KR20060101095A - Red grape extract with skin wrinkle improvement effect - Google Patents

Abstract

The present invention relates to a skin wrinkle improver using an extract obtained by high temperature treatment of red grapes. Since the red grape extract of the present invention promotes the production of collagen precursors and inhibits the activity of fibroblasts, the red grape extract is effective in preventing and improving skin wrinkles.

Red Grape, Collagen, Skin Wrinkle Improvement

Description

RED GRAPE EXTRACT WITH SKIN WRINKLE IMPROVEMENT EFFECT}

The present invention relates to a red grape extract having an effect of improving skin wrinkles and a skin wrinkle improving agent using the same, and more particularly, a cosmetic and a pharmaceutical for improving skin wrinkles, and a red food extract that can be used as one component of a functional food for improving skin wrinkles. Relates to grape extract.

In recent years, the increase in the elderly population due to the extension of life expectancy is increasing the share of aging research in medicine and biology. All living things age with age. Aging is the gradual deterioration of an organism's ability to adapt to environmental changes over time. The same is true for skin. Wrinkles on the skin are aging-related phenomena that indicate various structural changes due to deterioration of biological functions. That is, it is classified into intrinsic aging that occurs with age and photoaging that occurs due to damage caused by external factors, particularly ultraviolet rays, in addition to endogenous aging. There may be many external factors, but the effect of ultraviolet rays in the sun's rays is greatest. In endogenous aging, characteristic clinical skin findings include fine wrinkles, atrophy of the dermis, and a decrease in the subcutaneous fat layer, and in photoaging, rough and deep wrinkles appear, abnormal elastic fibrous layer accumulates, and the skin becomes leathery and loose. Lose.

It is certain that ultraviolet rays cause wrinkles, but the mechanism of action is not yet clear. Ultraviolet rays cause modifications of nucleic acids and proteins and oxidation of lipids, altering chromosomes of cells, damaging cell membranes, and causing various gene expression levels. These various changes can also be caused by UV light directly altering biomolecules or mediating reactive oxygen species. Ultraviolet irradiation causes various clinical changes, including inflammatory reactions such as erythema and edema, skin blackening, and degeneration of dermal intercellular substances. Many studies are underway to determine which of these many reactions is deeply correlated with the development of wrinkles. .

As mentioned above, in photoaging, changes in the dermis are usually noticeable. Therefore, wrinkle generation is also considered to be due to dermal changes. In particular, significant changes are the excessive accumulation of amorphous elastic tissue in the outer dermis and the reduction of collagen fibers in the dermis. Many researchers have tried to curb wrinkles and reduce wrinkles, but they lack understanding of how wrinkles develop. Wrinkle remodeling studies have reported results related to the occurrence of wrinkles, such as decreased synthesis or degradation of collagen in the dermis, damage to the epidermal basement membrane, and decreased epidermal metabolic activity. The development of wrinkles is accepted as a comprehensive effect of the various biochemical and clinical changes that produce ultraviolet radiation.

Conventionally, collagen was sometimes formulated into cosmetic products, but when collagen is applied to the skin surface as a cosmetic product, there is a problem that the percutaneous absorption of collagen, which is a polymer, is difficult, and its function cannot be sufficiently expected. In addition, there was a method of injecting collagen directly into the dermis of the skin, but this method also did not provide a solution to improve skin wrinkles by side effects.

Recently, retinoic acid, a protein derived from an animal placenta (Japanese Patent Laid-Open No. 8-231370), and the like have been known as substances which promote collagen synthesis. However, retinoic acid is unstable, so the formulation technology has been limited and irritated to the skin. Therefore, there are limitations in terms of safety, and the animal placenta-derived protein has a fatal shortcoming that can be used to extract cows with mad cow disease.

In addition, alpha-hydroxy acid (AHA) and various vitamin A derivatives (retinoids), which have been confirmed to be effective in the human body, have been developed and used in cosmetics. To date, clinically evident efficacy has been demonstrated with these substances and sunscreens. In Europe, cosmetics have been reported and advertised for wrinkles since around 1990. By 1993, the term cosmetics and functional cosmetics was introduced to improve skin condition such as ceramide, AHA and retinol.

In the cosmetics market as well as in the dietary supplements market, the importance of functional foods related to beauty of the skin is increasing more and more, and the consumer's interest in improving skin wrinkles is becoming more and more fragmented. Almost all conventional cosmetic companies have developed cosmetics as a product for improving skin wrinkles, but this is limited to cosmetics, and it was not possible to obtain the effect of skin wrinkle improvement through ingestion. In addition, considering that “Eating Cosmetics” is faster than “Coating Cosmetics,” research and development on “Eating Cosmetics” is urgent.

And, it is not known yet regarding the skin wrinkle improvement effect of the red grape extract.

The present inventors have continually researched to find a new natural material for skin wrinkle improvement as described above. When the extract obtained by treating red grapes at a high temperature of 85 ° C. or higher is used for skin wrinkle improvement, It has been found that there is an effect of inhibiting the degradation of collagen. The present invention provides a cosmetic composition for improving skin wrinkles containing red grape extract as an active ingredient.

