KR102124255B1 - Skin-lightening Composition Using an Extract of Adlay Bran or Fermentation Product Thereof - Google Patents

Abstract

The present invention discloses a composition for skin whitening using Yulmu rice bran extract or a fermented product thereof having melanin biosynthesis inhibitory activity and anti-inflammatory and antioxidant activity.

Description

Skin-lightening Composition Using an Extract of Adlay Bran or Fermentation Product Thereof}

The present invention relates to a composition for skin whitening using a rice bran extract or fermentation product of Yulmu ( Coix lacryma-jobi L. var. mayuen (Roman.) Stapf).

With the development of medical technology and the improvement of living standards, people's desire for beauty becomes stronger and the perception that appearance itself is a social competitiveness is increasing, so the preference for skin whitening etc. is increasing and the cosmetics market is expanding (J. Korean Soc. Food Sci. Nutr., 42(3):355 (2013).

Skin occupies about 16% of the human body and is directly in contact with the external environment, and plays an important role in protecting the human body from external harmful factors such as temperature, humidity, and ultraviolet rays. The skin consists of epidermis, dermis, and subcutaneous tissue under the keratin. The epidermis is a thin protective layer composed mainly of keratinocytes made of keratin, immune cells such as melanocytes that produce and secrete melanin, and langerhans cells, and perceptual cells (Merkel's copuscles). It prevents external stimulation and invasion of pathogens and regulates body temperature and maintains moisture and lipid components.

Melanin is a phenolic polymer natural pigment widely present in living organisms. It is synthesized from melanosomes in melanocytes in the basement layer of the epidermis and transferred to surrounding keratinocytes to show human skin color. An abnormally low amount of melanin causes skin lesions such as vitiligo, and, conversely, when overproduced, acquired hyperpigmentation such as melansma, postinflammatory melanoderma, and solar lentigo. It can cause a variety of skin diseases, including.

The synthesis of melanin is achieved by making melanosomes from melanocytes present in the base layer of the skin, and is collectively referred to as melanin production pathway (melanogenesis). Melanin production is based on tyrosine, one of amino acids, as a substrate, and DOPA (3,4) by tyrosinase, tyrosinase related protein-1 (TRP-1), and tyrosinase related protein-2 (TRP-2). dihydroxyphenylalanine) and then converted to dopa quinone, then undergoes a non-enzymatic reaction, a spontaneous oxidation process, and then melanin is synthesized by polymerization with amino acids or proteins (Korean J. FOOD SCI. TECHNOL., 2000. 32(3):736; Journal of Life Science, 2013. 23(12):1445; Pigment Cell Res. 1999. 12(1):4-12.). Recently, studies on inhibition of melanin synthesis using proteins such as tyrosinase, TRP-1 and TRP-2, as well as MITF (microphtalmia associated transcription factor) have been conducted. The main intracellular signaling pathway is the cAMP/PKA (cyclic monophosphate/protein kinase A) pathway, cAMP promotes the expression of MITF via PKA, CREB1 (cAMP responsive element binding protein 1), and MITF is important in the melanin synthesis process. It promotes the transcription of tyrosinase, TRP-1, TRP-2 as a transcription regulator (Pharmazie. 2015 Oct;70(10):646-9.; Int J Mol Sci. 2015 Oct 13;16(10):24219- 42; Curr Pharm Biotechnol. 2015;16(12):1111-9.).

Since tyrosinase plays the most important role as the first step of producing melanin, studies on whitening agents mainly inhibit tyrosinase activity inhibitors or tyrosinase substrate antagonists to regulate tyrosinase activity. Research has been conducted in the direction of developing (Kor. J. Pharmacogn., 44(3):220, 2013).

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

In particular, natural substances often have antioxidant activity, and antioxidant activity inhibits the oxidation of dopa quinone in the process of melanin production, thereby showing a whitening activity, and thus increases melanin production as well as inhibition of melanogenesis. It can lead to an increase in activity (J. Am. Acad. Dermatol. 3: 99-105, 1994).

The present invention discloses the activity of inhibiting melanin biosynthesis, antioxidant activity, and the like, of the extract of Yulmu rice bran and its fermentation.

An object of the present invention is to provide a composition for skin whitening using the extract of yulmu gangmi and its fermentation products.

