KR101382113B1 - Skin external composition containing white ginseng polysaccharide extract - Google Patents

Abstract

The present invention relates to a composition for external application for skin having skin wrinkle improvement, skin elasticity enhancement and skin anti-aging effect, and more particularly, containing white ginseng polysaccharide extract having a molecular weight of 30,000 or more and less than 100,000 as an active ingredient, human skin keratinocytes The present invention relates to an external composition for skin having an effect of improving skin wrinkles, enhancing skin elasticity and preventing skin aging by increasing the expression of mitochondrial electron transfer enzymes.

White ginseng, polysaccharide, molecular weight, mitochondrial electron transfer enzyme, skin cell activity, skin wrinkle improvement

Description

Skin external composition containing white ginseng polysaccharide extract

The present invention relates to a composition for external application for skin having skin wrinkle improvement, skin elasticity enhancement and skin anti-aging effect, and more particularly, containing white ginseng polysaccharide extract having a molecular weight of 30,000 or more and less than 100,000 as an active ingredient, human skin keratinocytes The present invention relates to an external composition for skin having an effect of improving skin wrinkles, enhancing skin elasticity and preventing skin aging by increasing the expression of mitochondrial electron transfer enzymes.

Mitochondria in cells synthesize chemical energy of nutrients, ATP, which is carried out through a system called electron transport system, and an enzyme that plays an important role in the system is an electron transport system enzyme. When the nutrient's chemical energy is transferred to the electrons, electrons with high energy are created, and these electrons are exchanged by various electron transfer enzymes and release their energy. At this time, proton gradient is made by using the emitted energy and ATP is synthesized using this slope. Mitochondria use the energy of electrons to transport protons and to differentiate proton concentrations inside and outside the inner membrane. This difference is called proton slope. When the proton passage of the inner membrane is opened in this state of concentration difference, the proton flows from the high concentration to the low concentration due to the difference in concentration, and synthesizes ATP using the energy of this flow.

There are many electron transport enzymes in the mitochondrial inner membrane (cristae), the energy of the electron is consumed when electrons are transferred between them and the protons are transported out of the mitochondrial inner membrane. Enzymes located in the inner mitochondrial membrane and their action is shown in FIG. Looking at this in more detail, the first electron transferase is flavin mononuleotide (FMN). It is an enzyme immobilized on the inner membrane of the mitochondria and receives two electrons and two protons from NADH, the product of the process before the electron transport system. The flavin mononucleotide transports two protons from the NADH out of the inner membrane and two electrons to the next electron transfer enzyme, Coenzyme Q (CoQ). The former coenzyme Q binds to two protons inside the intima, becomes CoQH ₂ , and then transports the two combined protons out of the intima. At the same time, the two received electrons are transferred to cytochrome b. The electrons transferred to cytochrome b are again transferred in the order of cytochrome c1-cytochrome c-cytochrome a-cytochrome a3 and then transferred to oxygen in the last cytochrome a3 to form water. Proton gradient is formed based on the mitochondrial lining by the proton transported in the above process. The electron transport system has four functional complexes: complex I (ubiquinone oxidoreductase), complex II (succinate: ubiquinone oxidoreductase), complex III (ubiquinol: cytochrome C oxidoreductase) and complex IV (cytochrome C: oxidoreductase). In the transfer of electrons to oxygen molecules in a stepwise manner from complex I to complex IV, hydrogen ions exit the mitochondrial substrate, forming a membrane potential difference, and hydrogen ions to the mitochondrial substrate by F0 / F1-ATP synthase (complex V). As it reflows, ATP is produced in ADP. This process is called oxidative phosphorylation (OXPHOS).

Accordingly, the present inventors have improved the expression of mitochondrial electron transfer system enzymes of human skin keratinocytes by the molecular weight of more than 30,000 to less than 100,000 of white ginseng polysaccharide extract improves the activity of skin cells to improve skin wrinkles, improve skin elasticity and prevent skin aging The present invention has been found to impart an effect.

