KR101926179B1 - Cosmetic composition containing canavalia lineata extract for moisturizing and anti-wrinkle effects on the skin - Google Patents

Abstract

The present invention relates to a cosmetic composition for improving skin wrinkles and skin moisturizing, which comprises an extract of sea urchin soybean as an active ingredient. More specifically, the present invention relates to a cosmetic composition for skin wrinkle improvement and skin moisturizing which has a wrinkle-reducing effect due to inhibition of MMP-1 expression and a skin moisturizing effect due to promotion of AQP-3 expression by containing ethanol extract of virgin soybean as an active ingredient. The seedling soybean extract of the present invention exerts excellent skin wrinkle-improving effect through effectively inhibiting the expression of MMP-1. In addition, it exerts an excellent moisturizing effect by effectively increasing the expression of AQP-3.

Description

Technical Field [0001] The present invention relates to a cosmetic composition for improving skin wrinkles and moisturizing cosmetic composition containing soybean extract,

The present invention relates to a cosmetic composition for improving skin wrinkles and skin moisturizing, which comprises an extract of sea urchin soybean as an active ingredient. More specifically, the present invention relates to a cosmetic composition for skin wrinkle improvement and skin moisturizing which has a wrinkle-reducing effect due to inhibition of MMP-1 expression and a skin moisturizing effect due to promotion of AQP-3 expression by containing ethanol extract of virgin soybean as an active ingredient.

Skin is an important institution that protects the human body from the external environment and plays a variety of physiological functions such as barrier function, body temperature control, excretion, etc., which prevent the internal moisture and useful components from leaking out. In general, however, as the age increases, these functions are gradually weakened and the aging of the skin progresses. In particular, the lipid composition and the content of the lipid layer, which functions as a skin barrier, At this time, as the skin moisture content decreases, the skin becomes dry, wrinkles are formed, and spots, freckles, pigmentation and various skin lesions are induced.

In addition, skin moisture content is about 70% in the dermis, but it decreases as it goes to the epidermis and becomes about 10-30% in the outermost layer. In general, the normal stratum corneum contains about 30% of water or water-soluble ingredients, and moisture of the stratum corneum must be properly maintained to maintain elasticity and flexibility of the skin.

Also, there is a high concentration of Natural Mosituring Factors (NMF) in the keratinocytes of a healthy person, which functions to help the moisture retention of the skin. Despite these functions of the skin, however, due to the exclusion of skin constituents due to physical and chemical stimuli such as extrinsic aging, endogenous aging and friction, shaving and cleansing by external environment such as wind, low temperature, polluted atmosphere and ultraviolet rays Which causes the moisture balance to collapse, causing skin dryness. In order to prevent this, the necessity of a skin softener and a moisturizer is important, and the demand therefor is also increasing.

Therefore, a number of products for moisturizing purposes have been marketed and studies have been actively carried out. However, as products having moisturizing effect to date, lipid components such as ceramide which can increase water retention in the stratum corneum, Fatty acids and lipid complexes (J. Invest. Dermatol., 5, pp. 731-740, 1994).

On the other hand, Aquaporin (AQP) is a type of water channel proteins that prevents bursting of cells and prevents the movement of ions or solutes, . It has been reported that 13 AQPs (AQP-0 to AQP-12) exist in mammals, and AQP-3 has been reported to be expressed in the basal layer of keratinocytes. Therefore, it can be expected that a skin moisturizing effect can be obtained by using AQP-3 activated in the skin layer and using a substance that transfers moisture deep into the skin.

In addition, factors such as collagen, elastin fiber, and hyaluronic acid, which are extracellular matrix, are involved in the formation of wrinkles in the skin. MMP (matrix metalloproteinase), a metalloproteinase, is an enzyme that degrades extracellular matrix collagen and elastin, and is known as an enzyme that specifically degrades collagen type 1. When exposed to ultraviolet light, MMP-1 is produced more in the body than usual, and MMP-1 binds to collagen and cleaves collagen, causing skin aging. More specifically, the collagen proteins specifically bind to the 'MMP-1 hemopexin domain', thereby solving the tertiary structure of the collagen, and the zinc of the MMP-1 catalytic domain After bonding with the ions, they are cleaved.

On the other hand, interest in cosmetics has been increasing in recent years in order to maintain and improve the elasticity of skin as interest in appearance has increased. Therefore, studies for strengthening the skin barrier through skin moisturizing and wrinkle improvement have been actively carried out. In particular, there is a need for research to find a material that activates moisturizing factors in the skin by an intrinsic solution, Is increasing.

