KR102052522B1

KR102052522B1 - Anti-oxidating composition containing Panax ginseng polysaccharides and Green tea polysaccharides

Info

Publication number: KR102052522B1
Application number: KR1020130109335A
Authority: KR
Inventors: 이옥찬; 김동현; 황경환; 염명훈; 조준철
Original assignee: (주)아모레퍼시픽
Priority date: 2013-09-11
Filing date: 2013-09-11
Publication date: 2019-12-06
Also published as: KR20150030093A

Abstract

The present invention relates to an antioxidant composition containing ginseng polysaccharide and green tea polysaccharide, and more particularly, to a composition for reducing skin wrinkles and enhancing elasticity by providing an excellent antioxidant effect by using a mixture of ginseng polysaccharide and green tea polysaccharide. will be.

Description

Antioxidant composition containing Panax ginseng polysaccharides and Green tea polysaccharides

Reactive oxygen species (ROS) are involved in the aging of the skin is well known by many studies. The oxygen oxidative theory of aging was proposed by Harman in 1956, suggesting that oxidative damage to bioconstituents is caused by various free radicals, which are generated by incidental metabolic processes, and these damages accumulate and lead to aging and death. will be. Damages caused by free radicals are handled by the body's defense ability, but since the defense is not 100% complete, some of the free radicals are adversely affected. This adverse action occurs slowly and, in some cases, for years or even throughout life, and accumulates, degrading the function of cells and tissues in the skin, which causes disease and aging.

Expression of inflammation is a series of reactions to maintain homeostasis in the body, but clinically, redness, swelling, fever, pain and dysfunction may appear. Histologically, the site of inflammation can be seen in the infiltration of neutrophils, monocytes, neutrophils, monocytes, as well as necrosis of cells, expansion of capillaries and edema. However, these phenomena do not occur simultaneously, but appear in a sequence of orders. In recent years, many chemical mediators (chemical mediators) and their actions are known. The process of inflammation is divided into three stages: hemodynamic changes due to changes in blood flow and blood vessel size, exudation of inflammatory cells, and proliferation of connective tissue. Among these, the part being studied in relation to ROS is hemodynamic variation.

In inflammation, platelet activating factor (PAF) is secreted to increase the permeability of blood vessels, increase nitric oxide synthesis, expand blood vessels, express adhesion molecules and participate in the attachment and migration of white blood cells. In addition, during acute inflammation, the surface properties of endothelial cells change. There is a lot of research that relates the above series of reactions and ROS.

As such, antioxidants are essential in living organisms. If you consume antioxidants as food or supplements, it is not easy to reach your skin. The skin surrounds the outermost part of the human body and is a peripheral organ that is in direct contact with the external environment and harmful factors such as ultraviolet rays, but is not directly related to life. Therefore, when the body lacks vitamins or antioxidants, the skin becomes deficient first. easy. In other words, from the point of view of the human body and organs such as the five organs to supply the necessary ingredients first to survive and then send the remaining to the skin. Therefore, it is necessary to give antioxidant power to the skin.

Therefore, if the anti-oxidation effect is implemented in the skin, it will satisfy the customers' demand for skin anti-oxidation for cosmetics and will be very popular in various effects on skin aging, especially antioxidant, skin wrinkle and elasticity improving effect.

Thus, the present inventors attempted to find a substance having excellent antioxidant efficacy among natural extracts to solve the above problems, and when using the ginseng polysaccharide derived from ginseng and green tea polysaccharide derived from green tea at the same time, it provides excellent antioxidant power by synergistic effect. It was confirmed that this can be done, and this invention was completed.

Therefore, an object of the present invention is to provide a composition that can provide excellent antioxidant effect by using both ginseng polysaccharide and green tea polysaccharide simultaneously, and consequently delay skin aging.

In order to achieve the above object, the present invention provides a skin external composition or pharmaceutical composition for antioxidant containing ginseng polysaccharide and green tea polysaccharide as an active ingredient.

The composition of the present invention provides excellent antioxidant efficacy by containing ginseng polysaccharide and green tea polysaccharide, thereby reducing skin wrinkles and enhancing elasticity and consequently delaying skin aging.

