Cosmetic composition for improving skin aging

Abstract

This invention relates to a composition containing protoplasts of ginseng as an active ingredient, which improves aging caused by the deepening of wrinkles or loss of firmness in the skin. Specifically, the present invention contains protoplasts of ginseng as an active ingredient and can improve aging caused by deep wrinkles or loss of firmness of the skin. Furthermore, by including one or more components selected from the group consisting of phloretin, oryzanol, tranexamic acid, and pyridoxine, the stability of the composition can be enhanced and the effect of improving skin wrinkles and firmness can be strengthened by suppressing the clumping or cell membrane defects of the ginseng protoplasts in the composition.

Description

Technical Field

[0001] The present invention relates to a composition containing ginseng protoplasts as an active ingredient and capable of improving aging due to deep wrinkles or loss of firmness of the skin.

Background Art

[0002] The skin is in direct contact with the external environment and functions to protect against temperature, humidity, antigens, ultraviolet rays, etc. However, due to physical and chemical stimuli such as external pollutants, ultraviolet rays, and stress, the function of the skin deteriorates. With aging, cell proliferation and immune activity decrease, and damage to skin cells cannot be quickly prevented, leading to an overall deterioration of skin components, resulting in the appearance of skin wrinkles, lack of firmness, and keratinization.

[0003] Skin aging can be broadly divided into intrinsic aging and extrinsic aging. Intrinsic aging is natural aging, which clinically features loss of firmness, roughening of skin texture, the appearance of deep wrinkles, and pigmentation. Extrinsic aging is an aging phenomenon caused by external environmental factors such as ultraviolet rays, reactive oxygen species, and stress. Recently, with the rapid industrialization, the impact of powerful oxidants generated by air pollution and increased UV due to ozone layer depletion has been increasing.

[0004] Oxidants such as these reactive oxygen species induce the destruction of connective tissues such as skin collagen, inhibit the function of cell membranes, promote DNA mutations, denature proteins, and deform cell signal transmitters in humans, inducing the formation of skin wrinkles, pigmentation, etc., and causing skin aging. To prevent such skin damage, cosmetics or skin protectants containing various skin protection components such as retinol, retinoids, vitamin C, flavonoids, tocopherol, and coenzyme Q10 have been developed.

[0005] Recently, in order to develop cosmetics that are relatively less toxic and environmentally friendly, natural ingredients are being extracted or fermented to search for bioactive substances with properties such as whitening, anti-aging, anti-wrinkle, antioxidant, and anti-inflammatory effects, and these are then used as ingredients in cosmetics. [Overview of the Initiative] [Problems that the invention aims to solve]

[0006] The object of the present invention is to provide a cosmetic composition containing protoplasts of ginseng as an active ingredient that can improve aging caused by deep wrinkles or loss of firmness of the skin.

[0007] Another object of the present invention is to provide a cosmetic composition that can be used with ginseng protoplasts together with phloretin, oryzanol, tranexamic acid, or pyridoxine to further enhance skin-improving effects. [Means for solving the problem]

[0008] To achieve the above objective, the present invention provides a cosmetic composition containing protoplasts of ginseng.

[0009] In one embodiment of the present invention, the protoplasts of the ginseng species are derived from ginseng or wild ginseng, preferably from wild ginseng.

[0010] In one embodiment of the present invention, a cosmetic composition containing the protoplasts of the ginseng species exhibits an effect of improving skin wrinkles or firmness.

[0011] In one embodiment of the present invention, the composition may further contain one or more components selected from the group consisting of phloretin, oryzanol, tranexamic acid, and pyridoxine. The composition according to the present invention, further containing phloretin, oryzanol, tranexamic acid, or pyridoxine, exhibits the effect of improving skin wrinkles or firmness.

[0012] Another object of the present invention is to provide a method for improving skin condition, comprising the step of treating the skin with protoplasts of ginseng.

[0013] Another object of the present invention is to provide a method for improving skin wrinkles or firmness, comprising the step of treating the skin with protoplasts of ginseng.

[0014] Another object of the present invention is to provide uses for ginseng protoplasts for producing cosmetic compositions that improve skin condition.

[0015] Another object of the present invention is to provide uses for protoplasts of ginseng species for producing cosmetic compositions that improve skin wrinkles or firmness.

[0016] Another object of the present invention is to provide a method for improving the condition of the skin, comprising the step of treating the skin with protoplasts of ginseng; and one or more components selected from the group consisting of phloretin, oryzanol, tranexamic acid, and pyridoxine.

