KR102045293B1 - Skin-lightening Composition Using an Extract of Shindari - Google Patents
Skin-lightening Composition Using an Extract of Shindari Download PDFInfo
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- KR102045293B1 KR102045293B1 KR1020170027010A KR20170027010A KR102045293B1 KR 102045293 B1 KR102045293 B1 KR 102045293B1 KR 1020170027010 A KR1020170027010 A KR 1020170027010A KR 20170027010 A KR20170027010 A KR 20170027010A KR 102045293 B1 KR102045293 B1 KR 102045293B1
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Abstract
The present invention discloses a composition for skin whitening using fifty-legged and oak tree petal mixed extracts exhibiting concentration-dependent melanin production inhibitory activity when treated with B16F10 cells, a mouse melanoma cell line stimulated with α-MSH, a melanogenesis inducing agent. .
Description
The present invention relates to a composition for skin whitening using fifties and oak petals mixed extract.
The skin is the largest organ that protects the interior of our body from a variety of external environments and is the outermost organ of our body. These skins contain keratinocytes, which inhibit the invasion of foreign substances, Langerhans cells and macrophages, which are responsible for skin immunity, fibroblasts, which maintain skin tissue, and pigment production. There are melanocytes (melanocytes) and the like.
Melanin is a black pigment synthesized in melanocytes and is a protein complex that serves to protect skin cells from the external environment such as ultraviolet rays. However, the overproduction of melanin can cause blemishes, freckles, and even skin cancer in the human body (JP Ortonne and JJ Nordlund, The pigmentary system: physiology and pathophysiology, eds. JJ Norlund, RE Boissy, VJ Hearing, RA King and JP Ortonne, 489, Oxford University Press, New York, 1998).
Such melanin is produced by the action of tyrosinase and tyrosinase related protein present in melanocytes. Specifically, tyrosine is oxidized by tyrosinase and is converted into dopa (DOPA, 3,4-dihydroxyphenylalanine), followed by dopaoxidase to oxidize the dopa to be dopaquinone. After conversion, it is produced by the action of tyrosinase-related proteins TRP-1 (5,6-dihydroxy indole-2-carboxylic acid oxidase) and TRP-2 (dopachrome tautomerase). Tyrosinase, TRP-1 and TRP-2 are promoted by the transcription factors microphthalmia-associated transcription factor (MITF), which is known to inhibit the expression of extracellular signal-regulated kinase (ERK) (DS). Kim, et al., J Cell Sci. 2003; 116: 1699-706).
Since tyrosinase is involved as the first step in producing melanin, the research on whitening agents has been mainly focused on finding tyrosinase inhibitors (Kor. J. Pharmacogn., 44 (3): 220, 2013). Most of the whitening agents currently used are such tyrosinase inhibitors, and hydroquinone or its glycosides, arbutin (β-D-glucopyranoside) and kojic acid, all inhibit tyrosinase inhibitory activity. Having whitening agents. Hydroquinone or its glycoside, arbutin (β-D-glucopyranoside), is a competitive inhibitor of tyrosinase activity, and kojic acid chelates tyrosinase active sites in tyrosine. It is a substance that inhibits the process of dopa and dopaquinone (Korean J. Plant Res., 26 (5): 526, 2013; J. Korean Soc.Food Sci.Nutr., 41 (2): 156, 2012) .
However, all of the whitening agents are synthetic whitening agents, and these synthetic whitening agents have side effects such as dermatitis, erythema, vitiligo, and burns for a long time, and in some countries such as Taiwan, allowable concentrations can be used without prescription from medical professionals. Recently, researches to find a whitening material from natural products with relatively little irritation or side effects on human skin have been actively conducted due to such safety issues of synthetic whitening agents.
