KR102310366B1 - Skin external composition for anti-aging comprising cyanidin-3-o-glucoside and natto gum - Google Patents

Abstract

The present invention relates to an anti-aging composition for external application for skin containing cyanidin glucoside and natto gum as active ingredients, and more particularly, to a composition for external application for skin that exhibits a collagen-degrading enzyme inhibitory effect by mixing and containing cyanidin glucoside and natto gum. .
When the composition for external application for skin according to the present invention is used, it has excellent antioxidant effects such as effectively removing free radicals harmful to the skin, and suppressing the expression and activity of collagen degrading enzymes to strengthen skin elasticity and improve wrinkles It has an excellent anti-aging effect.

Description

TECHNICAL FIELD [0001] Skin external composition for anti-aging comprising cyanidin-3-o-glucoside and natto gum as active ingredients

The present invention relates to an anti-aging composition for external application for skin containing cyanidin glucoside and natto gum as active ingredients, and more particularly, to a composition for external application for skin that exhibits a collagen-degrading enzyme inhibitory effect by mixing and containing cyanidin glucoside and natto gum. .

As the human body's primary barrier, the skin protects organs from external environmental stimuli such as temperature and humidity changes, ultraviolet rays and pollutants, and plays an important role in maintaining body homeostasis such as body temperature regulation. However, excessive physical and chemical stimuli, stress, and nutritional deficiencies from the outside reduce the normal function of the skin and promote skin aging such as loss of elasticity, keratinization, and wrinkle formation.

In order to prevent this phenomenon and maintain healthy and elastic skin, cosmetics containing physiologically active substances obtained from various plants and microorganisms are manufactured to maintain the unique function of the skin and activate skin cells to effectively prevent skin aging. Efforts have been made to contain it. However, most of the existing cosmetic raw materials had various problems such as insufficient efficacy or causing skin side effects.

Cyanidin-3-O-glucoside, an active ingredient of the composition of the present invention, is a type of anthocyanin, also called chrysanthemin, and has a structure in which cyanidin is linked to carbon 3 of glucose by a glycosidic bond. have It is a color pigment contained in berry fruits and grains, and is known for its antioxidant effect. Recently, research results have been reported that cyanidin glucoside is effective in preventing cancer or heart disease (Cooke et al. , International journal of cancer 119.9 (2006): 2213-2220; Serraino et al., Life sciences 73.9 (2003): 1097-1114). However, it is not yet known whether cyanidin glucoside has an anti-aging effect by inhibiting the action of collagen degrading enzymes to maintain skin elasticity and improve wrinkles.

Another active ingredient of the composition of the present invention, natto gum, is a highly viscous material obtained when soybean is fermented with a gram-positive bacterium called Bacillus subtilis. Recently, cosmetics containing natto gum, which act as a thickener and have a strong moisturizing effect, have been released. However, like cyanidin glucoside, natto gum has an anti-aging effect by inhibiting the action of collagen-degrading enzyme to maintain skin elasticity and improve wrinkles, or whether natto gum has an antioxidant or anti-aging effect of cyanidin glucoside. Whether or not it improves is not yet known.

As a result of diligent efforts by the present inventors to discover natural substances that can maximize the anti-aging effect of the skin, a composition for external application for skin containing a mixture of cyanidin glucoside and natto gum inhibits radical generation and effectively inhibits the expression of collagen degrading enzymes By doing so, it was found that the antioxidant effect and anti-aging effect were very excellent, and the present invention was completed.

Cooke et al., International journal of cancer 119.9 (2006): 2213-2220 Serraino et al., Life sciences 73.9 (2003): 1097-1114

It is an object of the present invention to provide a composition for external application for skin that contains cyanidin glucoside and natto gum as active ingredients to suppress the generation of organic radicals in the skin and suppress the expression of collagen degrading enzymes, thereby exhibiting an anti-aging effect.

In order to achieve the above object, the present invention provides an anti-aging composition for external application for skin containing cyanidin glucoside and natto gum as active ingredients.

In addition, the present invention provides a composition for external application for skin for improving skin elasticity containing cyanidin-3-O-glucoside and natto gum as active ingredients.

In addition, the present invention provides a composition for external application for skin wrinkle improvement containing cyanidin-3-O-glucoside and natto gum as active ingredients.

When the composition for external application for skin according to the present invention is used, there is an excellent antioxidant effect such as effectively removing free radicals harmful to the skin, and the expression and activity of collagenase is suppressed to strengthen skin elasticity and improve wrinkles It has an excellent anti-aging effect such as becoming.

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. In general, the nomenclature used herein is those well known and commonly used in the art.