The composition of the present invention provides an excellent antioxidant effect by containing ginseng polysaccharide and green tea polysaccharide as active ingredients.

The ginseng (Panax ginseng CA Meyer) polysaccharide used in the present invention is a pectin-like substance having a molecular weight of 34,600, and contains about 60% of galacturonic acid. Galactose and the like constitute a side chain. Ginseng used in the present invention is a plant belonging to the genus Ogapi and ginseng, ginseng in the present invention can be used without limitation in its kind and annual root, can be used both ginseng and dried. In addition, ginseng in the present invention can be used without limitation in the area, specifically, the root of ginseng can be used.

In addition, the green tea polysaccharide used in the present invention is derived from green tea, and unlike the polysaccharides found in other plants, it is an acidic polysaccharide group, and is produced by combining sugar components and amino acids made through photosynthesis. Green tea in the present invention can be used without limitation in the type and cultivation days, it is possible to use both dried or not dried green tea. In addition, the green tea in the present invention can be used without limitation, specifically, the leaves of green tea can be used.

Ginseng polysaccharide and green tea polysaccharide used in the present invention may be prepared by a method known in the art, the method is not particularly limited.

Specifically, after preparing ginseng root or green tea leaf extract with water at 38-42 ° C., and concentrating it, removing impurities with a filter, and then adding ethanol and hot air drying, green tea polysaccharide and ginseng polysaccharide can be obtained. .

The composition of the present invention may contain a ginseng polysaccharide and a green tea polysaccharide in an amount of 0.001 to 10% by weight based on the total weight of the composition, respectively, preferably a ginseng polysaccharide and green tea polysaccharide to 0.005 to 9.5% by weight, respectively , 0.01-9%, 0.03-8.5%, 0.05-8%, 0.07-7.5%, 0.09-7%, 0.1-6.5%, 0.3-6%, 0.5-5.5% or 0.7 It may be contained in an amount of ˜5 wt%.

In the present invention, ginseng polysaccharide and green tea polysaccharide are used by mixing in the range of 0.1 to 10: 1 by weight. In addition, in order to maximize the synergistic effect of the combination of the two components, preferably used to mix the ginseng polysaccharide and green tea polysaccharide in the range of 0.5 to 9.5: 1, 1 to 9: 1 or 1.5 to 8.5: 1 by weight ratio Can be.

The composition of the present invention may be formulated as an external composition for skin, in particular a cosmetic composition, and may be formulated containing a cosmetically or dermatologically acceptable medium or base. In addition, the compositions of the present invention may be provided in all formulations suitable for topical application, for example, emulsions obtained by dispersing the oil phase in a solution, aqueous phase, emulsions obtained by dispersing the aqueous phase in oil phase, suspensions, solids, gels, powders, It may be provided in the form of a paste, foam or aerosol composition. Compositions of such formulations may be prepared according to conventional methods in the art.

In addition, the composition according to the present invention may include other ingredients in addition to the above-mentioned materials within a range not impairing the main effect, preferably a synergistic effect on the main effect. In addition, the composition according to the present invention may further include a moisturizer, an emulsifier, a UV absorber, a preservative, a fungicide, an antioxidant, a pH adjuster, organic and inorganic pigments, flavoring, cooling agent or limiting agent. The blending amount of the above components can be easily selected by those skilled in the art within the range that does not impair the object and effect of the present invention, the blending amount may be 0.01 to 5% by weight, specifically 0.01 to 3% by weight relative to the total weight of the composition. .

In addition, the compositions of the present invention may be formulated as pharmaceutical compositions. When the composition according to the present invention is applied to a pharmaceutical product, it may be formulated as an oral or parenteral dosage form in the form of a solid, semi-solid or liquid by adding a commercially available inorganic or organic carrier to the active ingredient used in the present invention. , The pharmaceutical composition according to the invention can be administered orally, parenteral, rectal, topical, transdermal, intravenous, intramuscular, intraperitoneal, subcutaneous.

