KR20080090160A - Extract of rhodotus palmatus cultured broth and cosmetic composition comprising the same for providing anti-wrinkle activity - Google Patents

Abstract

A cosmetic composition comprising the extract of Rhodotus palmatus cultured broth medium is provided to promote growth of fibroblasts and increase the amount of collagen produced from the fibroblasts, so that the composition is useful to improve skin wrinkles. A cosmetic composition for improving skin wrinkles comprises 0.01-10 wt.% of the extract of Rhodotus palmatus cultured broth medium which is prepared by homogenizing the Rhodotus palmatus cultured broth medium, adding 40% ethanol into the homogenized Rhodotus palmatus cultured broth medium, agitating the mixture at room temperature to 40 deg. C for 24 hours, and filtering the extracted mixture, and is formulated as soft and nutrition lotion, nutrition cream, massage cream, essence or pack.

Description

Extract of Rhodotus palmatus Cultured Broth and Cosmetic Composition Comprising the Same for Providing Anti-wrinkle Activity}

The present invention relates to an extract of the creatinine creased red mushroom liquid culture and wrinkles cosmetic composition containing the same. More specifically, by using mushrooms capable of liquid culture among the mushrooms growing in Korea, it is possible to provide a substance that can improve skin wrinkles and to improve skin wrinkles by activating the function of fibroblasts, thereby suppressing and improving wrinkle occurrence. The present invention relates to an excellent liquid extract of the nettle wrinkled red mushroom and anti-wrinkle cosmetic composition containing the same.

Cosmetic products, historically known as products representing women in general, are not only simple expression tools of beauty, but also daily necessities, and are located close to life regardless of age or gender. Interest in 'functional' products that expect special effects or efficacy is also increasing.

"Cosmeceutical", which is a compound word of cosmetic and pharmaceutical, means functional makeup according to Korean law (Article 2, Paragraph 2 of the Cosmetic Act), a product that helps to whiten the skin, and a product that helps to wrinkle the skin. It is a product that helps to burn the skin finely or protect the skin from UV rays. In general, such functional cosmetics are interpreted as cosmetics that affect the structure or function of the body as well as cosmetic effects and prevent and treat, and in particular, as the proportion of functional cosmetics on the market increases, There is an active research to seek and use it effectively on the skin.

Natural materials used in functional cosmetics are the most widely used herbs (herb) including medicinal plants, mushrooms that are used a lot of medicinal plants are spotlighted as a good natural materials. For example, Korean Patent Application No. 1999-4497 discloses a moisturizing cosmetic composition containing beta-1,6-branched beta-1,3-glucan, which is a polysaccharide of Schizophyllum commune , and is disclosed in Grifola frondosa . Polysaccharides also have the effect of moisturizing, irritating, and inhibiting photoaging and whitening (Lee et al., Anti-irritation and moisturizing effects of exoploysaccharide produced by Grifola frondosa.J. Of the Society of Cosmetic Soientists of Korea 31: 35- 41, 2005). In addition, Korean Patent Application No. 2003-19746 discloses a natural preservative-free cosmetic composition containing Dictyophora echinovolvata (mantle mushroom) extract, and Korean Patent Application No. 2001-32706 discloses that Agaricus blazei (Agaricus mushroom) extract is applied to cosmetics. A composition of cosmetics having an anti-edema effect and an inhibitory effect on edema formation is disclosed.

In addition, the skin aging effect, especially wrinkle improvement, Korean Patent Application No. 2003-73323 discloses an anti-wrinkle cosmetic composition containing Inonotus obliquus (chaga) extract, Korean Patent Application No. 2001-10171 Phellinus linteus (situation mushroom) ), Anti-cancer, other skin whitening effect, such as skin wrinkle improvement, blemishes, freckles improvement, anti acne, antioxidant effect is disclosed. In addition, Korean Patent Application No. 2003-24042 discloses the use of Hericium erinaceus fruiting body extract to prevent skin aging and whitening, to reduce the damage of skin, and to use Cordyceps sp., Cordyceps sp., Coriolus hirsutus . (Lee et al., Screening of traditional herbal medicines to develop new materials for anti-aging and anti-wrinkle in the skin, J Soc. Cosmet. Scientists Korea 31: 147-) 152, 2005).