In an embodiment of the present invention, as a result of measuring DPPH oxidation inhibitory efficacy by mixing equal amounts of cyanidin glucoside and natto gum, cyanidin glucoside or natto gum was used alone, or delphinidin-3-O-glucoside and natto gum were mixed. It was confirmed that it exhibited a much better antioxidant effect than when mixed and used. In addition, as a result of measuring the inhibitory effect of collagen-degrading enzyme by mixing cyanidin glucoside and natto gum in equal amounts, the anti-aging effect was much better than when cyanidin glucoside or natto gum was used alone or when delphinidin glucoside and natto gum were mixed. was able to confirm

Accordingly, in one aspect, the present invention relates to an anti-aging composition for external application for skin containing cyanidin-3-O-glucoside and natto gum as active ingredients.

In another aspect, the present invention relates to a composition for external application for skin for improving skin elasticity or for improving skin wrinkles, comprising cyanidin-3-O-glucoside and natto gum as active ingredients.

The cyanidin glucoside of the present invention has the structure of the following formula (1), and can be directly extracted from a plant rich in cyanidin glucoside using a conventional method in the art, or purified cyanidin glucoside can be purchased and used after extraction. may be

[Formula 1]

In the present invention, the natto gum may be produced by inoculating and fermenting activated Bacillus subtilis or Bacillus licheniformis into soybeans. Natto gum is a viscous material produced on the surface of soybeans when soybeans are inoculated with Bacillus subtilis or Bacillus licheniformis and fermented at a certain temperature. After purifying and crystallizing the natto gum, it can be used in the composition of the present invention in the form of a powder. As the type of soybean fermented to produce natto gum, soybean is the most preferable, but is not limited thereto.

When a composition in which natto gum is mixed with cyanidin glucoside is used, the negative charge of polygamma glutamic acid contained in natto gum interacts with the positive charge of cyanidin glucoside to stabilize the structure of cyanidin glucoside, thereby preventing the antioxidant and anti-oxidation of cyanidin glucoside. It is presumed to further enhance the aging effect.

In the present invention, the composition for external application for skin containing cyanidin glucoside and natto gum as active ingredients inhibits the expression of collagen-degrading enzymes that reduce skin elasticity by decomposing collagen in skin tissues, thereby improving anti-aging effects, particularly improving skin wrinkles, and It may be characterized in that it exhibits an effect of improving skin elasticity. Also, in the present invention, "collagen degrading enzyme" and "collagenase" are used as the same meaning.

Also, in the present invention, the cyanidin glucoside and natto gum may be mixed in a weight ratio of 1:1 to 1:10, preferably 1:1. When mixed in a weight ratio within the above range, it is appropriate to maximize the effect of inhibiting the expression of collagen degrading enzymes of cyanidin glucoside and natto gum, respectively, and the stability of the composition can be satisfied.

In addition, in the present invention, the cyanidin glucoside is contained in an amount of 0.01 to 5% by weight based on the total weight of the composition, and the natto gum is contained in an amount of 0.01 to 5% by weight based on the total weight of the composition. If the total of the two active ingredients is 0.02% by weight or less, the effect of inhibiting collagenase expression is weak, and if it is 10% by weight or more, there is a problem in that the viscosity of the composition is excessively high.

When the composition for external application for skin according to the present invention is formulated in the form of a cosmetic, softening lotion, astringent lotion, nourishing lotion, eye cream, nourishing cream, massage cream, cleansing cream, cleansing foam, cleansing water, powder, essence or pack, etc. It may be formulated in the form of, and the formulation is not particularly limited. In addition, the composition according to the present invention is a fatty substance, an organic solvent, a solubilizer, a thickening agent, a gelling agent, an emollient, an antioxidant, a suspending agent, a stabilizer, a foaming agent, a fragrance, a surfactant, water, an ionic or non Ionic emulsifiers, fillers, sequestering agents, chelating agents, preservatives, vitamins, blocking agents, wetting agents, essential oils, dyes, pigments, hydrophilic or lipophilic actives, lipid vesicles or any other ingredients commonly used in cosmetics; It may contain adjuvants commonly used in the field of cosmetology or dermatology. The adjuvant is introduced in an amount generally used in the field of cosmetology or dermatology. In addition, the composition of the present invention may contain a skin absorption promoting substance in order to increase the skin improvement effect.