On the other hand, shien-dari is a low-alcoholic beverage made by fermenting yeast in rice, barley rice, or rice that has just started to rest. It is also called a new moon or a single drink. When it is difficult to store leftover rice, it is derived from using it. When the rice becomes fermented and the shape is not recognizable, it is sifted and eaten raw or boiled. It fermented for a day or two in summer and 5 or 6 days in winter. The short fermentation period is about 1% lower, and the acidity and sweetness are strong. Sheen is similar to makgeolli with barley (or rice), yeast, and water, but is similar to yogurt than makgeolli due to its short fermentation and low alcohol content.
Recently, as interest in health has increased, interest in the fifties using fermented bacteria beneficial to the body has increased. The effects of fifty-legged legs on intestinal health improvement and dementia improvement are known and attempts are being made to apply them to new fields such as cosmetics as well as food using useful ingredients. Known studies include changes in composition during the production of fifty legs (Kim SC. 1998. Master thesis.Jeju National University of Korea. Soc Food Sci Nutr 28 (5): 1017-1021), Characterization of Sheen Legs Prepared with Citrus Fruits (Kim HS. 2011. Master thesis.Jeju National University of Korea, 40p), Characterization of Sheen Legs with Carrots (Kim S et al., 2015. Korean J Food Cook Sci. 31 (1) 009-017), and the method of manufacturing fifty-legged legs with improved presentation odor (Korean Patent No. 1396825).
The present invention discloses the use of fifty-legged extract as a skin whitening agent based on melanin production inhibitory activity.
It is an object of the present invention to provide a composition for skin whitening using a mixed foot and oak petal extract.
Other and specific objects of the present invention will be presented below.
The present invention, as confirmed in the following Examples and Experimental Examples, concentration-dependent melanin when the fifty-legged and oak petal extract is treated with B16F10 cells, a mouse melanoma cell line stimulated with α-MSH, a melanogenesis inducer It was completed by confirming that the production inhibitory activity is not exhibited, especially cytotoxicity. In particular, when the mixture of each extract was treated with α-MSH, a melanogenesis-inducing mouse melanoma cell line, B16F10 cells, the concentration of the melanin inhibitory activity was higher than that of the fifties extract and the oak petal extract. By confirming that the tendency to show clearly is completed.
The present invention is provided on the basis of the experimental results, the skin whitening composition of the present invention is characterized in that it comprises a sheen leg extract and oak petal extract as an active ingredient.
As used herein, the term "extract" refers to water, methanol, ethanol, butanol lower alcohols having 1 to 4 carbon atoms, methylene chloride, ethylene, acetone, hexane, ether, chloroform, ethyl acetate, butyl acetate, N, N -Obtained using supercritical extraction solvents such as dimethylformamide (DMF), dimethylsulfoxide (DMSO), 1,3-butylene glycol, propylene glycol or a leaching thereof, carbon dioxide, pentane, etc. It means an extract or a fraction obtained by fractionating the extract, and the extraction method may be any method such as cold needle, reflux, heating, ultrasonic radiation, supercritical extraction in consideration of the polarity, degree of extraction, and degree of preservation of the active substance. . In the case of the fractionated extract, the fraction obtained by suspending the extract in a specific solvent and mixing and standing with a solvent having a different polarity, adsorbing the extract on a column filled with silica gel or the like, and then using a hydrophobic solvent, a hydrophilic solvent, or a mixed solvent thereof It is meant to include fractions obtained by mobile phase. In addition, the meaning of the extract includes a concentrated liquid extract or solid extract in which the extraction solvent is removed in a manner such as freeze drying, vacuum drying, hot air drying, spray drying, and the like. Preferably it refers to an extract obtained by using water, ethanol or a mixed solvent thereof as the extraction solvent, more preferably an extract obtained by using a mixed solvent of water and ethanol as the extraction solvent.
In addition, in the present specification, the "active ingredient" means a component that can exhibit the desired activity alone or in combination with a carrier which is itself inactive.
In addition, in the present specification, "skin whitening" may be understood as an increase in skin brightness reduced due to excessive production of melanin, specifically, improvement (reduction of symptoms) such as blemishes, freckles, and mushrooms caused by excessive production of melanin, By treatment, prevention, expression inhibition or delay.