[0017] Another object of the present invention is to provide a method for improving skin wrinkles or firmness, comprising the step of treating the skin with protoplasts of ginseng; and one or more components selected from the group consisting of phloretin, oryzanol, tranexamic acid, and pyridoxine.

[0018] Another object of the present invention is to provide uses for protoplasts of ginseng and one or more components selected from the group consisting of phloretin, oryzanol, tranexamic acid, and pyridoxine for producing cosmetic compositions that improve skin condition.

[0019] Another object of the present invention is to provide uses for protoplasts of ginseng and one or more components selected from the group consisting of phloretin, oryzanol, tranexamic acid, and pyridoxine, for producing cosmetic compositions for improving skin wrinkles or firmness.

[0020] In one specific embodiment, the composition can suppress the formation of fine wrinkles, and for example, by suppressing the expression of BNC2, a gene associated with the formation of fine wrinkles, it can show an effect of improving skin wrinkles or firmness.

[0021] In one specific embodiment, the composition can suppress deep wrinkles, and for example, by promoting the expression of PPARGC1B, a gene that suppresses deep wrinkles, it exhibits an effect of improving skin wrinkles or firmness.

[0022] In one specific embodiment, the composition can promote collagen formation, for example, by promoting the expression of COL1A1 and COL3A1, which are genes associated with collagen formation, thereby showing an effect of improving skin wrinkles or firmness.

[0023] In one embodiment of the present invention, the protoplasts of the ginseng species may be present in an amount of 0.00001 to 1% by weight, for example, 0.00001 to 0.005% by weight, 0.00005 to 0.01% by weight, 0.0001 to 0.05% by weight, 0.0002 to 0.1% by weight, 0.0004 to 0.5% by weight, 0.0005 to 1% by weight, 0.1 to 1% by weight, or 0.5 to 1% by weight.

[0024] In this invention, the term "protoplast" refers to the protoplasm enclosed in a cell membrane, obtained by removing the cell wall from bacteria or plant cells, and by decomposing or removing the cell wall components with an enzyme-like substance. Using protoplasts allows for achieving skin improvement effects that differ from those achieved by using extracts of the protoplast material (ginseng in this invention) or the protoplast material itself.

[0025] In this invention, the term "phloretin" refers to a dihydrochalcone compound, a polyphenol compound reported to be present not only in apples but also in pears and other fruits, primarily found in the peel rather than the flesh. It is believed to have effects such as skin whitening, anti-inflammatory properties, and UV protection.

[0026] The term "oryzanol" in the present invention is a mixture of ferulic acid esters of sterols and triterpene alcohols. Gamma oryzanol can be obtained from rice bran or the aqueous ethanol fraction after decomposition with hexane or acetone from the endosperm using aqueous ethanol at room temperature. It is said to have effects such as alleviating menopausal symptoms.

[0027] The term "tranexamic acid" in the present invention does not directly participate in the process of melanin production, but serves to block the process in which plasminogen, which is involved in intercellular signal transduction outside melanocytes, changes into plasmin. It suppresses the stimulation of melanocytes and reduces the activity of tyrosinase. Anti-inflammatory, anti-allergic, and hemostatic effects have also been reported.

[0028] The term "pyridoxine" in the present invention is one of the vitamins belonging to the vitamin B group. It acts in the metabolic processes of carbohydrates and proteins, produces neurotransmitters, and is useful for the production of hemoglobin and antioxidant effects. When pyridoxine is deficient, symptoms such as dermatitis, angular cheilitis, stomatitis, interstitial or peripheral neuropathy, nausea, dizziness, depression, and anemia are known to appear.

[0029] In one embodiment of the present invention, one or more components selected from the group consisting of phloretin, oryzanol, tranexamic acid, and pyridoxine may be contained at 0.00001 to 1% by weight. For example, it may be 0.00001 to 0.00005% by weight, 0.00005 to 0.0001% by weight, 0.0001 to 0.0005% by weight, 0.0005 to 0.001% by weight, 0.001 to 0.005% by weight, 0.005 to 0.01% by weight, 0.01 to 0.05% by weight, 0.05 to 0.1% by weight, 0.1 to 0.5% by weight, or 0.5 to 1% by weight.

[0030] Within the relevant range, the effect of preventing protoplast formation or cell membrane defects in ginseng is the most excellent, which results in the most excellent synergistic effect of the composition on improving skin wrinkles or firmness, and may also increase the stability of the composition.

