KR102215164B1 - Composition for preventing or improving skin aging containing acetic acid 16-hydroxy-octadeca-9,17-diene-12,14-diynyl ester - Google Patents

Abstract

The present invention relates to a cosmetic composition for anti-aging, and more specifically, contains 16-hydroxy-octadeca-9,17-diene-12,14-diynyl ester acetic acid as an active ingredient to reduce skin wrinkles and skin elasticity. It increases, and also relates to a composition capable of preventing or improving skin photoaging due to ultraviolet rays.

Description

Composition for preventing or improving skin aging containing acetic acid 16-hydroxy-octadeca- containing 16-hydroxy-octadeca-9,17-diene-12,14-diynyl ester acetic acid 9,17-diene-12,14-diynyl ester}

The present invention relates to a cosmetic composition for anti-aging, and more specifically, contains 16-hydroxy-octadeca-9,17-diene-12,14-diynyl ester acetic acid as an active ingredient to reduce skin wrinkles and skin elasticity. It increases, and also relates to a composition capable of preventing or improving skin photoaging due to ultraviolet rays.

Human skin is the primary barrier of the human body, protecting the internal organs from stimuli of the external environment such as changes in temperature and humidity, ultraviolet rays, and pollutants. As we age, it is affected by various internal and external factors. Undergoing change. In other words, internally, the secretion of various hormones that regulate metabolism decreases, the function of immune cells and the activity of cells decrease, thereby reducing the biosynthesis of immune proteins and biological components necessary for the living body, and externally, due to the destruction of the ozone layer. Due to the increase in the amount of ultraviolet rays reaching the surface of the sun's rays and increasing environmental pollution, free radicals and active harmful oxygen increase, so that the skin thickness decreases, wrinkles increase, and elasticity decreases. The complexion of the skin becomes dull, skin troubles frequently occur, spots, freckles and age spots also increase, the complexion deteriorates and the skin tone darkens, and various changes occur.

In order to prevent changes in the skin condition caused by such internal and external factors, and to maintain a healthy skin condition, the skin condition is improved by using physiologically active substances obtained from various known animals, plants, microorganisms, etc. in addition to cosmetics. There has been an effort for it.

On the other hand, in recent years, as a kind of component separated from natural materials, there is increasing interest in components extracted from ginseng, especially polyyne components. Among them, it has been found that panaxinol, panaxidol, panaxtriol, and the like, as polyacetylene components, have cytotoxicity against cancer cells and thus have anticancer effects. However, studies on the polyacetylene component are still insignificant, and in particular, notable research results have been reported on the efficacy exhibited in relation to improving skin conditions other than the anticancer effect.

Korean Patent Application Publication No. 10-2012-0083943 (published on July 27, 2012)

Accordingly, the inventors of the present invention were studying to find a substance that exhibits anti-aging effects, etc., while 16-hydroxy-octadeca-9,17-diene-12,14-diynyl ester acetic acid has anti-aging and improving effects, so that the skin wrinkles and It was found that it can improve skin elasticity and the like, and the present invention was completed.

Accordingly, an object of the present invention is to provide a composition capable of preventing or improving skin aging by containing 16-hydroxy-octadeca-9,17-diene-12,14-diynyl ester acetic acid.

In order to achieve the above object, the present invention is an anti-aging composition containing 16-hydroxy-octadeca-9,17-diene-12,14-diynyl ester acetic acid, which is a compound represented by the following formula (1) as an active ingredient Provides.

The composition of the present invention can effectively prevent or improve skin aging through excellent antioxidant power by containing 16-hydroxy-octadeca-9,17-diene-12,14-diynyl ester acetic acid.

The composition according to the present invention is 16-hydroxy-octadeca-9,17-diene-12,14-diynyl ester acetic acid (acetic acid 16-hydroxy-octadeca-9,17-diene-12,14-diynyl ester) By containing as an active ingredient, it can prevent or improve skin aging.

16-hydroxy-octadeca-9,17-diene-12,14-diynyl ester acetic acid used in the present invention is a compound having the structure of the following formula (1), has a molecular weight of 316, and (Z)-16 Also known as -hydroxy-9,17-octadecadiene-12,14-diynyl acetate ((Z)-16-hydroxy-9,17-octadecadiene-12,14-diynyl acetate).