Accordingly, it is an object of the present invention to provide an external composition for skin for improving skin wrinkles, enhancing skin elasticity and preventing skin aging.

In order to achieve the above object, the present invention provides a skin external composition comprising a white ginseng polysaccharide extract having a molecular weight of 30,000 to less than 100,000 as an active ingredient.

White ginseng polysaccharide extract having a molecular weight of 30,000 or more and less than 100,000 according to the present invention was able to increase the expression of mitochondrial electron transfer enzymes of human dermal keratinocytes than red ginseng polysaccharide extract of the same molecular weight and white ginseng polysaccharide extract of different molecular weight. By increasing the energy activity of the skin keratinocytes, it was possible to give effects such as skin regeneration, skin activity enhancement, skin elasticity enhancement, skin aging prevention and complexion improvement.

The present invention relates to an external preparation composition for skin comprising a white ginseng polysaccharide extract having a molecular weight of 30,000 to less than 100,000 as an active ingredient.

The present invention also relates to an external preparation composition for promoting the expression of mitochondrial electron transfer system enzymes in skin cells comprising the white ginseng polysaccharide extract having a molecular weight of 30,000 to less than 100,000 as an active ingredient.

The present invention also relates to a skin external preparation composition for improving skin wrinkles comprising a white ginseng polysaccharide extract having a molecular weight of 30,000 or more and less than 100,000 as an active ingredient.

The present invention also relates to a skin external preparation composition for enhancing skin elasticity, the white ginseng polysaccharide extract having a molecular weight of 30,000 to less than 100,000 as an active ingredient.

The present invention also relates to a skin external preparation composition for preventing skin aging comprising white ginseng polysaccharide extract having a molecular weight of 30,000 or more and less than 100,000 as an active ingredient.

Hereinafter, the present invention will be described in more detail.

As used herein, the term "extract" refers to a substance obtained by extracting a component contained therein from a natural product, regardless of the method of extraction or the kind of the component. For example, it is a broad concept including both extracting a component dissolved in a solvent from a natural product using water or an organic solvent, or extracting only a specific component such as oil, for example.

As used herein, the term "skin" refers to a tissue covering the body surface of an animal, and includes the scalp and hair as well as a tissue covering the body surface such as a face or a body.

White ginseng used in the present invention is made from 4 to 6 years old ginseng as a raw material, peeled off mostly and dried in the sun as it is a round-shaped ginseng processed to have a water content of 14% or less. The color is pale yellow white. Unlike red ginseng processing method, it is dried without solar natural drying or heat and other methods.

On the other hand, red ginseng is a pale yellow brown or light brown ginseng, which is dried by steaming steamed 6-year-old ginseng without peeling.

The external preparation composition for skin according to the present invention is characterized by containing an extract having a molecular weight of 30,000 or more and less than 100,000 as an active ingredient among the white ginseng polysaccharide extracts separated by molecular weight. Compositions containing white ginseng polysaccharide extracts having a molecular weight of 30,000 to less than 100,000 as an active ingredient increase the mRNA expression of mitochondrial electron transfer enzymes in the keratinocytes to activate skin cells, enhance skin elasticity, and further prevent skin aging. It has an effect. Among the white ginseng polysaccharide extracts, the extract having a molecular weight of 30,000 or more and less than 100,000 is more effective than the red ginseng polysaccharide extract of the same molecular weight.

The external preparation composition for skin according to the present invention contains white ginseng polysaccharide extract having a molecular weight of 30,000 or more and less than 100,000, based on the total weight of the composition, at 0.1 to 10% by weight.

