KR102062311B1 - Method for Producing Agaricus Blazei Extract, Extract and Cosmetic Composition Using the Same - Google Patents

Abstract

The present invention relates to a cosmetic composition for improving skin condition comprising white mushroom extract as an active ingredient, having a skin whitening, wrinkle and skin elasticity improving effect.

Description

Method for producing white mushroom extract, extract and cosmetic composition using the same {Method for Producing Agaricus Blazei Extract, Extract and Cosmetic Composition Using the Same}

The present invention relates to a method for producing a white mushroom extract, a white mushroom extract using the same, and a cosmetic composition containing the white mushroom extract as an active ingredient.

Aging is largely divided into natural aging or endogenous aging and external aging. Natural aging is caused by genetic causes, so it is difficult to control, but external aging is caused by environmental causes, and thus artificial regulation is easy. Therefore, studies to prevent external aging have been continued, and in particular, research on the prevention of wrinkle formation caused by exogenous photoaging due to prolonged ultraviolet exposure has been focused (Gilchrest BA, J. Am. Acad. Dermatol., 1989: 21: 610-613). The clinical characteristics of the exogenous skin aging, photoaging, is that the skin becomes rough and elastic, irregular pigmentation occurs, and deep wrinkles increase.

Active ingredients for skin wrinkle improvement or anti-aging currently being developed cannot be used for some cosmetic ingredients or are very unstable, and are difficult to transfer to the skin, requiring a special stabilization system and delivery system, and improving skin wrinkles. There is a problem that is not visible, etc. Recently, attention has been gradually focused on skin protection agents containing retinoids. The retinoid is used as a means to solve photoaging phenomena such as wrinkles, skin thickening, sagging, and elasticity, which are a result of the accumulation of sunlight. However, retinoids are very unstable compounds and are sensitive to ultraviolet rays, moisture, heat, and oxygen, and easily cause chemical changes. Therefore, research is focused on developing active ingredients derived from natural products to solve them.

The present inventors have studied to find a substance derived from natural products that can improve the skin condition, and as a result, the effect of inhibiting the activity of the white mushroom extract elastase, tyrosinase and collagenase (collagenase) Confirmed that the present invention was completed.

1. Republic of Korea Patent No. 10-0535875

One object of the present invention is to provide a cosmetic composition having an effect of improving wrinkles, skin whitening or skin elasticity.

In order to achieve the above object, an aspect of the present invention provides a cosmetic composition comprising the extract of Agaricus blazei as an active ingredient.

White mushrooms belong to the fungi fold fungi, Agaricus mushrooms, and also called spiritual mushrooms. It is rich in various nutrients such as protein, fat, fiber, vitamins, inorganic salts and nucleic acids, and is known to be effective in various adult diseases including cancer.

According to one embodiment of the invention, the white mushroom extract may be prepared according to conventional methods known in the art. Specifically, after washing, drying and grinding white mushrooms, the white mushroom extract can be prepared using a conventional solvent under the conditions of a normal temperature and pressure.

The solvent may be selected from the group consisting of distilled water, alcohol having 1 to 4 carbon atoms, ethyl acetate, chloroform, diethyl ether, dichloromethane, hexane, acetone, and mixtures thereof, preferably distilled water.

According to an embodiment of the present invention, as a cosmetic composition comprising the extract of Agaricus blazei as an active ingredient,

The white mushroom extract,

Washing and drying the white mushrooms;

Put dried white mushrooms into the extraction tank, 8 to 10 times the weight of purified water to the total weight of the white mushrooms, and then extracted for 5 to 9 hours at 110 to 125 ℃, recovering the first extract from the extraction tank Doing;

After additionally adding 8 to 10 times the weight of purified water relative to the total weight of the white mushrooms remaining in the extraction tank, the first extract was recovered, and extracted for 5 to 9 hours at 110 to 125 ℃, the second extract from the extraction tank Recovering; And

It may be obtained by the extraction method comprising the step of mixing the recovered first extract and the second extract and filtered with a filter.

