KR20190102824A - A composition for skin whitening comprising extracts of tenebrio molitor - Google Patents

Abstract

The present invention relates to a composition containing a fermented extract of Tenebrio molitor. The composition comprising the fermented extract of Tenebrio molitor of the present invention as an active ingredient has a tyrosinase inhibitory effect and melanin production inhibitory effect, thereby having a skin whitening effect. In addition, the present invention has no cytotoxicity and skin side effects, and thus can be safely used in cosmetic, pharmaceutical and food compositions.

Description

A composition for skin whitening comprising brown gourd extract as an active ingredient {A COMPOSITION FOR SKIN WHITENING COMPRISING EXTRACTS OF TENEBRIO MOLITOR}

The present invention relates to a composition containing a brown mealworm extract, and more particularly, to a cosmetic, pharmaceutical composition and food composition for skin whitening containing a brown mealworm extract or brown rice wine fermentation extract as an active ingredient.

Generally, the eyes, hair, and skin color of a person are determined by the concentration and distribution of melanin in the skin, and also by the type of melanin, but are also affected by various environmental and physiological factors such as ultraviolet rays and stress. Melanin is a series of enzymatic enzymes that begins by oxidizing tyrosine, a type of amino acid in melanocytes in the basal layer of the epidermis, into oxidized by merosinase tyrosinase in the melanocytes of melanocytes. Produced through oxidation and non-enzymatic oxidation. The production and synthesis of melanin is a representative influence factor, ultraviolet rays, and other genetic factors, environmental factors, hormones, foods, pharmaceuticals and other chemicals. This melanin absorbs a certain amount of ultraviolet light to protect the skin and maintain body temperature, but when severely irritated, over-produced melanin deposits, causing freckles, senile spots, blemishes, brown or sunspots, sunburn, and wounds. Alternatively, hyperpigmentation and phototoxic reactions may occur after inflammation due to dermatitis.

Recently, Asian women prefer white and clean skin, and this is an important criterion of beauty. Therefore, the development of whitening agents for the treatment of abnormal skin pigmentation and the fulfillment of beauty needs has been actively made. At present, in the development of a whitening agent for skin beauty, a method of reducing the generated melanin pigment by decolorization and a method of inhibiting the activity of tyrosinase, an enzyme that forms melanin pigment, are known. However, since whitening agents using tocopherols and vitamins used to reduce the melanin pigment are known to have a slight decolorizing effect of the melanin pigment, many studies have been conducted on inhibitors that inhibit the production of melanin pigment.

In order to improve skin whitening or hyperpigmentation, conventionally, inhibitory activity against tyrosinase such as hydroquinone, ascorbic acid, kojic acid, glutathione and cysteine Has been used. However, hydroquinone exhibits a predetermined whitening effect, but has a problem of limiting the amount of the compound to a very small amount due to severe skin irritation. Ascorbic acid is easily oxidized, causing problems such as discoloration and discoloration. There is a disadvantage that the manufacturing process of the cosmetic is complicated by the instability in the silver solution. In addition, thiol-based compounds such as glutathione and cysteine not only have a characteristic unpleasant odor, but also have problems with transdermal absorption, and glycosides and derivatives thereof have high polarity, making it difficult to use as a component of a cosmetic composition. In addition, vitamin C has a problem in that it is easily oxidized in the aqueous solution state does not have a lasting effect.

Therefore, research and development of functional cosmetics having a skin whitening effect are actively progressing in the cosmetic field, and in particular, researches using natural materials are continued to avoid toxicity or irritation to the skin.

On the other hand, "Tenebrio molitor" is an insect of the coleoptera family, about 15mm long, dark brown and glossy. Brown goose larvae are called worms (mealworms) and are used for feeding insects or pets. The reason why worms are used for pets is because they are relatively short in metamorphosis, so they can easily experience insects and are very clean insects that feed grain. to be. Caterpillars molt 9-20 times depending on food and temperature until they become pupa. After two to three weeks of pupae, it becomes an adult, and when it is the first allegory, it becomes light brown but gradually turns black. The imago is nocturnal, hiding in a hole during the day, and active at night. It is usually overwintered as a larva in a grain bag near Gaeng, and then transforms into a pupa and adult in spring.

