KR20040040787A - cosmetic composition involved Berberin Glycyrrhizinate - Google Patents

Abstract

PURPOSE: Provided is a cosmetic composition containing Berberin Glycyrrhizinate as an active ingredient. The composition is useful for cosmetics having excellent anti-aging, skin protection and skin whitening effects, therapeutics for treating over-pigmentation, and food additives for inhibiting browning. CONSTITUTION: A cosmetic composition is characterized by containing as an active ingredient, 0.0001-20.0 wt.% of berberin glycyrrhizinate which is an ion-pair precipitate of berberine chloride with ammonium glycyrrhizinate, sodium glycyrrhizinate or potassium glycyrrhizinate. The cosmetic composition is used for skin whitening.

Description

Cosmetic composition involved Berberin Glycyrrhizinate}

The present invention relates to a cosmetic composition comprising berberine glycyrizinate as an active ingredient, and relates to a cosmetic composition containing berberine glycyrizinate with respect to the total dry weight of the cosmetic composition.

Herbal remedies using herbal medicines may use a single herbal medicine, but most of them are characterized by using a combination of two or more herbal medicines according to the patient's condition as premise, extract, granule, pill, and so on. The drug efficacy, mechanism of action, and action body components of individual herbal medicines are often unclear, and the behavior of herbal ingredients during the preparation of whole herbal medicines is characterized by the elution rate, precipitation of components, holding of herbal residues, adsorption, volatilization, decomposition, and chemical reaction between components. It is difficult to expect a certain mechanism of action as well as its mechanism of action due to solubilization (precipitation by solubilization, salting out, ion-pair or complex formation), and it is difficult to separate and quantify the active ingredient. Is holding. In addition, due to the lack of research on the components of traditional drugs, their application is limited to medicines, so if they study the components of these traditional drugs, or herbs, and apply them to other useful fields, new added value can be created.

Noguchi et al. Lost the strong delicacy and antimicrobial activity of berberine in the mixed solution of rhubarb and licorice, and the decrease of the content of berberine and glycyrrhizinate has been attributed to the berberine glycyrrhizinate It is reported that it is due to the removal of sediment, and the properties of this compound are revealed. Kim et al. Reported increasing the activity of Na ⁺ , K ⁺ -ATPase. In order to improve the bioavailability of berberine preparations, Taekjeon et al. Synthesized berberine tannins, which added tannin acid to methanol solution of berberine chloride to remove the strong bitter taste of hydrochloride. Maintaining high to increase the intestinal sterilization and branch effect. Thus, berberine glycyrizinate was already studied in the late 1960s, but the research was limited to medicines, and it was difficult to realize the practical use of the substance itself. Berberine glycyrizinate has not been studied since the 1970s, even in Japan, where research began for the first time due to the limitation of practical use. The research is insignificant.

Recently, research and development on the theme of the three functional cosmetics, such as whitening, wrinkle improvement, sun protection, etc. are showing significant progress by the basic research on skin and the development of bioactive substances according to biotechnology. In particular, in the whitening research, the development of a substance that controls and inhibits melanin production by inhibiting tyrosinase and using melanin metabolic pathways using herbal extracts and safe natural mixtures, and thus, the whitening cosmetics market is being activated. . A total of 216 patent applications for whitening cosmetics were published by August this year, of which 1,175 were filed by Koreans. Patent application for whitening cosmetics began in 1985 and has been increasing rapidly since 1995. The main components of the whitening cosmetics were natural extracts (57%), followed by 27% of new organic compounds, and 16% of vitamin C derivatives, kojic acid derivatives and retinol derivatives. On the trend of whitening cosmetics patents using natural extracts of domestic companies, foreign companies mainly applied for organic compounds until the early 90s, but as they entered the late 90s, they focused on new whitening ingredients through natural products. Natural extract whitening cosmetic ingredients research is expected to settle in the global trend.

Human skin has a primary defense against the harmful environment of the outside and also has an immune function to normalize it quickly if it is irritated. A representative example of the harmful environment for the skin is ultraviolet rays and melanin, which mainly determines the skin color, is a substance that helps the primary skin protection action. This melanin is composed of black melanin and tan melanin, and its quantity and ratio vary depending on race, sex, individual and skin area. White skin becomes black, blemishes and blemishes develop because of increased melanin in the skin. This melanin is produced by melanin-forming cells located in the bottom layer of the skin's epidermis.