According to one embodiment of the present invention, the white mushroom extract may be obtained by an extraction method which additionally performs the step of vacuum concentration of the filtered extract after the step of filtering with the filter.

According to one embodiment of the present invention, the step of concentrating in vacuum may be concentrating to 5 brix or more.

On the other hand, the white mushroom extract includes not only the extract by the solvent extraction method described above, but also an extract that has undergone a conventional purification process. Fractions obtained through various additional purification methods, such as, for example, 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 may also be included in the extract of the present invention. In addition, the white mushroom extract may be prepared in a powder state by an additional process such as distillation under reduced pressure and freeze drying or spray drying. Specifically, the white mushroom extract may be prepared in powder form through sterilization and spray drying, and may be prepared in powder form using a spray dryer after sterilization for 10 to 40 minutes at a temperature of 70 to 90 ℃. have.

According to one embodiment of the present invention, the first and second extracts are obtained by extraction being performed at 110 to 125 ° C, 115 to 125 ° C, 115 to 121 ° C, 118 to 121 ° C, or 120 to 121 ° C. When extracted at a temperature of less than 110 ℃ may not be sufficiently extracted active ingredients having wrinkle improvement, skin whitening, skin elasticity enhancement, skin irritation, or moisturizing effect, extracted at a temperature above 125 ℃ In this case, the active ingredient may be denatured.

According to one embodiment of the invention, the first and second extracts are extracted for 5 to 9 hours, 6 to 9 hours, 5 to 8 hours, 6 to 8 hours, 7 to 8 hours, or 6 to 7 hours. It may be obtained by performing the extraction, if the extraction is performed for less than 5 hours, the active ingredient may have not been extracted enough to improve wrinkles, whiten skin, enhance skin elasticity, reduce skin irritation, or moisturize, 9 hours If the extraction is carried out in excess of the extracted components may be modified to reduce the wrinkles, skin whitening, skin elasticity, skin irritation, or moisturizing effect according to an embodiment of the present invention.

According to one embodiment of the present invention, the first extract and the second extract may be obtained by performing the extraction under high pressure conditions, the pressure in the extraction tank in which the extraction is performed 1 to 10atm, 1 to 8atm, 1 to 6 atm, 1 to 5 atm, 1.1 to 5 atm, 1.1 to 4 atm, 1.1 to 3 atm, 1.1 to 2.1 atm, 1.1 to 1.5 atm, 1.1 to 1.2 atm, or 1.2 to 1.5 atm.

According to one embodiment of the invention, the filter may be one having a pore size of 0.5 to 2.0 μm, 0.5 to 1.5 μm, 0.5 to 1.0 μm, or 1 μm.

According to one embodiment of the invention, the cosmetic composition may be to inhibit the activity of at least one enzyme selected from the group consisting of elastase, tyrosinase and collagenase.

According to one embodiment of the invention, the cosmetic composition may have one or more effects selected from the group consisting of wrinkle improvement, skin whitening, skin elasticity enhancement, skin irritation relief, and moisturizing effect.

In the present invention, "whitening" means whitening the skin, and means all actions of inhibiting or inhibiting melanin's skin deposition by inhibiting the synthesis of melanin.

According to one embodiment of the invention, the composition may be to inhibit the activity of tyrosinase (tyrosinase).

In the present invention, "tyrosinase" is a metal-containing protein containing a pair of copper ions in the active site, and L-tyrosine is subjected to hydroxylation reaction in the production path of melanin, and 3,4-dihydroxy phenylalanine ( L-DOPA), which is then oxidized to phenylalanine-3,4-quinone (dopaquinone) and then synthesized into melanin via several intermediates such as dopachrome, indole-5,6-quinone, etc. Means enzymes involved in.

Barley mushroom extract according to an embodiment of the present invention has a concentration-dependent tyrosinase inhibitory activity, the composition according to an embodiment of the present invention may have a whitening activity.

In the present invention, "elastase" means an enzyme having a role in catalyzing the hydrolysis reaction of elastin involved in skin elasticity.

