KR102114682B1 - Wrinkle improvement cosmetic containing ginsenoside Rg3, Rg5 and Rk1 - Google Patents

Abstract

The present invention relates to a cosmetic composition for improving wrinkles containing ginsenosides Rg3, Rg5, and Rk1, more specifically, containing ginsenosides Rg3, Rg5, and Rk1, wherein the water-soluble Rg3 (S) isomer is fat-soluble Rg3 (R ) It is related to a cosmetic composition that is more effective in improving wrinkles caused by stress stimulation by ultraviolet rays and infrared rays, characterized in that it contains more than isomers.
According to the present invention, ginsenosides can be separated and purified by an enzymatic conversion method rather than a normal boiling or boiling method to include three or more times the water-soluble Rg3 (S) isomer than the fat-soluble Rg3 (R) isomer. In addition, the cosmetic composition for improving wrinkles containing ginsenosides Rg3, Rg5, and Rk1 of the present invention inhibits skin aging by inhibiting 11β-HSD1, a stress hormone related to skin aging due to photoaging, and MMP-1, a matrix proteolytic enzyme. It can have a preventive or improvement effect.

Description

Wrinkle improvement cosmetic containing ginsenoside Rg3, Rg5 and Rk1

The present invention relates to a cosmetic composition for improving wrinkles containing ginsenosides Rg3, Rg5, and Rk1, more specifically, containing ginsenosides Rg3, Rg5, and Rk1, wherein the water-soluble Rg3 (S) isomer is fat-soluble Rg3 (R ) It is related to a cosmetic composition that is more effective in improving wrinkles caused by stress stimulation by ultraviolet rays and infrared rays, characterized in that it contains more than isomers.

Physical stimulation from the outside causes a physiological stress response in the body by stress hormones such as the brain and corticotrophin-releasing hormone (CRH), glucocorticoids, and epinephrine. Skin is a major sensory organ that senses stress from external irritants, and the representative irritant is ultraviolet rays, and skin that is frequently exposed to ultraviolet rays causes initial skin aging. In addition, ultraviolet rays repeatedly activate stress hormones, causing harm to the skin. This leads to severe skin side effects such as disruption of dermal fibrous tissue, as in the case of excessive application of steroids to the skin. UV stimulation is known to be the leading cause of glucocorticoids activity, such as cortisol, in human skin, and, in addition, increases the expression of 11β-hydroxysteroid dehydrogenase type 1 (11β-HSD1), an enzyme in the human body that activates cortisol. The increased activity and expression of 11β-HSD1 by UV light is closely related to skin aging, and thus has been noted as a new target for suppressing skin aging.

Wrinkles and skin elasticity, skin sagging and dryness appearing on aged skin are mostly due to changes in matrix proteins present in the dermis. Since the dermis plays a role in supporting the strength and shape of the skin within the skin, if morphological changes occur in this area as aging progresses, it plays a decisive role in wrinkle formation and skin sagging. Matrix metalloproteinases (MMPs) are representative enzymes that degrade these matrix proteins, and MMPs activity in the skin is increased even after a single UV irradiation, and it is a major factor in promoting skin aging by significantly degrading collagen and other matrix proteins in the skin. do.

Recently, as a new point of view on the cause of photoaging, research results have been published that the heat generated by infrared rays affects skin aging. The wavelength range of infrared rays is 750 nm to 1 mm, of which near infrared rays, which occupy 750 to 3000 nm, are converted into thermal energy when reaching the skin, thereby increasing the skin temperature. For example, when heat stimulation is applied to skin cells, expression of matrix metalloproteinase (MMP), an enzyme that breaks down collagen, increases, and collagen synthesis decreases.

In the meantime, as a material for wrinkle improvement, vitamin C, α-tocopherol, retinol, and derivatives thereof have been used in cosmetics and pharmaceuticals, but these are disadvantageous in that odor or chemical stability is poor when formulated in formulations. have.

In order to overcome these drawbacks, the development of a cosmetic composition for wrinkle improvement using materials derived from natural products has recently been spotlighted and requires its discovery.

Thus, the present inventors tried to overcome the problems of the prior arts, and as a result of researching hard to find out that the role of 11β-HSD1 activated by ultraviolet rays and infrared rays is related to photoaging, and to discover substances that improve them, anti-aging Compared and selected based on the expected ginsenosides, the mixture of ginsenosides Rg3, Rg5, and Rk1 inhibits the increase in 11β-HSD1 and MMP-1 expression, thereby preventing or improving skin aging. Upon confirmation, the present invention has been completed.

Therefore, the main object of the present invention is to suppress wrinkles caused by stress stimulation by ultraviolet and infrared rays by inhibiting the expression of 11β-HSD1 and the expression of MMP-1, ginsenosides Rg3, Rg5, which can effectively improve wrinkle formation. And it is to provide a cosmetic composition for improving wrinkles containing Rk1.

