comprising the same - Google Patents

Abstract

The present invention relates to a melanin pigment biosynthesis inhibitor and / or composition for skin whitening comprising soybean embryo extract. Soybean embryo extract of the present invention has an excellent inhibitory effect on melanin pigment biosynthesis.

Soybean embryo, tyrosinase activity inhibition, melanin pigment biosynthesis inhibition, whitening effect

Description

Soybean embryo extract and composition for skin whitening comprising same {EXTRACTS OF SOY HYPOCOTYL WITH EFFECTS ON SKIN WHITENING AND COMPOSITION COMPRISING THE SAME}

1 is a table measuring the efficacy of inhibiting tyrosinase activity of soybean embryos and soybean fraction extract according to the present invention.

Figure 2 is a graph measuring the effect of soybean embryo and soybean fraction extract according to the present invention on the proliferation of basal layer melanocytes of the epidermis.

Figure 3 is a graph measuring the efficacy of soybean embryos and soybean fraction extract according to the present invention inhibit the biosynthesis of melanin pigment in the basal layer melanocytes (melanocyte) of the epidermis.

The present invention relates to a skin whitening preparation comprising a solvent extract of soybean embryos having an effect of inhibiting melanin pigment biosynthesis by inhibiting the activity of tyrosinase.

In addition to human skin, pigments, such as hair, are made from melanocytes of the epidermal layer of the epidermis in order to adapt to dangerous natural environments such as sunlight, and spread to the stratum corneum of the skin or the hair. In humans, the pigment of the skin is black, which is called melanin. The production of melanin is made from the amino acid tyrosine in transparent cells called melanocytes in the basal epidermis. Ultraviolet rays are the main factor in this process. On the basis of this mechanism, the cosmetic world has raised the control of skin pigment generation as a big task from the past and many related studies are in progress. Tyrosine, a raw material of melanin, is oxidized by oxygen, and the enzyme tyrosinase acts as an accelerator. Thus, it has been used as a tyrosinase inhibitor of various enzymes (such as arbutin) or a strong antioxidant (vegetable polyphenol). , Persimmon leaf extracts, etc.) have been used as a cosmetic whitening material.

In particular, soybean, one of the traditional foods in Korea, is generally used as a source of high-quality protein and lipids, and has been widely used for fermented foods such as soybean paste, soy sauce, and non-fermented foods such as tofu, bean sprouts, and soy flour. As a food ingredient, it has recently been spotlighted as a functional food that can prevent adult diseases including aging and skin whitening.

Meanwhile, soybean hulls and embryos produced as a by-product of soybean are almost discarded. According to the US Soybean Association data, about 1.5 million tons of soybeans were imported and used in 2003, resulting in about 45,000 tons of soybean hulls and soybean embryos being discarded by-products.

Antihyperlipidemic Effect of Soybean Embryo (Lee Hong-soo et al., Effect of Soybean Embryo on the Components of Serum Lipids in a High Cholesterol Diet, Journal of the Korean Society of Food Science and Nutrition, 2000) , 2001), but there are no reports that the embryo extracts of soybeans used as food have a superior effect on skin whitening effect than soybeans or halves.

Therefore, by extracting the active ingredient from the soybean embryo, which has been discarded, it was intended to be applied to food whitening, anti-aging and food or cosmetic materials including the same, which can be safely ingested and applied without a separate purification process.

Thus, the inventors of the present invention, while searching for a skin whitening substance derived from natural foods that ensure human safety, soybean embryo extract exhibits melanin synthesis inhibitory effect by inhibiting tyrosinase activity in melanocytes (melanocyte), skin whitening effect By discovering this superiority, the present invention has been completed.

Therefore, an object of the present invention is to provide a soybean embryo extract which is a foodstuff for whitening and cosmetics through melanin biosynthesis inhibition.

The present invention is characterized by a melanin pigment biosynthesis inhibitor comprising soybean embryo extract.

Hereinafter, the present invention will be described in detail.

The inventors of the present invention, while studying the inhibitors of melanin pigment biosynthesis derived from natural foods to ensure the safety of the human body, confirmed that the extract of soybean embryos have an excellent effect on the skin whitening effect by inhibiting melanin pigment biosynthesis, based on this The invention was completed.

Therefore, the present invention relates to a composition for inhibiting melanin pigment biosynthesis comprising soybean embryo extract as an active ingredient. In addition, the extract is a water extract or a solvent extract.

In relation to the extract, conventional extraction methods known to those skilled in the art may be used in the preparation of a composition for inhibiting melanin pigment biosynthesis using soybean embryos. Preferably, the solvent is acetone, ethanol, or methanol, and more preferably water extraction or ethanol extraction, but is not limited thereto. In addition, the extract preferably contains 20 to 80% of a solvent such as ethanol, acetone, or methanol.