Excessive ultraviolet rays, aging, stress, and childbirth activate the tyrosinase enzyme in melanocytes to produce melanin pigments, which are delivered to keratinocytes that form the outer layer of the skin in the form of small particles called melanosomes. . As a result, many melanosomes are distributed on the skin epidermis and the skin color becomes black. However, keratinocytes, which make up most of the skin's epidermis, are newly replaced every 28 days, so the melanin that remains undecomposed in the fate with fate of the keratinocytes becomes thinner. If the factors that cause excessive production of melanin pigments remain, they develop into blemishes and blemishes.

In general, it is important to determine the black and white color of a person's skin color by various external products (ultraviolet rays, medications, irritant skin external preparations) and internal factors (hormones, inflammatory factors, cytokines). It is biosynthesized in the melanosomes in pigmented cells called melanocytes present. In other words, these factors are activated after mitosis of melanocytes. In activated melanocytes, tyrosinase synthesis is promoted and melanin production is enhanced and transported out of the epidermis.

Melanin is a phenolic biopolymer with a complex of black pigment and protein, which is found in browning or feathers, feathers, skin, hair, eyes, etc. that occur when the cut surfaces of apples, potatoes, and bananas are exposed to the air. However, if melanin is excessively produced, blemishes, freckles, etc. are formed on the skin, skin aging is promoted, skin cancer may also be caused, and foods may also lower the quality of vegetables, fruits and fish.

Melanin is synthesized by using tyrosine in the substrate as a substrate. First, tyrosinase generates dopaquinone, and a copolymer melanin is produced through autooxidation and enzymatic reaction from dopaquinone. The melanin thus produced is transferred to keratinocytes through the melanosomes and comes out to the skin surface during the keratinization process.

The concept of whitening began to be introduced to consumers since the release of cosmetics containing vitamin C in the 60s and 70s, and the whitening product became a full-fledged whitening product in 83 years when the derivatives that stabilized unstable vitamin C were developed. Koji-san in 1988, arbutin in 1989, and AHAs, which eliminate keratin and promote the regeneration of skin cells in the mid-90s, introduced various methods to block inflammation-related signaling. Raw materials are being developed one after another. Particularly in the latter stages, the development of dermatological research that inhibits melanin-forming intracellular signaling or extracellular signaling receptors is the mainstream approach to converting signaling. The first-generation whitening raw materials currently widely used are raw materials for inhibiting tyrosinase activity such as kojic acid, arbutin and vitamin C, and skin induced by free radicals in the epidermis, which are increased by UV irradiation as the second-generation whitening raw materials. Licorice, baekryepi, aloe, etc. that function to eliminate the inflammatory mediators are the mainstream.

Currently, 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 mainly used. However, existing tocopherols and vitamins used to reduce melanin pigment have a slight decolorizing effect of melanin pigment, and compounds such as hydroquinone have a strong decolorizing effect, but they have skin sensitization, which may cause skin allergy. In addition, kojic acid is excellent in inhibiting the activity of tyrosinase, but because it denatures melanin pigment or melanocytes, it causes various skin irritation such as dermatitis, skin cancer. In addition, the whitening cosmetics containing kojic acid have a problem in formulation that is easily oxidized at a high temperature (45 ° C.) and discoloration occurs severely as time passes, thereby reducing the whitening effect. Recently, the extracts of lettuce extract and licorice from natural plants have been developed, but most plant extracts show relatively high inhibitory effect of tyrosinase activity at high concentrations, but little inhibitory effect at low concentrations.

Accordingly, it is an object of the present invention to provide a cosmetic composition effective for skin whitening by preparing a mixture of berberine glycinateate alone or a mixture of kojic acid, arbutin, oil-soluble licorice extract and the like previously known whitening effect.

Figure 1 is a schematic of the manufacturing method of the yellow licorice licorice coprecipitate.

Figure 2 shows the inhibitory effect of melanin biosynthesis in B16 melanoma cells of coarse licorice licorice coprecipitation (CGP means coprecipitation of the present invention).

Accordingly, the present invention provides a cosmetic composition comprising berberin glycyrrhizinate as an active ingredient.

The present invention also provides a cosmetic composition, wherein the berberine glycyrizinate is an ion-pair precipitate of berberine chloride and one of ammonium glycyrizinate, sodium glycyrizinate or potassium glycyrizinate.

The present invention also relates to berberin glycyrrhizinate derived from co-precipitate of one species of yellowish white or yellow lotus and licorice.