The skin whitening composition of the present invention may be prepared including skin whitening ingredients, skin wrinkle improvement ingredients, UV protection ingredients, skin moisturizing ingredients and the like known in the art in order to enhance and add skin related activities in addition to the active ingredient. . Specifically, skin whitening ingredients include arbutin, niacinamide, ascorbyl glucoside, alpha-bisabolol, oil soluble licorice (Glycyrrhiza) extract, and the like, and skin wrinkle improvement ingredients such as retinol and retinyl palmi Tate, adenosine, a polyethoxylated amide, etc., As a ultraviolet protection component, dromethazole, a dromethazole trisiloxane, the digaloyl tri oleate, the dimethco diethyl benzal malonate, the diehexyl buty Butami Complexes such as dotriazone, pine bark extract, phosphatidylserine, finger root extract powder, complex extracts such as red ginseng and cornus milk. The moisturizing ingredients include AP collagen degrading peptide, Collactive collagen peptide, N-acetylglucosamine, konjac potato extract, dandelion and other complex extracts, rice bran extract, corn germ extract, low molecular collagen peptide, herbal extract powder, phosphatidylserine, hyaluronic acid, etc. Can be mentioned. For other skin whitening ingredients, wrinkle improvement ingredients, UV protection ingredients, and skin moisturizing ingredients, the functional cosmetics code according to the Cosmetics Act ("Notice of functional cosmetics standards and test methods" of the KFDA) and the health functional foods of the Act on Health Functional Foods Reference may be made to the Code of Ethics (NFDA standards and standards). One or more of these ingredients may be included with the active ingredient in the skin whitening composition of the present invention.
In the composition for skin whitening of the present invention, the active ingredient may be included in any amount within a range that does not toxic to the human body depending on the use, formulation, etc., specifically, from 0.001% to 99.99% by weight of the total weight of the composition It may be included in weight percent.
The skin whitening composition of this invention can be grasped | ascertained as a cosmetic composition in a specific aspect.
Even when the composition of the present invention is identified as a cosmetic composition, the cosmetic composition may be classified into any product according to its use and law. Specifically, the cosmetic composition may be a functional cosmetic having a use such as skin whitening, a non-functional general cosmetic, or the like. . The product form can also be in any product form, specifically solutions, suspensions, emulsions, pastes, gels, creams, lotions, powders, soaps, surfactant-containing cleansing, oils, powder foundations, emulsion foundations, waxes It can be formulated into a foundation, spray, and the like. In a specific product form, it may be a softening lotion, a nourishing lotion, a nourishing cream, a massage cream, an essence, an eye cream, a cleansing cream, a cleansing foam, a cleansing water, a pack, a spray or a powder formulation.
The cosmetic composition of the present invention may include, in addition to the active ingredient, components conventionally used in the cosmetic composition, for example, conventional adjuvants such as stabilizers, solubilizers, surfactants, vitamins, pigments and pharmaceuticals, and carriers.
When the formulation of the present invention is a paste, cream or gel, animal oils, vegetable oils, waxes, paraffins, starches, trachants, cellulose derivatives, polyethylene glycols, silicones, bentonites, silicas, talc or zinc oxide may be used as carrier components. 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, in particular 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 the carrier component, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1 , 3-butylglycol 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, a liquid diluent such as water, ethanol or propylene glycol, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester, polyoxyethylene sorbitan ester, and microcrystals are used as carrier components. Castle cellulose, aluminum metahydroxy, bentonite, agar and the like can be used.
When the formulation of the present invention is a surfactant-containing cleansing, the carrier component is aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyltaurate, sarcosinate, fatty acid amide. 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 the manufacturing method of the cosmetic composition usually carried out in the art, except for including the active ingredient exhibiting skin whitening activity.
The skin whitening composition of this invention can be grasped | ascertained as a food composition in another specific aspect.