Korean Patent Laid-Open No. 10-2015-0101980 (Publication date: 2015.09.04) discloses that cumestrol; The present invention relates to a skin cosmetic composition comprising germinated soybean extract and germinated soybean extract containing gum arabic extract and germinated soybean extract containing gum arabic extract, Improvement of atopic dermatitis, alleviation of skin irritation, maintenance of skin homeostasis, alleviation or improvement of skin allergy, and cosmetics and foods. Korean Patent No. 10-0508528 (Registration Date: Aug. 8, 2005) relates to a skin cosmetic composition using a mixture of extracts of soybean curd and green tea extract. In particular, skin whitening effect and wrinkle prevention effect, antioxidative effect, hair growth effect, An anti-inflammatory effect, and an anti-acne effect. Also, there is provided a method for producing a soybean curd, comprising: a first step of preparing a soybean curd and a golden turf top by drying at room temperature until there is no change in weight; A second step of mixing and pulverizing the dried soybean paste and the gold discharge preparae at a ratio of 1: 1 by weight to obtain a mixed powder; The soybean paste and the gold-sending preliminary mixed powder obtained in the second step are added to any one aqueous solution selected from distilled water / 30-95% ethanol / methanol / 1,3-butylene glycol / 30% propylene glycol / , And heating for 48 hours under a water bath condition of 60 占 폚; And a fourth step of filtering the heated product obtained in the third step with 0.45 mu m filter paper.

It is an object of the present invention to develop a cosmetic composition containing an extract of a natural substance having excellent efficacy for improving skin wrinkles and moisturizing and containing it as an active ingredient.

The present invention relates to a method for the treatment of canvals spp . ) Extract as an active ingredient. The present invention also provides a cosmetic composition for improving skin wrinkles.

In the present invention, the above- spp . ) Is preferably selected from the group consisting of Canavalia lineata ).

In the present invention, the above- spp . ) Extracts can be prepared according to conventional methods known in the art, that is, using conventional solvents under the conditions of ordinary temperature and pressure. However, it is preferable that the extract is obtained by adding at least one extraction solvent selected from the group consisting of water, an anhydrous or hydro- lower alcohol having 1 to 4 carbon atoms, acetone, ethyl acetate and butyl acetate to a plant of Canavalia spp. More preferably 50% -90% ethanol aqueous solution as an extraction solvent, and most preferably 70% ethanol extract. The extract of the soymilk bean extract thus obtained is safe for preventing cell toxicity, and is effective for improving the wrinkles of the skin.

In the present invention, it is preferable that the above-mentioned Canavalia spp. Extract is contained in an amount of 0.00001-30.0% by weight based on the total weight of the cosmetic composition. When the content of the extract is less than 0.00001% by weight, the effect of improving the skin wrinkles is not exhibited. When the content of the extract is more than 30.0% by weight, the degree of improvement of the skin wrinkles is insignificant, And it is not economical.

In the present invention, the cosmetic composition may be prepared in any form conventionally produced in the art, and examples thereof include a solution, a suspension, an emulsion, a paste, a gel, a cream, a lotion, a powder, a soap, And may be any one selected from the group consisting of cleansing, oil, powder foundation, emulsion foundation, wax foundation and spray.

On the other hand, the invention in divers beans (Canavalia spp . ) Extract as an active ingredient. The present invention also provides a cosmetic composition for moisturizing the skin.

In the present invention, the above- spp . ) Is preferably a canvalia lineata .

In the present invention, the above-mentioned Canavalia spp. Extract can be produced according to a conventional method known in the art, that is, using a conventional solvent under the conditions of ordinary temperature and pressure. However, it is preferable that the extract is obtained by adding at least one extraction solvent selected from the group consisting of water, an anhydrous or hydro- lower alcohol having 1 to 4 carbon atoms, acetone, ethyl acetate and butyl acetate to a plant of Canavalia spp. More preferably 50% -90% ethanol aqueous solution as an extraction solvent, and most preferably 70% ethanol extract. The extract of the soymilk bean extract thus obtained is safe for skin moisturization because it does not show cytotoxicity.

In the present invention, it is preferable that the above-mentioned Canavalia spp. Extract is contained in an amount of 0.00001-30.0% by weight based on the total weight of the cosmetic composition. When the content of the extract is less than 0.00001% by weight, the skin moisturizing effect is not exhibited. When the content is more than 30.0% by weight, the skin moisturizing effect is insignificantly increased with respect to the content increase, It is not economical.

In the present invention, the cosmetic composition may be prepared in any form conventionally produced in the art, and examples thereof include a solution, a suspension, an emulsion, a paste, a gel, a cream, a lotion, a powder, a soap, And may be any one selected from the group consisting of cleansing, oil, powder foundation, emulsion foundation, wax foundation and spray.

The seedling soybean extract of the present invention exerts excellent skin wrinkle-improving effect through effectively inhibiting the expression of MMP-1. In addition, it exerts an excellent moisturizing effect by effectively increasing the expression of AQP-3.