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

The present inventors confirmed that in the following Examples and Experimental Examples, Yulmu rice bran extract and its lactic acid bacteria fermentation product have melanin biosynthesis inhibitory activity in B16F10 melanoma cells, and in addition, LPS (Lipopolysaccharide, Bacterial Lipid Polysaccharide) It was confirmed that the production of NO (nitrogen monoxide), which is an inflammation-inducing substance, was inhibited in raw264.7 cells of mouse macrophages that induced an inflammatory reaction. Also, it was confirmed that it has antioxidant activity according to the ABTS method.

Considering the above, the present invention, in one aspect, can be identified as a skin whitening composition containing the active ingredient, Yulmu rice bran extract, its lactic acid bacteria fermentation product or the extract of the fermentation, in another aspect, the present invention Yulmu rice bran extract, the lactic acid bacteria fermentation product or the extract of the fermentation product can be identified as an anti-inflammatory composition containing an active ingredient or an NO production inhibitor composition that causes an inflammatory reaction, and in another aspect, the present invention, The lactic acid bacteria fermentation product or the extract of the fermentation product can be identified as an antioxidant composition using as an active ingredient.

In the present specification, "Yulmu rice bran" is a by-product obtained in the process of painting Yulmu, and is generally composed of a seed coat and an embryo.

In addition, in the present specification, "extract" refers to the extract of Yulmu rice bran or its fermented product, which is the extraction target in the present invention, water, lower alcohol having 1 to 4 carbon atoms (methanol, ethanol, butanol, etc.), methylene chloride, ethylene, acetone, hexane , Ether, chloroform, ethyl acetate, butyl acetate, N,N-dimethylformamide (DMF), dimethylsulfoxide (DMSO), 1,3-butylene glycol, propylene glycol, or a mixture of these solvents. Extract, carbon dioxide, pentane, etc. refers to an extract obtained by using a supercritical extraction solvent or a fraction obtained by fractionating the extract, and the extraction method is cold immersion, reflux, warming, ultrasound in consideration of the polarity, extraction degree, and preservation degree of the active substance. Any method such as spinning or supercritical extraction can be applied. In the case of fractionated extracts, the fractions obtained by suspending the extracts in a specific solvent and mixing and policing with a solvent having a different polarity, adsorb the crude extract on a column filled with silica gel, etc., and then add a hydrophobic solvent, a hydrophilic solvent or a mixed solvent thereof. It means that it contains the fraction obtained as a mobile phase. In addition, the meaning of the extract includes a concentrated liquid extract or a solid extract in which the extraction solvent is removed by a method such as freeze drying, vacuum drying, hot air drying and spray drying. Preferably, it is an extract obtained using water, methanol or a mixed solvent thereof as an extraction solvent, more preferably an extract obtained using a mixed solvent of water and methanol as an extraction solvent.

In addition, in the present specification, "active ingredient" refers to a component that can exhibit a desired activity alone or itself can exhibit activity with an inactive carrier.

In addition, in this specification, "fermented substance" means a culture obtained by inoculating and culturing lactic acid bacteria in Yulmu rice bran, which is a target for fermentation. This culture may be used as it is, concentrated under reduced pressure and/or lyophilized to be used in a liquid or powder form, or extracted using water, ethanol, etc. in the manner as described above, fractionated the extract, or the extract or fraction It can be used by concentration.

When preparing the fermentation product, a carbon source of lactic acid bacteria, which is an inoculum microorganism, may be added to the medium, and such a carbon source is well known in the art, and thus, any of those known in the art can be used. Usually, oligosaccharide, lactose, glucose, fructose, Sugar, molasses, dextrose, and mixtures thereof will be used. The carbon source may be added in an arbitrary range in consideration of intended fermentation time, fermentation degree, and the like. When added, it will be added in a range of 0.1 parts by weight to 5 parts by weight based on 100 parts by weight of Ulmu rice bran powder to be fermented.

When producing the fermentation product, the lactic acid bacteria, which are inoculated microorganisms, have no particular limitation, Lactobacillus sp. , Streptococcus sp. , Pediococcus sp. , Leuconostoc Both genus Lactobacillus ( Leuconostoc sp. ) and Bifidobacterium sp. may be used, specifically Lactobacillus delbrueckii , Lactobacillus bulgaricus , Lactobacillus bulgaricus , Lactobacillus casei Lactobacillus casei ), Lactobacillus brevis , Lactobacillus acidophilus , Lactobacillus rhamnosus , Lactobacillus plantarum , Lactobacillus plantarum , P. ( Pediococcus pentosaceus ), Pediococcus cerevisiae , Lactococcus lactis , Leuconostoc citreum , Leconococococcus mesenteroides , Bifidobacterium bifidobacterium ( Bifidobacterium bifidum ) may be used. Preferably, when considering the following examples, Lactobacillus plantarum is used among Lactobacillus sp .