[Formula 1]

The composition according to the present invention contains 16-hydroxy-octadeca-9,17-diene-12,14-diynyl ester acetic acid in an amount of 0.001 to 50% by weight, preferably 0.01 to 30% by weight, based on the total weight of the composition. Preferably, it may be contained in an amount of 0.1 to 10% by weight. If the content of the 16-hydroxy-octadeca-9,17-diene-12,14-diynyl ester acetic acid is less than 0.001% by weight, the efficacy and effect of the component is weak, and if it exceeds 50% by weight, skin safety Or because there is a problem in the formulation.

16-hydroxy-octadeca-9,17-diene-12,14-diynyl ester Since acetic acid is a component with excellent antioxidant power, 16-hydroxy-octadeca-9,17-diene-12,14-diene The composition of the present invention containing nil ester acetic acid provides excellent antioxidant power. Accordingly, the composition of the present invention can effectively protect or improve endogenous aging, which is aging according to an increase in age, and exogenous aging (ie, photoaging) affected by free radicals generated by external factors such as ultraviolet rays and cigarette smoke. Thereby, it is possible to reduce skin wrinkles and improve skin elasticity.

In addition, the composition of the present invention can promote skin regeneration.

In addition, the composition of the present invention can prevent the generation of skin irritation due to its excellent antioxidant power.

The composition of the present invention may be formulated as a composition for external application for skin, in particular, a cosmetic composition, and may be formulated containing a cosmetically or dermatologically acceptable medium or base. In addition, the composition of the present invention may be provided in any formulation suitable for topical application, for example, a solution, an emulsion obtained by dispersing an oil phase in an aqueous phase, an emulsion obtained by dispersing an aqueous phase in an oil phase, a suspension, a microemulsion, a microcapsule, It may be provided in the form of microgranular or ionic (liposome) and nonionic vesicle dispersants, solids, gels, powders, pastes, foams or aerosol compositions. Specifically, the composition of the present invention may be provided in the form of a cream, skin, lotion, powder, ointment, spray or conceal stick. Compositions of such formulations can be prepared according to conventional methods in the art.

In addition, the composition according to the present invention may contain other ingredients that can give synergistic effect to the main effect, preferably within a range that does not impair the main effect in addition to the above-described substances. In addition, the composition according to the present invention may further include a moisturizing agent, an emollient agent, an ultraviolet absorber, a preservative, a sterilizing agent, an antioxidant, a pH adjuster, an organic and inorganic pigment, a fragrance, a cooling agent or a limiting agent. The blending amount of the ingredients can be easily selected by a person skilled in the art within a range that does not impair the object and effect of the present invention, and the blending amount may be 0.01 to 5% by weight, specifically 0.01 to 3% by weight, based on the total weight of the composition. .

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

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 may include injections, drops, ointments, lotions, sprays, suspensions, emulsions, and suppositories. The composition according to the present invention can be easily formulated as an active ingredient by conducting it according to a commercial method, and in this case, surfactants, excipients, coloring agents, spices, preservatives, stabilizers, buffers, suspensions, and other commonly used auxiliary agents can be appropriately used. have.

The dosage of the active ingredient of the pharmaceutical composition of the present invention will vary depending on the age, sex, weight, pathological condition and severity of the subject to be administered, the route of administration, or the judgment of the prescriber. Determination of an appropriate dose based on these factors is within the level of those skilled in the art, and its daily dose is, for example, 0.1 mg/kg/day to 100 mg/kg/day, more specifically 5 mg/kg/day to 50 mg/kg. /It can be work, but is not limited thereto.

Hereinafter, the present invention will be described in more detail through examples. These examples are for illustrative purposes only, and it will be apparent to those of ordinary skill in the art that the scope of the present invention is not construed as being limited by these examples.

[Reference Example 1] Preparation of 16-hydroxy-octadeca-9,17-diene-12,14-diynyl ester acetic acid

16-hydroxy-octadeca-9,17-diene-12,14-diynyl ester acetic acid for testing the efficacy of the composition of the present invention was purchased and used from Cosvision (Korea).

