KR101854158B1 - Skin External Composition Containing Extracts of Soybean Flower - Google Patents

Abstract

The present invention relates to a composition for external application for skin containing a bean curd extract, and more particularly to a composition for inhibiting skin antioxidation, skin aging, skin moisturizing effect and skin whitening effect by containing a bean curd extract.

Description

[0001] The present invention relates to a skin external composition containing a soybean extract,

The present invention relates to a composition for external application for skin containing a bean curd extract, and more particularly to a composition for external application for skin which is excellent in skin antioxidation, anti-aging, whitening and moisturizing by containing a bean curd extract.

BACKGROUND OF THE INVENTION [0002] Human skin undergoes changes due to various internal and external factors as it ages. In other words, internally, secretion of various hormones that regulate metabolism is reduced, the function of immune cells and the activity of cells are lowered, and the biosynthesis of immune proteins and biocompatible proteins necessary for the living body is reduced, and ozone layer destruction As the amount of ultraviolet rays reaching the surface of the sunlight increases and the environmental pollution is further intensified, free radicals and active noxious oxygen are increased, thereby decreasing the thickness of the skin, increasing the wrinkles and decreasing the elasticity Skin color is also dull, skin troubles occur frequently, and spots, freckles, and black spots also increase.

As the aging progresses, the content and arrangement of collagen, elastin, hyaluronic acid, and glycoprotein, which constitute the skin, are changed or decreased, and oxidative stress due to free radicals and active noxious oxygen is experienced. Cox-2 (cyclooxygenase), an enzyme that produces proinflammatory cytokines, known to cause inflammation in most of the cells that make up the skin by aging or ultraviolet light The biosynthesis of matrix metalloproteinase (MMP), which is an enzyme that decomposes the skin tissue by these inflammatory factors, is increased, and nitric oxide (NO) production by iNOS (inducible nitric oxide synthase) . In other words, decrease of cellular activity due to natural endogenous aging and decrease of biosynthesis of substrate material by microinflammation, increase of stress due to various harmful environment, increase of active oxygen species by sunlight, As the degradation and denaturation are accelerated, the skin substrate is destroyed and thinned, and various symptoms of skin aging appear. Therefore, it is a reality that many researches have been conducted on active ingredients that can prevent and improve such aging phenomena.

On the other hand, active oxygen generated by various physical, chemical, and environmental factors such as in vivo enzymatic system, reductive metabolism, chemical agents, pollutants and photochemical reaction, etc. is non-selective, irreversible It is known to cause various diseases including cancer or aging cells by destructive action. In addition, lipid peroxides and various peroxides such as lipid peroxides produced as a result of lipid peroxidation by these active oxygen also cause oxidative destruction to cells and cause various dysfunctions and cause various diseases. Accordingly, antioxidants such as free radical scavengers or peroxidation-inhibiting substances are expected to be used as agents for inhibiting or treating various diseases caused by these oxides.

On the other hand, soybean (Glycine Max.) Is used as a health functional food utilizing various processed foods such as edible oil, tofu, soy milk, and soy isoflavone, which is a functional ingredient of soybeans. In recent years, however, research has been devoted to studying various parts such as leaves and roots to diversify the use of soybeans. As a result of the study of the soybean part, the present invention uses the extract of the soybean flour as an external composition for skin, and found excellent antioxidation, anti-aging, whitening and moisturizing effect without irritating the skin.

The inventors of the present invention have confirmed through repeated and various experiments that it is possible to provide a sufficient amount of antioxidative effect, anti-aging effect, whitening effect and moisturizing effect on the skin of a bean curd extract.

Accordingly, an object of the present invention is to provide a composition for external application for skin, which contains an extract of a bean curd plant as an active ingredient.

In order to achieve the above object, the present invention provides an antioxidant composition for external application for skin, which comprises a extract of a bean curd as an active ingredient.

The present invention provides a composition for external application for skin for anti-aging comprising a extract of a bean curd as an active ingredient.

Further, the present invention provides a composition for external application for skin for whitening comprising a soybean flour extract as an active ingredient.

The present invention provides a composition for external application for skin for moisturizing, which comprises a bean curd extract as an active ingredient.

The composition of the present invention not only exhibits antioxidant and anti-aging effects, but also exhibits a whitening effect and a moisturizing effect by containing a soybean extract, and thus can be used in various fields in the field of cosmetics or medicine.

The composition for external application for skin according to the present invention contains a bean curd extract as an active ingredient.