In addition, the present invention is to provide a composition for skin whitening comprising soybean embryo extract as an active ingredient, which has melanin pigment biosynthesis inhibitory effect. The skin whitening composition may be used as a pharmaceutical, processed food, functional food, food additives, functional beverages, beverage additives or cosmetic raw materials.

The soybean embryo extract is preferably included in an amount of 0.01 to 10% by weight based on the whole composition. If it is less than 0.01, the present invention does not have the desired effect and if it exceeds 10% by weight, there is no dose-dependent presence for the effect, so the above range is preferable.

The composition comprising the extract of soybean embryo having the melanin pigment biosynthesis inhibitory effect as an active ingredient is preferably oral administration and skin application, when used as a medicament can be prepared by a known method in the pharmaceutical field, as the form It can be used in oral preparations such as powders, granules, tablets, capsules, pills, troches, liquid solutions, suspensions, emulsions, and syrups. When used as food, the extract in the group consisting of soybean embryo extract can be prepared as a food as it is, or mixed with various ingredients used in conventional food. As the form of the food to be prepared, it can be prepared in all kinds of foods such as solid foods, creamy or jam-like semi-liquid foods, gel foods, beverages, etc., and are commonly used in the form of capsules, granules, tablets, drinks, etc. Any substrate can be used to prepare soft drinks, juices, coffee, tea, liqueurs, milk, whey beverages, lactic acid bacteria beverages, candies, chewing gum, chocolate, gummi, yogurt, ice cream, puddings, water yolks and the like. When used as a cosmetic, the composition of the soybean embryo extract may be prepared as it is, or by adding various excipients used in the conventional cosmetics. The cosmetics may be prepared in the form of creams, suspensions, oil liquids, aqueous solutions, powders, and the like. Lotion, skin, sunscreen, powders, emulsions, aerosols, ointments, etc., may be used using any substrate that is commonly used. Can be used as a skin external preparation.

1, 2, and 3, it can be seen that the extract of soybean embryos inhibits tyrosinase activity, thereby inhibiting melanin pigment biosynthesis in the basal layer melanocytes of the epidermis.

In conclusion, the extract of soybean embryos showed the effect of inhibiting tyrosinase activity, indicating the effect of inhibiting melanin pigment biosynthesis.

When using a melanin pigment biosynthesis inhibitor comprising an extract of soybean embryos according to the present invention as a food and cosmetic composition, the skin whitening effect can be obtained by safely ingesting and applying without a separate purification process for removing toxicity. In addition, it is possible to separate the material showing the high melanin pigment biosynthesis inhibitory effect with a simple separation process, which lowers the production cost compared to the conventional melanin pigment biosynthesis inhibitors, and recycles the large amount of soybean embryos discarded during soybean processing. This can improve economics.

Hereinafter, more specific examples are presented to help the understanding of the present invention, but the following examples are merely to illustrate the invention, but the scope of the present invention is not limited to the following examples, within the scope of protection described in the claims It will be apparent to those skilled in the art that various modifications are possible.

Example  1: Preparation of Soybean Extract

     Soybean varieties are yellow soybean US No. 1 (not a single variety of soybeans), and soybean fractions (soybeans, embryos, halved soybeans, and husks) are obtained from shells, embryos, and soybeans by stripping during soymilk production. A half-half was obtained by the half-red process. 20 g of each of the soybean fractions (soybeans, embryos, halved soybeans, husks) was taken, and 70% ethanol (Duksan, Korea) aqueous solution was added about 10 times and refluxed for 6 hours using a stirrer at room temperature. The extract thus obtained was filtered through a filter paper (whatmann 41), and then evaporated at 40 ° C. for 2 hours, and then freeze-dried soybean, embryo, half-half, and bark extract (hereinafter, “soybean, embryo, half-half, husk extract”). Was obtained).

Example  2: Study of Whitening Effect of Soy Glycoside

1) Evaluation of Whitening related Enzyme Activity of Soybean Extract

Melanin is a phenolic substance widely distributed in the natural world, and its overproduction in the human body is known to form spots, freckles, and promote skin aging. Melanin biosynthesis is mainly caused by the action of tyrosinase, which measures the degree to which tyrosinase inhibits soybean extract.