These brown mealworms have been reported to be very effective as a protein source as the highest protein content and low fat content among insects, and are known to exhibit increased anticancer activity, especially when treated with liver cancer cell lines with anticancer substances.

As a food patent associated with such brown gourds, Korean Patent Registration No. 10-1752253 discloses an insect fish cake composition using a brown gourd and a method for preparing the same. In Korean Patent Registration No. 10-1746111, swallowing food using brown gourd and its It describes a manufacturing method, and the patent related to the use of brown gourds describes a composition for preventing or treating diabetes comprising brown gourd larvae or extracts thereof as an active ingredient in Korean Patent Registration No. 10-1651908. No. 10-2016-0041110 discloses a composition for the prevention or treatment of rheumatoid arthritis comprising a suspension of brown mealworms as an active ingredient, and the Republic of Korea Patent Publication No. 10-2014-0134331 is an extract of brown mealworms or brown mealworms Obesity prevention or containing a suspension of Discloses a therapeutic composition, and Korean Patent Publication No. 10-2015-0050712 describes an anticancer composition comprising an extract or a fraction thereof as an active ingredient, Korean Patent Registration No. 10-1424125 Reported a composition for treating inflammatory diseases comprising larvae, and Korean Patent Registration No. 10-1404566 has reported a composition for preventing or treating dementia, including brown gourd extract.

However, there has been no report on the skin whitening effect using brown mealworms.

Therefore, the present inventors have continued to develop new natural substances having skin whitening effect and low human side effects, and the fact that brown gourd extract or brown gourd fermented extract shows excellent skin whitening effect without cytotoxicity. The present invention was completed by finding.

Therefore, the technical problem to be solved in the present invention is to provide a material having excellent skin stability while having a skin whitening effect.

In order to solve the above technical problem, the present invention provides a composition for skin whitening, characterized in that it comprises a brown gourd extract or fermented extract of brown gourd as an active ingredient.

Preferably, the composition comprising the brown rice wine fermentation extract in the present invention may be a cosmetic composition, a pharmaceutical composition or a food composition.

The term "extract" of the present invention refers to a formulation prepared by squeezing a crude drug into an appropriate leachate and evaporating the leachate, but not limited thereto, and extracting the extract obtained by the extraction treatment, the dilution or concentrate of the extract, and drying the extract. It can be a dried product obtained by these, these adjustment products, or a refined product.

Extract in the present invention can be prepared using common extraction methods, separation and purification methods known in the art. The extraction method may be a hot water extraction, hot water extraction, cold needle extraction, reflux cooling extraction or ultrasonic extraction, but is not limited thereto.

According to a specific embodiment of the present invention, the brown gourd extract may be extracted with brown gourd larvae or shells using an organic solvent to obtain useful components contained in the brown gourd larvae.

According to a specific embodiment of the present invention, the fat and fat components contained in the brown rice wine can be obtained by freeze-drying or hot-air drying the larvae and then extracting at 10 to 20 ° C. using water or an organic solvent. At this time, the organic solvent is methanol, ethanol, isopropanol, butanol, ethylene, acetone, hexane, ether, chloroform, ethyl acetate, butyl acetate, dichloromethane, N, N-dimethylformamide (DMF), dimethyl sulfoxide (DMSO) , 1,3-butylene glycol, propylene glycol or a mixed solvent thereof, preferably ethanol can be used.

In the present invention, the brown gourd fermented extract is freeze-dried or hot air dried the brown gourd larvae and then extracted using an organic solvent to remove fats and oils contained in the brown gourd larvae, and then lyophilized using a yeast strain It can be obtained by fermentation followed by extraction with water or an organic solvent.

According to a specific embodiment of the present invention, the fermentation is inoculated with the yeast strain to brown larva larvae from which the fat and fat components are removed, and then 60 to 80 hours, preferably at 20 to 30 ° C, preferably 23 to 27 ° C. Incubate for 70 to 75 hours. If the fermentation temperature is less than 20 ℃ fermentation does not occur sufficiently or takes a long time to fermentation is inefficient, if the fermentation temperature is more than 30 ℃ may cause the activity of the yeast strain is lowered enough fermentation may not occur. In addition, when the fermentation time is less than 60 hours, the fermentation extract of brown gourd does not occur sufficiently, so that the prepared brown gourd fermentation extract cannot exert sufficient skin whitening effect, and when the fermentation time exceeds 80 hours, It is uneconomical because there is no difference between fermentation and fermentation efficiency.

