KR20030089598A - Composition for external application to the skin having free radical scavenging effects or protective effects for cellular anti-oxidation system - Google Patents

Abstract

PURPOSE: Provided is a composition of dermatoiogics having free radical scavenging effects or protective effects from the cellular anti-oxidation system. Therefore, it prevents and retards skin aging. CONSTITUTION: A composition of dermatoiogics is characterized by containing one of blueberry, cranberry or blackberry extracts in the amount of 0.001-30 wt.%. The extract has free radical scavenging effect and protection effects from the cellular anti-oxidation system.

Description

Composition for external application with free radical scavenging ability or protection against intracellular antioxidant system {Composition for external application to the skin having free radical scavenging effects or protective effects for cellular anti-oxidation system}

The present invention relates to an external composition for skin having free radical scavenging activity or intracellular antioxidant protection. More specifically, the present invention contains one or more extracts of blueberry, raspberry, cranberry or blackberry extracts to eliminate free radicals and inhibit the damage of antioxidant defenses such as superoxide dismutase, catalase or glutathione. The present invention relates to an external preparation composition for skin, having an intracellular antioxidant protection ability resulting from the prevention and delaying of skin aging.

The relationship between aging and free radicals is well known in many studies. Free radicals occur during normal metabolic processes, resulting in oxidative damage to biological components and the accumulation of these damages leading to aging and death. The fact is known. Free radicals produced during normal metabolism are antioxidant enzymes such as superoxide dismutase or catalase, glutathione, ascorbic acid (vitamin C) and alpha present in vivo. Although it is removed by antioxidant defense factors such as tocopherol (α-tocopherol, vitamin E) or ubiquinone (ubiquinone), but not completely removed. Free radicals, which have not been removed, accumulate in cells and tissues in large amounts, damaging DNA and proteins in the cells, leading to genetic diseases such as cancer, Down syndrome, Alzheimer's disease, etc., as well as progressing aging by destroying cells and tissues.

Since the skin is exposed to the atmosphere, not only intrinsic aging but also photoaging caused by daylight and ultraviolet rays is performed. Such sunlight and ultraviolet rays generate a lot of free radicals when they come in direct contact with the skin, and the antioxidant defenses of the skin are damaged by these free radicals, thereby increasing wrinkles and relaxing the skin, thereby accelerating skin aging.

To date, most studies on antioxidants have been conducted in vitro using 1,1-diphenyl-2-picryl hydrazyl (DPPH). It has been formulated to confirm the free radical scavenging ability, and it has not yet reached the in vivo test stage to measure the actual protective effect by measuring the defense factor activity of the antioxidant system present in skin cells or skin.

Therefore, the present inventors have studied extracts of various substances in order to search for substances capable of protecting not only free radical scavenging but also antioxidant protective agents present in skin cells that are reduced in activity by ultraviolet rays. Blueberry, raspberry, cranberry or blackberry extracts belonging to the berry family not only eliminate free radicals, but also superoxide dismutase, catalase and glutathione in skin cells known to decrease their activity by ultraviolet light. The present invention has been found to have antioxidant activity such as protecting the activity of antioxidant defense factors and inhibiting damage.

Accordingly, an object of the present invention is to provide an anti-aging and anti-aging skin composition containing blueberry, raspberry, cranberry or blackberry extract having free radical scavenging ability or intracellular antioxidant protection.

1 is a result of measuring superoxide dismutase (SOD) activity on human skin cell lines of blueberry, raspberry, cranberry or blackberry extract.

2 is a result of measuring catalase activity on human skin cell lines of blueberry, raspberry, cranberry or blackberry extract.

3 is a result of measuring glutathione (GSH) activity of human skin cell lines of blueberry, raspberry, cranberry or blackberry extract.

Figure 4 is a result of measuring the superoxide dismutase activity on the hairless mouse skin of blueberry, raspberry, cranberry or blackberry extract.

FIG. 5 shows the results of measuring catalase activity on hairless mouse skin of blueberry, raspberry, cranberry or blackberry extract. FIG.

6 is a result of measuring glutathione activity on hairless mouse skin of blueberry, raspberry, cranberry or blackberry extract.

The present invention relates to an external preparation composition for skin having free radical scavenging ability or intracellular antioxidant protection, and is characterized by containing at least one extract of blueberry, raspberry, cranberry or blackberry extract.

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

Blueberry, raspberry, cranberry or blackberry extracts having excellent free radical scavenging ability or intracellular antioxidant protection are as follows.