Another object of the present invention is to provide a method for separating and purifying ginsenosides that can selectively include more water-soluble Rg3 (S) isomers than fat-soluble Rg3 (R) isomers.

According to one aspect of the present invention, in the cosmetic composition containing ginsenosides Rg3, Rg5, and Rk1, wrinkle improvement characterized in that the water-soluble Rg3 (S) isomer is contained more than the fat-soluble Rg3 (R) isomer. Provide a cosmetic composition for.

In the present invention, the wrinkles are characterized by being caused by stress stimulation by ultraviolet rays and infrared rays. The term stress stimulation of the present invention refers to that harmful stimulation by internal and external elements causes wrinkles by increasing stress hormone (glucocorticoid) in the skin. In the present invention, it means that the stress is stimulated by ultraviolet rays and infrared rays.

Ginsenoside refers to an active ingredient extracted from ginseng and the like, and has been known to have anti-cancer, atopic, anti-inflammatory, and anti-aging effects. However, as in the present invention, the ginsenoside Rg3, Rg5, and Rk1 complex is caused by ultraviolet and infrared rays. There is no known effect on wrinkle improvement caused by stress stimulation.

In the present invention, the ginsenoside is characterized by being obtained by enzymatic decomposition of ginseng or red ginseng extract. In the present invention, ginsenosides can be separated and purified by an enzymatic conversion method rather than a conventionally known boiling or boiling method to include three or more times the water-soluble Rg3 (S) isomer than the fat-soluble Rg3 (R) isomer. Preferably, the enzymatic decomposition is characterized in that it is decomposed with beta-glycosidase (β-glycosidase) derived from Aspergillus sp.

In the present invention, the ginseng or red ginseng extract can be extracted with any solvent used conventionally, preferably methanol, ethanol, propyl alcohol, butyl alcohol, glycerin, propylene glycol, butylene of water, anhydrous or hydrous Characterized in that it is extracted with one or more solvents selected from the group consisting of glycols, or mixed solvents thereof.

In the present invention, the extract can be extracted by any extraction method conventionally used, and the extraction method using an extraction solvent is a conventional plant extraction method, for example, hot water extraction, reflux cooling extraction, ultrasonic extraction, super Methods such as critical extraction can be used. The above-described extract may be in the form of a concentrate or in the form of lyophilized powder.

In the present invention, the ginsenoside is characterized in that it is obtained by passing the result of the enzymatic decomposition through a column filled with a methacrylic resin to remove the passing liquid and then desorbing it with ethanol. In the present invention, a complex of ginsenosides Rg3, Rg5, and Rk1 can be obtained by enzymatic decomposition and separation and purification through a methacrylic column.

In the present invention, the complex of ginsenoside Rg3, Rg5, and Rk1 is characterized by containing 0.01 to 10% by weight based on the total weight of the composition. If it is less than 0.01% by weight in the above range, it is not preferable because there is a fear that the photoaging inhibitory effect will be weak, and conversely, exceeding 10% by weight may not be desirable in terms of economy.

In the present invention, the composition is characterized by having a wrinkle improvement effect by inhibiting 11β-HSD1 (11β-hydroxysteroid dehydrogenase type 1) and MMP-1 (Matrix metalloproteinase-1) activity.

According to the experimental example of the present invention, UVB or IRA was irradiated, and it was confirmed that the group treated with the ginsenoside Rg3, Rg5, and Rk1 complex inhibited the activity of 11β-HSD1 and MMP-1 than the group not treated with the group. . As a result of this, the ginsenoside Rg3, Rg5, Rk1 complex of the present invention can exhibit wrinkle improvement effect by inhibiting the expression of MMP-1 at the same time as relieving stress in the skin by inhibiting 11β-HSD1 activity Can be seen.

In the present invention, the composition may be prepared in any formulation conventionally prepared in the art, preferably skin lotion, skin softener, skin toner, astringent, lotion, milk lotion, moisture lotion, nutrition lotion, massage It is characterized by being one or more formulations selected from formulations consisting of cream, nourishing cream, moisturizing cream, hand cream, foundation, essence, nourishing essence, pack, soap, cleansing foam, cleansing lotion, cleansing cream, body lotion and body cleanser.

As described above, according to the present invention, ginsenosides can be separated and purified by an enzymatic conversion method rather than a normal boiling or boiling method, so that the water-soluble Rg3 (S) isomer can be included three times more than the fat-soluble Rg3 (R) isomer. In addition, the cosmetic composition containing ginsenosides Rg3, Rg5, Rk1 of the present invention as an active ingredient effectively suppresses the expression of 11β-HSD1 and MMP-1 over-expressed by ultraviolet rays and infrared rays to relieve stress in the skin and wrinkles It shows an excellent effect in improving.