Enzyme was used mushroom tyrosinase (Sigma T-7755) and substrate was L-tyrosine. The enzyme concentration in the reaction system was 33.3 µg / ml, and the substrate was 0.5 mM. The reaction rate was measured as a change in absorbance per minute at 475 nm. Tyrosinase IC ₅₀ value of the inhibitory activity of the soybean extract are shown in Figure 1 is calculated and compared to (IC ₅₀ values in the present invention, the enzymatic activity of tyrosinase refers to the soybean extract concentration capable of inhibiting 50%) It was.

According to the results, as a result of comparing the four soybean extracts, the effects of inhibiting tyrosinase activity involved in melanin production were found in the order of embryo> half-half> soybean> soybean hull.

2) Toxicity Testing of Melanoma Producing Cells:

Toxicity to melanin-producing cells was measured using the solvent extracts of soybeans, embryos, halves, and husks obtained in the above examples.

As a melanin-producing cell line, melanoma cell B16BL6 was sold from the Bank of Korea Cell Line. Culture of the cell line was performed by adding 10% FBS to DMEM (Dulbeco's Modified Egles Medium), a medium containing 100 IU / ml of penicillin and 100 μg / ml of streptomycin, and 5% CO. ₂ , 37 ° C. incubator (Forma Scientific Co., Marjetta, OH, USA). The medium compositions used for cell culture were all used by GIBCO BRL (Grand Island, NY, USA).

The cultured melanoma cell B16BL6 cell line was dispensed with 1 × 10 ⁴ cells in a 96 well plate, and the solvent extract extracted in the above example was added at a concentration of 0.125, 0.25, 0.5, 1 mg / ml for 72 hours. Incubated for

In order to measure the degree of toxicity of melanocyte-producing cells, an MTT assay was performed according to a known method (Mosmann, J. Immunol . Method , 65: 55-63, 1983). Cultured cells were washed twice with phosphate buffer. 3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyltetrazolium bromide (3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyl tetrazolium in 96 well plates bromide; MTT, Sigma) was added and reacted at 37 ° C. for 4 hours. The supernatant was removed by centrifuging the 96 well plate at 800 g for 25 minutes at room temperature. 0.1 ml of dimethylsulfoxide was added to the blue formazan product and reacted at 80 rpm for 30 minutes on a shaker, followed by a microplate reader (BioRad, Hercules, CA, USA). Absorbance at 570 nm was measured using. Three repeated experiments were performed according to this example, and cell proliferation rate by soybean extract was calculated in% and is shown in FIG. 2.

As a result, it was confirmed that the results of comparing the four soybean extracts did not affect the cell proliferation at the sample concentration of 0.125 ~ 1mg / ml.

 3) Evaluation of anti aging (whitening efficacy) using cell line

Skin aging causes melanin biosynthesis and increase in wrinkles, blemishes, and freckles. To prevent this, a test for whitening substance is performed on melanoma, a skin cancer cell. In this study, anti-aging (whitening efficacy) was also evaluated on melanoma cells.

Samples were dissolved at 100 mg / ml in 10% DMSO in DDW and each sample was filtered with a 0.2 um filter. On day 1, dispense 2 ml of 5x10 ⁴ melanoma cells / ml in a 6-well plate. On day 2, 20 μg of the sample was administered so that the content of DMSO was below 0.1% after medium exchange, and 20 μg of 10% DMSO was administered to the control. After 3 days of medium exchange, 20 μg of the sample was administered so that the DMSO content was below 0.1%, and Control was administered 20 μg of 10% DMSO. On Day 5, the cells were washed twice with PBS, treated with Trypsin-EDTA, and the number of cells was measured. 1 ml of 1N NaOH was added to the cell pellet and incubated for 1 h at 60 ° C. After Vortexing, the absorbance at Abs 475 and 280 nm was measured. The content was obtained.

According to the results, as a result of comparing the melanin biosynthesis inhibitory effect of the four soybean extracts appeared in the order of embryo> half-half> soybean skin> soybean as shown in the graph shown in FIG. In summary, it can be seen that the embryo extract among the four soybean extracts is remarkably excellent in the whitening effect by inhibiting melanin biosynthesis.

As described above, the embryo extract of the soybean extract had an excellent inhibitory effect on melanin pigment biosynthesis. In addition, the extract can be used as a food grade, can be obtained safely without the separate purification process to remove the toxicity to obtain melanin pigment biosynthesis inhibitory effect, and by separating the material showing the skin whitening effect by a simple separation process As the goods can be produced, the cost of production can be lowered compared to the existing skin whitening agent, and in particular, the economical efficiency can be improved by recycling the enormous amount of soybean embryos discarded during processing.

Claims (5)

delete  A cosmetic composition for skin whitening comprising 0.01 to 10% by weight of ethanol extract of soybean embryo as an active ingredient based on the whole composition. delete delete delete

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