The food composition of the present invention may be prepared in any form, for example, beverages such as tea, juice, carbonated beverages, ionic beverages, processed oils such as milk, yogurt, gums, rice cakes, sweets, bread, sweets, noodles, and the like. It can be prepared as a dietary supplement, such as foods, tablets, capsules, pills, granules, liquid, powder, flaky, paste, syrup, gel, jelly, bar. In addition, the food composition of the present invention can distinguish any product as long as it conforms to the enforcement regulations at the time of manufacture and distribution in the legal and functional divisions. For example, it may be a health functional food according to the Act on Health Functional Foods, or soy milk, fermented beverages, special purpose foods, etc. according to each food type in the Food Code of the Food Sanitation Act (KFDA notice, standards and standards of food).
The food composition of the present invention may include food additives in addition to the active ingredient. Food additives are generally understood as substances that are added to, mixed with, or infiltrated in the manufacture, processing, or preservation of foods, and because they are taken daily with food for a long time, their safety should be ensured. The Food Additives Code of the Food Sanitation Act (KFDA Notification, Food Additives Standards and Standards) defines the food additives that are guaranteed to be safe by dividing them into chemical synthetics, natural additives, and mixed preparations.
These food additives may be divided into sweeteners, flavors, preservatives, emulsifiers, acidulants, thickeners, and the like in terms of their functionalities.
Sweeteners may be used in amounts that give the food a suitable sweet taste, and may be natural or synthetic. Preferably, a natural sweetener is used. 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 the taste or aroma, both natural and synthetic. It is the case of using a natural thing preferably. In addition to flavors, the use of natural ones can be combined with nutritional purposes. The natural flavor may be obtained from apples, lemons, citrus fruits, grapes, strawberries, peaches, and the like, or may be obtained from green tea leaves, round leaves, jujube leaves, cinnamon, chrysanthemum leaves, jasmine and the like. In addition, ginseng (red ginseng), bamboo shoots, aloe vera, ginkgo, etc. may be used. Natural flavors can 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.
Sodium sorbate, sodium sorbate, potassium sorbate, calcium benzoate, sodium benzoate, potassium benzoate, EDTA (ethylenediaminetetraacetic acid), etc. may be used as a preservative, and as an emulsifier, acacia gum, carboxymethylcellulose, xanthan gum, Pectin etc. can be mentioned, As acidic acid, acid, malic acid, fumaric acid, adipic acid, phosphoric acid, gluconic acid, tartaric acid, ascorbic acid, acetic acid, phosphoric acid, etc. can be used. The acidulant may be added so that the food composition is at an appropriate acidity for the purpose of inhibiting the growth of microorganisms in addition to the purpose of enhancing the taste.
As the thickener, suspending implementers, sedimenters, gel formers, swelling agents and the like can be used.
In addition to the food additives described above, the food composition of the present invention may include a bioactive substance or minerals known in the art for the purpose of supplementing and reinforcing the functionality and nutritional properties and ensuring the stability as a food additive.
Examples of such physiologically active substances include catechins, vitamin B1, vitamin C, vitamin E, vitamin B12 and the like, tocopherol, dibenzoylthiamine, and the like contained in green tea. Examples of the minerals include calcium preparations such as calcium citrate and magnesium stearate. Magnesium preparations such as iron, iron preparations such as iron citrate, chromium chloride, potassium iodine, selenium, germanium, vanadium, zinc and the like.
In the food composition of the present invention, the food additive as described above may be included in an amount that can achieve the purpose of addition according to the product type.
Regarding other food additives that may be included in the food composition of the present invention, reference may be made to food or food additives.
The composition for skin whitening of the present invention can be regarded as a pharmaceutical composition in specific embodiments.