The present invention is to extract the red grapes through a high temperature treatment of 85 ℃ or more, to provide a red grape extract for skin wrinkle improvement to increase the synthesis of collagen and inhibit the decomposition of collagen.

The present invention also provides a cosmetic composition for improving skin wrinkles containing red grape extract as an active ingredient.

In addition, the present invention, the cosmetic composition for improving skin wrinkles is at least one selected from the group consisting of retinol, retinol palmitate, polyethoxylated retinamide, adenosine, kainetin, cocoon extract, isoflavones and mixtures thereof It provides a cosmetic composition for improving skin wrinkles, further comprising the component.

The present invention also provides a health functional food for improving skin wrinkles comprising red grape extract and food acceptable food additives.

The present invention also includes a red grape extract and a pharmaceutically acceptable carrier, and provides a red grape extract for skin wrinkle improvement that increases the synthesis of collagen and inhibits the degradation of collagen.

Red grape extract of the present invention can be obtained by extracting the active ingredient of grapes for 8 hours at a high temperature of 85 ^° C or more red grapes.

One embodiment of the present invention uses a red grape extract, and provides a cosmetic composition for skin whitening containing the red grape extract. When preparing cosmetics using red grape extract, at least one component selected from the group consisting of retinol, retinol palmitate, polyethoxylated retinamide, adenosine, kinetin, cocoon extract, isoflavones and mixtures thereof It may further include.

Cosmetic formulations are not particularly limited, but may be formulated as solutions, suspensions, emulsions, pastes, gels, creams, lotions, powders, soaps, surfactant-containing cleansing, oils, powder foundations, emulsion foundations, and sprays. Those skilled in the art can easily adopt a suitable carrier according to the type of formulation.

As another aspect of use of the red grape extract, the present invention provides a functional food for skin wrinkle improvement comprising the red grape extract and food additives.

Here, 'health functional food' is a food manufactured and processed in the form of tablets, capsules, powders, granules, liquids, pills, etc., using raw materials or ingredients with useful functionalities as defined in the Act on Health Functional Foods. (Refer to Article 3 No. 1 of the same law), and 'functionality' refers to obtaining useful effects on health uses such as nutrient control or physiological effects on the structure and function of the human body. In other words, the health functional food means that it can be usefully used for health use of healthy people or semi-healthy people, not for the treatment of patients' diseases.

Efforts to improve skin wrinkles can be obtained by simply eating foods containing red grape extract or applying it to the skin.However, for convenience of use, it is recommended to use it as a functional food having a formulation such as tablets, dragees, capsules, and drinks. desirable.

In addition, the health functional food of the present invention may be prepared in the form of a general food containing food additives, such as food acceptable with red grape extract components, for example, beverages, medicine, kimchi, yogurt, milk, ice cream It may also be prepared in the form of a bread, rice cake and noodles. Here, the 'food additive' refers to an additive used in the manufacturing, processing or preservation of foods by adding, mixing, infiltration, etc. in the food.

Experimental Example 1: Effect of promoting the production of collagen precursor (procollagen) in fibroblast cells (Human)

Fibroblasts were cultured in Medium Essential Medium Eagle (MEM) (Sigma-Aldrich, USA) medium supplemented with 10% Fetal Bovine Serum (FBS) and attached with trypsin-EDTA (Gibco, USA) solution. Remove the cells. After washing twice with phosphate buffered saline (PBS, phosphate buffered solution), 10% FBS + 90% MEM medium was added to prepare a cell suspension of 10 ⁵ cells / ml. Cells were attached by adding 1 ml of the above suspension per well to 24 well tissue culture plates (Corning, USA) and leaving it for 14 hours. After confirming the adherence of the cells, the medium was removed and the medium was added to the drug by concentration in each well (red grapes obtained in Example 1; 0.05, 0.1 0.5, 1, 2 mg / ml, Retinoic acid (Sigma, USA); 0.001, 0.01, 0.1, 1, 10, 100 μM) was added 2 ml per well and incubated for 24 hours at 37 ℃, 5% CO ₂ air mixing incubator (Vision, Korea). After 24 hours, the supernatant was removed and measured with a commercial kit (Procollagen Type IC-Peptide EIA Kit) (Takara, Japan). To briefly introduce the experimental method, dilute the supernatant of the cell medium with the diluent solution contained in the product at a ratio of 1:10, and then place the sample in a 96-well tissue culture plate coated with an antibody (procollagen type I carboxy-terminal peptide). Or 100 μl of standard solution was added and reacted at 37 ° C. for 2 hours. After the reaction, the solution was removed, washed three times with phosphate buffered saline (PBS, phosphate buffered solution), and 100 μl of antibody-POD conjugate solution was added to each well, followed by reaction at 37 ° C. for 1 hour. After the reaction, the solution was removed, washed three times with phosphate buffered saline (PBS, phosphate buffered solution), and 100 μl of substrate solution (containing hydrogen peroxide and tetramethylbenzidine) was added to each well. The reaction was carried out for 15 minutes. After the reaction, 100 μl of 1 NH ₂ SO ₄ solution was added to each well to stop the reaction and measured at 450 nm. The results are shown in Table 1 below.