In addition, the culture after inoculation of lactic acid bacteria in consideration of the addition amount of the fermented raw material Yulmu rice bran extract to the medium, the addition or absence of a carbon source, the intended degree of skin whitening activity, etc., the temperature condition of 15℃~45℃, especially 30℃~ At a temperature of 40° C., it may be performed for 10 hours or more, particularly 1 to 5 days.

The composition for skin whitening of the present invention may be prepared by including skin whitening ingredients, skin wrinkle improving ingredients, UV protection ingredients, skin moisturizing ingredients, etc., which are well known in the art, in order to reinforce/add skin-related activities in addition to the active ingredients. . Specifically, examples of skin whitening ingredients include arbutin, niacinamide, ascorbyl glucoside, alpha-bisabolol, oil soluble licorice (Glycyrrhiza) extract, and retinol, retinyl palmimi as skin wrinkle improvement ingredients. Tate, adenosine, polyethoxylated amide, and the like, and examples of UV protection components include drometrizole, drometrizoletrisiloxane, digaloyl trioleate, dimethycodiethyl benzalmalonate, and diethylhexylbutami And complexes such as dotria zone and pine bark extract, phosphatidylserine, fingerroot extract powder, and complex extracts such as red ginseng and cornus. In addition, as a moisturizing ingredient, AP collagen enzyme degrading peptide, Collactive collagen peptide, N-acetylglucosamine, konjac potato extract, dandelion and other complex extracts, rice bran extract, corn germ extract, low molecular collagen peptides, ground extract powder, phosphatidylserine, hyaluronic acid, etc. Can be heard. For other skin whitening ingredients, wrinkle improvement ingredients, UV protection ingredients, and skin moisturizing ingredients, the functional cosmetics exhibition under the Korean Cosmetics Act (「Functional Cosmetic Standards and Test Methods」 by the Ministry of Food and Drug Safety), the health of the Korean Health Functional Food Act You can refer to the Functional Foods Code ("Health Functional Food Standards and Standards" at the Ministry of Food and Drug Safety). These ingredients may be included in the composition for skin whitening of the present invention together with one or more of its active ingredients.

The composition of the present invention may include the active ingredient in any amount (effective amount) as long as it can exhibit skin whitening effect, anti-inflammatory effect, etc. according to the use, formulation, blending purpose, etc., the typical effective amount is the total weight of the composition It will be determined within the range of 0.001% to 15% by weight as a reference. "Effective amount" means intended medical and cosmetic effects, such as skin whitening effect, anti-inflammatory effect, etc., when the composition of the present invention is administered to a mammal, preferably a human subject, to which it is applied, according to the recommendation of a medical expert or the like. Refers to the amount of active ingredients contained in the composition of the present invention, which can represent the like. Such an effective amount can be determined empirically within the ordinary skill in the art.

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

The food composition of the present invention can be produced in any form, such as tea, juice, carbonated beverages, beverages such as ionic beverages, processed oils such as milk, yogurt, gums, rice cakes, sweets, breads, cookies, noodles, etc. Food, tablets, capsules, pills, granules, liquids, powders, flakes, pastes, syrups, gels, jelly, bars, etc. can be prepared as health functional food formulations.

In addition, the food composition of the present invention can be classified into any product as long as it complies with the enforcement regulations at the time of manufacturing and distribution in terms of legal and functional classification. For example, it is a health functional food pursuant to the Korean Act on Health Functional Food, or a confectionary, soybean, tea, or beverage according to each food type in the Food Fair of the Korean Food Sanitation Act (「Food Standards and Standards」) , Special purpose foods, and the like.

The food composition of the present invention may include a food additive in addition to the active ingredient. Food additives can be generally understood as substances added to foods, mixed or infiltrated in the manufacture, processing, or preservation of foods, and their safety must be ensured because they are consumed daily and for a long time with foods. Food additives in accordance with national laws governing the manufacture and distribution of food ("Food Sanitation Act" in Korea) are limited in terms of ingredients or functions in terms of food additives with guaranteed safety. According to the Korea Food Sanitation Act, the food additives are classified as chemical synthetic products, natural additives, and mixed preparations in terms of ingredients. It is divided into sweeteners, flavoring agents, preservatives, emulsifiers, acidulants, and thickeners.