The yeast strains are, for example, Saccharomyces cerevisiae strain, S. ellipsoideus , Saccharomyces obayanus , Saccharomyces pas S. pastorianus , S. florentinus , and the like, and Saccharomyces cerevisiae strain is preferably used.

According to a specific embodiment of the present invention, the extraction of brown meal fermentation is carried out using water or an organic solvent such as water, methanol, ethanol, isopropanol, butanol, ethylene, acetone, hexane, ether, chloroform, ethyl acetate, butyl acetate, Hot water, cold extraction, ultrasonic extraction, reflux cooling extraction using dichloromethane, N, N-dimethylformamide (DMF), dimethyl sulfoxide (DMSO), 1,3-butylene glycol, propylene glycol or a mixed solvent thereof It may be carried out by a method such as, specifically, in the present invention can be extracted by heating, non-heating or cold needle fermented brown gourd using water or 60 to 80% alcohol.

In addition, the brown rice wine fermented extract may be obtained through a conventional purification process in addition to the extraction method by the extraction solvent. Obtained by various additional purification methods, such as separation using an ultrafiltration membrane having a constant molecular weight cut-off value, separation by various chromatography (manufactured for separation according to size, charge, hydrophobicity or affinity). It is also possible to obtain a brown rice wine fermentation extract through the fraction.

The brown rice wine fermentation extract of the present invention may be all extracts, fractions, purified products, dilutions thereof, concentrates, or dried products obtained at each stage of extraction, fractionation, or purification (separation, fractionation).

In the present invention, the "active ingredient" alone means a component that can exhibit the desired activity or itself can exhibit activity with a carrier that is not active.

In the composition of the present invention, the brown rice bran extract or brown rice bran fermentation extract may comprise 0.005 to 50% by weight, more preferably 0.01 to 30% by weight, most preferably 0.1 to 10% by weight, based on the total weight of the composition. Can be. In this case, if the content of brown rice wine fermentation extract is less than 0.005% by weight, the skin whitening effect of the present invention cannot be obtained. If the content exceeds 50% by weight, the effect is not proportional to the increase in content, which may be inefficient. And there is a problem that the stability of the formulation is not secured.

The composition containing the brown rice bran extract or brown rice bran fermentation extract of the present invention exhibits a skin whitening effect and has little cytotoxicity as a natural substance.

According to one embodiment of the present invention, it provides a cosmetic composition comprising a brown gourd extract or brown gourd fermented extract as an active ingredient.

The cosmetic composition according to the present invention includes ingredients commonly added to cosmetic compositions in addition to the brown gourd extract or brown gourd fermentation extract as active ingredients, such as conventional auxiliaries such as antioxidants, stabilizers, solubilizers, vitamins, pigments and flavorings, and Carriers may be further added.

The cosmetic composition of the present invention may be prepared in any formulation commonly prepared in the art, for example, solutions, suspensions, emulsions, pastes, gels, creams, lotions, powders, soaps, surfactant-containing cleansing , Oils, powder foundations, emulsion foundations, wax foundations and sprays, and the like, but are not limited thereto. More specifically, it may be prepared in the form of a nourishing cream, astringent lotion, a flexible lotion, a lotion, an essence, a nourishing gel or a massage cream.

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

Toss, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used as the carrier component when the formulation of the present invention is a powder or a spray, and 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 Fatty acid esters of, 3-butylglycol oil, glycerol aliphatic ester, polyethylene glycol or sorbitan.

When the formulation of the present invention is a suspension, liquid carrier diluents such as water, ethanol or propylene glycol, suspending agents such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, microcrystals Castle cellulose, aluminum metahydroxy, bentonite, agar or tracant gum can be used.

When the formulation of the present invention is a surfactant-containing cleansing, the carrier component is an aliphatic alcohol sulfate, an aliphatic alcohol ether sulfate, a sulfosuccinic acid monoester, an isethionate, an imidazolinium derivative, a methyltaurate, a sarcosinate, a 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.

According to one embodiment of the present invention, there is provided a pharmaceutical composition comprising the brown gourd extract or brown gourd fermented extract as an active ingredient.