TABLE 1

As a result of this experiment, retinoic acid increased concentration-dependently from 0.01 ~ 1μM to 13 ~ 64% compared to the untreated control group, and then decreased again at high concentration (100μM. It was the highest (39% increase compared to the control) and showed a sharp decrease at more than 1 mg / ml, which was more effective at low concentration (0.05 ~ 0.1mg / ml) than high concentration.

Experimental Example 2: Inhibitory Effect of MMP-1 (Matrix Metalloproteinase-1) on Fibroblast Cells and Humans

Fibroblasts were cultured in Medium Essential Medium Eagle (MEM) (Sigma-Aldrich, USA) medium supplemented with 10% Fetal Bovine Serum (FBS) and attached with trypsin-EDTA (Gibco, USA) solution. Remove the cells. After washing twice with phosphate buffered saline (PBS, phosphate buffered solution), 10% FBS + 90% MEM medium was added to prepare a cell suspension of 10 ⁵ cells / ml. Cells were attached by adding 1 ml of the above suspension per well to 24 well tissue culture plates (Corning, USA) and leaving it for 14 hours. After confirming the adherence of the cells, the medium was removed and the medium was added to each well by the concentration of the drug (red grapes obtained in Example 1; 0.05, 0.1, 0.5, 1, 2 mg / ml, retinoic acid (Sigma, USA) 0.001, 0.01, 0.1, 1, 10, 100 μM) was added 2 ml per well and incubated in 37 ° C., 5% CO ₂ air incubator (Vision, Korea) for 24 hours. After 24 hours, the supernatant was removed and measured with a commercial kit (Matrix Metalloproteinase-1, Human, Biotrak ELISA Kit) (Amersham, USA). To briefly introduce the experimental method, 100 μl of the standard solution and the sample were added to a 96-well tissue culture plate coated with antibody (mouse anti-MMP-1) in the cell supernatant, and reacted at 20-25 ° C. for 2 hours. . After the reaction, the solution was removed, washed four times with phosphate buffered saline (PBS, phosphate buffered solution), and 100 μl of antiserum was added to each well, followed by reaction at 20-25 ° C. for 2 hours. After the reaction, the solution was removed, washed four times with phosphate buffered saline (PBS, phosphate buffered solution), and 100 μl of peroxidase conjugate was added to each well, followed by reaction at 20-30 ° C. for 1 hour. After the reaction, the solution was removed, washed four times with phosphate buffered saline (PBS, phosphate buffered solution), and immediately, each substrate was prepared with 100 (3,2 ', 5,5'-Tetramethylbenzidine, TMB substrate) solution. 1 µl each was added and reacted at 15-30 ° C for 30 minutes. After the reaction was added to 100 μl of 1 NH ₂ SO ₄ solution to stop the reaction was measured at 450 nm.

The results are shown in Table 2 below.

TABLE 2

                    MMP-1 (ng / ml) sample concentration (μM, mg / ml) Retinoic Acid Rose Number (Example 1) 0 (unsampled control) 3.0 ± 0.1 0.001 μM 2.77 ± 0.12 - 0.01 μM 2.61 ± 0.12 - 0.05 μM 2.41 ± 0.22 0.1 μM 2.64 ± 0.04 2.20 ± 0.12 0.5 mg / ml - 1.82 ± 0.09 1 μM, mg / ml 2.81 ± 0.05 1.83 ± 0.16 2 mg / ml - 1.64 ± 0.07 10 μM 2.58 ± 0.09 - 100 2.77 ± 0.06 -

The results of this experiment showed that the retinoic acid was lower in all concentration groups than the control group. In the case of red grapes, it was found that the concentration was lowered. Therefore, retinoic acid, which is said to be effective in wrinkle formation, and red grapes are expected to be effective in preventing and improving skin wrinkles.

As described above, the red grape extract of the present invention was found to have an effect of promoting the production of collagen precursors and inhibiting the activity of fibroblasts. Red grape extract of the present invention will be effective in preventing and improving skin wrinkles.

Claims (5)

An extract obtained by treating red grapes at a high temperature of 85 ° C. or higher, wherein the red grape extract for improving skin wrinkles increases collagen synthesis and inhibits collagen degradation. A cosmetic composition for improving skin wrinkles comprising the red grape extract of claim 1 as an active ingredient. The method of claim 2, wherein the cosmetic composition for improving skin wrinkles is selected from the group consisting of retinol, retinol palmitate, polyethoxylated retinamide, adenosine, kainetin, cocoon extract, isoflavones and mixtures thereof The cosmetic composition for skin wrinkle improvement further containing the above component. Health functional food for improving skin wrinkles comprising red grape extract of claim 1 and food additives. The red grape extract of claim 1, comprising the red grape extract and a pharmaceutically acceptable carrier, which increases the synthesis of collagen and inhibits the degradation of collagen.