The composition of the present invention preferably contains the extract of the sprout extract in an amount of 0.001 to 10% by weight based on the total weight of the composition. If the content of the active ingredient is less than 0.001% by weight, the antioxidant, anti-aging, whitening and moisturizing effect are insignificant. If the content is more than 10% by weight, no significant increase in the effect is observed.

For example, in the extraction process of the bean curd extract of the present invention, the washed and dried trehalized soybean flour is put into water or an organic solvent and extracted, followed by filtration and centrifugation of the filter cake to separate the residue and the filtrate. The resulting filtrate is concentrated under reduced pressure to give a bean leaf extract. The organic solvent usable in the present invention may be selected from ethanol, methanol, butanol, ether, ethyl acetate, chloroform or a mixed solvent of these organic solvents and water. In view of the safety of raw materials, Of ethanol. After obtaining the extract using a solvent, the extract can be obtained by cooling, heating and filtering at room temperature by a conventional method known in the art, or the solvent can be further evaporated, spray dried or freeze-dried.

The composition of the present invention may be a composition for external application for skin, more specifically,

Can be used as a composition for external application for skin for antioxidation, which can provide an excellent antioxidative effect by inhibiting the oxidation of DPPH which is an organic radical. In addition, the composition of the present invention can be used as an anti-aging external composition for skin, which effectively inhibits the expression of collagenase in the skin, thereby reducing collagen degradation in the skin, thereby enhancing skin elasticity and improving wrinkles. In addition, the composition of the present invention can be used as a composition for external application for skin whitening, and it can provide an excellent whitening effect by inhibiting the production of melanin. Furthermore, the composition of the present invention can be used as a composition for external application for moisturizing skin, which can enhance the skin barrier function and induce differentiation of dermal keratinocytes. Therefore, it can be effectively used as a composition for external application for skin, which prevents or improves skin dryness or psoriasis caused by incomplete epidermal differentiation.

When the composition for external application for skin according to the present invention is formulated in the form of a cosmetic composition, it may be formulated into cosmetic formulations such as softening lotion, convergent lotion, nutritional lotion, eye cream, nutritional cream, massage cream, cleansing cream, cleansing foam, cleansing water, powder, essence, And the formulations thereof are not particularly limited. In addition, the composition according to the present invention can also be used as a lubricant, an organic solvent, a solubilizer, a thickening agent, a gelling agent, a softening agent, an antioxidant, a suspending agent, a stabilizer, a foaming agent, a fragrance, The active ingredient may be combined with any other ingredients commonly used in cosmetics, such as ionic emulsifiers, fillers, sequestering agents, chelating agents, preservatives, vitamins, blockers, wetting agents, essential oils, dyes, pigments, hydrophilic or lipophilic active agents, And may contain adjuvants commonly used in the same cosmetic or dermatological fields. Such adjuvants are introduced in amounts commonly used in the cosmetics or dermatological fields. In addition, the composition of the present invention may contain a skin absorption promoting substance to increase the skin improving effect.

In addition, the composition according to the present invention may be a pharmaceutical composition. The pharmaceutical composition may further contain a preservative, a stabilizer, a wetting or emulsifying accelerator, a pharmaceutical adjuvant such as a salt and / or a buffer for controlling osmotic pressure, and other therapeutically useful substances.

Hereinafter, the configuration and effects of the present invention will be described in more detail with reference to test examples. However, these test examples are provided for illustrative purposes only in order to facilitate understanding of the present invention, and the scope and scope of the present invention are not limited by the following examples.

[Example 1] Production of a soybean flour extract

The soybean flour was washed with purified water and dried. 100 g of the soybean flour powder which had been made into hyaluronic acid was placed in 1 liter of 70% ethanol aqueous solution, boiled for 12 hours in an extractor equipped with a cooling condenser, and then extracted with 300 mesh filter plug. After aging for 7 days at 0 ° C, filter with Wattmann filter paper 2. Add the above extract to a 3 liter separatory funnel, add 1 liter of ethylacetate to it, shake it well and decant it completely, then separate it into two layers and take an upper layer (ethyl acetate layer). The lower layer (water layer) is extracted twice more with a separatory funnel. The separated upper layers were combined and concentrated under reduced pressure at 50 ° C using a distillation apparatus equipped with a cooling condenser and dried to obtain 8.91 g of a dry weight.

[Test Example 1] Antioxidant effect test (DPPH test)

To examine the antioxidative effect of the extract of the bean sprouts produced in Example 1, the reduction of the organic radical DPPH (1,1-diphenyl-2-picryl hydrazyl) (Antioxidant is oxidized). The antioxidant activity was evaluated by comparing the DPPH antioxidant effect with the change of the absorbance. That is, the degree of decrease in the absorbance of DPPH was suppressed to that of the soybean flour extract obtained in Example 1 compared to the control, and the concentration showing an absorbance of 50% or less as compared with the absorbance of the control group was evaluated as an effective antioxidant concentration Respectively.