Formulations for oral administration include tablets, pills, granules, capsules, powders, fine granules, powders, emulsions, syrups, pellets and the like. In addition, the formulation for parenteral administration includes injections, drops, ointments, lotions, sprays, suspensions, emulsions, suppositories, and the like. The composition according to the present invention can be easily formulated by carrying out according to a conventional method, and at this time, surfactants, excipients, coloring agents, spices, preservatives, stabilizers, buffers, suspending agents, and other commercially available auxiliaries can be suitably used. have.

The dosage of the active ingredient of the pharmaceutical composition of the present invention will vary depending on the age, sex, weight, pathology and severity of the subject to be administered, the route of administration or the judgment of the prescriber. Determination of a suitable dose based on these factors is within the level of one of skill in the art and its daily dosage may be, for example, 0.1 mg / kg / day to 100 mg / kg / day, more specifically 5 mg / kg / day to 50 mg / kg. May be, but is not limited to.

Hereinafter, the present invention will be described in more detail with reference to Examples. These examples are only for illustrating the present invention, and it will be apparent to those skilled in the art that the scope of the present invention is not to be construed as being limited by these examples.

Reference Example 1 Preparation of Ginseng Polysaccharide

1 kg of dried white ginseng was pulverized with a blender, and 10 liters of purified water was added thereto, stirred and extracted with 38-42 ° C. hot water for 24 hours, and then deposited at 15 ° C. for 1 day. Thereafter, the residue and the filtrate were separated through filter cloth filtration and centrifugation. The separated filtrate was concentrated under reduced pressure to 1/10 the amount of purified water. To this was added dropwise ethanol in a concentrated amount of 4 times and hot air dried to obtain 65 g of ginseng polysaccharide.

Reference Example 2 Preparation of Green Tea Polysaccharide

1 kg of the first processed green tea leaves were pulverized with a blender, and then 10 liters of purified water was added thereto, stirred and extracted with hot water at 38-42 ° C. for 24 hours, and then deposited at 15 ° C. for 1 day. Thereafter, the residue and the filtrate were separated through filter cloth filtration and centrifugation. The separated filtrate was concentrated under reduced pressure to 1/10 the amount of purified water. To this was added dropwise four times the concentrated ethanol and hot air dried to obtain 72g of green tea polysaccharide.

Example 1 Preparation of a Mixture of Ginseng Polysaccharide and Green Tea Polysaccharide

The ginseng polysaccharide prepared in Reference Example 1 and the green tea polysaccharide prepared in Reference Example 2 were mixed at a ratio of 1: 3.

Example 2 Preparation of Mixture of Ginseng Polysaccharide and Green Tea Polysaccharide

The ginseng polysaccharide prepared in Reference Example 1 and the green tea polysaccharide prepared in Reference Example 2 were mixed at a ratio of 1: 1.

Example 3 Preparation of Mixture of Ginseng Polysaccharide and Green Tea Polysaccharide

The ginseng polysaccharide prepared in Reference Example 1 and the green tea polysaccharide prepared in Reference Example 2 were mixed at a ratio of 3: 1.

Test Example 1 Inhibitory Effect of Reactive Oxygen Species Production

The keratinocytes (F1eratinocyte) isolated from the skin tissue of the person into a 5 × 10 ⁴ are in each well of a 24-well plate was attached for 24 hours. After 16 hours, the materials of Examples 1 to 3 and Reference Examples 1 and 2 were each treated with 1%. At this time, the control did not process anything for comparison. After 2 hours, the culture solution was removed, and 100 µl of phosphate buffered saline (PBS) was added to each well. The keratinocytes were irradiated with UV 30 mJ / cm 2 using an ultraviolet B (UVB) lamp (Model: F15T8, UV B 15 W, SanF1yo DennF1i, Japan), and then PBS was removed and each cell was prepared with keratinocyte culture solution 200. Μl was added. Here, the test materials of Examples 1 to 3 and Reference Examples 1 to 2 were again treated, and the amount of reactive oxygen species increased by ultraviolet stimulation at regular time intervals was quantified. The amount of ROS was quantified by referring to Tan's method for measuring the fluorescence of DCF-DA (dichlorofluorescin diacetate) oxidized by ROS (Tan et al., 1998, J. Cell Biol. Vol. 141, pp1423-1432). Table 1 shows the results of calculating the ratio of the vehicle-treated control group ROS.