FIG. 1 is a graph showing the survival rate (line graph) and the inhibition rate (bar graph) of MMP-1 expression of human dermal fibroblast neonatal (HDFn) treated with 70% Adenosine (AD) was used at a concentration of 100 μM.
FIG. 2 is a graph showing the promotion rate of AQP-3 expression in keratinocytes (HaCaT) treated with 70% ethanol extract of sea cucumber bean. FIG. 2A is a result of measuring mRNA expression level of AQP-3 FIG. 2 (B) shows the result of measuring the protein expression level of AQP-3. As a positive control, 10 nM retinoic acid (RA) was used.

Hereinafter, the contents of the present invention will be described in more detail in the following examples, but the scope of the present invention is not limited to the following examples, but includes modifications of equivalent technical ideas.

[ Example  One: Dairy bean  Preparation of extract]

In this Example, soybean seed extract was prepared. Dried soybean beans were crushed to prepare bean ground bean pulverized products. 1 kg of dried bean pulverized beans were put into a flask, and 10 L of 70% (v / v) ethanol was added thereto, followed by cooling at room temperature for 3 days. Thereafter, filtration and concentration with a filter paper (Whatman No.1 filter paper, Maidstone, UK) were carried out to obtain 110.3 g of soybean seed extract.

[ Example  2: Dairy bean  The effect of the extract on cell viability and MMP -1 expression in mice.

 (One) Dairy bean  Effect of Extract on Cell Survival Rate

In this Example, the effect of the seedling extract prepared in Example 1 on cell survival was examined.

Human dermal fibroblast neonatal (HDFn) was inoculated in a 96-well plate at a concentration of 2 × 10 ⁴ cells / well, and then stabilized in a 5% CO ₂ incubator at 37 ° C. for 24 hours . After that, 12.5, 25, 50 and 100 μg / mL of sea urchin soybean extract was added to each cell for 24 hours, and 100 μM of adenosine (AD) was further treated as a positive control. Then, 5 μg / mL of '3- (4,5-dimethylthiazol-2-yl) -2-5-diphenyltetrazolium bromide' (MTT) reagent was treated for 2 hours in each cell. After 2 hours, the supernatant was removed, 150 μl of DMSO was added per well, and the formed formazan was measured for absorbance at a wavelength of 540 nm using a spectrophotometer.

As a result, it was found that the soybean seed extract did not significantly affect cell survival (FIG. 1). FIG. 1 is a graph showing the survival rate (line graph) and the inhibition rate (bar graph) of MMP-1 expression of human dermal fibroblast neonatal (HDFn) treated with 70% Adenosine (AD) was used at a concentration of 100 μM.

 (2) Dairy bean  The extract MMP -1 expression

In this Example, the effect of the soybean extract prepared in Example 1 on the expression of MMP-1 was examined.

Human dermal fibroblast neonatal (HDFn) was inoculated into a 24-well plate at a concentration of 1 × 10 ⁵ cells / well and then stabilized in an incubator at 37 ° C. and 5% CO ₂ for 24 hours . The cells were then rinsed twice with PBS and the cells were irradiated with UVA at 6 J / cm ² in the presence of PBS. After the irradiation, the PBS was removed and the culture medium supplemented with 0.2% FBS was replaced with 100 μM adenine (AD) (100 μg / ml) Lt; / RTI > After 48 hours, only the supernatant was obtained and the absorbance was measured at 450 nm using a spectrophotometer using a human total MMP-1 ELISA kit (human total MMP-1 Cat. No. DY901, R & D systems).

As a result, it was confirmed that MMP-1 was inhibited about 40% from the lowest concentration of 12.5 μM in adipocyte-treated cells. In addition, it was confirmed that MMP-1 was inhibited up to 80% in cells treated with 100 μg / mL of soybean extract (FIG. 1).

FIG. 1 is a graph showing the survival rate (line graph) and the inhibition rate (bar graph) of MMP-1 expression of human dermal fibroblast neonatal (HDFn) treated with 70% Adenosine (AD) was used at a concentration of 100 μM.

Taken together, it was confirmed that the seedling soybean extract of the present invention effectively inhibited the expression of MMP-1 without affecting the survival rate of the cells.

