KR20040015469A - Cosmetic composition containing extract from phellinus linteus cultivated in rice - Google Patents

Abstract

PURPOSE: Provided is a dermatologic composition containing the extract of Phellinus linteus cultivated in rice, as a medium. The composition is useful for prevention of skin aging, skin whitening, and treatment of skin damage. CONSTITUTION: A dermatologic composition for prevention of skin aging, skin whitening, and treatment of skin damage comprises 0.0001-20% of the extract of Phellinus linteus cultivated in rice, based on the total weight of the composition. It is formulated into emulsion, nutritive cream, mask pack, massage cream and the like.

Description

Skin external preparation composition containing extract of Situation mushroom rice (Shangmi rice) {COSMETIC COMPOSITION CONTAINING EXTRACT FROM PHELLINUS LINTEUS CULTIVATED IN RICE}

The present invention relates to an external composition for skin containing the extract of Situation mushroom rice (condiments of rice), and in particular, the hot water extract of the situation mushroom rice as a component of the external preparation for skin due to its excellent anti-aging, whitening effect, skin damage relief and wound healing effect The present invention relates to an external preparation composition for skin containing an extract extracted with ethanol extract, 1,3-butylene glycol, propylene glycol and water or a mixed solution of these extracting solvents.

Conventional natural situation mushrooms are difficult to breed in nature and rarely grow native to mulberry trees, but the situation mushrooms or mycelium grown by artificial cultivation methods using various culture conditions are used, but the intake requirements and dietary methods are not easy. On the side. The situation mushroom rice is a situation mushroom composition in which the state mushroom mushroom spawn is completely covered with the entire surface of the grains by cultivating the progeny of the situation mushroom mycelium and then using rice, which is an edible grain, as a culture medium. It is an effective invention that is easy and has pharmacological various functions of situation mushrooms. It is registered as a Korean patent. (1002250500000, 1002049840000, 1002049820000, inventor: Sung Jae-mo)

Skin external preparations generally include cosmetics, medicines and quasi-drugs, and have been using raw materials extracted from various natural products as conventional external preparations, and are particularly derived from natural products having excellent efficacy in preventing skin aging, whitening, skin damage and wound healing. Extracted raw materials include ginseng, matsutake mushrooms, wild situation mushrooms, artificially cultivated situation mushrooms, brown rice and other rice-derived extracts, green beans, and upper limbs. These natural extracts have been widely used as raw materials for cosmetics and medicines.

An object of the present invention is to provide an extract of a situation mushroom rice (salt rice) excellent in the skin aging prevention, whitening effect, skin damage relief and wound healing effect and excellent safety and external skin composition containing the same. In addition, it is applicable to cosmetics and when applied to cosmetics to prepare a situation mushroom rice extract having the functions of antioxidant effect, whitening effect, wrinkle relief and skin damage recovery.

Sichuan mushroom rice extract of the present invention for the above purpose is 50-100% ethanol extract, 20-100% 1,3-butylene glycol extract, hydrothermal extract and the like of hydrothermal extract 2 A precipitate precipitated by addition of 4 to 4 times ethanol and these extracts can be used as a single extract or mixed. In addition, the extract may be applied as a skin external preparation composition, wherein the content of the situation mushroom rice extract is preferably 0.0001% to 20% by weight ratio.

In the present invention, the patent registration in Korea (1002250500000, 1002049840000, 1002049820000, inventor: Seong Jae-mo) hot water extract and 50-100% ethanol extract, 20-100 obtained from the situation mushroom rice produced in Daejeon material production plant of the licensee % 1,3-butylene glycol extract and the like can be used as a skin external preparation and skin aging prevention, whitening effect, skin damage alleviation and wound healing effect has been found to complete the present invention.

In order to obtain the extract of the situation mushroom rice of the present invention, 50 to 200 times the weight of the situation mushroom rice, water, ethanol, methanol, 1,3-butylene glycol, hexane, etc. by adding a polar, non-polar solvent, extraction by room temperature or warming Was used. In addition, in order to increase the extraction yield, ultrasonic waves were added 4 to 10 times for 30 minutes or extracted by adding sodium hydroxide, acetic acid, hydrochloric acid, and the like. Preferably, however, the hydrothermal extract, 50-100% ethanol extract, 20-100% 1,3-butylene glycol extract and the like are not limited to these extraction solvents. On the other hand, the present invention also includes a protein polysaccharide precipitated by adding ethanol 2 to 4 times the volume of the extract to the hot water extract.

In the present invention, as described in detail below, the antioxidant effect on reactive oxygen species (ROS) and reactive nitrogen species (RNS), the antioxidant effect of keratinocytes (melanocytes) Inhibition of pigment production, measurement of absorbance in the UV region, and collagen production in fibroblasts were tested by various test methods to determine whether the situational mushroom rice extract actually prevented skin aging, whitening, skin damage and wound healing. Was identified.