Test substance UVB 30 mJ / cm ² Time since survey 0hr 2hr 3hr Vehicle 100 244 287 UVB + Vehicle 100 325 381 UVB + Example 1 100 272 311 UVB + Example 2 100 267 304 UVB + Example 3 100 263 297 UVB + Reference Example 1 100 279 319 UVB + Reference Example 2 100 283 326

In the results of Table 1, the ginseng polysaccharide and the green tea polysaccharide used in the present invention inhibits the production of ROS known to cause skin cell damage by ultraviolet rays, and is particularly a mixture of the ginseng polysaccharide and the green tea polysaccharide. In addition, it is possible to confirm that the antioxidant activity is very excellent by inhibiting the generation of ROS very effectively, and inhibiting the generation of ROS to a level almost similar to the case of not irradiating UV light after the stimulation of UV light.

Therefore, it can be seen that the mixture of ginseng polysaccharide and green tea polysaccharide used in the present invention can help to prevent and improve skin aging by inhibiting oxidation and defending the production of skin irritation.

Test Example 2 Measurement of Collagenase Expression Inhibition Efficacy

Inhibition of collagenase production of Examples 1 to 3 and Reference Examples 1 to 2 was measured by comparing with tocopherols and EGCG as controls.

Human fibroblasts (Human Dermal Fibroblasts, neonatal (HDFn)) in 96-well plates containing DMEM (Dulbecco's Modified Eagle's Media) medium containing 2.5% fetal calf serum (lot # 617766, Cascade Biologics Inc.). , Portland, Oregon (OR) USA) was added to 5,000 cells / well (well), and cultured until 90% growth. After culturing in serum-free DMEM medium for 24 hours, the serum-free DMEM medium was treated with the test materials of Examples 1 to 3 and Reference Examples 1 to 2 at a molar concentration of 10 ^-4 for 24 hours. Was collected.

The collected cell culture medium was measured for collagenase production using a commercially available collagenase measuring instrument (Amersham Pharma Co., Ltd., USA). First, the cultured cells were placed in 96-well plates uniformly coated with primary collagenase antibodies, and antigen-antibody reactions were performed in a thermostat for 3 hours.

After 3 hours, the secondary collagen antibody conjugated with the chromophore was placed in a 96-well plate and reacted again for 15 minutes. After 15 minutes, the coloring stimulant was added, causing color development at room temperature for 15 minutes, and 1M sulfuric acid was added again to stop the reaction (color development). The color of the reaction solution was yellow and the degree of yellow color was different according to the progress of the reaction.

The absorbance of the yellow-colored 96 well plate was measured at 405 nm using an absorbance meter, and the degree of synthesis of collagenase was calculated by Equation 1 below. At this time, the reaction absorbance of the prepared cell culture medium of the group not treated with the composition was used as a control.

The results of measuring the collagenase expression inhibitory effect of the test substances in the human fibroblasts are shown in Table 2 below the degree of collagenase expression, which is prepared by comparing the collagenase expression level of the untreated group to 100. .

matter Collagenase expression level (%) Untreated group 100 Tocopherol (positive control) 76 EGCG (positive control) 63 Example 1 66 Example 2 67 Example 3 68 Reference Example 1 79 Reference Example 2 77

Looking at Table 2, it can be seen that the mixture of ginseng polysaccharide and green tea polysaccharide used in the present invention inhibits collagenase expression in vitro.

In addition, as shown in Table 1, each of Examples 1 to 3 is not only superior to collagenase inhibitory effect than Reference Examples 1 to 2, but also significantly higher collagenase expression than tocopherol, a positive control group. It can be seen that the effect.

Test Example 3 Procollagen Production Promoting Effect Experiment

The procollagen producing ability of Examples 1 to 3 and Reference Examples 1 to 2 was measured by comparing with vitamin C.