[0031] The present invention also relates to protoplasts of ginseng; and The present invention provides a cosmetic composition comprising one or more components selected from the group consisting of phloretin, oryzanol, tranexamic acid, and pyridoxine.

[0032] Protoplasts are affected by many components (emulsifiers, polyols, oils, etc.) contained in cosmetic compositions and can easily rupture or deform. (See Korean Registered Patent No. 10-2243502) In the composition according to the present invention, the combination with one or more of the above components can prevent clumping or cell membrane defects of ginseng protoplasts, thereby increasing the stability of ginseng protoplasts. Cell membrane defects refer to phenomena such as the cell membrane of ginseng protoplasts cracking, rupturing, or being destroyed by physical or chemical action, causing the ginseng protoplasts to deviate from their initial or normal state.

[0033] Furthermore, combining it with the aforementioned components can synergistically enhance the skin wrinkle-reducing or firmness-improving effects of ginseng protoplasts.

[0034] In one embodiment of the present invention, the mass ratio of ginseng protoplasts to components in the composition may be 1:0.0001 to 1:1, for example, 1:0.0001 to 1:0.001, 1:0.001 to 1:0.01, or 1:0.01 to 1:1.

[0035] In one embodiment of the present invention, the protoplasts of the ginseng species; and A cosmetic composition containing one or more ingredients selected from the group consisting of phloretin, oryzanol, tranexamic acid, and pyridoxine exhibits an effect of improving skin wrinkles or firmness.

[0036] In one specific embodiment, the composition can suppress the formation of fine wrinkles, and for example, it exhibits an effect of improving skin wrinkles or firmness by suppressing the expression of BNC2, a gene associated with the formation of fine wrinkles.

[0037] In one specific embodiment, the composition can suppress deep wrinkles, and for example, by promoting the expression of PPARGC1B, a gene that suppresses deep wrinkles, it exhibits an effect of improving skin wrinkles or firmness.

[0038] In one specific embodiment, the composition can promote collagen formation, and for example, by promoting the expression of COL1A1 and COL3A1, which are genes associated with collagen formation, it can show an effect of improving skin wrinkles or firmness. [Effects of the Invention]

[0039] The present invention provides a composition containing ginseng protoplasts as an active ingredient that can improve aging caused by deep wrinkles or loss of firmness in the skin. Furthermore, by including one or more components selected from the group consisting of phloretin, oryzanol, tranexamic acid, and pyridoxine, the stability of the composition can be increased and the effect of improving skin wrinkles and firmness can be enhanced by suppressing the clumping or cell membrane defects of the ginseng protoplasts in the composition. [Brief explanation of the drawing]

[0040] [Figure 1] This photograph shows the morphological maintenance efficacy of protoplasts when ginseng protoplasts are combined with phloretin, oryzanol, tranexamic acid, and / or pyridoxine according to the present invention. [Figure 2] This photograph shows the shape-retaining efficacy of protoplasts in a cosmetic formulation combining ginseng protoplasts with phloretin, oryzanol, tranexamic acid and / or pyridoxine according to the present invention. [Modes for carrying out the invention]

[0041] Since the present invention is subject to various modifications and can take on various forms, the specific embodiments and descriptions described below are for the sole purpose of aiding the understanding of the invention and do not limit the invention to any particular form of disclosure. The scope of the present invention should be understood to include all modifications, equivalents, or substitutes that fall within the spirit and technical scope of the invention. [Examples]

[0042] The present invention will be described in more detail below through examples and comparative examples. These examples are solely for the purpose of illustrating the present invention, and therefore the scope of the invention is not limited by these examples.

[0043] Experimental Example 1. Regulatory function of wrinkle-related genes in protoplasts of ginseng. To confirm whether the protoplasts of ginseng according to the present invention can regulate wrinkle-related genes, we measured the inhibitory or promoting activity of BNC2, a gene associated with fine wrinkle formation; PPARGC1B, a gene associated with deep wrinkle formation; and COL1A1 and COL3A1, genes associated with collagen formation.

[0044] [Cell culture] Human dermal fibroblasts (Hs68) were purchased from ATCC. The purchased cells were cultured in a medium prepared by mixing DMEM (Dulbecco Modified Eagle Medium, Thermo Fisher Scientific) with 10% FBS (Fetal Bovin Serum) and 1% Antibiotics (Penicillin streptomycin) in a 37°C, 5% CO2 incubator.

