KR100893912B1 - Anti-wrinkle Cosmetics comprising Paeonia lactiflora and Gastrodia elata extracts as an effective component - Google Patents

Abstract

The present invention relates to a functional cosmetic for wrinkle improvement comprising peony and cheonma extract as an active ingredient,

Peony extract and cheonma extract is (a) preparing peony root and cheonma dry powder cut into sizes of 0.5 ~ 1.0cm, respectively; (b) adding 8-12 times distilled water to the peony root and cheonma dry powder, extracting the hydrothermal extract for 210 to 270 minutes and 450 to 510 minutes at a temperature of 90 to 97 ° C., and then filtering the filtrate with filtrate; (c) concentrating the filtrate under reduced pressure at a temperature of 50-55 ° C. and then cooling to room temperature; (d) adding ethanol such that the material produced in step (c) becomes an ethanol solution in a range of 70 to 80%; (e) leaving the material produced in step (d) in a range of 3 to 7 ° C. for 210 to 270 minutes, and then lyophilizing the filtrate; It is characterized in that it is manufactured through.

Peony, celestial horse, wrinkle, cosmetic

Description

Anti-wrinkle Cosmetics comprising Paeonia lactiflora and Gastrodia elata extracts as an effective component

The present invention relates to a functional cosmetic for wrinkle improvement comprising peony and cheonma extract as an active ingredient.

Skin aging can be largely divided into endogenous aging and photoaging, which is a skin aging phenomenon that can be prevented by avoiding exposure to ultraviolet rays. However, most of the functional cosmetics for improving wrinkles, including functional cosmetics that block skin aging by blocking ultraviolet rays, rely on imports of the raw materials, and thus, a large portion of the expenses are accounted for.

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However, peony, a natural herbaceous plant, is a high-value medicinal plant, and its main ingredient, peoniflorin, is known to have a central inhibitory effect, vasodilation, and antioxidant activity. In addition, in the case of cheonma as a herbal medicine that has been widely used for the treatment of bouts of affairs or to improve the blood symptoms, the cultivation method has been developed in recent years has a foundation that can be produced in large quantities.

An object of the present invention created to solve the above problems is as follows.

First, to provide a functional cosmetics for wrinkle improvement with the functionality of wrinkle improvement using peony and cheonma extract as an active ingredient.

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Second, through the study of bioactive new materials using natural materials in Korea, we will increase the economic utility value of natural resources and realize high added value of these resources.

In order to solve the above problems, the present invention provides a functional cosmetic for wrinkle improvement comprising peony and cheonma extract as an active ingredient.

Here, the peony extract and cheonma extract of the present invention comprises the steps of (a) preparing peony root and cheonma dry powder cut into 0.5 ~ 1.0cm size; (b) adding 8-12 times distilled water to the peony root and cheonma dry powder, extracting the hydrothermal extract for 210 to 270 minutes and 450 to 510 minutes at a temperature of 90 to 97 ° C., and then filtering the filtrate with filtrate; (c) concentrating the filtrate under reduced pressure at a temperature of 50-55 ° C. and then cooling to room temperature; (d) adding ethanol such that the material produced in step (c) becomes an ethanol solution in a range of 70 to 80%; (e) leaving the material produced in step (d) in a range of 3 to 7 ° C. for 210 to 270 minutes, and then lyophilizing the filtrate; It is manufactured through.

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According to the present invention as described above through the functional cosmetics for improving wrinkles, including peony and cheonma extract as a natural ingredient as an active ingredient, it is possible to realize a high value-added of these resources with an increase in economic utility value for domestic natural resources.

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The present invention is an anti-wrinkle functional cosmetics containing peony extract and cheonma extract as an active ingredient, look at the extraction method and embodiment of the peony and cheonma extract, with respect to the efficacy of the functional cosmetic for wrinkle improvement comprising the same This will be verified through an experimental example.

■ Manufacture of Peony and Cinnamon Extract

Peony is one of the five basic herbal medicines in Korea, along with Cheongung, Angelica, Chihwang, and Astragalus. Peonyniflorin, the main bioactive substance of peony root, is used as an indicator for evaluating the quality of peony. If peony is used as a material for food or cosmetics, hot water extraction should be used, which has many advantages in safety, extraction equipment, and manufacturing cost.

In the case of Chunma, gastrodin, benzyl alcohol, vanillin, etc., which are the main bioactive substances, are easily extracted from water, and thus hot water was extracted using these properties.