However, mushrooms used to prevent wrinkles and skin aging among the cosmetics developed using mushrooms have only been used as a mixture extract of chaga fruiting body and mycelium culture only in Korean Patent Application No. 2003-73323. In the case of fruiting bodies, the ingredients change depending on the culture conditions and habitats. Therefore, in order to industrialize, materials that can suppress wrinkles through liquid mycelium culture that can control the culture conditions, the yield, and the potency can be controlled. There is a need for development. In addition to the commonly known mushrooms, there is a need to develop industrializable materials by utilizing various mushrooms native to Korea.

The present invention has been made in view of the above-mentioned problem and made by the request as described above, an object of the present invention to provide a substance that can improve the skin wrinkles using mushrooms that can be cultured in the liquid mushrooms native to Korea It is.

In addition, another object of the present invention is to provide a wrinkle improving cosmetic composition that can improve the wrinkles of the skin through the activation of the function of the fibroblasts to suppress and improve the occurrence of wrinkles.

To achieve the above object, the extract of the cultivation of the creased red mushrooms according to the first aspect of the present invention is characterized in that it is effective for improving wrinkles as an anti-wrinkle material.

Preferably, the liquid extract of the nettle wrinkled red mushroom is homogenized the entire culture solution containing the nettle wrinkled pink mushroom mycelium, soaked in ethanol to 20 to 40% by weight, and stirred at room temperature to 40 ℃ for at least 24 hours After extraction, it is characterized in that the ethanol extract obtained by filtration.

More preferably, the ethanol extract is characterized in that the dry powder.

In addition, the wrinkle improvement cosmetic composition according to the second aspect of the present invention for achieving the above object is characterized in that it contains the extract of the creatinine creatine mushroom culture described above.

Preferably, the extract of the creased red fuchsia mushroom liquid culture is characterized in that it comprises 0.01 to 10% by weight based on the total weight of the cosmetic composition.

More preferably, the cosmetic composition is characterized in that it has a formulation selected from the group consisting of supple and nourishing longevity, nourishing cream, massage cream, essence and pack.

Hereinafter, the present invention will be described in detail with reference to embodiments of the present invention. These examples are only for illustrating the present invention in more detail, it will be apparent to those skilled in the art that the scope of the present invention is not limited by these examples in accordance with the gist of the present invention. .

As used in the present invention, the "Wrinkles of Red Flemish Mushroom" is a genus of Rhodotus spp., Which occurs in the alveolar tree in spring and autumn, and particularly in the elm tree, and is distributed in the northern temperate hemisphere. The whole area is pink to pink, and the surface of the lampshade is often smooth or meshed with wrinkles, the wrinkles are slightly coarse and pink to pink, the stem is short, and the spores are 6 to 8㎛ and are approximately spherical.

In the composition according to the present invention, "Molecular wrinkle pink mushroom liquid culture extract" is homogenized the whole culture medium containing the strain of the strain of red mesh mushroom after aeration using a liquid medium, and then 5 times (by weight of mycelia) 40% Ethanol was added and extracted at room temperature to 40 ° C. for at least 24 hours, followed by filtration and ethanol extract.

In addition, in the composition according to the present invention, the ethanol extract can be dried and used as a powder. However, the present invention is not limited to the nettle wrinkled red mushroom liquid culture extract of the present invention includes those obtained by all known processing means used for processing into a usable form.

There has been no report on the physiological activity of foods, medicines and cosmetics of the nettle wrinkled mushrooms contained in the present invention. Therefore, the present inventors confirmed that it is effective in the improvement of skin wrinkles by using a liquid culture extract of the nettle wrinkled red mushroom as follows.

 Confirmation of the improvement of the skin wrinkles of the nettle wrinkled red mushroom liquid culture extract of the present invention comprises the step of treating the extract to human fibroblasts.

The present inventors used NHF (Normal Human Fibroblast), a kind of fibroblasts, to search for a material that can effectively suppress the death of fibroblasts and a decrease in collagen production function, which is a major cause of skin wrinkles. It was used for the experiment.