[0045] [Analysis of gene expression levels] To analyze the activity of BNC2, PPARGC1B, COL1A1, and COL3A1 mRNA, human dermal fibroblasts were dispensed into 6-well plates at a rate of 1.5 × 10⁵ cells / well and cultured for 24 hours at 37°C in a 5% CO₂ incubator. Subsequently, the culture medium was changed to serum-free media containing 0.01% (wt%) of ginseng protoplasts, and the cells were cultured under the same conditions for another 24 hours. After that, mRNA was obtained using an RNA-spin total RNA extraction kit (Intron Biotechnology, 17211), and cDNA was synthesized using a cDNA synthesis kit (Philekorea Technology, ET21100). The expression levels of each gene were analyzed by qPCR using the synthesized cDNA and taqman probe.

[0046] When the culture medium was changed to serum-free media that does not contain the active ingredient, this was set as the negative control group. The results of the negative control group were set to 100%, and the relative increase or decrease (percentage increase or decrease) of the results of the evaluated group was calculated and is shown in Table 1 below.

[0047] [Table 1]

[0048] As shown in Table 1, it was confirmed that wild ginseng protoplasts reduced the expression of a gene (BNC2) associated with the formation of fine wrinkles, while promoting the expression of a gene (PPARGC1B) associated with the formation of deep wrinkles, a gene (COL1A1) associated with the formation of collagen type 1, and a gene (COL3A1) associated with the formation of collagen type 3.

[0049] Therefore, wild ginseng protoplasts can be used as an excellent anti-aging material.

[0050] Experimental Example 2. Regulatory Function of Wrinkle-Related and Collagen-Related Genes by Types of Korean Ginseng To compare the wrinkle-related gene and collagen-promoting regulatory functions of protoplasts depending on the type of ginseng according to the present invention, examples with the compositions shown in Table 2 below were prepared.

[0051] [Table 2]

[0052] Next, we measured the inhibitory or promoting activity of BNC2, a gene associated with the formation of fine wrinkles, PPARGC1B, a gene associated with the formation of deep wrinkles, and different types of collagen.

[0053] [Cell culture] The procedure was carried out in the same manner as in Experiment Example 1.

[0054] [Analysis of gene expression levels] The BNC2, PPARGC1B, COL1A1, and COL3A1 mRNA activity effects of Examples 2 and 3 were analyzed in the same manner as in Experimental Example 1. Furthermore, a negative control group was established, as in Experimental Example 1, and the effects of each evaluated target were calculated and shown in Table 3.

[0055] [Table 3]

[0056] As can be seen in Table 3, ginseng protoplasts were also confirmed to be a useful material in terms of BNC2 inhibition, PPARGC1B enhancement, and collagen I and III gene strengthening effects. When comparing Tables 1 and 3, wild ginseng protoplasts showed superior BNC2 inhibition, PPARGC1B enhancement, and collagen I and III gene strengthening effects compared to ginseng protoplasts.

[0057] Therefore, wild ginseng protoplasts can be used as a more effective anti-aging ingredient than Korean ginseng protoplasts.

[0058] Experimental Example 3. Regulatory function of wrinkle-related genes by additional components. To compare the wrinkle-related gene function when protoplasts of ginseng according to the present invention are used in combination with additional components, examples and comparative examples with the compositions shown in Table 4 below were prepared.

[0059] [Table 4]

[0060] Next, we measured the inhibitory or promoting activity of PPARGC1B, a gene associated with the formation of deep wrinkles.

[0061] [Cell culture] The procedure was carried out in the same manner as in Experiment Example 1.

[0062] [Gene expression level analysis] The PPARGC1B mRNA activity effects of Comparative Example 1, Comparative Example 2, Example 4, Example 5, and Example 6 were analyzed in the same manner as in Experimental Example 1. Furthermore, a negative control group was established, as in Experimental Example 1, and the effects of each evaluated target were calculated and shown in Table 5.

[0063] [Table 5]

[0064] As shown in Table 5, it was confirmed that when the additional component and the ginseng protoplast were treated simultaneously (Example 6), the expression of the deep wrinkle gene (PPARGC1B) was synergistically promoted.

[0065] Experimental Example 4. Regulatory function of collagen-related genes by additional components. To compare the regulatory function of collagen-related genes when protoplasts of ginseng according to the present invention are used in combination with additional components, examples and comparative examples with the compositions shown in Table 6 below were prepared.

[0066] [Table 6]

[0067] Next, we measured the promoting activity of COL1A1, a gene involved in collagen formation.

[0068] [Cell culture] The procedure was carried out in the same manner as in Experiment Example 1.