In one embodiment of the present invention, the white mushroom extract has a concentration-dependent elastase inhibitory activity, the composition according to one embodiment of the present invention may have a wrinkle improving activity.

The composition according to one embodiment of the present invention may be to inhibit collagen fiber damage.

Collagen fiber is a fibrous tissue composed of collagen and is present in connective tissues such as bone, cartilage, skin, and tendons. When the skin is exposed to ultraviolet rays, the collagen fibers involved in the elasticity of the skin are damaged, thereby aging the skin.

In one embodiment of the present invention, the white mushroom extract has a concentration-dependent collagen fiber damage inhibitory activity, the composition according to one embodiment of the present invention may have a wrinkle improving activity.

The cosmetic composition of the present invention may include components commonly used in cosmetic compositions in addition to including the white mushroom extract as an active ingredient, and include conventional auxiliaries such as stabilizers, solubilizers, vitamins, pigments and flavorings, and And a carrier.

On the other hand, the cosmetic composition of the present invention can be prepared in any formulation commonly prepared in the art, for example, solutions, suspensions, emulsions, pastes, gels, creams, lotions, powders, soaps, surfactants- It may be formulated with containing cleansing oil, powder foundation, emulsion foundation, wax foundation, spray, and the like, but is not necessarily limited thereto.

According to one embodiment of the invention, the cosmetic composition is a flexible lotion, gel, water-soluble liquid, milk lotion, cream, essence, skin toner, oil-in-water emulsion, water-in-oil Formulations selected from the group consisting of emulsions, cleansing foams, cleansing water, packs, body oils, body lotions, oil-in-water makeup bases, water-in-oil makeup bases, and foundations.

In case the formulation of the cosmetic composition of the present invention is a paste, cream or gel, the carrier component is animal oil, vegetable oil, wax, paraffin, starch, tracant, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide. This can be used.

In addition, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used as a carrier component when the formulation of the cosmetic composition of the present invention is a powder or a spray, and in particular, in the case of a spray, additionally chlorofluoro Propellants such as rohydrocarbon, propane / butane or dimethyl ether.

In addition, when the formulation of the cosmetic composition of the present invention is a solution or emulsion, a solvent, a solubilizing agent or an emulsifying agent is used as the carrier component, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate. Fatty acid esters of propylene glycol, 1,3-butylglycol oil, glycerol aliphatic ester, polyethylene glycol or sorbitan may be used.

In addition, when the formulation of the cosmetic composition of the present invention is a suspension, a liquid diluent such as water, ethanol or propylene glycol, ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester, and polyoxyethylene sorbitan ester are used as carrier components. Suspending agents, microcrystalline cellulose, aluminum metahydroxy, bentonite, agar or tracant and the like can be used.

In addition, when the formulation of the cosmetic composition of the present invention is a surfactant-containing cleansing agent, as a carrier component, an aliphatic alcohol sulfate, an aliphatic alcohol ether sulfate, a sulfosuccinic acid monoester, an isethionate, an imidazolinium derivative, a methyltaurate, and sarco Cinates, fatty acid amide ether sulfates, alkylamidobetaines, fatty alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, lanolin derivatives or ethoxylated glycerol fatty acid esters, and the like.

White mushroom extract according to an embodiment of the present invention has an excellent effect of inhibiting the activity of elastase, tyrosinase and collagen degrading enzymes can be usefully used as an active ingredient of the cosmetic composition for skin improvement.

Figures 1a to 1c shows the results of confirming the extraction yield of the white mushroom extract according to the change of extraction conditions: Figure 1a shows the extraction yield according to the extraction temperature (60, 80, 90, 100 or 120 ℃), Figure 1b Shows the extraction yield according to the extraction time (6, 8, 10 or 12 hours), Figure 1c shows the extraction yield according to the ethanol content (0, 30, 50 or 70%).
Figure 2 schematically shows a process of manufacturing white mushroom extract according to an embodiment of the present invention.
Figure 3 shows the results of measuring the elastase inhibitory activity of the white mushroom extract.
Figure 4 shows the results of measuring the collagenase inhibitory activity of the white mushroom extract.
Figure 5 shows the expression level of MMP-1 measured when the white mushroom extract in human skin fibroblasts.
Figure 6 shows the expression of type I collagen measured in the treatment of white mushroom extract in human skin fibroblasts.
Figure 7 shows the results of measuring the tyrosinase inhibitory activity of the white mushroom extract.