Fibroblasts were cultured after dispensing in 96 well plates at a concentration of 1 x 10 ⁴ cells / well to measure the fibroblast proliferation effect of the liquid culture extract of the oleander wrinkled red mushroom. This was treated with a liquid concentration extract of the nettle wrinkled red mushrooms and cultured in 37 ℃, 5% CO ₂ incubator for 24 hours. After incubation, the MTT reagent was treated, the color was induced, and the absorbance was measured at 540 nm using an ELISA microplate reader (ELx800, Bio-tec, USA). As a result, it was confirmed that the growth of fibroblasts of about 109-134% when treated with 0.001-0.1% by weight of the liquid culture extract of the nettle wrinkled red mushroom.

In addition, the present inventors divided the fibroblasts by 1 x 10 ⁴ cells / well in 96well plate and incubated for 24 hours in DMEM medium to see the collagen synthesis effect to assay the wrinkle improvement effect of the skin, stepwise dilution in the medium One strain of Wrinkle Flemish Mushroom was added and incubated in a CO ₂ incubator for 24 hours. After completion of the culture, the collagen was measured using Procollagen Type I C-epetide EIA (Takara Bio Inc., MK101) to produce collagen more than 120 ng / ml when treated with 0.1% by weight. As a result, it could be seen that the treatment of the liquid culture extract of the mesh wrinkled red mushrooms increased the collagen production amount of fibroblasts.

Although an Example and a comparative example are given to the following and it demonstrates in detail, the technical scope of this invention is not limited to these Examples.

EXAMPLE

Example 1: Preparation of the extract of the creatinine creatine mushroom

Wrinkles of Red Finch on Potato dextrose agar (PDA, Difco Co.) The mycelium is cut out uniformly with a cork border of 7 mm in diameter, and homogenized 9 to 10 fragments of this mycelium, and then inoculated into 100 ml potato dextrose broth (PDB, Difco Co.) at a speed of 120 rpm at 25 ° C. Primary seed cultures were carried out for 15 days. 20% of the homogenized seed was inoculated again into the PDB (pH 5.0) 500 ml baffle flask, and the second seed was incubated at 25 ° C. and 120 rpm for 5 to 25 days.

For the preparation of the liquid culture extract of the oleander wrinkled red mushroom, after homogenizing all of the culture medium containing mycelia after aeration, 5 times 40% ethanol was added and extracted over 24 hours at room temperature to 40 ° C. After filtration, the filtrate was concentrated under reduced pressure or lyophilized to a minimum solid content of 2% or more at 50 ° C. to obtain a extract of “Ferrisian creased light pink mushroom liquid culture.

Example 2 Effects of Extracts from Cultured Wrinkle Fuchsia Mushrooms on Human Fibroblast Proliferation

In the process of differentiating human fibroblasts, the effect of the extracts from the liquid culture extract of Fructus oleracea on each concentration was measured and the effects on the differentiation of fibroblasts were measured. In order to investigate the effect of the extract of the cultivation of Fructus oleracea on the growth of human fibroblasts, cell proliferation was measured by MTT assay. Human fibroblasts, NHF, were cultured in DMEM containing 10% FBS, 1% penicillin, and streptomycin. When the cells were grown to 80 ~ 90% in the culture dish, 700 μl of trypsin mixture was added and then incubated at 37 ° C. for 3 minutes to separate the cells, and then used in the experiment while passaged at a ratio of 1: 3. MTT assay, which was used to measure the growth of human fibroblasts, was carried out by the extracts of the creatinine creased red mushrooms. The mitochondria dehydrogenase in living cells reacted with MTT in dark blue MTT- formazan crystal. To form a was based on the principle. That is, NHF was inoculated into a 96 well plate at a concentration of 1 x 10 ⁴ cell / well, and then incubated in a 37 ° C., 5% CO ₂ incubator for 24 hours so that the cells could be attached to the plate bottom. After removing the culture medium used for the culture, dilution of the creatinine creatine mushroom liquid culture extract in a new medium by concentration, and then divided into 200ul, and incubated for 24 hours in 37 ℃, 5% CO ₂ incubator. After incubation, the medium containing the test substance was removed, and 200 μl of MTT reagent diluted in the medium at 0.5 mg / ml was dispensed per well, and the light of the 96 well plate was blocked for 4 hours at 37 ° C. and 5% CO ₂ incubator. Incubated for When the incubation is completed, the medium containing the MTT reagent is removed, 100 μl of DMSO (dimethyl sulfoxide) is added to dissolve the MTT-formazan crystal, and 20 μl of Sorenson's buffer (0.1 M NaCl, 0.1 M glycine, pH 10.5) is treated at room temperature. Color development was induced for 5 minutes and then absorbance was measured at 540 nm using an ELISA microplate reader (ELx800, Bio-tec, USA). The viability of cells through the MTT experiment was calculated according to the following Equation 1, and the results of the treatment of the extract of the cultivation of the creatinine hydrangea liquid culture on the growth of human fibroblasts are shown in Table 1 below.