The composition for external application for skin according to the present invention may include other ingredients that can give a synergistic effect to the main effect, preferably within a range that does not impair the main effect, in addition to the above substances. In addition, the composition for external application for skin according to the present invention may further include a moisturizer, an emollient, a UV absorber, a preservative, a disinfectant, an antioxidant, a pH adjuster, an organic and inorganic pigment, a fragrance, a cooling agent or a limiting agent. The blending amount of the component can be easily selected by those skilled in the art within the range that does not impair the purpose and effect of the present invention, and the blending amount may be 0.01 to 5 wt%, specifically 0.01 to 3 wt%, based on the total weight of the composition have.

Hereinafter, the present invention will be described in more detail through examples. These examples are only for illustrating the present invention, and it will be apparent to those of ordinary skill in the art that the scope of the present invention is not to be construed as being limited by these examples. Accordingly, the substantial scope of the present invention will be defined by the appended claims and their equivalents.

[Reference Example 1]

Cyanidin-3-O-glucoside for testing the efficacy of the composition of the present invention was purchased from Sigma and used.

[Reference Example 2]

As a comparative example of the composition of the present invention, one of anthocyanins, delphinidin glucoside (delphinidin-3-O-glucoside) was purchased from Sigma and used.

[Reference Example 3]

Natto gum for testing the efficacy of the composition of the present invention was prepared and used in the following manner.

(1) Heat sterilization process:

Soybeans were soaked in water for about 10 to 15 hours, thoroughly washed with ionized water (primary distilled water), and then sterilized by heat in an autoclave at 100 to 121°C for about 2 hours.

(2) Cultivation process:

The sterilized soybeans were cooled to 40°C, and the activated Bacillus subtilis inoculum was inoculated with 5-10% of the total soybean mass, and then cultured at 35-40°C for about 36 hours. ^{The inoculum was used for inoculating Bacillus subtilis at a concentration of 10 7-8} Cells/ml in 8 g/ℓ of a nutrient medium (Neutrient Brooth).

(3) Extraction and purification process:

An appropriate amount of ionized water was added to the cultured soybean culture solution, and the mixture was sufficiently stirred to dissolve the pungent substance on the soybean surface in the ionized water, followed by filtration to obtain a transparent extract.

(4) Crystallization process:

An appropriate amount of a solvent such as methanol, ethanol, or acetone was added to the clear extract to reduce solubility in water to obtain a white precipitate. Then, the solvent contained in the obtained precipitate was evaporated to dryness to obtain natto gum powder.

[Preparation of Example 1-2 and Comparative Example 1-2]

Examples 1 and 2 were prepared by mixing the cyanidin glucoside of Reference Example 1 and the natto gum prepared in Reference Example 3 in the weight ratio shown in Table 1 below, but the total weight was the same.

In addition, as a comparative example to Examples 1 and 2, delphinidin glucoside of Reference Example 2 and the natto gum prepared in Reference Example 3 were mixed in the weight ratio shown in Table 1 below, but the total weight was both as in Examples 1 and 2 Comparative Examples 1 and 2 were prepared in the same manner.

(Unit: % by weight) Example 1 Example 2 Comparative Example 1 Comparative Example 2 Cyanidin Glucoside 50 25 - - Delphinidin Glucoside - - 50 25 natto gum 50 75 50 75 total 100 100 100 100

[Test Example 1] Antioxidant activity test

In order to investigate the antioxidant effect of Examples 1 and 2, which is a mixture of cyanidin glucoside and natto gum, the organic radical DPPH (1,1-diphenyl-2-picrylhydrazyl; 1,1-diphenyl-2-picryl) hydrazyl), the DPPH oxidation inhibitory efficacy was compared with those of Comparative Examples 1 and 2 in which cyanidin glucoside alone, natto gum alone, and delphinidin glucoside and natto gum were mixed.

190 μl of 100 μM (in ethanol) DPPH solution and cyanidin glucoside (Reference Example 1), natto gum (Reference Example 3), Examples 1, 2, Comparative Examples 1 and 2, and the synthetic antioxidant Trolox ( positive control) at the same concentration of 10,000 ppm, each diluted so that the final reaction concentrations are 500 ppm, 250 ppm, 125 ppm, 62.5 ppm, 31.25 ppm, and 15.63 ppm, and then add 10 μl each to make a reaction solution 37 After reacting at ℃ for 30 minutes, absorbance was measured at 540 nm. The analysis results are shown in Table 2 below, and IC ₅₀ means the sample concentration when the absorbance is reduced by 50% by the added sample.

test substance IC ₅₀ (ppm) Trolox (positive control) 43.3 Cyanidin Glucoside 231.5 Delphinidin Glucoside 287.5 natto gum no effect Example 1 39.2 Example 2 108.5 Comparative Example 1 58.6 Comparative Example 2 143.5