1 is a result of Experimental Example 1 measuring the effect on ginsenoside Rg3, Rg5, Rk1 treatment on 11β-HSD1 expression in human fibroblasts exposed to UVB.
2 is a result of Experimental Example 1 measuring the effect on cortisol expression when ginsenosides Rg3, Rg5, and Rk1 are treated on human fibroblasts exposed to UVB.
3 is a result of Experimental Example 1, which measured the effect of ginsenosides Rg3, Rg5, and Rk1 on MMP-1 expression in human fibroblasts exposed to UVB.
4 is a result of Experimental Example 1 measuring the effect on ginsenoside Rg3, Rg5, Rk1 treatment on type 1 procollagen expression in human fibroblasts exposed to UVB.
5 is a result of Experimental Example 2, which measured the effect on ginsenoside Rg3, Rg5, and Rk1 treatment on 11β-HSD1 expression in human fibroblasts exposed to IRA.
6 is a result of Experimental Example 2 measuring the effect on cortisol expression when ginsenosides Rg3, Rg5, and Rk1 are treated on human fibroblasts exposed to IRA.
7 is a result of Experimental Example 2, which measured the effect of ginsenosides Rg3, Rg5, and Rk1 on MMP-1 expression in human fibroblasts exposed to IRA.
8 is a result of Experimental Example 2, which measured the effect on ginsenoside Rg3, Rg5, and Rk1 treatment on type 1 procollagen expression in human fibroblasts exposed to IRA.
9 is a chromatograph obtained by testing the sample of Example 1 according to the HPLC method.

Hereinafter, the present invention will be described in more detail through examples. Since these examples are only for illustrating the present invention, the scope of the present invention is not to be construed as being limited by these examples.

Example 1: Preparation of samples

1 kg of ginseng or red ginseng is put into 10 L of 50% alcohol (ethanol) solution, extracted (80 ° C, 48 hours), the extract is separately taken, and the alcohol is removed using a vacuum concentrator, then concentrated to a water content of 35%, total 492 g. After dissolving the concentrate (492 g) in a 10 L buffer solution (Acetic aicd-Na ₂ HPO ₄ buffer, pH 3.0, 100 mM), Aspergillus sp . After the resulting β-glycosidase was added and reacted at 70 ° C., the supernatant was passed through a column filled with 2L of methacrylic resin (Diaion HP2MG) to remove the passing solution, and then desorbed with 10L ethanol to concentrate the alcohol (ethanol) layer under reduced pressure. To obtain 51 g of the powder shown in Table 1 (ginsenoside Rg3, Rg5, Rk1, etc.). The obtained extract powder was dissolved in DMSO (Dimethyl sulfoxide) at a concentration of 0.1 μg / ml and 1 μg / ml as a solubilizer, and used in the following experimental examples.

In addition, after dissolving the powder in methanol, a stainless column filled with octadecylsilylated silica gel was tested according to the HPLC method at a flow rate of 1.0 mL / min with water: acetonitrile = 18: 20 to 15: 85, a gradient mobile phase. A chromatograph as shown in FIG. 9 was obtained, and the content (%) of each ginsenoside shown in Table 2 was calculated by calculating as follows.

[Calculation process]

As shown in Table 2, the sample according to Example 1 contains ginsenosides Rg3, Rg5, and Rk1, but ginsenoside Rg3 (S) has three times more content (%) than ginsenoside Rg3 (R). You can see a lot of the above.

Experimental Example 1: Ginsenoside Rg3, Rg5, Rk1 of UVB by photo-aging stress suppression effect

Culture of 6 well cells of human fibroblasts to confirm the inhibitory effect of 11β-HSD1 and MMP-1 messenger RNA expression induced by UVB of ginsenosides Rg3, Rg5, and Rk1 in human fibroblasts using Example 1 prepared above. The plate was inoculated at a density of 4X10 ⁵ and then incubated for 24 hours in a 37 ° C, 5% CO ₂ incubator. After irradiating UVB with 15mJ, extract or 11β-HSD1 inhibitor PF915275 (hereinafter, PF) was added as a control, incubated for 24 hours, and cells were recovered, and 1 ml of trizol (RNA iso, DAKARA, Japan) was added to isolate RNA. . After quantifying RNA at 260 nm using an ultraviolet detector, cDNA was synthesized (Reverse Transcriptase Mix, ELPIS biotech, Korea). RT-PCR was assayed by using a PCR machine (Step One Plus, Applied Biosystems, USA) and adding Cyburgrin (SYBRGreen supermix, Applied Biosystems, USA) with 11β-HSD1 primer and cDNA. Primer and reaction conditions were as shown in Table 3 below, and the expression levels of 11β-HSD1 and MMP-1 genes were corrected for β-actin genes.