[Test Example 1] Effect of inhibiting the production of reactive oxygen species (ROS)

Keratinocytes (cell line: HaCaT; source: ATCC) isolated from human epidermal tissue are cultured in 24-well cells with keratinocyte growth media (Clonetics, BioWhittacker, MD, USA) Put 5×10 ⁴ in each well of the plate and attached for 24 hours. After removing the culture solution, 100 µl of phosphate buffered saline (PBS) was added to each well. After irradiating the keratinocytes with UV rays of 30mJ/cm2 using an ultraviolet B (UV B) lamp (Model: K5T8, UV B 15W, Sankyo Dennki, Japan), the PBS was removed and the keratinocyte culture solution was added to each well. 200 μl was added. Here, as a test substance, 16-hydroxy-octadeca-9,17-diene-12,14-diynyl ester acetic acid was treated with 2 μg/ml, and the reactive oxygen species (ROS) increased by UV stimulation at a certain time ) Was quantified. At this time, for comparison, the amount of reactive oxygen species was also measured for those that were not treated with the test substance (no treatment) and were not stimulated with ultraviolet rays and those that were stimulated with ultraviolet rays without treatment with the test substance. The amount of ROS was quantified by referring to Tan's method, which measures the fluorescence of dichlorofluorescin diacetate (DCF-DA) oxidized by ROS (Tan et al., 1998, J. Cell Biol. Vol. 141, pp1423-1432), The ratio of the control group to ROS is shown in Table 1 below.

Time elapsed after irradiation with UVB 30mJ/cm ² 0 hours 2 hours 3 hours No treatment 100 244 287 UVB + no treatment 100 325 381 UVB+16-hydroxy-octadeca-9,17-diene-12,14-diynyl ester acetic acid 100 283 330

In the results of Table 1, 16-hydroxy-octadeca-9,17-diene-12,14-diynyl ester acetic acid effectively inhibits the production of ROS, which is known to cause skin cell damage by ultraviolet rays, and has excellent antioxidant efficacy. It can be seen that it represents.

This suggests that the 16-hydroxy-octadeca-9,17-diene-12,14-diynyl ester acetic acid used in the present invention can inhibit oxidation and prevent aging.

[Test Example 2] Elastase activity inhibition efficacy measurement

The ability of 16-hydroxy-octadeca-9,17-diene-12,14-diynyl ester acetic acid to inhibit the elastase activity was measured by comparing with EGCG. The elastase and substrate used were commercially purchased from Sigma-Aldrich, USA (Cat. No. E0127).

The inhibitory effect of elastase activity was tested by the following test method.

In a 96-well plate prepared with 10 mg/L Tris-HCL buffer (pH 8.0), 16-hydroxy-octadeca-9,17-diene-12,14-diynyl ester acetic acid (200 μL) and 20 of Reference Example 1 above. 50 µL of a µg/mL elastase-type III solution was mixed. EGCG 250 μM was used as a positive control, and distilled water was used for the untreated group, which is a negative control. Then, 0.4514 mg/mL N-SUCCINYL-ALA-ALA-ALA-p-NITROANILIDE 100 µL prepared in the above buffer was added, followed by reaction 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 and corrected.

The method of calculating the inhibitory effect of elastase activity is as shown in Equation 1 below, and the results are shown in Table 2 below.

A: Absorbance at a wavelength of 415 nm when no test substance was added and enzyme was added

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

C: absorbance at a wavelength of 415 nm in the addition of the test substance and the addition of enzyme

D: Absorbance at a wavelength of 415 nm with the addition of the test substance and no enzyme

compound Degree of inhibition (%) Untreated group 0 EGCG 65 16-hydroxy-octadeca-9,17-diene-12,14-diynyl ester acetic acid 55

As shown in Table 2, it can be seen that 16-hydroxy-octadeca-9,17-diene-12,14-diynyl ester acetic acid exhibits an excellent degree of inhibition of elastase activity.

[Test Example 3] Collagenase (MMP-1) inhibitory ability

The ability of the 16-hydroxy-octadeca-9,17-diene-12,14-diynyl ester acetic acid of the present invention to inhibit collagenase production was measured in comparison with retinoic acid.