The reaction solution was prepared by adding 190 μl of 100 μM (in ethanol) DPPH solution and 10 μl of the bean sprout extract and the positive control of Example 1 obtained above, reacting at 37 ° C. for 30 minutes, and then measuring the absorbance at 540 nm. Trolox, a synthetic antioxidant widely used as a positive control, was used. The DPPH analysis results of each substance are shown in Table 1 below, and IC ₅₀ means the sample concentration when the absorbance was reduced by 50% by the added sample.

DPPH analysis result Test substance IC ₅₀ (ppm) Trolox (positive control) 43 Soybean extract 47

As can be seen from the above Table 1, the extract of the bean sprouts of Example 1 showed similar efficacy effects compared with Trolox, which is a synthetic antioxidant used as a positive control. Therefore, it was found that the extract of the bean curd according to the present invention is excellent in antioxidative effect.

[Test Example 2] Anti-aging effect test (measurement of inhibitory effect on expression of collagenase)

The inhibitory effect on the collagenase production of the bean curd extract obtained in Example 1 was compared with that of tocopherol (Comparative Example 1) and EGCG (Comparative Example 2). Tocopherol and EGCG are antioxidants that are known to regenerate epidermal cells of the skin and prevent aging of the skin.

The test was carried out by placing human fibroblasts at 5,000 cells / well in a 96-well microtiter plate containing 2.5% fetal bovine serum-containing DMEM (Dulbecco's Modified Eagle's Media) And cultured until it grew to about 90%. After that, the cells were cultured in serum-free DMEM medium for 24 hours. Then, 100 μg / ml concentration of the bean curd extract of Example 1 dissolved in serum-free DMEM medium, tocopherol and EGCG were treated for 24 hours at a concentration of 10 ^-4 mol , And cell culture fluid was collected.

The collected cell culture medium was measured for collagenase production using a commercially available collagenase measuring device (Amersham Pharmacia, USA). First, the cell culture solution collected in a 96-well plate uniformly coated with the primary collagenase antibody was added and the antigen-antibody reaction was performed in a thermostatic chamber for 3 hours.

After 3 hours, the second collagen antibody bound to the chromophore was placed in a 96-well plate and reacted for another 15 minutes. After 15 minutes, the color development inducing substance was added and coloration was induced at room temperature for 15 minutes. When the reaction (color development) was stopped by adding 1 M sulfuric acid again, the color of reaction solution was yellow and the degree of yellow was different according to the degree of reaction progress.

The absorbance of a yellow 96-well plate was measured at 405 nm using a spectrophotometer and the degree of synthesis of collagenase was calculated by the following equation (1). At this time, the reaction absorbance of the cell culture obtained from the untreated group was used as a control. That is, the degree of expression of collagenase in the untreated group was set at 100, and the degree of expression of collagenase in the group treated with the test substance was determined, and the results are shown in Table 2 below.

Test substance Expression level of collagenase (%) Untreated group 100 Tocopherol (Comparative Example 1) 78 EGCG (Comparative Example 2) 62 Soybean extract 67

The lower the degree of expression of collagenase, the higher the ability to inhibit the expression of collagenase and the less collagen decomposition in the skin, resulting in a smaller amount of wrinkles. In Table 2, the bean curd extract was in vitro In vitro , the expression of collagenase was effectively inhibited and the inhibitory effect of collagenase on the expression of tocopherol was better than that of tocopherol, which is known as an antioxidant. Therefore, it was found that the extract of the bean curd according to the present invention effectively inhibited the expression of collagenase, thereby reducing collagen decomposition in the skin and thus exhibiting excellent anti-aging effect.

[Test Example 3] Whitening effect test (measurement of inhibitory effect on melanin formation using mouse pigment cells)

The ability of the soybean flour extract obtained in Example 1 to inhibit melanin formation was compared with that of hydroquinone.