When the composition for skin whitening of the present invention is identified as a pharmaceutical composition, the target disease may be understood as skin hyperpigmentation, and specific examples of such hyperpigmentation include freckles, senile spots, liver spots, blemishes, brown or sunspots, and sun pigments. Half, cyanic melasma, hyperpigmentation after use of drugs such as calcium antagonists, sequelae following sclerotherapy, gravidic chloasma, melanoma in women taking oral contraceptives, Post-inflammatory hyperpigmentation, phototoxic reactions or other similar small fixed pigmented lesions, including, but not limited to, wounds or dermatitis, including scratches and burns.
The pharmaceutical compositions of the present invention may be prepared in oral or parenteral formulations according to the route of administration by conventional methods known in the art, including pharmaceutically acceptable carriers in addition to the active ingredient. "Pharmaceutically acceptable" here means that the subject of application (prescription) is not toxic as far as adaptable without inhibiting the activity of the active ingredient.
When the pharmaceutical composition of the present invention is prepared in an oral dosage form, powders, granules, tablets, pills, dragees, capsules, solutions, gels, syrups, suspensions, wafers according to methods known in the art with suitable carriers It may be prepared in a formulation such as. Examples of suitable pharmaceutically acceptable carriers include sugars such as lactose, glucose, sucrose, dextrose, sorbitol, mannitol and xylitol, starch such as corn starch, potato starch, wheat starch, cellulose, methylcellulose, ethylcellulose, Celluloses such as sodium carboxymethyl cellulose and hydroxypropyl methyl cellulose, polyvinyl pyrrolidone, water, methyl hydroxybenzoate, propyl hydroxybenzoate, magnesium stearate, mineral oil, malt, gelatin, talc, polyol, vegetable Yu etc. can be mentioned. If formulated, it may be formulated to include diluents and / or excipients, such as fillers, extenders, binders, wetting agents, disintegrants, surfactants, if necessary.
When the pharmaceutical compositions of the present invention are prepared in parenteral formulations, they may be formulated in the form of injections, transdermal administrations, nasal inhalants and suppositories with suitable carriers according to methods known in the art. When formulated as an injection, suitable carriers include sterile water, ethanol, polyols such as glycerol or propylene glycol, or mixtures thereof. Preferably, PBS (phosphate buffered saline) containing Ringer's solution or triethanol amine or sterile water for injection , Isotonic solutions such as 5% dextrose, and the like can be used. When formulated as a transdermal administration, it may be formulated in the form of an ointment, cream, lotion, gel, external solution, pasta, linen, aerosol and the like. Nasal inhalants can be formulated in the form of aerosol sprays using suitable propellants, such as dichlorofluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide, etc. The suppository bases are witepsol, twin (tween) 61, polyethylene glycols, cacao butter, laurin paper, polyoxyethylene sorbitan fatty acid esters, polyoxyethylene stearate, sorbitan fatty acid esters and the like can be used.
Regarding the formulation of pharmaceutical compositions, it is known in the art and specific reference may be made to Remington's Pharmaceutical Sciences (19th ed., 1995) and the like. The document is considered 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, depending on the condition, body weight, sex, age, severity of the patient and route of administration. It can range from / kg. Administration can be done once a day or divided into several times. Such dosage should not be construed as limiting the scope of the invention in any aspect.
As described above, according to the present invention can provide a composition for skin whitening using a mixed foot and oak petal extract.
The composition for skin whitening of the present invention may be commercialized as food, cosmetics, medicines, etc., and may have relatively little side effects when compared to synthetic skin whitening agents in that the fifty-leg has been used for food.
Figure 1 is a cytotoxic result of B16F10 cells, a mouse melanoma cell line such as fifty-legged extract.
2 is a result showing that the fifty-legged extract and the like have melanin production inhibitory activity in B16F10 cells, a mouse melanoma cell line stimulated with α-MSH, a melanogenesis-stimulating agent.
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> Flower Petal Extract and Sheen Leg Extract Preparation
Example 1 Preparation of Oak Flower Petal Extracts
Chamkkot Tree (Rhododendron weyrichii) then was added to 70% ethanol of 10 times the weight of the petal powder extracted at room temperature for 24 hours and filtered with a filter paper. The filtrate was concentrated under reduced pressure and lyophilized to obtain an oak extract.