The external preparation composition for skin according to the present invention may be formulated in the form of a cosmetic composition in one embodiment of the present invention, such cosmetic composition may be, for example, a cosmetic. In this case, the external preparation composition for skin according to the present invention contains a cosmetically or dermatologically acceptable medium or base. These are all formulations suitable for topical application, for example emulsions, suspensions, microemulsions, microcapsules, microgranulocytes or ionic (liposomes) obtained by dispersing an oil phase in solutions, gels, solids, pasty anhydrous products, aqueous phases and It may be provided in the form of a nonionic vesicle dispersant or in the form of a cream, skin, lotion, powder, ointment, spray or cone stick. These compositions may be prepared according to conventional methods in the art. The composition according to the invention can also be used in the form of a foam or in the form of an aerosol composition further containing a compressed propellant.

In addition, the composition for external application for skin according to the present invention is a fatty substance, an organic solvent, a dissolving agent, a thickening agent, a gelling agent, an emollient, an antioxidant, a suspending agent, a stabilizer, a foaming agent, a fragrance, a surfactant, water, an ionic type or Nonionic emulsifiers, fillers, metal ion sequestrants, chelating agents, preservatives, vitamins, blockers, wetting agents, essential oils, dyes, pigments, hydrophilic or lipophilic active agents, lipid vesicles or any other ingredients commonly used in cosmetics It may contain adjuvants conventionally used in the cosmetic or dermatology field. Such adjuvants are introduced in amounts commonly used in the cosmetics or dermatological fields.

The external preparation composition for skin according to the present invention is not particularly limited in formulation, and may be appropriately selected according to the purpose. For example, softening cream (skin lotion and milk lotion), nourishing cream, essence, nourishing cream, massage cream, pack, gel, essence, eye cream, eye essence, cleansing cream, cleansing foam, cleansing water, pack, powder, It may be prepared in a formulation such as body lotion, body cream, body oil or body essence, but is not limited thereto.

In one embodiment of the present invention, the topical skin composition may be used in the face, especially around the eyes, around the mouth, cheeks, forehead, neck, hands or feet, but is not limited thereto.

In addition, the external preparation composition for skin according to the present invention may be formulated as a herbal medicine.

In Chinese medicine, Gi is interpreted as ① fine and fine substance with fluid and rich nutrients that make up the human body and maintain the vital activity of human body, and ② physiological function of the cog tissue. ① is described as a basic substance of ② and ② is expressed as a function of ①. Dividing these groups according to their origin, they are inhaled by the parents' regular routines, that is, the genetic predispositions and the nutrients ingested by the digestion and absorption of food, and the respiratory tracts of the lungs. It can be divided into fresh air coming from the air. Here, the controllable part of the food is related to food, and thus Bogi means 도 (helping) 氣 (energy), which can lead to energy generation by nutrient intake. This leads to an increase in cellular metabolism and developmental energy, which, according to dermatological mechanisms, can be observed by increasing intracellular mitochondrial activity.

Hereinafter, the present invention will be described more specifically with reference to examples and test examples. These examples are provided only for understanding the contents of the present invention, and the scope of the present invention is not limited to these examples, and modifications, substitutions, and insertions commonly known in the art may be performed. This is also included in the scope of the present invention.

Example 1 Preparation of White Ginseng and Red Ginseng Polysaccharide Extract

The sliced white and red ginseng were sufficiently dried at 60 ° C. for 2 days. The moisture content of dried white and red ginseng slices is less than 5%. The dried white and red ginseng sections were then ground into fine particles using a grinder. It is appropriate to maintain the distribution of the particle size at about 2 to 5 mm when pulverizing into fine particles. Thereafter, the white and red ginseng pulverized fine powder was adjusted to a concentration of 15%, dispersed in water, and then extracted with stirring slowly at room temperature for 24 hours. The extract was separated by centrifugation and filtered using a 1 μm membrane. Then concentrated at about 55 ℃ until the concentrated volume is 1/10 of the initial volume. Ethanol was slowly added to the concentrate to obtain purified white and red ginseng polysaccharide particles. At this time, the required volume of ethanol is required to the level that the ethanol concentration is about 85%. After recovering the white and red ginseng polysaccharide precipitated and dried for 24 hours at 40 ℃ to recover the final white ginseng and red ginseng polysaccharide powder and then less than 10,000, less than 10,000 more than 30,000, more than 30,000 less than 100,000 and more than 100,000 Each extract was fractionated by ultrafiltration.