Blueberry (blueberry) extract is preferably used to extract from the fruit of Vaccinium angustifolium , a shrub of the dicotyledonous plant Rhododendron. Blueberry is a fruit belonging to the berry group together with strawberry, blackberry, bilberry, and cranberry, and contains a large amount of effective substances useful for the physiological function of the human body such as vitamin A, vitamin C, niacin, anthocyanin pigment, potassium and calcium. It contains.

Raspberry (rasberry) extract is preferably used to extract from the fruit of Rubus idaeus , a dicotyledon rose rosaceae raspberry fruit tree.

Cranberry (cranberry) extract is preferably used to extract from the fruit of Vaccinium macrocarpon , an evergreen juniper of the Azalea family dicotyledon.

Blackberry (blackberry) extract is preferably used to extract from the fruit of Rubus vilosus , a perennial plant of the monocotyledonous lily of the valley .

Blueberry, raspberry, cranberry or blackberry extract provided by the present invention can be extracted by a conventional extraction method, and the type and content of the extraction solvent may be optionally selected according to the purpose .

The external preparation composition for skin provided in the present invention contains one or more extracts of the blueberry, raspberry, cranberry or blackberry extract as described above.

The external preparation composition for skin provided in the present invention may contain 0.001 to 30% by weight of blueberry, raspberry, cranberry or blackberry extract, preferably 0.01 to 20% by weight, based on the total weight of the external preparation for skin. Can be.

The external preparation composition for skin provided by the present invention contains one or more extracts of blueberry, raspberry, cranberry or blackberry extracts to eliminate free radicals occurring on the skin and to protect the intracellular antioxidant system, thereby acting on free radicals. It is effective in preventing and delaying aging of the skin caused by oxidation.

The external preparation composition for skin provided in the present invention is used for the purpose of eliminating free radicals or protecting an intracellular antioxidant system, and is not particularly limited in the formulation thereof. For example, softening water, nourishing cosmetics, massage cream, nutrition It may be a skin external preparation composition having a formulation of a cream, a pack, a gel or a skin sticking type cosmetic.

In addition, in the external composition of the skin of each formulation, other components than the above components can be appropriately selected and blended by those skilled in the art without difficulty according to the formulation or purpose of use of the external skin preparation.

Hereinafter, the present invention will be described in more detail with reference to Examples and Test Examples, but the scope of the present invention is not limited thereto.

Preparation Example 1 Preparation of Blueberry, Raspberry, Cranberry, Blackberry Extract

Blueberries, raspberries, cranberries, and blackberries were purchased from the market, and each dried 100g of dry weight was put into a flask, and extracted by chilling with 2ℓ of butylene glycol for 3 days, followed by filtration to remove debris. Berry and blackberry extracts were prepared, respectively.

<Test Example 1: Determination of free radical scavenging activity of blueberry, raspberry, cranberry, blackberry extract>

The free radical scavenging activity of the blueberry, raspberry, cranberry or blackberry extract of Preparation Example 1 was measured as follows. The measuring method is 1,1-diphenyl-2-picryl-hydrazyl (DPPH) method, and the reagents are all US Sigma (St. Louis, MO, USA) ) Was used. Since this reagent exists as a relatively stable free radical, it is primarily used as an in vitro method in confirming free radical scavenging action.

The concentrations of blueberry, raspberry, cranberry or blackberry extract used for the measurement of DPPH free radical scavenging activity were 1% by volume, 0.1% by volume, and 0.01% by volume, respectively. Extracts of the above concentrations were placed in 96 well plates, respectively, to which DPPH prepared in 100 μM ethanol solution was added so that the total volume of the solution was 200 μl. After leaving for 30 minutes at 37 ℃ absorbance was measured by 520nm ELISA reader. Free radical scavenging activity (%) was calculated by the following equation.

A: absorbance of control wells not treated with the extract of the present invention

B: Absorbance of Experimental Wells Treated with Extracts of the Invention

The results are shown in Table 1 below.

content Vitamin c Blueberry Extract Raspberry Extract Cranberry Extract Blackberry Extract 50 ppm 79.7 - - - - 1 volume% - 85.6 80.3 75.2 78.6 0.1% by volume - 54.3 48.9 37.8 45.3 0.01% by volume - 25.7 16.7 12.7 22.1 NOTE The data above is an average of four measurements.

As shown in the results of Table 1, blueberry, raspberry, cranberry or blackberry extract has excellent free radical scavenging activity, such as vitamin C, it can be seen that this free radical scavenging activity is concentration dependent.