In order to examine the antioxidant effects of physiological mushrooms, physiological mushroom rice is extracted with various polar and non-polar solvents, and the extracts are reactive oxygen species (ROS) and harmful nitrogen species that cause skin aging. By measuring the antioxidant effects on reactive nitrogen species (RNS), we tried to provide a more convincing scientific basis for the application of physiological mushroom rice as a bioactive substance. In particular, research on harmful nitrogen is significant in that it seeks effective blocking of pollutants by automobile exhaust gas in addition to their pathophysiological meaning.

Xanthine, DTPA (diethylenetriaminepentaacetic acid), cytochrome-seed and catalase were absorbed after adding xanthine, DTPA (diethylenetriaminepentaacetic acid) in phosphate buffer to find out the antioxidant effect of superoxide radical among active oxygen species. After taking the spectrum and put xanthine oxidase (xanthine oxidase) was absorbed for about 4-5 minutes at 550nm. The increase in absorbance at 550 nm indicates the production of superoxide radicals, and the change in absorbance at 550 nm was measured over time before the xanthine oxidase was added. If the extract is capable of scavenging superoxide radicals, the increase in absorbance at 550 nm decreases, which is calculated as a relative activity for superoxide dismutase (SOD), indicating that there is 47% superoxide radical scavenging activity compared to SOD. Came out.

Nitrogen monoxide, nitrogen dioxide, and peroxynitrite are the ones that are most concerned with the health of humans among the active nitrogens that have recently been of much interest. Nitrogen monoxide and nitrogen dioxide are problematic substances in urban areas with high air pollution, and peroxynitrite plays an important role in inflammatory reactions. This study investigated how well the situation mushroom rice extract removes the active nitrogen related to skin aging and wound healing and examined whether it is useful for diseases such as skin aging and human aging and skin inflammation.

Nitric oxide (NO) reacts very quickly with superoxide radicals produced in vivo to produce peroxynitrite, which is also the oxidation of glutathione, a bioreducing agent, and the nitrification of tyrosine. Cause back. Peroxynitrite is prepared by reacting isoamylnitrite with hydrogen peroxide and stored at high pH. When you want to generate peroxynitrite in situ, add SIN-1 (3-morpholinosydnonimine). do. In this test, the degree of phenol phosphorylation by peroxynitrite was measured by absorbance, and the inhibitory effect of the situation mushroom rice extract was found to be about 53%.

From the results of the above test, the antioxidant and active nitrogen effects of the situation mushroom rice extracts were examined, and as a result, it was confirmed that the extract has an effective effect on skin aging or inflammation related to aging or disease, and thus the extract prevents skin aging and wounds. It was found that it can be utilized as an external composition for skin to be applied to healing and the like, and the effect is excellent.

Under oxidative stress in cells, the expression of NF-kB is increased to cause expression of genes involved in aging, etc., using an analytical method that quantitatively measures the expression level of NF-kB in human skin keratinocytes. Among mushroom rice extracts, candidate substances that can induce anti-oxidant effects in the skin were treated to determine whether the test substance acts on the skin cells. In particular, the treatment of UV light significantly increased the expression level of NF-kB in this system. When the test material was treated, it was confirmed whether the test material actually inhibited the expression of NF-kB in the cells. This analytical system for measuring the expression of NF-κB is very sensitive to the quantitative measurement of NF-κB expression in terms of time and concentration of sample material by measuring by fluorescence measurement using a fluorescent substrate, and also a simple measurement method. Therefore, it can be effectively used to test whitening, anti-inflammatory or antioxidants from various natural products and organic compounds.

The activity of the reporter enzyme was measured to confirm whether the human vector HaCaT cells transfected with the expression vector expressed the transcriptional reporter enzyme (SEAP) due to NF-κB activity and secreted out of the cell. First, transfected cells (1 × 10 6 cells) were incubated for 20 hours by diluting Dulbeccos modification of eagles medium (DMEM) medium containing geneticin (500 μg / ml) in 6-well plates. The cells were washed twice with phosphate-buffered saline, fresh medium was added, and each sample (20 ㎍ / ml) was added and incubated for 2 hours. To remove endogenous alkaline phosphatase activity, the culture was heated at 65 ° C. for 5 minutes and used immediately for analysis or stored at −20 ° C. 20 μl medium was taken at 0 hours and after 2 hours, the cells were irradiated with ultraviolet (40 mj / cm 2). After 6 hours of incubation, 20 μl of medium was taken, cut off at 65 ° C. for 6 minutes, and then frozen. After the frozen medium was dissolved at room temperature, it was placed in a 96-well plate, and the same amount of SEAP buffer (20 µl) was added and shaken. After 13 minutes at 37 ° C, 2 times of SEAP buffer was added and shaken. The plate was placed in the dark and 10 mM fluorescent substrate 4-methylumbelliferyl phosphate (MUP) (10 μl) was added. The plate is wrapped in foil and left at 37 ° C. for 1 hour. This plate was measured for fluorescence in a fluorometer. Fluorescence from the digest of SEAP / MUP is excited at a wavelength of 360 nm and the amount of light emitted at 449 nm was measured using a 96-well plate fluorometer (Molecular Devices, F max). . As a result, NF-κB activity was increased after UV irradiation in the transfected HaCaT cell line, and the change in NF-κB activity (100%) before and after UV irradiation (5%) and kojic acid ( 8%) was compared with the results after treatment. On the basis of 20μg / ml, NF-κB activity was found to be valuable in terms of future product development compared to kojic acid. In particular, the situation mushroom rice extract was confirmed to affect the regulation mechanism of NF-κB was found to be effective in whitening substances, antioxidants, and anti-inflammatory drugs.