In addition, in order to confirm the effect of inhibiting the expression of cortisol induced by UVB of ginsenosides Rg3, Rg5, and Rk1 in human fibroblasts using the prepared Example 1 and type 1 procollagen expression inhibitory effect, human fibroblasts were placed in a 6-well cell culture dish. After inoculation at a density of 4X10 ⁵ , cultured for 24 hours in a 37 ° C, 5% CO ₂ incubator. After irradiating UVB with 15 mJ, the extract and PF, an 11β-HSD1 inhibitor, were added as a control to further culture for 24 hours and to collect the medium. The recovered medium was centrifuged at 12,000 rpm for 10 minutes to remove impurities and cortisol ELISA (Enzyme-linked immunosorbent assay) kit (R & D system, USA) and type 1 procollagen EIA (Enzyme immuno assay) kit (DAKARA, Japan) were used. By essay.

As a result, as shown in Figures 1 to 4, ginsenosides Rg3, Rg5, Rk1 concentration-dependently increased by UVB 11β-HSD1, cortisol, inhibition of MMP-1 and the recovery efficacy of type 1 procollagen was confirmed. In the figure, (-) is not irradiated with UVB, (+) is irradiated with UVB, PF is irradiated with UVB and PF is added, Ginsenoside is irradiated with UVB, and then ginsenosides Rg3, Rg5, Rk1 It is added.

Experimental Example 2: Ginsenoside Rg3, Rg5, Rk1 by IRA effect of photoaging stress suppression

Using the prepared Example 1 to describe the mRNA expression inhibitory effect of 11β-HSD1 and MMP-1 induced by IRA of ginsenosides Rg3, Rg5, Rk1 in human fibroblasts described in Experimental Example 1 above IRA was investigated by culturing human fibroblasts in the same manner as the method, and after adding an experimental sample (Example 1) or PF, the cells were further cultured for 24 hours to confirm the expression levels of 11β-HSD1 and MMP-1 messenger RNA. The primers and reaction conditions are as shown in Table 3 above, and the expression levels of 11β-HSD1 and MMP-1 genes were corrected for β-actin genes.

In addition, in order to confirm the inhibitory effect of ginsenoside Rg3, Rg5, Rk1 induced cortisol and type 1 procollagen expression, human fibroblasts were cultured in the same manner as described in Experimental Example 1 above to investigate IRA. After the experimental sample (Example 1) or PF was added, the cells were further cultured for 24 hours to confirm the cortisol and type 1 procollagen protein expression levels.

As a result, as shown in FIGS. 5 and 7, ginsenosides Rg3, Rg5, Rk1 showed 11β-HSD1 and MMP-1 messenger RNA inhibitory effects, and cortisol inhibition and type 1 procollagen as shown in FIGS. 6 and 8. It showed recovery efficacy. In the figure, (-) is not irradiated with IRA, (+) is irradiated with IRA, PF is irradiated with IRA, PF is added, Ginsenoside is irradiated with UVB, and ginsenosides Rg3, Rg5, Rk1 are added. Did.

Formulation Example 1: Preparation of toner

The toner agent containing the material obtained in Example 1 was prepared according to the compositional components and compositional ratios shown in Table 4 below.

Formulation Example 2: Preparation of lotion agent

The lotion agent containing the material obtained in Example 1 was prepared according to the compositional components and composition ratios shown in Table 5 below.

Formulation Example 3: Preparation of cream

The cream containing the material obtained in Example 1 was prepared by a conventional method according to the composition components and composition ratios shown in Table 6 below.

Claims (6)

In the cosmetic composition containing ginsenosides Rg3, Rg5, and Rk1, the ginsenoside is enzymatically decomposed ginseng or red ginseng extract with beta-glycosidase (β-glycosidase) derived from Aspergillus sp. A cosmetic composition for improving wrinkles, which is obtained and contains more water-soluble Rg3 (S) isomer than fat-soluble Rg3 (R) isomer.
delete delete The cosmetic composition for improving wrinkles according to claim 1, wherein the ginsenoside is obtained by passing a product of the enzymatic decomposition through a column filled with a methacrylic resin to remove a passing liquid and then desorbing it with ethanol.
The cosmetic composition for improving wrinkles according to claim 1, wherein the cosmetic composition has a wrinkle improvement effect by inhibiting 11β-HSD1 (11β-hydroxysteroid dehydrogenase type 1) and MMP-1 (Matrix metalloproteinase-1) activity. The method of claim 1, wherein the cosmetic composition is a skin lotion, skin softener, skin toner, astringent, lotion, milk lotion, moisture lotion, nutrition lotion, massage cream, nutrition cream, moisture cream, hand cream, foundation, essence, nutrition essence , Pack, soap, cleansing foam, cleansing lotion, cleansing cream, body lotion, and a cosmetic composition for improving wrinkles, characterized in that it is one formulation selected from the formulation consisting of a body cleanser.

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