The cells were cultured in DMEM supplemented with 10% fetal bovine serum, 100 nM 12-O-tetradecanoyl (10 μM) Acetate, and 1 nM cholera toxin at 37 ° C in the presence of 5% CO ₂ . The cultured Mel-Ab cells were detached with 0.25% trypsin-EDTA, and the cells were cultured at a concentration of 10 ⁵ cells / well (cells / well) in a 24-well plate. And cultured. As the test substance, hydroquinone and the bean leaf extract of Example 1 were used at a concentration of 10 ppm, respectively. At this time, the hydroquinone was used as a positive control. Then, the culture solution was removed, washed with PBS (phosphate buffered saline), and the cells were dissolved with 1 N sodium hydroxide. The absorbance at 400 nm was measured, and the inhibition rate of melanin production was calculated according to the following formula (2) 3 (Dooley's method).

Test substance Melanin production inhibition rate (%) Untreated group 100 Hydroquinone (positive control) 44.2 Soybean extract 47.5

As shown in Table 3, it was confirmed that the soybean flour extract of the present invention showed a similar inhibition rate of melanin formation to that of hydroquinone, which is a known whitening substance. Thus, it was found that the extract of the bean curd according to the present invention effectively inhibited the production of melanin in the skin, thereby excelling in the whitening effect.

[Test Example 4]

In order to compare the usability of kojic acid, which is a known whitening substance, with the extract of bean sprouts used as an active ingredient in the present invention, the degree of irritation such as stinginess and burning was tested in 15 panels sensitive to stimuli such as stinging and burning .

0.5 ml of kojic acid (purchased from YM chemical) and the extract of the bean sprouts obtained in Example 1 were applied to the subject at random, and rubbed and scratched to give a score of 0 to 3.0 in units of 0.1 point. The results are shown in Table 4 below.

<Evaluation Criteria>

0 to 0.4: No stimulation

0.5 to 1.0: slight irritation

1.1 to 2.0: normal irritation

2.1 ~ 3.0: Severe irritation

division Kojic acid Soybean extract Stinginess 0.86 0.21 Burning 0.47 0.16 Average 0.62 0.19

As can be seen from Table 4, in the case of kojic acid, it was found that there was some degree of stinging and burning, and there was a feeling of irritation to the extent that it could feel a slight feeling. On the other hand, the bean curd extract used in the present invention showed almost no irritation because it can hardly feel both the burning and stinging.

Therefore, it was confirmed that the extract of the bean curd of the present invention has no irritating sensation unlike the kojic acid, and can provide a better feeling of use.

[Test Example 5] Skin moisturizing test (Differentiation induction test of human keratinocytes)

In order to confirm the skin barrier function and skin moisturizing ability of the extract of Example 1, the keratinocyte cells cultured in primary culture were adhered to the bottom of a culture flask, and then 50 mg / ml and 100 mg / ml were added to the culture medium and cultured for 5 days until the cells were grown to 80 to 90% of the floor area. The cells were harvested and washed with PBS (phosphate buffered saline). The cells were washed with 10 mM Tris-HCl buffer (Tris-HCl buffer, pH 7.4) containing 2% SDS (sodium dodecyl sulfate) and 20 mM DTT (Dithiothreitol) , pH 7.4), sonication was performed for 3 minutes, and the mixture was boiled for 10 minutes. This was centrifuged at 1200 rpm for 30 minutes, and the separated precipitate was suspended in 1 ml of PBS and the absorbance at 340 nm was measured.

Separately, a part of the solution after the sonication was taken to measure the protein content, and was used as a standard for evaluation of the degree of cell differentiation. The test results are shown in Table 5 below, in which low calcium (0.03 mM) and high calcium (1.2 mM) treatment groups were used as negative / positive control groups and low calcium concentration test substances were added.

Test substance Concentration used The ability to differentiate in keratinocytes (%) Control group Low Ca &lt; ^{2 +} & gt ^; (0.03 mM) 100 High Ca ^{2 +} (1.2 mM) 206 Soybean extract 100 mg / ml 152 50 mg / ml 126

As shown in Table 5, when the amount of CE (cornified envelope) produced during keratinocyte differentiation was measured to compare cell differentiation promoting effects, The extract of the bean curd according to the present invention promoted differentiation in keratinocytes. Therefore, it was found that the extract of the bean curd according to the present invention enhances the skin barrier function and improves the skin moisturizing ability.

Claims (7)

A cosmetic composition comprising flower extract of soybean (Glycine Max.). The cosmetic composition according to claim 1, which is contained in an amount of 0.001 to 10% by weight based on the total weight of the composition. The cosmetic composition according to claim 1 or 2, wherein the composition is for enhancing skin elasticity. The cosmetic composition according to claim 1 or 2, wherein the composition is for improving skin wrinkles. The cosmetic composition according to claim 1 or 2, wherein the composition is for whitening. The cosmetic composition according to claim 1 or 2, wherein the composition is for moisturizing. delete