Test Example 2: Determination of Antioxidant Protective Effects of Blueberry, Raspberry, Cranberry and Blackberry Extracts on Human Skin Cell Lines>

The cell line used in this experiment was developed by Dr. Human keratinocytes HaCaT cell line distributed from Fusenig. Dispense the cell lines into 6.0 × 10 ⁶ cells in a 150 × 150 culture plate and incubate for 1 day. The concentration of blueberry, raspberry, cranberry or blackberry extract in DMEM (Dulbecos modified eagles medium, GIBCO BRL, Life Technologues) medium containing bovine serum (FBS) was 1%, 0.1%, and 0.01% by volume. The aliquots were dispensed into the treated media and incubated at 37 ° C. and 5% CO ₂ for 1 day. 1X phosphate buffer solution (PBS, phosphate buffered saline, Sigma, St. Louis, MO, USA) was used for washing after incubation. After incubation, UV light was treated at 30 mJ / cm 2 using an ultraviolet B (UVB) lamp (Model: F15T8, UVB 15W, Sankyo Dennki, Japan) and incubated for 1 day at 37 ° C. and 5% CO _2. It was then harvested and used to measure antioxidant defenses. As a positive control for comparing the results, the concentration of vitamin C was 10 μM.

Superoxide dismutase (SOD), catalase and glutathione (GSH) were measured as antioxidant defense factors, respectively. Superoxide dismutase is described by McCord's method [ J. Biol. Chem. (1969) 244: 6049-6055], and catalase was determined by Budhuin's method [ Biochem. J. (1964) 92: 179-184], and glutathione was measured according to Akerboom's method [ Methods in Enzymology (1981) 77: 373-382].

The activity of the intracellular antioxidant system was expressed in% based on 100% of the activity of the control group not irradiated with UVB and calculated by the following equation.

A: activity of the control group

B: Activity of UVB treated group and each experimental group

In addition, the results of measuring the protective effect on the intracellular antioxidant system of the experimental group treated with blueberry, raspberry, cranberry or blackberry extract compared to the group irradiated with UVB was expressed in% and calculated by the following equation (Fig. 3). 1, 2, 3).

A: activity of the control group

B: activity of UVB treatment group

C: activity of each experimental group

The results are shown in Table 2 and FIGS. 1, 2, and 3.

Experimental group content Superoxide dismutase Catalase Glutathione Control 100 100 100 UVB treatment group 57.6 75.3 67.7 Vitamin c 10 μM 82.6 87.8 86.5 Blueberry extract 1 volume% 83.7 86.7 85.2 0.1% by volume 83.7 77.8 84.9 0.01% by volume 74.2 76.0 68.2 Raspberry extract 1 volume% 84.2 78.3 80.1 0.1% by volume 85.1 77.0 85.5 0.01% by volume 62.8 77.8 78.0 Cranberry Extract 1 volume% 78.7 80.6 80.4 0.1% by volume 73.5 78.3 80.3 0.01% by volume 64.1 76.7 71.2 Blackberry Extract 1 volume% 74.8 77.8 79.8 0.1% by volume 75.6 79.0 69.5 0.01% by volume 64.9 75.8 68.0 NOTE The data above is an average of four measurements.

As can be seen from Table 2 and FIGS. 1, 2, and 3, blueberry, raspberry, cranberry or blackberry extracts have antioxidant activity reduced by UV light, such as vitamin C, which is known for its antioxidant effect. It can be expected to exhibit antioxidant efficacy and further anti-aging activity by protecting against.

<Test Example 3: Determination of antioxidant system protective effect of blueberry, raspberry, cranberry or blackberry extract on hairless mouse skin>

The antioxidative protective effect of blueberry, raspberry, cranberry or blackberry extracts on the deactivation of antioxidant defenses of UVB-irradiated skin using about 10 weeks old female hairless mouse The experiment was conducted.

To prepare a cream containing the blueberry, raspberry, cranberry or blackberry extract (Examples 1 to 8), and used as a sample, as shown in Table 3, to compare the results 5% by weight of vitamin C Cream containing (Comparative Example 2) and cream containing neither Blueberry, Raspberry, Cranberry or Blackberry extract or Vitamin C (Comparative Example 1) were prepared.