Here, peony extract and cheonma extract are prepared by (a) preparing peony root and cheonma dry powder cut into 0.5 ~ 1.0cm size; (b) adding 8-12 times distilled water to the peony root and cheonma dry powder, extracting the hydrothermal extract for 210-270 minutes and 450-510 minutes at a temperature of 90-97 ° C., and then filtering the filtrate with filtrate; (c) concentrating the filtrate under reduced pressure at a temperature of 50-55 ° C. and then cooling to room temperature; (d) adding ethanol such that the material produced in step (c) becomes an ethanol solution in a range of 70 to 80%; (e) leaving the material produced in step (d) in a range of 3 to 7 ° C. for 210 to 270 minutes, and then lyophilizing the filtrate; It is characterized in that it is manufactured through.

[ Example ]

1. Preparation of Peony Extract

In order to produce a peony extract, 3000 ml of distilled water was added to a sample cut into 300 g of peony root in a size of 0.5 cm to 1 cm according to an embodiment of the present invention, followed by attaching a reflux condenser and extracting hot water at 95 ° C. for 4 hours and filtering it. do. The filtrate is concentrated in a vacuum condenser until the total amount is 200ml at 50 ~ 55 ° C, cooled to room temperature, and ethanol is added to make 75% ethanol solution. Next, it is stirred and left for 24 hours at 5 ° C. to remove the resulting precipitate. By freeze-drying the filtrate thus obtained to produce a peony extract containing the useful material of the peony. (Figure 1).

2. Preparation of Cheonma Extract

Chunma extract was produced in the same order as in Fig. 1 as peony extract. Here, cheonma dry powder was used, and hot water was extracted for 8 hours using heat-stable properties compared to peony.

3. Extraction yield of peony and cheonma extract

Extraction yield of peony and cheonma extract obtained through the above embodiment is shown in Table 1. The final yield of peony and cheonma extract is 26.5% and 52.5%, respectively.

Table 1

300 g of sample extract Final yield (%) Volume (g) Brix Peony 2.890 2.75 26.5 Cheonma 2.790 5.64 52.5

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■ Anti-aging effect of wrinkles functional cosmetics

Skin aging is caused by activating collagenase, a type of matrix metalloproteinase (MMP), which destroys cell walls through the destruction of collagen (collagen) or extracellula matrix (ECM). In particular, ultraviolet rays reduce the amount of collagen and at the same time increase the amount of MMP-1 to form skin wrinkles (Fig. 2). Therefore, in the present invention, the degree of synthesis of collagen, which is the most important component of ECM, and the degree of synthesis of MMP-1 (collagenase-1) or MMP-13 (collagenase-3) using human epithelial cells (CCD986SK) and keratinocytes (HaCaT). In addition, after irradiation of UVA (long wavelength UV), the degree of MMPs activity of peony and cheonma extract were measured.

Before the experiment, human epithelial cells, CCD986SK and keratinocytes, HaCaT, were cultured under certain conditions.

Experimental Example 1 Collagen Synthesis Promoting Effect of Peony and Chunma Extract

1. Experimental method

In order to investigate the synergistic effect of collagen, the most important component of ECM, after treatment with CCD986SK cells and HaCaT cells, the peony and cheonma extract were treated, and after 24 hours, the supernatant was recovered and 200 μl was mixed with 1 ml of Sircol red solution. It was. The mixture was mixed once every 5 minutes while reacting at room temperature for 30 minutes and then centrifuged at 10,000 rpm for 10 minutes. This supernatant was discarded and the precipitate was dissolved in 1 ml of 0.4N NaOH and measured at an absorbance of 550 nm.

2. Experimental Results

As can be seen in Figures 3 and 4, it can be seen that the synthesis of collagen is accelerated when the peony and cheonma extract are treated.

Experimental Example 2 Collagen Synthesis Promoting Effect Using ELISA

1. Experimental method

Enzyme-linked immunosorbent assay (ELISA) was used to measure the effect of promoting collagen synthesis. That is, the CCD986SK cells were passaged in 48 well plates, treated with peony and cheonma extract, and after 24 hours, the supernatant was collected, and then, 100 µl was dispensed into 96 well plates, and the antigens were attached and washed for 24 hours. In addition, 100 μl of a primary antibody (procollagen type I N-terminal propeptide; PICP), which is used as an index of synthesis of type I collagen, was added to each well for 2 hours at room temperature and washed. After diluting the secondary antibody (anti-mouse), 100 μl of each well was reacted at room temperature for 2 hours, and then washed again to make TEM (3,3 ', 5,5'-Tetramethyl-benzidine; TMB) was aliquoted into each well. After the color reaction, 50N of 2N H ₂ SO _{4 was} added thereto to stop the substrate reaction. Absorbance was measured at 450 nm with ELISA reader.

2. Experimental Results

Treatment of peony and cheonma extracts on CCD986SK cells and the effect of promoting collagen synthesis by ELISA using PICP antibody were confirmed that the synthesis of collagen is promoted in peony and cheonma extract (FIG. 5).