Hereinafter, one or more embodiments will be described in more detail with reference to Examples. However, these examples are intended to illustrate one or more embodiments, but the scope of the present invention is not limited to these embodiments.

Example 1 Preparation of White Mushroom Extract

Example 1-1. Establishment of extract preparation conditions

In order to confirm the extraction yield according to the extraction conditions were tested as follows.

Agaricus blazei was purchased from Kim Sang Min Agaricus (Jeju, South Korea) to remove adherents, and then washed with clean fresh water and dried at room temperature. The dried white mushrooms were added to the extraction tank, and hot water or ethanol was added twice to 8 to 10 times the weight of the white mushrooms. At this time, the extraction temperature was changed to 60 ℃, 80 ℃, 90 ℃, 100 ℃, 120 ℃, extraction time was changed to 6 hours, 8 hours, 10 hours, 12 hours. Ethanol was used 0% (hot water), 30%, 50% and 70%.

As a result, as shown in Figure 1a it was confirmed that the extraction yield increases as the extraction temperature increases. In addition, as shown in FIG. 1B, when only the extraction time was changed at the same extraction temperature (120 ° C.), the extraction yield was found to be constant within the error range regardless of the extraction time.

When extracted twice at 70 ° C. for 6 hours using ethanol as the extraction solvent, the extraction yield was increased as the ethanol concentration is increased as shown in FIG. The control (hot water extraction) extracted twice for 8 hours at 120 ° C. showed an extraction yield of 51.4%. However, the ethanol extract, unlike hot water extract, was found to be insoluble in higher concentrations, and even when not concentrated, insoluble components were precipitated as ethanol evaporated.

In addition, since the use of ethanol extract as a raw material of food or cosmetics may affect the formulation of the final product, hot water was used as an extraction solvent in subsequent experiments.

Example 1-2. White Mushroom Extract

The dried white mushrooms were added to the extraction tank, and purified water was added to 8 to 10 times the weight of the white mushrooms. The extract was recovered by extracting for 6 hours to 8 hours under conditions of 121 ° C. and 1.2 to 1.5 atm, and purified water of 8 to 10 times the weight of white mushrooms was further added to the extraction tank. The secondary extract was recovered by extracting for 6 hours to 8 hours under conditions of 121 ° C. and 1.2 to 1.5 atm. The collected primary and secondary extracts were mixed, filtered through a filter having a pore size of 1.0 μm (Rainbow, South Korea), and a hydrothermal extract (hereinafter referred to as extract 1) extracted at 121 ° C. was obtained.

The filtered extract was concentrated in vacuo to 5 brix or more. After recovering the concentrate of the white mushroom extract, it was confirmed that it is safe by examining the presence, foreign matter, water, general bacteria and E. coli.

The manufacturing method of the white mushroom extract according to the present embodiment is shown in Figure 2 in order. In addition, the same method as above except that the temperature was changed to 60 ℃ (hereinafter, extract 2), 80 ℃ (hereinafter, extract 3), 90 ℃ (hereinafter, extract 4), 100 ℃ (hereinafter, extract 5). White mushroom extract 2 to 5 was prepared.

Example 2. Analysis of Phenolic Content of White Mushroom Extract

The phenolic content of the white mushroom extract 1 prepared in Example 1-2 was analyzed.

10 ml of tertiary distilled water was added to 1 ml of the white mushroom hot water extract (extract 1) extracted at 121 ° C., and 2 ml of Folin-Ciocalteu phenol solution was added and mixed. After reacting at room temperature for 5 minutes, 2 ml of 20% sodium carbonate solution was added, and further reacted at room temperature for 2 hours. When the blue mixture was produced, the absorbance was measured at 680 nm, and the quantitative curve was checked using catechin, which is an indicator substance. Experimental results indicate the phenol content contained in mg unit per 1g of white mushroom extract.