The pharmaceutical composition according to the present invention includes a pharmaceutically acceptable carrier in addition to the brown rice bran extract or brown rice bran fermentation extract. Pharmaceutically acceptable carriers included in the pharmaceutical compositions of the present invention are those commonly used in the preparation of lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia rubber, calcium phosphate, alginate, gelatin, Calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methyl cellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil, and the like It doesn't happen. In addition to the above components, the pharmaceutical composition of the present invention may further include a lubricant, a humectant, a sweetener, a flavoring agent, an emulsifier, a suspending agent, a preservative, and the like. Suitable pharmaceutically acceptable carriers and formulations are described in detail in Remington's Pharmaceutical Sciences (19th ed., 1995).

The pharmaceutical composition of the present invention may be administered orally or parenterally, and is preferably applied by topical application by parenteral administration, more preferably by application.

Suitable dosages of the pharmaceutical compositions of the present invention vary depending on factors such as the formulation method, mode of administration, age, weight, sex, morbidity, food, time of administration, route of administration, rate of excretion and response to response of the patient. Can be. The dosage of the pharmaceutical composition of the present invention is in the range of 0.001-100 mg / kg on an adult basis. In addition, in the case of external preparations, it is recommended to continue the application for 1 month to 5 times a day in an amount of 1.0 to 3.0 ml on an adult basis. However, the dosage does not limit the scope of the present invention.

The pharmaceutical compositions of the present invention may be prepared in unit dosage form or in multidose form by formulating with a pharmaceutically acceptable carrier and / or excipient, according to methods which can be easily carried out by those skilled in the art. It may be prepared by incorporation into a container. In this case, the formulation may be in the form of a solution, suspension, syrup or emulsion in an oil or an aqueous medium, or may be in the form of an exir, powder, powder, granule, tablet or capsule, and may further include a dispersing or stabilizing agent.

In addition, according to one embodiment of the present invention, it provides a food composition comprising brown gourd extract or brown gourd fermented extract as an active ingredient.

The food composition according to the present invention, as an active ingredient, as well as brown gourd extract or brown gourd fermentation extract, as well as ingredients commonly added in food preparation, for example, proteins, carbohydrates, fats, nutrients, seasonings and flavoring agents It may include.

Examples of such carbohydrates are monosaccharides such as glucose, fructose and the like; Disaccharides such as maltose, sucrose, oligosaccharides and the like; And sugars such as conventional sugars such as polysaccharides such as dextrin, cyclodextrin and the like and xylitol, sorbitol, erythritol. As the flavoring agent, natural flavoring agents (tauumatin, stevia extract (for example rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be used.

For example, when the food composition of the present invention is prepared with a drink, in addition to the brown edible extract or brown edible fermented extract of the present invention, citric acid, liquid fructose, sugar, glucose, acetic acid, malic acid, fruit juice, tofu extract, jujube extract, licorice extract, etc. May be further included.

On the other hand, brown gourd extract or brown gourd fermentation extract of the present invention is a natural substance, harmless to the human body, there is little toxicity and side effects, so can be used safely even for long-term use, especially in the cosmetics, pharmaceutical and food compositions as described above It can be safely applied.

As such, the composition comprising the brown gourd extract or brown gourd fermentation extract of the present invention as an active ingredient has a skin whitening effect by not only tyrosinase inhibitory effect but also melanin production inhibitory effect. In addition, there are no cytotoxic and skin side effects, so it can be safely used in cosmetics, pharmaceutical and food compositions.

Figure 1 shows the measurement of tyrosinase inhibitory activity of brown rice wine extract.
Figure 2 is a graph showing the viability (%) of mouse melanoma melanocytes (Mouse melanoma melanocyte, B16F0 B16F0) cells by brown rice wine fermentation extract.
Figure 3 is a graph showing the tyrosinase inhibitory activity by the α-MSH induction of brown rice wine extract.
Figure 4 is a graph showing the change in melanin content due to tyrosinase inhibitory activity by the α-MSH induction of brown rice wine extract.
Figure 5 is a graph showing the cell survival rate by brown rice wine fermentation extract.

Hereinafter, examples and the like will be described in detail to help understand the present invention. However, embodiments according to the present invention can be modified in many different forms, the scope of the invention should not be construed as limited to the following examples. Embodiments of the present invention are provided to more fully describe the present invention to those skilled in the art.