[Equation 1]

Where A is the sample absorbance and B is the control absorbance.

TABLE 1

Treatment concentration 0.01 mg / mL 0.1 mg / mL 1.0mg / mL Cell growth rate (%)   109.06 ± 18.83 116.82 ± 17.95 133.93 ± 13.69

Result: Mean ± Standard Deviation (Repeat = 10)

As can be seen in Table 1, as a result of confirming the effect on the growth of fibroblasts cultured extract of the creased red filamentous mushroom mushrooms in the concentration of 0.01mg / mL and 0.1mg / mL, 1.0mg / mL 109, The cell proliferation rate of 117, 134% was shown to promote skin wrinkles by promoting fibroblast proliferation.

Example 3 Effects of Extracts from Cultured Wrinkle Fuchsia Mushrooms on Collagen Production

In order to examine the effect of wrinkle improvement on the skin, the effect of the extract of the liquid culture extract of Fructus oleracea on the collagen production of fibroblasts was measured. The mesh wrinkled mushrooms culture liquid extract used here is prepared in Example 1. To determine the effect, collagen production of human fibroblasts was measured.

First, in order to examine the collagen synthesis effect to test the skin's wrinkle improvement effect, human fibroblasts were dispensed in 96well-plate at 1 x 10 ⁴ cell / well and DMEM medium containing 10% FBS and 1% Penicillin-streptomycin 24 After incubation for a time, stepwise dilution of the creatinine liquid culture extract was added to the medium and incubated in a CO ₂ incubator for 24 hours. After completion of the culture, the culture was measured for collagen using Procollagen Type I C-epetide EIA (Takara Bio Inc., MK101). Standard concentration curves included in the collagen measurement kit were prepared, and after adjusting the amount of total protein contained in each medium, the amount of collagen increase was calculated in Table 2 with the control group as 0.

TABLE 2

Treatment concentration 0.01 mg / mL 0.1 mg / mL 1.0 mg / mL collagen increase (ng / ml) 14.3 ± 1.1 52.7 ± 3.6 127.1 ± 10.3

Result: Mean ± Standard Deviation (Repeat = 10)

As can be seen from the effect of the treatment of the extract of the mantle wrinkled red mushroom liquid culture, which is the result of Table 2, the result of confirming the effect of the mantle wrinkled red mushroom liquid culture extract on the production of fibroblast collagen 0.01 At the mg / mL, 0.1 mg / mL and 1.0 mg / mL concentrations, collagen of 14, 53, and 127 ng / ml was produced more than the control group, respectively, indicating that skin wrinkles could be improved.

Example 4 Measurement of Enhancement Effect on Skin Elasticity

In order to measure the effect of medicinal plant extracts on skin elasticity, nutrition creams were prepared to measure skin elasticity improvement effect. To this end, the nutrition cream of Preparation Examples 1 to 3 and Comparative Preparation Example 1 were prepared by mixing in a conventional manner using the ingredients and contents shown in Table 3 below. Preparation Examples 1 to 3 were added to the nettle wrinkled red mushroom extract to 0.01, 1.0 and 10% by weight, respectively, relative to the total weight of the cosmetic composition, Comparative Preparation Example 1 is a cosmetic composition is not added to the nettle wrinkled red mushroom extract .