[0069] [Analysis of gene expression levels] The COL1A1 mRNA activity effects of Comparative Example 1, Comparative Example 2, Example 4, Example 6, and Example 7 were analyzed in the same manner as in Experimental Example 1. Furthermore, a negative control group was established, as in Experimental Example 1, and the effects of each evaluated target were calculated and shown in Table 7.

[0070] [Table 7]

[0071] Furthermore, examples and comparative examples with the compositions shown in Table 8 below were prepared, and after evaluating COL3A1 using the same method as the evaluation for COL1A1, the effects of each evaluation target were calculated and are shown in Table 9.

[0072] [Table 8]

[0073] [Table 9]

[0074] As shown in Tables 7 and 9, it was confirmed that simultaneous treatment with the additional components and ginseng protoplasts synergistically promotes the expression of COL1A1 (Type I collagen gene) and COL3A1 (Type III collagen gene).

[0075] Experimental Example 5. Function of maintaining protoplast morphology by additional components. To confirm whether the morphology of the protoplast is maintained by the additional components according to the present invention, examples with the compositions shown in Table 10 below were prepared.

[0076] [Table 10]

[0077] Solutions were prepared by adding 10% (w / w) of water or dipropylene glycol (DPG) to each example contained in the isotonic solution. After storing these solutions at 4°C for 4 days, 200 µl was taken, concentrated 10-fold, and observed under a microscope. The results are shown in Figure 1.

[0078] As shown in Figure 1, in Example 4, when water or DPG was added, protoplast clumping occurred, and in particular, when DPG was added, cell membrane defects also occurred. On the other hand, in Example 6, when water or DPG was added, both protoplast clumping and cell membrane defects were significantly reduced.

[0079] Experimental Example 6. Function of maintaining protoplast morphology in cosmetic formulations. To confirm whether the combination of ginseng protoplast and additional components according to the present invention helps maintain the shape of the protoplast even in cosmetic formulations, we prepared example formulations and comparative formulations with the compositions shown in Tables 11 to 15 below.

[0080] [Manufacturing methods for example dosage forms and comparative dosage forms] The water-soluble components (water, polyol) and beneficial components (emulsifier, softener, additional components) listed in Tables 11 to 15 were mixed at 80°C. The beneficial components were then added to the water-soluble components and mixed, and the mixture was allowed to cool naturally to room temperature. Subsequently, ginseng protoplast (1 × 10⁶ / ml) was added to the mixture while stirring, and after stirring was complete, the mixture was stored at 4°C for 4 days. Next, the final mixture was centrifuged at 1300 rpm for 5 minutes, and after carefully removing all of the upper layer, 20 ul of PBS was added to the remaining pellet and observed under a microscope. The results are shown in Table 11 (comparative dosage form examples), Tables 12 to 15 (dosage form examples), and Figure 2.

[0081] [Table 11]

[0082] [Table 12]

[0083] [Table 13]

[0084] [Table 14]

[0085] [Table 15]

[0086] As shown in Tables 11-15 and Figure 2, in the case of comparative dosage forms, phenomena such as protoplast solidification and cell surface depression occurred. On the other hand, in the dosage form example according to the present invention, it was confirmed that the protoplasts were well maintained as single cells.

Claims

1. A cosmetic composition containing protoplasts of ginseng as an active ingredient.

2. The cosmetic composition according to claim 1, wherein the protoplasts of the ginseng species are derived from ginseng or wild ginseng.

3. The cosmetic composition according to claim 1, wherein the composition improves wrinkles or firmness of the skin.

4. The cosmetic composition according to claim 3, wherein the composition suppresses the formation of fine wrinkles or deep wrinkles.

5. Protoplasts of ginseng; and A cosmetic composition comprising one or more components selected from the group consisting of phloretin, oryzanol, tranexamic acid, and pyridoxine.

6. The cosmetic composition according to claim 5, wherein the weight ratio of the composition is 1:0.001 to 1:1, comprising protoplasts of ginseng and one or more components selected from the group consisting of phloretin, oryzanol, tranexamic acid, and pyridoxine.

7. The cosmetic composition according to claim 5, wherein one or more components selected from the group consisting of phloretin, oryzanol, tranexamic acid, and pyridoxine are included in an amount of 0.00001 to 1% by weight.

8. The cosmetic composition according to claim 5, wherein the composition suppresses the clumping of protoplasts or defects in the cell membrane of ginseng.

9. The cosmetic composition according to claim 5, wherein the composition improves wrinkles or firmness of the skin.

10. The cosmetic composition according to claim 9, wherein the composition suppresses the formation of fine wrinkles or deep wrinkles.

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