<Example 1> Brown mealworm extract preparation

The samples were lyophilized or hot-air dried with brown rice wine, or lyophilized with peeling shells, and extracted under four solvent conditions (50%, 70%, 99% ethanol and H ₂ O).

Extraction was performed at 80 ° C. by adding 2 L to 100 g of the sample. The supernatant and the precipitate were separated and extracted two times for 2 hours.

For comparison, 1 L of water was added to 100 g of the sample, which was then rotated at 4 ° C. for 24 hours to obtain a low temperature water extract.

The extract was used as a sample of this experiment while filtration and concentration, freeze-dried and stored in the refrigerator compartment.

<Example 2> brown rice wine fermented extract preparation

(1) Removal of fat components contained in brown mealworms

The brownish larvae were repeatedly extracted three times at 14 ° C. for 24 hours using hexane and filtered with a Wattman No. 2 paper filter to separate the oil and fat components. Brown skin larvae from which the fat and fatty components were removed were lyophilized.

(2) fermentation

Brown larvae from which the fat and fatty components were removed were powdered.

Saccharomyces cerevisiae strain (KCTC 17299) was purchased from the Bank of Korea Cell Line and was passaged for the third time.

Saccharomyces cerevisiae subcultured using the powdered brown mealworm larvae powder in place of peptone in the composition of yeast peptone dextrose medium (1000 mL) The strain (KCTC 17299) was inoculated in an amount of 10 μL and fermented at 25 ° C. at a stirring speed of 100 rpm for 72 hours.

(3) extraction

Test group 1 was added 1L of water from brown larva fermentation broth and extracted with hot water at 80 ° C., separated from supernatant and sediment two times for 2 hours, test group 2 was added 1L of water from brown larva fermentation broth 4 Cold extraction at ℃ ℃ was extracted twice each 12 hours, Test Group 3 was extracted at room temperature using 70% alcohol instead of 1L brown larva fermentation broth fermentation broth, Test Group 4 was 70% instead of 1L brown larva fermentation broth The alcohol was extracted using the same method as the extraction of hot water, and test group 5 was extracted in the same manner as the extraction of the hot water using alcohol instead of water in 1 L of brown larva fermentation broth.

The extracts, fermentation broth and strain culture solution obtained above were each filtered using a Wattman No. 2 paper filter and then concentrated using a rotary vacuum concentrator. The concentrated samples were filtered using a sterile membrane filter (0.45 μm) and then stored at -80 ° C.

All the following experiments were measured in 3 repetitions and the mean value was expressed as the standard deviation. The statistical significance of the experimental results was Student's t-test, and the statistical significance was determined when the p value was less than 0.05.

Test Example 1 Determination of Tyrosinase Inhibitory Activity of Brown Rice bran Extract

Tyrosinase inhibitory activity was measured by Yagi et al. (Yagi et al., 1986). The reaction zone was prepared by adding 0.2 ml of mushroom tyrosinase (125 U / mL) to a mixture of 0.2 ml of 10 mM L-DOPA and 0.5 ml of 0.67 M sodium phosphate buffer (pH 6.8) and 0.1 ml of sample solution. The reaction was carried out for 10 minutes at and measured at DOPAchrome 475 nm wavelength generated in the reaction solution. Tyrosinase inhibitory activity was indicated by the absorbance decrease rate of the sample solution added and the non-added sample.

Figure 1 shows the measurement of tyrosinase inhibitory activity of brown rice wine extract. Here, as a result of measuring the mushroom tyrosinase inhibitory activity, the brown lyophilized lyophilized showed a high inhibitory effect when the brown lyophilized lyophilized, hot air dried and peeled shell lyophilized were compared. Showed the highest activity and hot water showed the next highest inhibitory activity. However, 70% and 99% ethanol extracts did not show low or inhibitory activity. As a result of comprehensive analysis, it was confirmed that the extraction of lyophilized brown mealworms with 50% ethanol was most effective.