[Test Example 1] Expression Control of Mitochondrial Electron Transfer System Active Enzymes

Step 1: Cell Line and Cell Culture

Human skin keratinocytes (Human Epidermal Neonatal Keratinocyte Cells) is to be recommended as a way to secure commercially bought from its loans (Lonza, Inc. Walkersville, MD, USA) and subculture, CO ₂ incubator (CO ₂ incubator) The cells were cultured at 37 ° C. under 5% CO ₂ conditions. Cell cultures were cultured according to the guidelines of Lonza, Inc. Walkersville, MD, USA. In 500 ml of KBM-2 (Clonetics CC-3103) medium, 2 ml of KGM-2 Burlet kit (BPE: Bovine pituitary extract), human epidermal growth factor (hEGF) 0.5 ml, Insulin 0.5 ml, Hydrocortisone 0.5 ml, Transferrin 0.5 ml, Epinephrine 0.5 ml and GA-1000 (Gentamycin Sulfate + Amphoricin-B ( Amphofericin-B)) 0.5 ml] was added and used.

<Step 2: Confirmation of expression regulation of mitochondrial electron transfer system active enzyme gene>

Human keratinocyte cultures were treated with 10 μM of white ginseng and red ginseng polysaccharide extracts fractionated by molecular weight in Example 1, and 30 μM of coenzyme Q10 was treated as a positive control. After 48 hours of incubation, the cells were washed twice with 10 ml of Phosphate Buffered Saline (PBS), and the cells were treated with Trizol reagent (Invitrogen, Carlsbad, Calif.). Total RNA in the isolate was isolated.

The isolated RNA was purified once more using Qiagen RNeasy kit (Valencia, Calif.) And then Agilent's BioAnalyzer 2100 model instrument (Agilent 2100 BioAnalyzer, Santa, Calif., USA). The quality and concentration of RNA were determined using Agilent Technologies of Clara. From the isolated RNA, cDNA was synthesized using Invitrogen's Superscript Reverse Transcriptase (RT) II kit, and the real time-reverse transcription polymerase chain reaction (Q-RT-PCR) was performed. The quantitative analysis of the gene expression changes of CYB belonging to complex III and ATP8 belonging to complex V in the mitochondrial electron transfer enzymes was quantitatively analyzed. Changes in gene expression patterns are based on the human Glyceraldehyde-3P-dehydrogenase (GAPDH, 4333764F, Applied Biosystems). Evaluation was performed using Forster City, California, USA.

First, the white ginseng polysaccharide extract was separated by molecular weight, followed by RT-PCR for mitochondrial enzymes CYB and ATP8, and the relative ratios of the control groups were shown in FIGS. 2A and 2B. As a result, the white ginseng polysaccharide extract having a molecular weight of 30,000 or more and less than 100,000 was confirmed to increase the expression amount of CYB and ATP8 most, and the efficacy was superior to that of the coenzyme Q10, which is a positive control group.

The PCR process is as follows. The denaturation temperature was 95 ° C. for 15 seconds and the annealing / extension temperature was 1 minute at 60 ° C. and this cycle was repeated a total of 50 times. In addition, the gene name, registration number and its sequence used in the PCR process are shown in Table 1 below.

Gene name (registration number) order MT-CYB (Hs02596867_s1) 5'-CGCCCTCGGCTTACTTCTCTTCCTT-3 '(SEQ ID NO: 1) MT-ATP8 (Hs02596863_g1) 5'-CGTATGGCCCACCATAATTACCCCC-3 '(SEQ ID NO: 2)

Based on the above results, RT-PCR was performed under the same conditions as above to measure the expression levels of CYB and ATP8 after treatment with white and red ginseng polysaccharide extracts having a molecular weight of 30,000 or more and less than 100,000, and the results are illustrated in FIGS. 3A and 3B. Shown in Here, it was confirmed that the white ginseng polysaccharide extract having a molecular weight of 30,000 or more and less than 100,000 according to the present invention increased the expression level of mitochondrial enzymes than the red ginseng polysaccharide extract having the same molecular weight.