As a result, as shown in Table 1, the total phenolic content of the white mushroom extract was found to be 898.96 mg / g.

extract Total Phenolic Content (mg / g) White Mushroom Extract 1 898.96 ± 42.66

Example 3 Inhibitory Effect of Elastase Activity

In the dermal tissue of the skin, collagen and elastin form a network structure, and when elastin is broken down by elastase, the network structure of the skin is broken up to form wrinkles on the skin. Therefore, in order to confirm the skin improvement effect of the white mushroom extract was tested for the inhibition of elastase activity.

Elastase hydrolyzes N-succinyl-Ala-Ala-Ala-p-nitroanilide to produce p-nitroaniline . 100 μl of 2 mM N-succinyl-Ala-Ala-Ala-p-nitroanilide solution was prepared using 0.1 M Tris-Cl buffer (pH 8.0), and this solution was added to retinol (positive control) or Example 1-. White mushroom extract 1 prepared in 2 was added at a concentration of 100 µg / ml and mixed lightly. 50 µl of elastase solution (0.1360 unit / ml, E7885, SIGMA USA) was added thereto, reacted at 37 ° C for 10 minutes, and absorbance was measured at 410 nm (Synergy2, BioTek Inc., USA). Elastase activity inhibition rate is calculated according to the following equation 1, it is shown in Table 2 and FIG.

As a result, as shown in Table 2 and Figure 3 it was confirmed that the elastase inhibitory activity of the white mushroom extract 1 is about 25% higher than the positive control retinol.

extract Elastase Inhibitory Activity (%) Retinol (positive control) 19.26 ± 1.00 White Mushroom Extract 1 34.35 ± 0.38

Example 4 Collagen Inhibitory Activity

Collagen is the main protein of all connective tissues such as skin, blood vessels, and muscles, and accounts for 70% to 80% of dry skin and is degraded by collagen degrading enzymes. In this study, primary screening of collagenase inhibitory activity of samples containing white mushroom extract using 4-Phenylazobenzyloxycarbonyl-Pro-Leu-Gly-Pro-D-Arg trifluoroacetate salt, which is used as a substrate of collagenase It was.

4 mM CaCl ₂ was added to 0.1 M Tris-HCl buffer (pH 7.5), and the substrate was dissolved with 4-phenyl azobenzyloxycarbonyl-Pro-Leu-Gly-Pro-D-Arg (Sigma, USA) at a concentration of 0.3 mg / ml. 125 µl of the solution, 75 µl of the collagen degrading enzyme (Sigma, USA) solution at a concentration of 0.2 mg / ml, and white mushroom extract 1 (100 µg / ml) prepared in Example 1-2 were added thereto, and left at room temperature for 20 minutes. . Ascorbic acid (100 µg / ml) was used as a positive control. Then, the reaction was stopped by adding 250 µl of 6% citric acid solution, and 1.5 ml of ethyl acetate was added to measure absorbance at 320 nm. To calculate the collagenase inhibitory activity (%) in accordance with Equation 2 shown in Table 3 and FIG.

As a result, as shown in Table 3 and Figure 4, it was found that the collagenase inhibitory activity of the white mushroom extract was about two times better than that of the ascorbic acid which is a positive control.

extract Collagen Inhibitor Inhibitory Activity (%) Ascorbic acid (positive control) 5.49 ± 2.14 White Mushroom Extract 1 11.88 ± 1.69

Example 5 Evaluation of MMP-1 Inhibitory Activity

Matrix metalloproteinase-1 (MMP-1), a collagen degrading enzyme, is one of the most influential enzymes in skin collagen decline.The active ingredient that inhibits MMP-1 can inhibit aging of the skin, improve skin wrinkles, It has a good effect on improving elasticity and preventing skin aging. Collagen degrading enzyme inhibitory ability of Example 4 was tested for a specific substrate defined in vitro, but in Example 5, by confirming the inhibitory activity of MMP-1 produced in human cells (HDF; Human dermal fibroblast), In-depth analysis of ADME (Absorption, Distribution, Metabolism) and metabolism (Excretion) of a sample to cells, taking into account the complex action of collagen synthesis-related factors not limited to specific substrates The purpose of this study was to investigate the anti-wrinkle activity.