Experimental Example 2 Cytotoxicity of Brown Rice Wine Extract

Cell viability was measured using CCK-8 (Cell Counting Kit-8) reagent (obtained from Dojindo Laboratories, Dojindo, Kumamoto, Japan). 96-well mouse melanoma melanin generation in cell plate after (Mouse melanoma melanocyte, B16F0 cells, American Type Culture Collection (Manassas, VA, USA) pre-sale receiving from) the inoculation with 1 X 10 ⁵ cells / well in each well the concentration Samples were treated at three concentrations of 10, 100 and 1000 μg / ml and incubated in a CO ₂ incubator for 24 hours. CCK-8 (obtained from Cell Counting Kit-8, Dojindo Laboratories, Dojindo, Kumamoto, Japan) solution was added and the absorbance was measured at 450 nm using a microplate reader (USA) 3 hours later.

Figure 2 is a graph showing the viability (%) of mouse melanoma melanocytes (Mouse melanoma melanocyte, B16F0) cells by the brown mealworm extract. As shown here, the brown gourd extract showed no viability at all concentrations and was not toxic.

Experimental Example 3 Tyrosinase Inhibitory Activity of Brown Rice bran Extract by α-MSH Induction

Tyrosinase inhibition activity was measured using the dopachrome method.

Mouse melanoma melanocyte (Mouse melanoma melanocyte) according to external stimulation to determine the effect of the brown gourd fermented extract on the activity of B16F0 intracellular tyrosinase activity as an external stimulant (α-Melanocyte stimulating hormone , α-MSH) 1000 μM was used.

The B16F0 cells were cultured at 5% CO ₂ in DMEM medium (Dulbecco's Modified Eagle's Medium) containing 10% heat-inactivated fetal bovine serum (FBS), penicillin (100 U / mL) and 2-mercaptoethanol (50 μM). Cultured at 37 ℃ was used.

Cells were dispensed at 1 × 10 ⁶ cells / well in ⁶ -well plates and treated with α-MSH as a positive control. After 1 hour, the samples were treated, and after 48 hours of incubation, cells were collected and dissolved by adding 1 mL of 0.1 M phosphate buffer (pH 6.5) containing 1% (W / V) Triton X-100. Subsequently, the supernatant obtained by centrifugation was used for tyrosinase activity and protein quantification, and the cell pellet was used for melanin quantification. Dispense 100 μL of the supernatant obtained after sample treatment into a 96-well plate, and add 100 μL of 0.1 M phosphate buffer (pH 6.5) to which 0.2 μg / mL 3,4-hydroxy-L-phenylalanine (L-DOPA) was added. After 2 hours incubation at 37 ℃ absorbance was measured at 490 nm. Each reaction was measured at least three times. Arbutin was used as a positive control.

Tyrosinase is an enzyme involved in the initial reaction, a rate-determining step in melanin synthesis, and represents an important regulatory step in melanin synthesis.Tyrosine hydroxylase activity converts tyrosinase to DOPA and DOPA oxidase which oxidizes DOPA to DOPAquinone. It has activity (SY Choi, et al., Korean journal of aesthetics and cosmetics society, vol. 14, no. 1, pp. 27-30, 2006).

Figure 3 is a graph showing the tyrosinase inhibitory activity by the α-MSH induction of brown rice wine extract. As shown here, the hot water extract of the peeled shell, the ethanol extract of the peeled shell, and the ethanol extract of the lyophilized brown mealworm showed inhibitory activity at a concentration of 1000 ug / ml, and the hot water extract of the lyophilized brown rice meal was inadequate. Could confirm. In other words, it was confirmed that all four samples showed inhibitory activity of tyrosinase.

<Test Example 4> melanin content of brown chopped extract

200 μL of a 1N NaOH solution containing 10% DMSO was added to the pellet obtained in the process of measuring the tyrosinase activity, and the resultant was completely dissolved in melanin at 90 ° C. for 30 minutes, and then absorbance was measured at 405 nm.

The melanin amount was obtained from a standard calibration curve prepared using synthetic melanin, and the melanin amount of the experimental group was calculated as a percentage of the melanin amount of the control group. Melanin amounts are expressed in μg / mg protein based on the protein concentration measured in each well.