Place human fibroblasts at 5,000 cells / well in 96-well plates containing DMEM (Dulbecco's Modified Eagle's Media) containing 2.5% fetal calf serum and incubate until 90% growth It was. Thereafter, the cells were incubated for 24 hours in serum-free DMEM medium, and then the test materials of Examples 1 to 3 and Reference Examples 1 and 2 dissolved in serum-free DMEM medium, and vitamin C for 10 hours at 10 ^-4 molar concentration, respectively. After treatment, cell culture fluid was collected. After 24 hours, the amount of free collagen in the medium was measured using the procollagen type-1 C-peptide EIA kit (MK101, Takara, Japan).

The degree of procollagen production in the untreated group was set to 100, and the degree of procollagen production in the group treated with the test substance was determined, and the results are shown in Table 3 below.

matter Procollagen production level (%) Untreated group 100 Vitamin c 121 Example 1 120 Example 2 120 Example 3 122 Reference Example 1 110 Reference Example 2 106

Looking at Table 3, it can be seen that the mixture of ginseng polysaccharide and green tea polysaccharide used in the present invention promotes production of procollagen in vitro.

In addition, Examples 1 to 3 is not only significantly superior to the effect of promoting the production of procollagen than Reference Examples 1 to 2, it can be confirmed that the effect is equivalent to the vitamin C of the positive control group.

From this, it can be seen that the mixture of ginseng polysaccharide and green tea polysaccharide used in the present invention has an excellent effect of promoting collagen production and has excellent collagen production ability, thus effectively preventing skin wrinkle formation and providing a wrinkle improvement effect. .

Test Example 4 Measurement of Elastase Activity Inhibition Effect

The elastase activity inhibitory ability of Examples 1 to 3 and Reference Examples 1 to 2 was measured by comparing with EGCG. The elastase and substrate used were purchased commercially from Sigma-Aldrich, USA (Cat.No. E0127).

Elastase activity inhibitory activity was tested by the following test method.

In a 96 well plate, 10 mg / L Tris-HCL buffer (pH 8.0) was mixed with Examples 1-3 and Reference Examples 1-2 (200 µL) and 50 µL of 20 µg / mL Elastase Type III solution, respectively. . 250 μM of EGCG was used as a positive control and distilled water was used as a non-treated group as a negative control. Thereafter, 100 µL of 0.4514 mg / mL N-SUCCINYL-ALA-ALA-ALA-p-NITROANILIDE prepared with the above buffer was added and reacted at 25 ° C for 15 minutes. After completion of the reaction, the absorbance at a wavelength of 415 nm was measured. A blank test was performed in the same manner to correct.

Calculation method of the elastase activity inhibitory activity is shown in Equation 2 below, the results are shown in Table 4.

A: Absorption at wavelength 415 nm with no test substance added and enzyme added

B: Absorbance at wavelength 415 nm with no test substance added and no enzyme added

C: absorbance at wavelength 415 nm at test substance addition and enzyme addition

D: absorbance at wavelength 415 nm with test substance added and without enzyme

compound Suppression degree (%) Untreated group 0 EGCG 65 Example 1 71 Example 2 70 Example 3 68 Reference Example 1 62 Reference Example 2 60

Referring to Table 4, the ginseng polysaccharide or green tea polysaccharide used in the present invention inhibits elastase activity, and in particular, Examples 1 to 3, in which ginseng polysaccharide and green tea polysaccharide are mixed, have an elastase inhibitory degree. It can be seen that it is remarkably superior to EGCG known as an activity inhibitor.

[Formulation Examples 1 to 3 and Comparative Formulation Examples 1 to 3]

Nutritional cream was prepared in a conventional manner according to the composition of Table 5 (unit: wt%).