 [Formulation Example 1] Nourishing cream

Nutritional cream was prepared in a conventional manner according to the composition shown in Table 2 (unit: wt%).

ingredient content Purified water Balance glycerin 3.0 Butylene glycol 3.0 Liquid paraffin 7.0 Beta Glucan 7.0 Carbomer 0.1 White ginseng polysaccharide extract with molecular weight of 30,000 to less than 100,000 3.0 Caprylic / capric triglyceride 3.0 Squalane 5.0 Cetearyl glucoside 1.5 Sorbitan stearate 0.4 Polysorbate 60 1.2 antiseptic Suitable amount incense Suitable amount Pigment Suitable amount Triethanolamine 0.1

Formulation Example 2 Massage Cream

To the massage cream was prepared in a conventional manner according to the composition shown in Table 3 (unit: wt%).

ingredient content Purified water Balance glycerin 8.0 Butylene glycol 4.0 Liquid paraffin 45.0 Beta Glucan 7.0 Carbomer 0.1 White Ginseng Polysaccharide Extract with Molecular Weight of 30,000 to 100,000 1.0 Caprylic / capric triglyceride 3.0 Wax 4.0 Cetearyl glucoside 1.5 Sesquioleic acid sorbitan 0.9 Vaseline 3.0 antiseptic Suitable amount incense Suitable amount Pigment Suitable amount paraffin 1.5

[Formulation Example 3] Pack

To prepare a pack in a conventional manner according to the composition shown in Table 4 (unit: wt%).

ingredient content Purified water Balance glycerin 4.0 Polyvinyl alcohol 15.0 Hyaluronic acid extract 5.0 Beta Glucan 7.0 Allantoin 0.1 White Ginseng Polysaccharide Extract with Molecular Weight of 30,000 to 100,000 0.5 Nonyl phenyl ether 0.4 Polysorbate 60 1.2 antiseptic Suitable amount incense Suitable amount Pigment Suitable amount ethanol 6.0

Figure 1 shows a schematic diagram of the electron transport system related enzymes present in the inner membrane of the mitochondria and the process through which ATP is formed.

Figures 2a and 2b is a graph showing the results of measuring the relative ratio to the control group by performing the RT-PCR for mitochondrial enzymes CYB and ATP8 after fractionation of white ginseng polysaccharide extract by molecular weight.

3a and 3b are graphs showing the results of measuring the expression levels of CYB and ATP8 after treatment with white ginseng and red ginseng polysaccharide extracts having a molecular weight of 30,000 or more and less than 100,000.

<110> Amorepacific corporation <120> Skin external composition containing white ginseng polysaccharide          extract <130> YPD / 200911-0074 <160> 2 <170> Kopatentin 1.71 <210> 1 <211> 25 <212> DNA <213> MT-CYB (Hs02596867_s1) <400> 1 cgccctcggc ttacttctct tcctt 25 <210> 2 <211> 25 <212> DNA <213> MT-ATP8 (Hs02596863_g1) <400> 2 cgtatggccc accataatta ccccc 25

Claims (6)

delete A skin external composition for preventing skin aging, containing white ginseng polysaccharide extract having a molecular weight of 30,000 or more and less than 100,000 as an active ingredient to promote enzyme expression in a mitochondrial electron transfer system in skin cells. The external preparation composition for skin according to claim 2, wherein the composition is for improving skin wrinkles. The external preparation composition for skin according to claim 2, wherein the composition is for promoting skin elasticity. delete The external preparation composition for skin according to claim 2, wherein the external preparation composition is formulated as a cosmetic.

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