Figure 4 is a graph showing the change in melanin content due to tyrosinase inhibitory activity by the α-MSH induction of brown rice wine extract. As shown here, it was confirmed that arbutin used as a positive control inhibited melanin activity in a concentration-dependent manner. The ethanol extracts of the lyophilized brown gourds inhibited melanin activity in a concentration-dependent manner, and the hot water extracts of the peeled shells, the ethanol extracts of the peeled shells, and the hot water extracts of the lyophilized brown meals were the highest at 1000 ug / ml. It was confirmed that the melanin inhibitory activity.

Experimental Example 5 Cytotoxicity of Brown Rice Wine Fermented Extract

Cell viability of the brown rice wine fermented extract was measured in the same manner as in Test Example 2.

In order to examine the effect of brown gourd extract on the viability of B16F0 cells, mouse-derived malignant melanoma cells, the cell viability was confirmed by treatment at three concentrations up to 10, 100 and 1000 ㎍ / ml.

Figure 5 is a graph showing the cell survival rate by brown rice wine fermentation extract.

As shown in Table 1, the brown rice wine fermentation extract was confirmed that there is no toxicity of the sample by confirming the survival rate of more than 98% even at high concentrations.

Test Example 6 Determination of Tyrosinase Inhibitory Activity of Brown Rice Wine Fermented Extracts

Tyrosinase inhibitory activity was measured using the same method as Test Example 3, and the results are shown in Table 2 below.

Brown Pain Cryogenic Water Extract Strain
Culture Brown meal fermented extract Fermentation
Culture 70% alcohol non-heating 70% alcohol heating Low temperature water extraction Hydrothermal extraction Inhibition rate 34.64 ± 0.02 46.18 ± 0.01 60.23 ± 0.01 58.82 ± 0.01 59.01 ± 0.01 55.92 ± 0.02 56.95 ± 0.02

As shown in Table 2, as a result of measuring the inhibitory effect of intracellular tyrosinase activity of the brown gourd fermented extract, the brown gourd cold water extract, which further inhibits the activity of tyrosinase in the brown gourd fermentation extract compared to the YPD strain culture extract Appeared. In addition, it was confirmed that tyrosinase activity inhibition was found to be 55% -60% in all five brown rice wine fermentation extracts. In addition, it can be seen that it shows a higher inhibitory effect than the arbutin (1000 μg / mL) used as a positive control group.

<Test Example 3> melanin content of brown rice wine fermented extract

Melanin content of the brown rice wine fermentation extract was measured using the same method as in Test Example 4 and the results are shown in Table 3 below.

Brown Pain Cryogenic Water Extract Strain
Culture Strain Fermentation Treatment Fermentation
Culture 70% alcohol non-heating 70% alcohol heating Low temperature water extraction Hydrothermal extraction Inhibition rate - 3.46 ± 0.10 61.93 ± 0.01 20.01 ± 0.01 49.55 ± 0.06 40.87 ± 0.05 -

As shown in Table 3, the results of measuring the melanin production inhibitory effect of the brown rice wine fermentation extract in the case of brown rice wine fermentation extract compared with no effect on inhibiting melanin production in the case of low temperature brown water extract, YPD strain culture extract It has been shown to inhibit melanin production by at least about 50%. Among them, the inhibition of melanin production of each of the four brown rice wine fermentation extracts was found to be 55% -60%. In particular, brown gourd fermented extract showed a melanin inhibitory effect similar to that of arbutin, compared to that of arbutin used as a positive control showed a 70% reduction rate at 1000 μg / mL.

Claims (6)

Cosmetic composition for skin whitening comprising a brown rice wine extract or brown rice wine fermentation extract as an active ingredient. The method of claim 1,
The brown gourd fermentation extract is obtained by extracting the brown gourd larvae with an organic solvent to remove fats and fats contained in the brown gourd larvae and then lyophilizing, fermenting with a yeast strain and extracting with water or an organic solvent. Cosmetic composition for skin whitening, characterized in that. The method of claim 2,
Cosmetic composition for skin whitening, characterized in that the yeast strain Saccharomyces cerevisiae strain. The method of claim 1,
The brown gourd extract or brown gourd fermented extract is contained in an amount of 0.005 to 50% by weight based on the total weight of the composition cosmetic composition for skin whitening. Pharmaceutical composition for skin whitening comprising brown rice wine extract or brown rice wine fermentation extract as an active ingredient. Food composition for skin whitening comprising brown rice wine extract or brown rice wine fermentation extract as an active ingredient.

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