The present invention is also characterized in that the berberine glycyrizinate contains 0.0001-20.0% by weight based on the total dry weight of the cosmetic composition.

If the content of berberine glycyrizinate is less than 0.0001% by weight, it is difficult to expect the efficacy, and if it is more than 20.0% by weight, the expected effect is not large and it is not economical.

In addition, the present invention is characterized in that the cosmetic composition is used as a skin lightening agent.

Furthermore, the present invention is characterized in that the cosmetic composition is selected from a lotion, gel, water-soluble liquid, cream, essence, oil-in-water (O / W) type and water-in-oil (W / O) type formulation.

Hereinafter, the present invention will be described in more detail with reference to Examples. However, these Examples are only illustrative description of the present invention and the scope of the present invention is not limited to these Examples.

Example 1 Preparation of Rhubarb Licorice Coprecipitate

Huang and licorice are commonly used in Korea as a sample. To prepare coprecipitates, licorice and sulfur are mixed with water and an organic solvent at a ratio of 1:10 and extracted at various temperatures. Preferably, the hydrothermal extraction at 80 ~ 90 ℃ considering the extraction solvent and the dry yield and berberine glycyrinate content according to the temperature. After equal amounts of the respective extracts were stabilized at 4 ° C. for 1 day, centrifuged and the precipitates were separated. The precipitates are recovered, recrystallized using alcohol, and freeze-dried to prepare coarse precipitates of sulfur and licorice. Preferably, 65-85% by weight of berberine glycyrizinate is contained in the coarse licorice colic.

Example 2: Cytotoxicity Test

Toxicity to mouse fibroblast L929 cells was measured by diluting the 10 mg / ml rhubarb licorice coprecipitate solution to each concentration. Fibroblast cells were purchased from a mouse-derived L929 cell line. Fibroblast cell lines were inoculated in DMEM medium containing 4.5 g / L glucose, 10% serum and 1% antibiotics and incubated at 37 ° C. in a 75 cm 2 T-flask. After 24 hours of incubation under 5% CO ₂ , cells were isolated by treatment with 0.05% Trypsin containing 0.02% EDTA, followed by incubation for 6 hours by 96-well-flask inoculation. At this time the cell number is 1 × 10 ⁴ cells / well. Here, 10 mg / ml rhubarb licorice coprecipitate was added to each concentration and incubated for 12 hours. After incubation, 10 μl of MTT (3- (4,5-dimethylthiazol-2-yl) 2,5-diphenyl tetrazolin bromide) solution (5 mg / ml) was added to the wells and incubated for 4 hours, followed by DMSO (dimethyl sulfoxide). After 100 μl of the solution was added, OD _{570 nm} absorbance was measured to compare cell viability. As shown in Table 1, the survival rate was found to be safe even at a concentration of 100 µg / ml or more. As a result of the above experiment, the toxicity of cells of coarse licorice colic was nontoxic or nontoxic.

Concentration (% by weight) 0.001 0.01 0.1 One MTT ₅₀ Survival rate (%) 100 100 91 61 > 1

Example 3 Comparative Study of Tyrosinase Enzyme Inhibition of Rhubarb Licorice Extract

In this example, the tyrosinase enzyme inhibition rate was measured for the rye licorice extract obtained in Example 1. All reagents except the above sample were from Sigma Co., Ltd. First, 1.15 mL of 0.1 M potassium-phosphate buffer (pH6.8) was placed in a test tube, and 0.1 mL of a 3 mM tyrosine solution dissolved in distilled water and 0.2 containing a sample were prepared. mL sample solution was added (the final concentration of each sample in the reaction solution was 2% (V / V)). In each test tube, 0.05mL of 2000unit / mL tyrosinase enzyme solution dissolved in 0.05M potassium-phosphate buffer solution was added and reacted in a 37 ° C constant temperature water bath for 10 minutes. At this time, only 0.2 mL of distilled water was added to the control A instead of the sample. In the control group B, 0.05 mL of distilled water was added instead of the tyrosinase enzyme solution. After the reaction, the absorbance at 475 nm was measured with a photospectrometer. The tyrosinase enzyme inhibition rate of each sample was calculated | required by substituting the said absorbance measurement value etc. to the following formula.

Inhibition Rate (%) = {(Reaction Absorption-Response Absorption of Control B) / Reaction Absorption of Control A} X100

sample 10 μg / mL 100 μg / mL 300 μg / mL 1000 μg / mL Tyrosinase enzyme inhibition rate (%) Arbutin 3.2 38 51 88 Yellow Licorice Coprecipitate 49 92 99.8 99.9

As a result, as shown in Table 2, the rye licorice licorice coprecipitate showed a much higher inhibitory effect of tyrosinase enzyme than the whitening agent arbutin generally used in the experimental concentration.