Sweeteners are used to impart a moderate sweetness to food, and both natural and synthetic can be used in the composition of the present invention. Preferably, a natural sweetener is used, and examples of the natural sweetener include sugar sweeteners such as corn syrup solids, honey, sucrose, fructose, lactose, and maltose.

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

As a preservative, sodium sorbate, sodium sorbate, potassium sorbate, calcium benzoate, sodium benzoate, potassium benzoate, EDTA (ethylenediaminetetraacetic acid) and the like can be used, and as emulsifiers, acacia gum, carboxymethylcellulose, xanthan gum, Pectin and the like, and as acidulant, arithmetic, malic acid, fumaric acid, adipic acid, phosphoric acid, gluconic acid, tartaric acid, ascorbic acid, acetic acid, phosphoric acid and the like can be used. The acidulant may be added so that the food composition has an appropriate acidity for the purpose of suppressing the growth of microorganisms in addition to the purpose of enhancing taste.

As a thickener, a suspending agent, a sedimentation agent, a gel-forming agent, a swelling agent, etc. can be used.

The food composition of the present invention may include physiologically active substances or minerals known in the art for the purpose of supplementing and reinforcing functional and nutritional properties in addition to the food additives as described above, and guaranteed stability as food additives.

Examples of such bioactive substances include catechins contained in green tea, vitamins such as vitamin B1, vitamin C, vitamin E, and vitamin B12, etc., and are published in Korea Health Functional Food Fair (Korea Food and Drug Administration "Health Functional Food Standards and Standards"). Functional ingredients include red ginseng, chlorella, chlorophyll-containing plants, spirulina, green tea extract, propolis extract, coenzyme Q10, squalene, and tomato extract. Minerals include calcium preparations such as calcium citrate, magnesium preparations such as magnesium stearate, And iron preparations such as iron citrate, chromium chloride, potassium iodide, selenium, germanium, vanadium, and zinc.

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

With regard to other food additives that may be included in the food composition of the present invention, it is possible to refer to national foods or food additives.

The composition of the present invention may be identified as a pharmaceutical composition in other specific embodiments. In particular, when the composition for skin whitening of the present invention is identified as a pharmaceutical composition, its use may be understood as a composition for improving skin hyperpigmentation.

The pharmaceutical composition of the present invention may be prepared in an oral dosage form or a parenteral dosage form according to the route of administration by a conventional method known in the art, including a pharmaceutically acceptable carrier in addition to the active ingredient. Here, the route of administration may be any suitable route including a local route, an oral route, an intravenous route, an intramuscular route, and direct absorption through mucosal tissue, and two or more routes may be used in combination. An example of a combination of two or more routes is when the drugs of two or more formulations according to the route of administration are combined, for example, when one drug is administered by intravenous route first and the other drug is administered by topical route secondarily.

Pharmaceutically acceptable carriers are well known in the art depending on the route of administration or formulation, and specifically refer to the pharmacopeia of each country, including "Korea Pharmacopoeia".

When the pharmaceutical composition of the present invention is prepared in an oral dosage form, powders, granules, tablets, pills, dragees, capsules, liquids, gels, syrups, suspensions, wafers according to methods known in the art with suitable carriers And the like. At this time, examples of suitable carriers include sugars such as lactose, glucose, sucrose, dextrose, sorbitol, mannitol, xylitol, starches such as corn starch, potato starch, wheat starch, cellulose, methylcellulose, ethylcellulose, sodium carboxymethylcellulose, Celluloses such as hydroxypropyl methylcellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, magnesium stearate, mineral oil, malt, gelatin, talc, polyol, vegetable oil, ethanol, green Serol, etc. are mentioned. In the case of formulation, if necessary, suitable binders, lubricants, disintegrants, colorants, diluents, etc. may be included. Suitable binders include starch, magnesium aluminum silicate, starch ferist, gelatin, methylcellulose, sodium carboxymethylcellulose, polyvinylpyrrolidone, glucose, corn sweetener, sodium alginate, polyethylene glycol, wax, etc., and lubricants include oleic acid Sodium, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride, silica, talcum, stearic acid, magnesium salts and calcium salts thereof, polydetylene glycol, etc., and starch and methyl cellulose as disintegrants , Agar, bentonite, xanthan gum, starch, alginic acid or its sodium salt. Moreover, as a diluent, lactose, dextrose, sucrose, mannitol, sorbitol, cellulose, glycine, etc. are mentioned.