ingredient Content (% by weight) Comparative Example 1 Comparative Example 2 Example 1 Example 2 Example 3 Example 4 Example 5 Example 6 Example 7 Example 8 glycerin 3.0 3.0 3.0 3.0 3.0 3.0 3.0 3.0 3.0 3.0 Butylene glycol 2.5 2.5 2.5 2.5 2.5 2.5 2.5 2.5 2.5 2.5 Propylene glycol 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 Squalene 5.0 5.0 5.0 5.0 5.0 5.0 5.0 5.0 5.0 5.0 Capric / Capric Triglycerides 3.0 3.3 3.3 3.3 3.3 3.3 3.3 3.3 3.3 3.3 Stearic acid 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 Carboxy Vinyl Polymer 0.3 0.3 0.3 0.3 0.3 0.3 0.3 0.3 0.3 0.3 Liquid paraffin 7.0 7.0 7.0 7.0 7.0 7.0 7.0 7.0 7.0 7.0 Beeswax 4.5 4.5 4.5 4.5 4.5 4.5 4.5 4.5 4.5 4.5 Cetostearyl alcohol 1.5 1.5 1.5 1.5 1.5 1.5 1.5 1.5 1.5 1.5 Polysorbate 60 1.5 1.5 1.5 1.5 1.5 1.5 1.5 1.5 1.5 1.5 Glycerin Monostearate 4.5 4.5 4.5 4.5 4.5 4.5 4.5 4.5 4.5 4.5 Sorbitan monosulfate 0.05 0.05 0.05 0.05 0.05 0.05 0.05 0.05 0.05 0.05 antiseptic Quantity Quantity Quantity Quantity Quantity Quantity Quantity Quantity Quantity Quantity Vitamin c - 5.0 - - - - - - - - Blueberry extract - - 1.0 0.1 - - - - - - Raspberry extract - - - - 1.0 0.1 - - - - Cranberry Extract - - - - - - 1.0 0.1 - - Blackberry Extract - - - - - - - - 1.0 0.1 Purified water Remaining amount Remaining amount Remaining amount Remaining amount Remaining amount Remaining amount Remaining amount Remaining amount Remaining amount Remaining amount Sum 100.0 100.0 100.0 100.0 100.0 100.0 100.0 100.0 100.0 100.0

The creams of Examples 1 to 8 and Comparative Examples 1 and 2 prepared according to the composition of Table 3 were applied to the back of the hairless mice once a day for 4 days. The amount of ultraviolet light B irradiated during the test period was 400mJ / cm 2 using Waldmann UV800 (Herbert Waldmann GmbH & E Phillips TL / 12 lamp emitting 280-305nm). The back skin of the hairless mouse was biopsied and used for analysis, and the analysis items were the same as described in Test Example 3 above. In addition, the result was computed by Formula (2) and (3) which were described in the said Test Example 3.

The results are shown in Table 4 and FIGS. 4, 5, and 6.

Experimental group and content SOD Catalase GSH Control 100 100 100 Comparative Example 1 UVB treatment group 78.2 81.5 73.1 Comparative Example 2 Vitamin C 5.0 wt% 89.5 90.1 92.2 Example 1 Blueberry Extract 1.0 wt% 93.2 88.7 93.5 Example 2 Blueberry Extract 0.1% by weight 81.3 84.2 87.3 Example 3 1.0% by weight of raspberry extract 87.6 86.7 79.6 Example 4 Raspberry Extract 0.0% by weight 79.9 86.7 83.1 Example 5 1.0% Cranberry Extract 85.3 87.2 83.6 Example 6 0.1% Cranberry Extract 82.1 84.3 76.7 Example 7 Blackberry Extract 1.0% by weight 88.9 89.2 76.2 Example 8 Blackberry Extract 0.1% by weight 84.3 82.1 75.9 NOTE The data above is an average of four measurements.

As can be seen from Table 4 and Figures 4, 5, 6, blueberry, raspberry, cranberry or blackberry extract is the antioxidant activity of the antioxidant reduced by UV, such as vitamin C is known for its antioxidant effect It is expected to show antioxidant efficacy and further anti-aging activity.

As described above, the external composition for skin composition of the present invention containing blueberry, raspberry, cranberry or blackberry extract is excellent in free radical scavenging or intracellular antioxidant protection effect, and thus has cosmetics having antioxidant and anti-aging effects. It is a very useful invention that can be manufactured.

Claims (5)

A composition for external application for skin, characterized in that it contains at least one extract of blueberry, raspberry, cranberry or blackberry extract. The method of claim 1, wherein the external composition for skin comprises at least one extract of the blueberry, raspberry, cranberry or blackberry extract in an amount of 0.001 to 30% by weight based on the total weight of the external composition of the skin. Skin external preparation composition. The composition for external application for skin according to claim 1, wherein the blueberry, raspberry, cranberry or blackberry extract has free radical scavenging ability or intracellular antioxidant protection. 4. The composition for external application for skin according to claim 3, wherein the intracellular antioxidant protection is attributable to the antioxidant protective activity of the blueberry, raspberry, cranberry or blackberry extract. Antioxidant or anti-aging skin external composition comprising at least one extract of blueberry, raspberry, cranberry or blackberry extract having free radical scavenging activity or intracellular antioxidant protection.