By measuring the amount of RNA expression of MMP-1 using qRT-PCR, whether the composition comprising the white mushroom extract according to an embodiment of the present invention inhibits the expression of MMP-1 and inhibits MMP-1. I looked at it. 50 μg of the pro-inflammatory cytokine TNF-α 10 ng / ml and the white mushroom extract 1 prepared in Example 1-2 for 12 to 24 hours using TRIzol reagent (Life Technologies, CA, USA) according to the manufacturer's instructions HDF cells after incubation at 37 ° C. and CO ₂ conditions for 24 hours after treatment to human dermal fibroblasts (Human dermal fibroblast, ATCC or less HDF cells, ATCC) at a concentration of / ml, 100 µg / ml or 200 µg / ml Total RNA was extracted from.

8 μl Molony Murine Leukemia Virus Reverse Transcriptase (M-MLV RT) 5 × buffer, 3 μl 10 mM dNTPs, 0.45 μl 40 U / μl RNase inhibitor, 0.3 μl 200 U / μl M-MLV RT (Promega, Madison, WI, USA) and 3.75 μl of 20 μM oligo dT (Bioneer, Daejeon, Korea) were used to synthesize cDNA by reverse transcription of 5 μg of total RNA using 40 μl of the reaction mixture.

2 μl of the reaction mixture consisting of 10 μl of RNaseH SYBR Green PCR master mix, 2 μl of primer, 7 μl of PCR-grade water (Takara, Mountain View, CA, USA) and 1 μl of cDNA was added to the Power SYBR Green PCR Master Mix. QRT-PCR was performed in a StepOnePlus Real-time PCR system with (Applied Biosystems). After 10 minutes at 95 ° C., 40 cycles of 15 seconds at 95 ° C. and 1 minute at 60 ° C. were performed. Fluorescence intensity was measured after every cycle. After normalizing the Ct (threshold cycle) value of each gene to the Ct value of β-actin, the expression level of each gene was analyzed by comparing the change in Ct value. The primer sequence of each gene used for the experiment is shown in Table 4.

Primer name order SEQ ID NO: MMP-1 Forward 5'-ATGCTGAAACCCTGAAGGTG-3 ' SEQ ID NO: 1 Reverse 5'-CTGCTTGACCCTCAGAGACC-3 ' SEQ ID NO: 2 Type I collagen
(Type I collagen) Forward 5'-CTTCCTGCGCCTGATGTCCA-3 ' SEQ ID NO: 3 Reverse 5'-CTCGTGCAGCCATCGACAGT-3 ' SEQ ID NO: 4 β-actin Forward 5'-GGATTCCTATGTGGGCGACGA-3 ' SEQ ID NO: 5 Reverse 5'-CGCTCGGTGAGGATCTTCATG-3 ' SEQ ID NO: 6

Figure 5 shows the RNA expression of MMP-1 measured when the white mushroom extract 1 in human skin fibroblasts. As shown in Figure 5, the expression of MMP-1 significantly increased more than three times in the treatment of TNF-α compared to the sample untreated group, and when treated with white mushroom extract 1 (described in AC on Figure 5) It was confirmed that the expression of increased MMP-1 is reduced to less than 50%.

Example 6 Measurement of Collagen Expression

By the method of Example 5-1, the relative expression amount of type I collagen was measured using qRT-PCR. The treated concentration of TNF-α used and white mushroom extract 1 prepared in Example 1-2 was also the same as in Example 5-1. Primer sequences used to determine the expression of type I collagen are described in Table 4 above.