As shown in Table 2, when natto gum was used alone, there was almost no antioxidant effect. However, in the case of Example 1, in which cyanidin glucoside and natto gum were mixed in a weight ratio of 1:1, an IC ₅₀ value similar to that of Trolox was displayed, and the antioxidant effect was significantly higher than that in the case of using cyanidin glucoside alone. could be seen to appear. In addition, in the case of Example 2, in which cyanidin glucoside and natto gum were mixed in a weight ratio of 1:3, the antioxidant effect was not as excellent as that of Example 1, in which 1:1 was mixed, but much better than when cyanidin glucoside was used alone. The effect was excellent. In addition, Examples 1 and 2 both had excellent antioxidant effects compared to Comparative Examples 1 and 2 in which delphinidin glucoside was mixed with natto gum. Therefore, it was confirmed that the composition of the present invention has an excellent antioxidant effect.

[Experimental Example 2] Anti-aging ability test

In order to examine the anti-aging effect of Examples 1 and 2, which is a mixture of cyanidin glucoside and natto gum, the inhibitory ability of each collagenase production was determined by mixing cyanidin glucoside alone, natto gum alone, delphinidin glucoside and natto gum. Comparative Examples 1 and 2, and compared with tocopherol or EGCG. Tocopherol and EGCG are substances known to have the function of preventing skin aging by regenerating epidermal cells of the skin.

Human fibroblasts (Cascade Biologics) (Portland, OR, USA) were placed in a 96-well microtiter plate containing DMEM (Dulbecco's Modified Eagle's Media) medium containing 2.5% by weight of fetal bovine serum. It was added to 5,000 cells/well, and cultured until about 90% growth. Thereafter, cultured for 24 hours in serum-free DMEM medium, cyanidin glucoside (Reference Example 1), natto gum (Reference Example 3), Examples 1, 2, Comparative Example 1, and 10 μg each of Comparative Example 2 was taken and treated at the same concentration of 50 ppm for 24 hours, and tocopherol and EGCG (Sigma Aldrich) were each treated at a concentration of 50 μM for 24 hours, and then the cell culture solution was collected.

Using a collagenase measuring instrument (Amersham Pharma, USA), the collagenase production level of the collected cell culture was measured. First, the collected cell culture solution was placed in a 96-well plate uniformly coated with the primary collagenase antibody, and the antigen-antibody reaction was performed in a thermostat (36° C.) for 3 hours.

After 3 hours, the chromophore-conjugated secondary collagen antibody was placed in a 96-well plate and reacted for 15 minutes again. After 15 minutes, add a color-generating substance to develop color at room temperature for 15 minutes, and then add 1M sulfuric acid to stop the reaction (color development), the color of the reaction solution is yellow. was confirmed.

The absorbance of a 96-well plate having a yellow color was measured at 405 nm using an absorber, and the degree of collagenase synthesis was calculated by Equation 1 below. At this time, the reaction absorbance of the cell culture solution collected from the untreated group was used as a control group. That is, the expression level of collagenase in the untreated group was set to 100, and in contrast, the expression level of collagenase in the group treated with the test substance was obtained, and the results are shown in Table 3 below.

test substance Collagenase expression level (%) untreated group 100 tocopherol 73 EGCG 59 Cyanidin Glucoside 78 Delphinidin Glucoside 84 natto gum 94 Example 1 51 Example 2 67 Comparative Example 1 61 Comparative Example 2 73

As shown in Table 3, collagenase expression was hardly inhibited when natto gum was used alone. However, in Example 1, in which cyanidin glucoside and natto gum were mixed in a weight ratio of 1:1, the expression of collagenase was more effectively inhibited in vitro than when cyanidin glucoside was used alone, and an anti-aging substance It was found that the ability to inhibit collagenase expression was superior to that of tocopherol and EGCG, which are known as In addition, in the case of Example 2, in which cyanidin glucoside or natto gum was mixed in a weight ratio of 1:3, the collagenase expression inhibitory effect was not as excellent as in Example 1, in which cyanidin glucoside was mixed alone. It was much more effective than when I used it. Furthermore, Examples 1 and 2 both had an excellent collagenase expression inhibitory effect compared to Comparative Examples 1 and 2 in which delphinidin glucoside was mixed with natto gum. Therefore, it was confirmed that the composition of the present invention has excellent anti-aging effects due to inhibition of collagenase expression, in particular, skin elasticity improvement and skin wrinkle improvement effects.

As described above in detail a specific part of the content of the present invention, for those of ordinary skill in the art, this specific description is only a preferred embodiment, and it is clear that the scope of the present invention is not limited thereby. something to do. Accordingly, it is intended that the substantial scope of the present invention be defined by the appended claims and their equivalents.