When the pharmaceutical composition of the present invention is prepared in a parenteral dosage form, it may be formulated in the form of injections, transdermal administrations, nasal inhalants and suppositories according to methods known in the art with suitable carriers. When formulated as an injectable agent, an aqueous isotonic solution or suspension may be used as a suitable carrier. Specifically, an isotonic solution such as PBS (phosphate buffered saline) containing triethanol amine, sterile water for injection, or 5% dextrose may be used. . When formulated as a transdermal dosage form, it can be formulated in the form of ointments, creams, lotions, gels, external solutions, pasta agents, linen agents, aerosols, and the like. For nasal inhalants, dichlorofluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide, etc. can be formulated in the form of an aerosol spray using a suitable propellant, and when formulated as a suppository, the carrier is Witthesol ( witepsol), tween 61, polyethylene glycols, cacao butter, laurin, polyoxyethylene sorbitan fatty acid esters, polyoxyethylene stearate, sorbitan fatty acid esters, and the like.

Regarding the specific formulation of the pharmaceutical composition, it is known in the art, and for example, see Remington's Pharmaceutical Sciences (19th ed., 1995) and the like. The above documents are regarded as part of this specification.

Preferred dosages of the pharmaceutical compositions of the present invention range from 0.001 mg/kg to 10 g/kg per day, preferably 0.001 mg/kg to 1 g per day, depending on the patient's condition, weight, sex, age, patient severity, and route of administration. /kg range. Administration can be done once a day or divided into several times. Such doses should not be construed as limiting the scope of the invention in any aspect.

In another specific aspect, the composition of the present invention can be identified as a cosmetic composition. In particular, when the anti-inflammatory composition of the present invention is identified as a cosmetic composition, its use may be understood as use for suppressing inflammatory skin troubles, alleviating inflammatory skin irritation, and the like.

Even when the composition of the present invention is identified as a cosmetic composition, the cosmetic composition can be classified into any product in accordance with its use, and in particular, functional cosmetics having non-functionality such as skin trouble improvement, atopic dermatitis improvement, etc. It may be a general cosmetic. As for the product form, any product form can be taken, specifically, solutions, suspensions, emulsions, pastes, gels, creams, lotions, powders, soaps, surfactant-containing cleansing, oils, powder foundations, emulsion foundations, waxes It can take the form of products such as foundations and sprays. In a specific product form, it may be flexible lotion, nutrition lotion, nutrition cream, massage cream, essence, eye cream, cleansing cream, cleansing foam, cleansing water, pack, spray or powder formulation.

The cosmetic composition of the present invention may include, in addition to its active ingredients, ingredients commonly used in cosmetic compositions, such as stabilizers, solubilizers, surfactants, vitamins, pigments and conventional auxiliary agents such as pigments and carriers.

When the formulation of the present invention is a paste, cream or gel, animal oil, vegetable oil, wax, paraffin, starch, trakant, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide, etc. may be used as a carrier component. Can be.

When the formulation of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used as a carrier component, and in the case of a spray, additionally chlorofluorohydrocarbon, propane /Propellant such as butane or dimethyl ether.

When the formulation of the present invention is a solution or emulsion, a solvent, solubilizer or emulsifier is used as a carrier component, specifically water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol, fatty acid ester of sorbitan, and the like can be used.

When the formulation of the present invention is a suspension, liquid diluents such as water, ethanol or propylene glycol as carrier components, ethoxylated isostearyl alcohol, suspensions such as polyoxyethylene sorbitol esters and polyoxyethylene sorbitan esters, microcrystals Sex cellulose, aluminum metahydroxide, bentonite, agar and the like can be used.

When the formulation of the present invention is a surfactant-containing cleansing, aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivatives, methyltaurate, sarcosinate, fatty acid amide as a carrier component Ether sulfates, alkylamidobetaines, aliphatic alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, lanolin derivatives or ethoxylated glycerol fatty acid esters and the like can be used.

The cosmetic composition of the present invention can be prepared according to a method of manufacturing a cosmetic composition that is commonly performed in the art, except that it contains an active ingredient that exhibits skin whitening activity and the like.