Figure 6 shows the RNA expression of type I collagen measured in the treatment of white mushroom extract 1 in human skin fibroblasts. As shown in FIG. 6, the expression of type I collagen was reduced by about 25% when TNF-α was treated compared to the sample untreated group, and when the white mushroom extract 1 (described as AC on FIG. 6) was treated. It was confirmed that the expression level of 1 collagen increased by about 1.5 times or more.

Example 7. Tyrosinase Inhibitory Activity

1.0 ml of 0.1 M potassium phosphate buffer (pH 6.8), 1.0 ml of 0.3 mg / ml L-tyrosine aqueous solution and 0.1 ml of 1250 unit / ml mushroom tyrosinase (sigma, USA) were mixed. The white mushroom extract 1 prepared in Example 1-2 was added at a concentration of 100 μg / ml, 200 μg / ml, or 400 μg / ml and reacted at 37 ° C. for 10 minutes. Thereafter, the absorbance of the reaction solution was measured at 480 nm (JASCO V-550, Japan), and tyrosinase inhibitory activity was calculated according to Equation 3 below and is shown in Table 5 and FIG. 7. As a positive control, kojic acid (nurukic acid) at a concentration of 50 µg / ml was used.

As shown in Table 5 and Figure 7 it was found that the white mushroom extract inhibits the activity of tyrosinase in a concentration-dependent manner. Addition of 200 μg / ml or 400 μg / ml of the mushroom extract showed higher tyrosinase inhibitory activity than koji acid, a positive control. White mushroom extract according to an embodiment of the present invention showed superior tyrosinase inhibitory activity than kojic acid when the concentration was increased even though it was not a purified single component.

extract Tyrosinase Inhibitory Activity (%) Koji-san (control) 95.59 ± 0.39 White Mushroom Extract 1 100㎍ / ㎖ 33.58 ± 1.55 White Mushroom Extract 1 200µg / ml 47.01 ± 1.97 White Mushroom Extract 1 400µg / ml 51.74 ± 1.14

Example 8. Preparation of Cosmetic Composition Comprising White Mushroom Extract

Example 8-1. Preparation of Nutritional Lotion

3.0 parts by weight of propylene glycol relative to 100 parts by weight of the total composition, 0.1 parts by weight of carboxypolymer, a preservative amount and a residual amount of purified water were mixed and stirred at 80 to 85 ° C., added to the manufacturing part, and then the emulsifier was activated. 1.0 part by weight, sorbitan sesquioleate 0.5 part by weight, liquid paraffin 10.0 part by weight, 1.0 part by weight of sorbitan stearate, 0.5 part by weight of lipophilic monostearate glycerine, 1.5 part by weight of stearic acid, glyceryl stearate / PEG-400 stearate 1.0 part by weight and 0.2 part by weight of triethanolamine were heated to 80 to 85 ° C. and emulsified. After emulsification, the mixture was thermally cooled to 50 ° C. while stirring using a stirrer, and a small amount of perfume was added thereto, followed by cooling to 45 ° C., followed by addition of a small amount of pigment, and the white mushroom extract prepared in Example 1-2 at 35 ° C. 1 to 5) 5.0 parts by weight of the mixture was mixed and then cooled to 25 ℃ and stored.

Example 8-2. Manufacture of nutritional cream

0.3 parts by weight of carboxypolymer, 5.0 parts by weight of butylene glycol, 3.0 parts by weight of glycerine and the remaining amount of purified water were added to the manufacturing part by stirring and mixing the remaining water, and then the emulsifier was activated. 2.0 parts by weight, cetyl alcohol 2.0 parts by weight, glyceryl monostearate 2.0 parts by weight, polyoxyethylene sorbitan monostearate 0.5 parts by weight, sorbitan sesquioleate 0.5 parts by weight, glyceryl monostearate / glyceryl stearate 1.0 parts by weight of polyoxyethylene stearate, 1.0 parts by weight of wax, 4.0 parts by weight of liquid paraffin, 4.0 parts by weight of squalane, and 4.0 parts by weight of caprylic / capric triglycerides after heating to 80 to 85 ° C. and then triethanolamine 0.5 Part by weight was emulsified. After emulsification, the mixture was cooled to 35 ° C. while stirring using a stirrer, and 5.0 parts by weight of the white mushroom extract (extracts 1 to 5) prepared in Example 1-2 was mixed and then cooled to 25 ° C. and stored.