Example 4 Inhibitory Effects of Melanin Production at the Cellular Level of Coarse Licorice Coprecipitate

Melanin produced by the coarse licorice coagulum obtained in Example 1, B16-F1 melanoma cells used in this example is a cell line derived from the mouse, cells that secrete a black pigment called melanin (used in this experiment) B16-F1 melanoma cells were distributed from ATCC (American Type Culture Collection, Accession No .: 6323). The cells were treated with a substance during the artificial culture to evaluate the degree of reduction of melanin black pigment.

The inhibition of melanin biosynthesis of B16-F1 melanoma cells was determined as follows. B16-F1 melanoma cells were suspended in DMEM (Dulbecco's Modified Eagle's Medium; Gibco, USA) containing 10% fetal bovine serum at a concentration of 5 × 10 ³ cells / ml. Suspension cells (5 ml) were placed in a tissue culture flask, and then 0.1% rhododendron licorice co-precipitate and 10% arbutin samples were added at various concentrations, and then cultured at 37 ° C. under 7% carbon dioxide-95% air. After incubation for 4 days, the melanin production was primarily determined by the color of the culture medium, and the cells were washed with phosphate buffer and trypsinized to separate the cells from the flask. The cells were collected in tubes and washed with phosphate buffer, centrifuged and melanin extracted. To this, 1 ml of sodium hydroxide solution (1N concentration) was added, boiled for 10 minutes to dissolve the melanin, and the absorbance was measured at 400 nm by using a spectrophotometer, indicating the amount of melanin produced as the absorbance of a unit cell count (10 ⁶ cells). The melanin production relative to the control group was calculated as the inhibition rate (%) and the results are summarized in Table 3, and the photograph is shown in FIG. As can be seen from the above results, the addition of 0.1% rhubarb licorice coprecipitate (CGP) and 10% arbutin, which is known as a conventional whitening agent, was compared with the inhibition of melanin pigment secretion. It can be seen that the increase by more than 100 times.

0.1 wt% 0.25 wt% 0.35 wt% 0.5 wt% Melanin biosynthesis inhibition rate (%) Coprecipitation of the Invention 66 71 83 N.D Arbutin 59 N.D 65 82

(N.D in the table means no measurement)

In the present invention, a mixture of coprecipitates of one of Coptidis rhizoma or sulfur white and extract of Glycyrrhizae radix and berberine glycyrizinate as a component of these coprecipitates is natural and has no side effects and inhibits tyrosinase enzyme activity and B16. Not only is it excellent in inhibiting melanin production in mouse melanoma cells, but also excellent in radical scavenging and lipid peroxidation, and it can be used to prevent skin aging, skin protection and whitening, cosmetics, blemishes, freckles, spots or hyperpigmentation in pregnancy. It is possible to provide a hyperpigmentation therapeutic agent and a food additive for suppressing browning of food.

Claims (7)

Cosmetic composition comprising berberin glycyrrhizinate as an active ingredient. The berberine glycyrrhizinate according to claim 1, wherein the berberin glycyrrhizinate is an ionic pair precipitate of berberine chloride and one of ammonium glycyrizinate, sodium glycyrizinate or potassium glycyrizinate. Cosmetic composition containing an active ingredient of siberate (berberin glycyrrhizinate). The cosmetic composition according to claim 1, wherein the berberine glycyrrhinate is derived from co-precipitation of one type of sulfur white or rhubarb and licorice, and a berberin glycyrrhizinate as an active ingredient. The cosmetic composition according to claim 1, wherein the berberine glycyrrhinate contains 0.0001-20.0% by weight relative to the total dry weight of the cosmetic composition, berberin glycyrrhizinate as an active ingredient. The cosmetic composition according to claim 1 or 2, wherein the cosmetic composition is used as a skin lightening agent, berberin glycyrrhizinate as an active ingredient. The cosmetic composition of claim 1 or 2, wherein the cosmetic composition is selected from a lotion, gel, water-soluble liquid, cream, essence, oil-in-water (O / W) type, and water-in-oil (W / O) type. A cosmetic composition comprising berberin glycyrrhizinate as an active ingredient. A food additive that suppresses browning of foods using berberin glycyrrhizinate as an active ingredient.