The tyrosinase activity inhibition test, the most important enzyme in the melanin synthesis reaction that determines human skin color, can provide results related to the whitening effect at the enzyme level. Human enzymes are best used as tyrosinase inhibitors, but human ones are not currently sold commercially and are difficult to extract. However, since most of them have similar effects in vivo, most of them are derived from mushrooms and were measured using enzymes of mushrooms in this test. 0.3% was used as the concentration of the mushroom extract, and the enzyme was measured as 2 units and the substrate was measured as 2mM of tyrosine. As a result, the situation mushroom rice extract also directly inhibited the enzyme activity, reaffirming its effect as a whitening agent.

Dermal pigment cells always produce a certain amount of melanin pigment, but melanin pigment production can be excessively increased due to external influences such as ultraviolet rays, which can lead to cosmetically unwanted results such as blemishes, freckles and spots. have. Therefore, the absorbance spectrum of the ultraviolet wavelength of the situation mushroom rice extract was shown in Figure 1. The results in Figure 1 shows that the absorbance value in the ultraviolet region of the situation mushroom extract is particularly excellent in the ultraviolet A region compared to koji acid. This is the result that the extract itself has an excellent ultraviolet absorbing ability is effective for the spots, freckles, spots caused by ultraviolet light.

Representative changes in the skin caused by aging and UV irradiation include degeneration of keratinocytes and fibroblasts, which are epidermal cells, weakening of the dermal-epidermal border, hyposynthesis of dermal collagen, and accumulation of denatured elastin proteins. It can be used to evaluate the efficacy effect. A test substance was added to the cells in culture to further incubate, and then the number of proliferated skin cells was counted to evaluate the efficacy of the effective substance. As this experiment is an evaluation of cell proliferation itself, it can be evaluated as an actual anti-aging substance. The cell lines used in the test were HaCAT cells and fibroblasts, which are keratinocyte cell lines. 10000 cells were attached to each well of 96-well, and the sample was dissolved in DMSO (dimethylsulfuroxide) so that the extract was 0.01% and 0.001%. As a result of the measurement, it was found that the proliferative effect on the fibroblasts of the situation mushroom rice extract was effective as a raw material for antiwrinkle and anti-aging cosmetics related to cell proliferation.

As a result of the situation mushroom rice extract obtained by the extraction method and the efficacy test for these extracts, the present invention provides a skin external preparation composition having a skin aging prevention, whitening effect, skin damage alleviation and wound healing effect, and preferably It may be contained in an amount of 0.0001% to 20% by weight of the total weight.

Hereinafter, the present invention will be described in more detail with reference to specific formulation examples of the skin cosmetic composition in the topical mushroom composition containing the situation mushroom rice extract of the present invention, but the present invention is not limited only to these formulation examples.

Formulation Example 1 Softening Lotion (Skin Lotion) Containing Situation Mushroom Rice Extract

0.1 2.5 2.0 2.0 0.2 0.3 10 0.3 to 100

Formulation Example 2 Nutrients (Milk Lotion) Containing Situation Mushroom Rice Extract

0.1 5.5 4.0 2.0 1.5 4.0 3.0 3.0 4.0 to 100

Formulation Example 3 Nutritional Cream Containing Situation Mushroom Rice Extract

0.2 2.5 2.5 1.5 1.5 3.0 2.5 1.5 0.8 to 100

As described above, by containing the extract of the situation mushroom rice (salt rice) in the external composition for skin such as cosmetics, the external composition for skin having an anti-aging, whitening effect, alleviation of skin damage and wound healing can be provided. .

Claims (3)

Skin external preparation composition having skin aging prevention, whitening effect, skin damage alleviation and wound healing effect, containing extract obtained from Situation mushroom rice [Claim 2] The cosmetic composition of claim 1, wherein the extract obtained from the situation mushroom rice is contained in an amount of 0.0001% to 20% based on the total weight of the composition, to prevent skin aging, whitening, skin damage, and wound healing. According to claim 1, wherein the cosmetic composition is a cosmetic composition having an anti-aging, whitening effect, skin damage relief and wound healing effect, characterized in that the formulation of the flexible cosmetics, nourishing cosmetics, nutrition cream, massage cream or pack