Example 9. Skin Stability Assessment

In order to evaluate the skin stability of the white mushroom extract of the present invention, the cream prepared in Example 8-2 was applied to the arms of 49 healthy men and women (24 males, 25 females) to carry out a human patch test. It was.

Specifically, after wiping the arm of the test subjects with 70% ethanol, the cream containing the white mushroom extract 1 to 5 prepared in Example 8-2 was attached to the closed for 24 hours using an adhesive tape of 20ul each. After 24 hours, the adhesive tape was removed, and the sample remaining on the skin with a gauze was lightly removed, and then the skin condition was observed, and again after 24 and 48 hours.

As a result, in the cream containing the white mushroom extract 1 to 5 was not confirmed the skin irritation, the composition according to one embodiment of the present invention was confirmed that it is stable without any irritation to the human skin.

So far I looked at the center of the preferred embodiment for the present invention. Those skilled in the art will appreciate that the present invention can be implemented in a modified form without departing from the essential features of the present invention. Therefore, the disclosed embodiments should be considered in descriptive sense only and not for purposes of limitation. The scope of the present invention is shown in the claims rather than the foregoing description, and all differences within the scope will be construed as being included in the present invention.

<110> UNIJEJU Co., Ltd. <120> Cosmetic Composition comprising Extract of Agaricus blazei <130> PN170370-P1 <150> KR 10-2017-0174568 <151> 2017-12-18 <160> 6 <170> KoPatentIn 3.0 <210> 1 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> MMP-1_Forward <400> 1 atgctgaaac cctgaaggtg 20 <210> 2 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> MMP-1_Reverse <400> 2 ctgcttgacc ctcagagacc 20 <210> 3 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Type I collagen_Forward <400> 3 cttcctgcgc ctgatgtcca 20 <210> 4 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Type I collagen_Reverse <400> 4 ctcgtgcagc catcgacagt 20 <210> 5 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> beta-actin_Forward <400> 5 ggattcctat gtgggcgacg a 21 <210> 6 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> beta-actin_Reverse <400> 6 cgctcggtga ggatcttcat g 21

Claims (5)

Washing and drying the white mushrooms;
Put dried white mushrooms in the extraction tank, 8 to 10 times the weight of purified water to the total weight of the white mushrooms, and then extracted for 5 to 9 hours at 115 to 121 ℃ and 1.2 to 1.5 atm, from the extraction tank Recovering the first extract;
After further adding 8 to 10 times the weight of purified water relative to the total weight of the white mushrooms remaining in the extraction tank in which the first extract was recovered, extracted for 5 to 9 hours at 115 to 121 ℃ and 1.2 to 1.5 atm, extraction Recovering the second extract from the tank; And
A method of producing white mushroom extract comprising mixing the recovered first extract and the second extract and filtering with a filter.
The method of claim 1, wherein the filter has a pore size of 0.5 to 1.5㎛ poultry mushroom extract manufacturing method.
Cosmetic composition comprising a white mushroom extract prepared by the method of claim 1.
The method of claim 3, wherein
The cosmetic composition is a composition having one or more effects selected from the group consisting of wrinkle improvement, skin whitening, skin elasticity enhancement, skin irritation relief, and moisturizing effect.
The method of claim 3, wherein
The cosmetic composition is a flexible lotion, gel, water-soluble liquid, milk lotion, cream, essence, skin toner, oil-in-water emulsion, water-in-oil emulsion, cleansing foam, cleansing water, The composition is a formulation selected from the group consisting of a pack, body oil, body lotion, oil-in-water makeup base, water-in-oil makeup base, and foundation.

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