In addition, the formulation examples of the composition for external application for skin including the composition of the present invention will be described in more detail below, but it will be clear that the scope of the present invention is not limited thereby.

[Formulation Example 1] Nutrient lotion

Nutrient lotion was prepared in a conventional manner according to the composition shown in Table 4 (unit: wt%).

ingredient content Purified water remaining amount glycerin 8.0 butylene glycol 4.0 Hyaluronic Acid Extract 5.0 beta glucan 7.0 Carbomer 0.1 Cyanidin Glucoside and Natto Gum (Example 1) 0.05 Caprylic/Capric Triglycerides 8.0 squalane 5.0 cetearyl glucoside 1.5 Sorbitan Stearate 0.4 cetearyl alcohol 1.0 triethanolamine 0.1

[Formulation Example 2] Nourishing cream

Nutrient creams were prepared in a conventional manner according to the composition shown in Table 5 below (unit: wt%).

ingredient content Purified water remaining amount glycerin 3.0 butylene glycol 3.0 liquid paraffin 7.0 beta glucan 7.0 Carbomer 0.1 Cyanidin Glucoside and Natto Gum (Example 1) 3.0 Caprylic/Capric Triglycerides 3.0 squalane 5.0 cetearyl glucoside 1.5 Sorbitan Stearate 0.4 Polysorbate 60 1.2 triethanolamine 0.1

[Formulation Example 3] Massage Cream

A massage cream was prepared in a conventional manner according to the composition shown in Table 6 below (unit: wt%).

ingredient content Purified water remaining amount glycerin 8.0 butylene glycol 4.0 liquid paraffin 45.0 beta glucan 7.0 Carbomer 0.1 Cyanidin Glucoside and Natto Gum (Example 1) 1.0 Caprylic/Capric Triglycerides 3.0 beeswax 4.0 cetearyl glucoside 1.5 Sorbitan Sesquioleate 0.9 Vaseline 3.0 paraffin 1.5

[Formulation Example 4] Pack

A pack was prepared in a conventional manner according to the composition shown in Table 7 (unit: wt%).

ingredient content Purified water remaining amount glycerin 4.0 polyvinyl alcohol 15.0 Hyaluronic Acid Extract 5.0 beta glucan 7.0 allantoin 0.1 Cyanidin Glucoside and Natto Gum (Example 1) 0.5 nonylphenyl ether 0.4 Polysorbate 60 1.2 ethanol 6.0

[Formulation Example 5] Ointment for external skin preparations

An ointment was prepared in a conventional manner according to the composition shown in Table 8 below (unit: wt%).

ingredient content Purified water remaining amount glycerin 8.0 butylene glycol 4.0 liquid paraffin 15.0 beta glucan 7.0 Carbomer 0.1 Cyanidin Glucoside and Natto Gum (Example 1) 1.0 Caprylic/Capric Triglycerides 3.0 squalane 1.0 cetearyl glucoside 1.5 Sorbitan Stearate 0.4 cetearyl alcohol 1.0 beeswax 4.0

Claims (8)

An external composition for anti-aging skin containing cyanidin-3-O-glucoside and natto gum as active ingredients. A composition for external application for skin for improving skin elasticity, comprising cyanidin-3-O-glucoside and natto gum as active ingredients. A composition for external application for skin wrinkle improvement comprising cyanidin-3-O-glucoside and natto gum as active ingredients. The composition for external application for skin according to any one of claims 1 to 3, wherein the natto gum is produced by inoculating soybeans with activated Bacillus subtilis or Bacillus licheniformis and fermenting them. . The composition for external application for skin according to any one of claims 1 to 3, wherein the composition exhibits an anti-aging effect by inhibiting the expression of collagen degrading enzyme. The composition for external application for skin according to any one of claims 1 to 3, wherein the cyanidin glucoside and natto gum are mixed in a weight ratio of 1:1 to 1:10. The method according to any one of claims 1 to 3, wherein the cyanidin glucoside is contained in an amount of 0.01 to 5% by weight based on the total weight of the composition, and the natto gum is contained in an amount of 0.01 to 5% by weight based on the total weight of the composition. A composition for external application to the skin, characterized in that. According to any one of claims 1 to 3, when the composition for external application for skin is formulated in the form of a cosmetic, softening lotion, astringent lotion, nourishing lotion, eye cream, nourishing cream, massage cream, cleansing cream, cleansing A composition for external application for skin, characterized in that it is formulated in any one form selected from the group consisting of foam, cleansing water, powder, essence and pack.