As described above, according to the present invention, a composition for skin whitening using Yulmu rice bran extract or its lactic acid bacteria fermentation product can be provided. The composition of the present invention can be commercialized as a food, drug or cosmetic.

1 is a result showing the melanin biosynthesis inhibition rate by treatment concentration of Yulmu rice bran extract (G77-1) and Yulmu rice lactic acid bacteria fermentation product (G77-2-2).

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

<Example> Preparation of sample

<Example 1> Preparation of Yulmu rice bran extract

20% by weight of 70% ethanol was added to the ulmu bran powder to leach for 24 hours, filtered with Whatman paper filter paper No. 2, and the filtrate was concentrated under reduced pressure to prepare a powdery ulmu bran extract Did.

<Example 2> Preparation of Yulmu rice lactic acid bacteria fermentation product

5 times the weight of purified water was added to the ulmu bran (adlay bran) powder, extracted at 80°C for 4 hours, and then cooled to 30 to 35°C. After inoculation with 3% (v/v) of the lactic acid bacteria ( Lactobacillus plantarum ), 3% (v/v) of the starter (10 ⁶ ∼ 10 ⁸ CFU/mL), the cells were cultured at 35-40° C. for 24 hours. Thereafter, the fermentation was terminated by heating at 90° C. for 10 minutes, and concentrated under reduced pressure to obtain a powdery fermentation product.

<Experimental Example> Skin whitening activity, anti-inflammatory activity and antioxidant activity of Yulmu rice bran extract

<Experimental Example 1> Skin whitening activity-melanin biosynthesis inhibitory activity

B16F10 melanoma cells were divided into 6 well plates with 1×10 ⁵ /well cell number, and after 24 hours of incubation, 100 nM α-MSH was treated to induce melanin synthesis, and samples were treated by concentration to incubate for 72 hours. . After washing twice with PBS, lysis buffer (100 mM sodium phosphate (pH6.8), 0.1 mM PMSF, 1% Triton X-100) was added, lysed at -80°C for 30 minutes, and the cells were collected. After reacting for 1 hour at 65°C with 1N NaOH containing 10% DMSO to dissolve the pellet, absorbance was measured at 405 nm. The inhibition rate of melanin biosynthesis (%) for each treatment concentration of the sample compared to the treatment group of α-MSH only is shown in FIG. 1, and IC ₅₀ , which is a concentration that inhibits melanin biosynthesis 50%, is shown in Table 1 below. The result is the result of three repeated experiments for each concentration.

Melanin biosynthesis inhibitory activity (IC ₅₀ , ㎍/ml) division Tyrosinase inhibition assay (㎍/ml)) Yulmu Rice Bran Extract 177.64 Yulmu Migang Fermentation 138.65 Albutin 399.26

As can be seen from Figure 1 and the above table, Yulmu rice bran extract and Yulmu rice lactic acid fermentation showed concentration-dependent melanin biosynthesis inhibitory activity, and IC ₅₀ of each sample appeared as 177.64 ㎍/ml and 138.65 ㎍/ml. Through this, it was found that the activity of inhibiting melanin biosynthesis was increased. As a positive control group, arbutin, a skin whitening agent listed in the Functional Cosmetics Code (Ministry of Food and Drug Safety, 「Functional Cosmetic Standards and Test Methods」) was used, and the IC ₅₀ of arbutin was 399.26 μg/ml.

<Experimental Example 2> Anti-inflammatory activity-NO production inhibitory activity evaluation

The anti-inflammatory efficacy was induced by NO synthesis of Raw264.7 cells, from which NO assay was analyzed to evaluate the anti-inflammatory efficacy. NO synthesis of Raw264.7 cells to evaluate the effectiveness was performed as follows.

The mouse macrophages, raw264.7 cells, were added with 10% FBS DMEM medium and cultured at 37°C and 5% CO ₂ . Raw264.7 macrophages were divided into 4×10 ⁴ cells/well cells in a 96 well plate, and treated with endotoxin after 24 hours to treat LPS (Lipopolysacharide) used as an NO synthesis mitogen at a concentration of 0.5 μg/ml.