TABLE 3

Raw material name Preparation Example 1 Preparation Example 2 Preparation Example 3 Comparative Production Example 1 Polysorbate-60 1.2 1.2 1.2 1.2 Sorbitan stearate 0.4 0.4 0.4 0.4 Stearic acid 0.5 0.5 0.5 0.5 Squalane 6.0 6.0 6.0 6.0 Cetostearyl alcohol 1.5 1.5 1.5 1.5 Cetyloctanoate 5.0 5.0 5.0 5.0 Carbomer 0.12 0.12 0.12 0.12 Triethanolamine 0.12 0.12 0.12 0.12 glycerin 3.0 3.0 3.0 3.0 Butylene Glycol 2.0 2.0 2.0 2.0 Wrinkle Fence Red Mushroom Extract 0.01 1.0 10 0 Spices A reasonable amount A reasonable amount A reasonable amount A reasonable amount antiseptic A reasonable amount A reasonable amount A reasonable amount A reasonable amount Purified water Remaining amount Remaining amount Remaining amount Remaining amount Total (% by weight) 100 100 100 100

In order to measure the effect of the medicinal plant complex extract on the skin elasticity of the cosmetic composition prepared in Preparation Examples 1 to 3 and Comparative Preparation Example 1, skin elasticity improvement effect measurement experiment was performed.

For this purpose, 20 healthy women over 20 years old were divided into 4 groups under the conditions of 22-24 ℃ and 55% relative humidity. In the cosmetic composition of Comparative Preparation Example 1 was measured the skin elasticity enhancing effect. After using the cosmetic composition for 12 weeks (2 times / day) around the eyes, skin elasticity was measured using a skin elasticity measuring instrument (Cutometer SEM575, Courage + Khazaka).

Skin elasticity measurement results (12 weeks) was calculated as ΔR5 value (Equation 2) as the average value for each test group of the skin elasticity measuring instrument, it is shown in Table 4 below. ΔR5 value represents a net elasticity coefficient, and closer to 1, the better the elasticity.

[Equation 2]

TABLE 4

Experimental group Skin elasticity enhancing effect (△ R5) A group 0.17 B group 0.19 C group 0.21 D group 0.07

As can be seen in Table 4, the skin elasticity in Group A (Preparation Example 1), Group B (Preparation Example 2), Group C (Preparation Example 3) containing a nettle wrinkled red mushroom extract, Group D ( It was confirmed that the increase of more than 142.2 ~ 200% compared to the skin elasticity in Comparative Preparation Example 1). Therefore, the wrinkle improvement cosmetic composition containing the nettle wrinkled red mushroom liquid culture extract according to the present invention can effectively promote skin elasticity.

In the present specification, only a few examples of various manufacturing and analysis experiments performed by the present inventors are described, but the technical idea of the present invention is not limited thereto, and may be variously modified and implemented by those skilled in the art. .

As described above, the extract of the creased red filamentous mushroom liquid culture according to the present invention promotes the proliferation of fibroblasts in connection with the improvement of skin wrinkles, and increases the amount of collagen produced by the fibroblasts to prevent wrinkles. It works.

Therefore, the wrinkle improvement cosmetic composition according to the present invention, which contains the extract of the creatinine creased red mushroom liquid as an active ingredient, can improve skin wrinkles through activation of fibroblast function, thereby having an excellent effect on suppressing and improving wrinkle occurrence.

Claims (6)

Wrinkle improvement material, Wrinkle liquid extract extract, characterized in that it is effective for wrinkle improvement. The method of claim 1, The liquid extract of the mesh wrinkled pink mushroom liquid is homogenized the entire culture solution containing the mycelium wrinkled pink mushroom mycelium, and then immersed and stirred by adding 5 times 40% ethanol (of mycelium weight) for 24 hours at room temperature to 40 ℃. After extracting the ethanol extract obtained by filtration after extracting, the extract of the creased white fuchsia mushroom liquid culture. The method of claim 2, The ethanol extract is characterized in that the dry powder, creased white fuchsia liquid culture extract. Wrinkle improvement cosmetic composition containing the liquid extract of the nettle wrinkled red mushroom of any one of Claims 1-3. The method of claim 4, wherein The creased creatine mushroom liquid culture extract is characterized in that it comprises 0.01 to 10% by weight relative to the total weight of the cosmetic composition, wrinkle improvement cosmetic composition. The method of claim 4, wherein The cosmetic composition is characterized in that it has a formulation selected from the group consisting of supple and nutritious longevity, nourishing cream, massage cream, essence and pack, wrinkle improvement cosmetic composition.