Ingredient Formulation Example 1 Formulation Example 2 Formulation Example 3 Comparative Formulation Example 1 Comparative Formulation Example 2 Comparative Formulation Example 3 Purified water To 100 To 100 To 100 To 100 To 100 To 100 Example 1 0.1 - - - - - Example 2 - 0.1 - - - - Example 3 - - 0.1 - - - Reference Example 1 - - - 0.1 - - Reference Example 2 - - - - 0.1 - Vegetable Cured Oil 1.50 1.50 1.50 1.50 1.50 1.50 Stearic acid 0.60 0.60 0.60 0.60 0.60 0.60 Glycerol Stearate 1.00 1.00 1.00 1.00 1.00 1.00 Stearyl alcohol 2.00 2.00 2.00 2.00 2.00 2.00 Polyglyceryl-10 Pentastearate & Behenyl Alcohol & Sodium Stearoyl Lactylate 1.00 1.00 1.00 1.00 1.00 1.00 Arakidil Behenyl Alcohol & Arakidil Glucoside 1.00 1.00 1.00 1.00 1.00 1.00 Cetylaryl Alcohol & Cetearyl Glucoside 2.00 2.00 2.00 2.00 2.00 2.00 PEG-100 Stearate & Glycerolate & Propylene Glycol 1.50 1.50 1.50 1.50 1.50 1.50 Caprylic / Carric Triglycerides 11.00 11.00 11.00 11.00 11.00 11.00 Cyclomedicon 6.00 6.00 6.00 6.00 6.00 6.00 Preservative, incense Quantity Quantity Quantity Quantity Quantity Quantity Triethanol amine 0.1 0.1 0.1 0.1 0.1 0.1

Test Example 5 Confirmation of Skin Elasticity Improvement Efficacy

In order to confirm the effect of improving skin elasticity in humans, the formulations of Examples 1 to 3 and Comparative Formulations 1 to 3 were evaluated as follows.

120 healthy women in their 30s and 40s were divided into 6 groups of 20 people and one composition was used for each group. After applying it to the face once a day for 12 weeks, the skin elasticity tester (Cutometer SEM 575, C + K) Electronic Co., Germany) was used to measure skin elasticity. The results are shown in Table 6 below. The results in Table 6 refer to the properties of skin viscoelasticity of the skin elasticity tester.

Test substance Skin elasticity effect Formulation Example 1 0.28 Formulation Example 2 0.26 Formulation Example 3 0.25 Comparative Formulation Example 1 0.19 Comparative Formulation Example 2 0.17 Comparative Formulation Example 3 0.10

As shown in Table 6, Formulation Examples 1 to 3 and Comparative Formulation Examples 1 to 2 containing the ginseng polysaccharide or green tea polysaccharide increased skin elasticity compared to the group to which the Comparative Formulation Example 3 was applied, especially ginseng polysaccharide and green tea. Groups to which Formulation Examples 1 to 3 containing a mixture of polysaccharides were applied can confirm that the skin elasticity was significantly increased.

As described above in detail the specific parts of the present invention, it is apparent to those skilled in the art that such specific description is merely a preferred embodiment, thereby not limiting the scope of the present invention. something to do. Thus, the substantial scope of the present invention will be defined by the appended claims and their equivalents.

Claims

A cosmetic composition for inhibiting skin wrinkles, comprising a mixture of ginseng polysaccharide and green tea polysaccharide as an active ingredient,
The mixing ratio of the ginseng polysaccharide and green tea polysaccharide in the mixture is 0.1 to 10: 1 based on the weight of the cosmetic composition for inhibiting skin wrinkles.

The cosmetic composition of claim 1, wherein the ginseng polysaccharide and the green tea polysaccharide are each contained in an amount of 0.001 to 10% by weight based on the total weight of the composition.

The cosmetic composition of claim 1, wherein the ginseng polysaccharide and the green tea polysaccharide are mixed and used in a ratio of 1: 3 to 3: 1 by weight.

delete

A cosmetic composition for improving skin elasticity containing a mixture of ginseng polysaccharide and green tea polysaccharide as an active ingredient,
The mixing ratio of the ginseng polysaccharide and green tea polysaccharide in the mixture is 0.1 ~ 10: 1 cosmetic composition for improving skin elasticity based on the weight.

The cosmetic composition for improving skin elasticity of claim 6, wherein the ginseng polysaccharide and the green tea polysaccharide are each contained in an amount of 0.001 to 10% by weight based on the total weight of the composition.

The cosmetic composition for improving skin elasticity of claim 6, wherein the ginseng polysaccharide and the green tea polysaccharide are mixed and used in a ratio of 1: 3 to 3: 1 by weight.

delete