Next, after incubation at 37°C and 5% CO ₂ for 1 hour and 30 minutes, the samples were processed by concentration and incubated for 24 hours. Next, for the anti-inflammatory efficacy evaluation, the NO assay was analyzed as follows. 1% sulfanilamide and 0.1% N-(1-naphtyl) ethylenediamine dihydrochloride reagent are mixed in a volume ratio of 1:1, and the medium supernatant cultured for 24 hours after sample treatment of the NO synthesis induction experiment of the mixed reagent and Raw264.7 cell 1: After mixing 50 µl each at a volume ratio of 1 and storing at room temperature for 10 minutes, ELISA was measured at 540 nm to measure the ability to inhibit NO production compared to the control group. The standard curve was prepared using NaNO ₂ .

The results are shown in Table 3 below with IC ₅₀ , a concentration that inhibits NO production by 50%. The result is the result of three repeated experiments for each concentration.

NO production inhibitory activity (IC ₅₀ , μg/ml) division NO assay (㎍/ml)) MTT assay (㎍/ml) Yulmu Rice Bran Extract 69.85 >100 Yulmu Migang Fermentation 62.01 >100 L-NMMA 19.14 (μM) >100 (μM)

The IC ₅₀ of Yulmu rice bran extract and Yulmu rice lactic acid lactic acid fermentation were 69.85 ㎍/ml and 62.01 ㎍/ml, respectively, indicating that the anti-inflammatory activity was not particularly enhanced through fermentation. L-NMMA (N-monomethyl-L-arginine) is a NO control inhibitor used as a positive control.

Meanwhile, the results of cytotoxicity evaluation according to the MTT method are shown in Table 3 above. There was no particular cytotoxicity even at the highest treatment concentration of 100 μg/ml.

<Experimental Example 4> Antioxidant activity-ABST method experiment

A 7 mM ABTS buffer and 2.45 mM potassium persulfate buffer (K ₂ S ₂ O ₈ ) were prepared, mixed with ABTS:K ₂ S ₂ O ₈ =2:1 ratio, and reacted under shading for 12-16 hours to form a cationic radical. After that, the stock buffer was diluted so that the OD value measured at 734 nm was 0.70±0.02. 100 µl of the buffer and 100 µl of the extract sample diluted by concentration were added and reacted at room temperature for 6 minutes. After the reaction, the absorbance at 734 nm was measured, and the ABTS cationic radical scavenging activity of the sample according to each treatment concentration was compared to the control-free group according to the following formula, while shown in FIG. 5, which corresponds to the scavenging activity of 50%. The concentration IC ₅₀ is shown in Table 5 below. Antioxidant BHA was used as a positive control. The result is the result of three repeated experiments for each concentration.

ABTS cationic radical scavenging rate (%) = (Control-sample) / Control X 100

Antioxidant activity according to ABTS method (IC ₅₀ , ㎍/ml) division ABTS assay (㎍/ml) Yulmu Rice Bran Extract 90.41 Yulmu Migang Fermentation 62.95 BHA 4.61

Antioxidant activity was found to be increased by fermentation as well as melanin production inhibitory activity. BHA was found to be 8.75 μg/mL. A synthetic antioxidant, BHA (Butyl hydroxyanisole), was used as a positive control.

Claims (15)

Water and ethanol mixed solvent extract of Yulmu rice bran having melanin production inhibitory activity, fermentation product by Lactobacillus plantarum of Yulmu rice bran having melanin production inhibitory activity, or water of the fermentation product having melanin production inhibitory activity A cosmetic composition for skin whitening comprising an ethanol mixed solvent extract as an active ingredient.
According to claim 1,
The mixed solvent is a composition characterized in that 70% ethanol.
Water and ethanol mixed solvent extract of Yulmu rice bran having melanin production inhibitory activity, fermentation product by Lactobacillus plantarum of Yulmu rice bran having melanin production inhibitory activity, or water of the fermentation product having melanin production inhibitory activity Food composition for skin whitening containing an ethanol mixed solvent extract as an active ingredient.
According to claim 3,
The mixed solvent is a composition characterized in that 70% ethanol.
Water and ethanol mixed solvent extract of Yulmu rice bran having melanin production inhibitory activity, fermentation product by Lactobacillus plantarum of Yulmu rice bran having melanin production inhibitory activity, or water of the fermentation product having melanin production inhibitory activity Pharmaceutical composition for improving skin hyperpigmentation including ethanol mixed solvent extract.
The method of claim 5,
The mixed solvent is a composition characterized in that 70% ethanol. delete delete delete delete delete delete delete delete delete

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