[ Example  3: Dairy bean  The extract AQP -3]

 (One) Dairy bean  The extract AQP -3 of mRNA  Identification of effects on expression

In this example, the effect of soybean extract on the mRNA expression of AQP-3 was examined. The keratinized soybean extract prepared in Example 1 was diluted to 25, 50, and 100 μg / mL, and the cells were cultured for 24 hours at a density of 2 × 10 ⁵ cells / mL. Treated and reacted again for 24 hours. After the reaction, cells were obtained using DEPC-treated cold PBS, and total RNA was isolated using TRIZOL reagent. Using 0.3 μg of the separated RNA, cDNA was synthesized at 42 ° C. for 60 minutes and at 94 ° C. for 5 minutes. PCR was performed using 3 μl of the synthesized cDNA and 10 μM of primer. The basic conditions of the PCR were pre-denaturing phase: 94 ° C for 5 minutes, 30 cycles of denaturation phase: 94 ° C for 30 seconds, annealing phase: 72 ° C for 1 minute, post-elongation phase at 72 ° C for 5 minutes The annealing temperature was varied between 54-58 ℃ for each primer. Thereafter, the PCR product was loaded into 2% agarose gel in an amount of 7 μl, and then electrophoresed at 50 V.

As a result, it was confirmed that mRNA expression of AQP-3 was significantly increased in cells having high treatment concentration of sea urchin soybean extract (FIG. 2 (A)).

FIG. 2 is a graph showing the promotion rate of AQP-3 expression in keratinocytes (HaCaT) treated with 70% ethanol extract of sea cucumber bean. FIG. 2A is a result of measuring the mRNA expression level of AQP-3 . As a positive control, 10 nM retinoic acid (RA) was used.

Thus, it was confirmed that the seedling soybean extract of the present invention elevated mRNA expression of AQP-3.

 (2) Dairy bean  The extract AQP -3 on protein expression

In this example, the effect of soybean extract on the protein expression of AQP-3 was examined.

The keratinocyte (HaCaT) was inoculated into a petri dish for culture, and then cultured in an incubator at 37 ° C and 5% CO ₂ until the cells grew to about 80% of the petri dish area for culture. Then, the cells were treated for 24 hours at a concentration of 25, 50, and 100 μg / mL, respectively. As a control, buffers were treated with 10 nM retinoic acid (RA) as a positive control. Then, proteins in the cells were lysed using a lysis buffer, proteins were quantitatively analyzed using the protein BCA method, and 20 μg of the protein was subjected to SDS-PAGE. Then, the membrane was transferred using a PVDF membrane, and then the PVDF membrane was treated with a buffer diluted 1: 1000 with anti-AQP-3 for 24 hours. The levels of protein expression levels were quantified and are shown in Table 1 and Figure 2 below.

FIG. 2 is a graph showing the promotion rate of AQP-3 expression in human keratinocytes treated with 70% ethanol extract of sea cucumber bean. FIG. 2 (B) is a result of measuring the protein expression level of AQP- As a control, 10 nM retinoic acid (RA) was used.

Sample AQP -3 protein expression level ( % ) Control (buffer) 100 The positive control (10 nM Retinoic acid) 194.9 Daughter soybean extract 25 μg / mL 299.1 Dairy soybean extract 50 μg / mL 311.8 Daughter soybean extract 100 μg / mL 281.5

Surprisingly, the AQP-3 protein expression rate (194.9%) in cells treated with retinoic acid (RA), a positive control, in all cells treated with soybean extract (25, 50, 100 μg / (299.1, 311.8, 281.5%, respectively).

Thus, from the above results, it can be concluded that the extract of the present invention is significantly superior to the protein and mRNA of AQP-3 And the results showed that the extract of the seedlings of the present invention had excellent skin moisturizing effect.

Claims (12)

A cosmetic composition for improving skin wrinkles, which comprises as an active ingredient an extract of Canavalia lineata obtained by cold-extraction at room temperature using a 70% aqueous ethanol solution as an extraction solvent.
delete delete delete The method according to claim 1,
The above-mentioned Canavalia lineata extract,
Wherein the composition is contained in an amount of 0.00001-30.0% by weight based on the total weight of the cosmetic composition.
The method according to claim 1,
In the cosmetic composition,
Is any one selected from the group consisting of solutions, suspensions, emulsions, pastes, gels, creams, lotions, powders, soaps, surfactant-containing cleansing, oils, powder foundations, emulsion foundations, wax foundations and sprays Wherein the cosmetic composition is a cosmetic composition for improving skin wrinkles.
A cosmetic composition for skin moisturizing, characterized by containing as an active ingredient an extract of Canavalia lineata obtained by cold-extraction at room temperature using a 70% aqueous ethanol solution as an extraction solvent.
delete delete delete The method according to claim 1,
The above-mentioned Canavalia lineata extract,
A cosmetic composition for skin moisturizing, characterized in that it is contained in an amount of 0.00001-30.0% by weight based on the total weight of the cosmetic composition.
The method according to claim 1,
In the cosmetic composition,
Is any one selected from the group consisting of solutions, suspensions, emulsions, pastes, gels, creams, lotions, powders, soaps, surfactant-containing cleansing, oils, powder foundations, emulsion foundations, wax foundations and sprays Wherein the cosmetic composition is a cosmetic composition for skin moisturizing.

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