<Example 2> Preparation of fifty-legged extract
10-fold weight of 70% ethanol was added to fifty-legged powder, and extracted at room temperature for 24 hours, followed by filtration through filter paper. The filtrate was concentrated under reduced pressure and lyophilized to obtain fifty-legged extract.
Experimental Example Whitening activity check
<1> Experiment method
<1-1> Cell culture
B16F10 cells, a mouse melanoma cell line, were purchased from the Korean Cell Line Bank, with 1% antibiotics (Gibco, USA) and 10% fetal bovine serum (FBS; Gibco, Grand Island, USA). The DMEM medium (Dulbecco's modified Eagle's medium) containing this was incubated in a 37 ℃, 5% CO 2 incubator, and passaged once every 4 days.
<1-2> Cytotoxicity Assessment
MTT analysis was performed to determine the effect of the sample of Example on cell growth. Live metabolism-rich living cells are water-soluble yellow MTT (3- (4,5-dimethylthiaxo-2-yl) -2,5-diphenyl tetrazolium bromide) (Sigma, MO, USA) by the dehydrogenase action of mitochondria in cells. ) To form a purple water-insoluble formazan.
B16F10 cells were added to a 96 well plate at 2.0 × 10 4 cells / mL using DMEM medium to which 10% FBS was added, and 18 hours of incubation were performed. Then 50 μl of MTT solution was added and reacted for 4 hours. After completely removing the culture medium and adding 200 µl of dimethylsulfoxide to completely dissolve the precipitate, the absorbance of 540 nm was measured using a microplate reader. The average absorbance value for each sample group was obtained, and cell growth rate was evaluated by comparing with the absorbance value of the control group.
<1-3> Measurement of melanin biosynthesis inhibitory activity
B16F10 cells incubated in DMEM medium were dispensed in a 24 well plate at 2.0 × 10 4 cells / well, and then cultured for 24 hours and treated with 100 nM of α-MSH (-melanocyte stimulating hormone) and the sample prepared in the same manner at the same time. Time incubation. Next, the cells were washed with phosphate buffer (pH 7.4), and cell lysis buffer (100 mM sodium phosphate (pH6.8), 0.1 mM PMSF, 1% Triton X-100) was added, and the cells were lysed at -80 ° C for 30 minutes. Next, 1N NaOH was added to the cell lysate and reacted at 80 ° C. for 1 hour to dissolve the precipitate, and the absorbance was measured at 405 nm using a spectrophotometer. Inhibition of melanin biosynthesis was indicated by the decrease in absorbance of the addition and no addition of the sample solution.
<2> Experiment result
<2-1> cytotoxicity measurement result
After treating the B16F10 cells with an example extract and incubating for 48 hours, the results of confirming cell viability using the MTT assay are shown in FIG. 1. Referring to [FIG. 1], it can be confirmed that all of the extract extracts do not exhibit cytotoxicity.
<2-1> melanin synthesis amount measurement result
In order to evaluate the possibility of using the extract as a natural whitening agent, B16F10 cells, which are mouse melanoma cells, were treated with the extract extract and α-MSH, which is a melanogenesis-inducing agent, to produce melanin induced by α-MSH. The degree of inhibition by was measured, and the results are shown in FIG. 2. Referring to Figure 2 it can be seen that the production of melanin induced by α-MSH decreases in a concentration-dependent manner by the example extract.
In particular, it can be seen that the effect of further reducing the case of the mixed extract (1: 1 in weight ratio) of the oak petal extract and fifty-legged extract of the example extract.
Claims (5)
The composition is a composition for skin whitening using a mixed foot and oak petals extract, characterized in that the cosmetic composition.
Said composition is a composition for skin whitening using a mixed leg and extract of oak and oak petals, characterized in that the food composition.
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