Experimental Example 3 Transformation

1. Experimental method

In order to measure the effect of collagen alpha2 (I) gene (COL1A2) transcriptional activity of peony and spermatozoa extracts, the promoter of COL1A2 was vectored upstream (luciferase) vector (pGL3-promoter) containing luciferase. upstream) to generate recombinant plasmids and allow luciferase to be expressed depending on the activity of the COL1A2 promoter. After transforming pGL3-COL1A2 using Lipofectamine to CCD986SK cells, the CCD986SK cells were passaged in 48 well plates, and then treated with peony and cheonma extract, and incubated for 24 hours. Next, the lysate was made, and only the supernatant was taken to carry out a reporter gene assay to measure the activity of luciferase.

2. Experimental Results

It can be seen that the transcriptional activity of the COL1A2 promoter was increased in both the extract and the peony extract (FIG. 6).

Experimental Example 4 Inhibitory Effect of Peony and Chunma Extract on Collagenase Activity

1. Experimental method

To investigate the inhibitory effect of MMP-1 and MMP-13 on collagenase activity, which is the most important component of ECM, after passage of CCD986SK cells and HaCaT cells in 48 well plates, peony and sperm extracts were treated 12 hours before PBS. UVA was irradiated after washing once. After 24 hours, the supernatant was taken and centrifuged for 5 minutes at 10,000 rpm. Next, add 50 µl of the supernatant and 1 mM AMP (4-aminophenylmercuric acetate; APMA) to a 96 well plate, and then mix well. The MMP-1 enzyme activity was reacted at 37 ° C. for 6 hours and the MMP-13 enzyme activity. Reacted for 40 minutes at 37 ° C. Finally, after adding and mixing the substrate solutions corresponding to MMP-1 and MMP-13, the fluorescence was measured at an interval of 5 minutes for 1 hour at an excitation wavelength of 340 nm and an emission wavelength of 490 nm.

2. Experimental Results

As shown in Figures 7 and 8, it can be seen that the activity of MMP-1 and MMP-13 is inhibited in a concentration-dependent manner of peony and cheonma extract (Figs. 7 and 8).

In other words, the functional cosmetics for wrinkle improvement of the present invention can improve wrinkles through collagen synthesis promoting effect and collagenase activity inhibiting effect.

While the invention has been described in connection with the preferred embodiment as mentioned above, various modifications and variations are possible without departing from the spirit of the invention. Therefore, the claims of the present invention include modifications and variations that fall within the true scope of the invention.

1 is a flow chart showing an embodiment of the preparation of peony and cheonma extract.

2 is a conceptual diagram showing the skin aging process.

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3 and 4 are graphs showing collagen synthesis promoting efficacy of peony and cheonma extract using CCD986SK cells and HaCaT cells, respectively.

Figure 5 is a graph showing the collagen synthesis promoting efficacy of peony and cheonma extract by ELISA.

Figure 6 is a graph showing the effect of peony and cheonma extract on luciferase activity.

7 and 8 are graphs showing the collagenase activity inhibitory effect of peony and cheonma extract using CCD986SK cells and HaCaT cells.

Claims (7)

To include peony extract and cheonma extract as an active ingredient, The peony extract is (a) cutting the peony muscle to a size of 0.5-1.0 cm; (b) adding 8-12 times distilled water to the peony root, extracting the hydrothermal solution for 210-270 minutes at a temperature of 90-97 ° C., and then filtering the filtrate with filtrate; (c) concentrating the filtrate under reduced pressure at a temperature of 50-55 ° C. and then cooling to room temperature; (d) adding ethanol such that the material produced in step (c) becomes an ethanol solution in a range of 70 to 80%; And, (e) leaving the material produced in step (d) in a range of 3 to 7 ° C. for 210 to 270 minutes, and then lyophilizing the filtrate; Functional cosmetics for wrinkle improvement comprising peony and cheonma extract as an active ingredient, characterized in that it is manufactured through. In claim 1, The cheonma extract is (a) preparing a dry horse powder; (b) adding 8-12 times distilled water to the cheonma dry powder, extracting the hot water for 450-510 minutes at a temperature of 90-97 ° C., and then filtering the filtrate with filtrate; (c) concentrating the filtrate under reduced pressure at a temperature of 50-55 ° C. and then cooling to room temperature; (d) adding ethanol such that the material produced in step (c) becomes an ethanol solution in a range of 70 to 80%; And, (e) leaving the material produced in step (d) in a range of 3 to 7 ° C. for 210 to 270 minutes, and then lyophilizing the filtrate; Functional cosmetics for wrinkle improvement comprising peony and cheonma extract as an active ingredient, characterized in that it is manufactured through. delete delete delete delete delete

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