Human fibroblasts in a 96-well microtiter plate containing DMEM (Dulbecco's Modified Eagle's Media) medium containing 2.5% fetal bovine serum (cell line: KF-4109; source: Klavosa) Was put so as to be 5,000 cells/well, and incubated in a CO ₂ 5%, 37°C incubator until about 70-80% growth. 16-hydroxy-octadeca-9,17-diene-12,14-diynyl ester acetic acid of Reference Example 1 was treated at a concentration of 30 μg/ml for 24 hours, and then a cell culture solution was collected.

A commercially available collagenase measuring instrument (Amer Sham Pharma, Catalog #: RPN 2610) was used to measure the collagenase production level of the collected cell culture solution. First, the collected cell culture solution was added to a 96-well plate uniformly coated with the primary collagenase antibody, and the antigen-antibody reaction was performed in a thermostat for 3 hours. After 3 hours, the secondary collagen antibody to which the chromophore was bound was added to a 96-well plate and reacted for 15 minutes. After 15 minutes, a color inducing substance (3,3',5,5'-tetramethylbenzidine, sigma) is added to induce color development at room temperature for 15 minutes, and then 1M sulfuric acid is added to stop the color reaction, and the color of the reaction solution turns yellow. And the degree of yellow was different depending on the degree of reaction progress.

The absorbance of the yellow 96-well plate was measured at 405 nm using an absorbance meter, and the degree of synthesis of collagenase was calculated by Equation 2 below, and the results are shown in Table 3 below. At this time, the reaction absorbance of the cell culture solution collected from the group not treated with the composition was used as a control.

Test substance Expression level (%) Untreated group 100 Retinoic acid 75 16-hydroxy-octadeca-9,17-diene-12,14-diynyl ester acetic acid 52

As shown in Table 3 above, it can be seen that 16-hydroxy-octadeca-9,17-diene-12,14-diynyl ester acetic acid exhibits an excellent degree of inhibition of collagenase expression.

Through these results, it can be confirmed that the 16-hydroxy-octadeca-9,17-diene-12,14-diynyl ester acetic acid of the present invention has an effect of inhibiting protease (MMP-1) as a substrate metal. have.

[Formulation Example 1 and Comparative Formulation Example 1]

A nutrient cream was prepared by a conventional method according to the composition in Table 4 below (unit: wt%).

Ingredients Formulation Example 1 Comparative Formulation Example 1 Purified water To 100 To 100 16-hydroxy-octadeca-9,17-diene-12,14-diynyl ester acetic acid 0.1 - Hydrogenated vegetable oil 1.50 1.50 Stearic acid 0.60 0.60 Glycerol stearate 1.00 1.00 Stearyl alcohol 2.00 2.00 Polyglyceryl-10 Pentastearate & Behenyl Alcohol & Sodium Stearoyl Lactylate 1.00 1.00 Arachidyl Behenyl Alcohol & Arachidyl Glucoside 1.00 1.00 Cetearyl Alcohol & Cetearyl Glucoside 2.00 2.00 PEG-100 stearate & glycerol oleate & propylene glycol 1.50 1.50 Caprylic/Carlic Triglyceride 11.00 11.00 Cyclomedicone 6.00 6.00 Preservatives, incense Appropriate amount Appropriate amount Triethanol amine 0.1 0.1

[Test Example 4] Checking the effect of improving skin elasticity

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

20 healthy women in their 30s to 40s were divided into two groups of 10 for each of the two groups of Formulation Example 1 and Comparative Formulation Example 1, and applied a nourishing cream once a day for 12 weeks on the left side of the face, and then a skin elasticity meter ( Cutometer SEM 575, C+K Electronic Co., Germany) was used to measure skin elasticity. The results are shown in Table 5 below. The result values in Table 5 are described as the ΔR8 value of Cutometer SEM 575, and the R8 value indicates the property of skin viscoelasticity.

Experimental product Skin elasticity effect Formulation Example 1 0.21 Comparative Formulation Example 1 0.10

As shown in Table 5, Formulation Example 1 containing 16-hydroxy-octadeca-9,17-diene-12,14-diynyl ester acetic acid of the present invention was compared to the group to which Comparative Formulation Example 1 was applied. The elasticity of the skin was increased even more.

Accordingly, it can be seen that the composition containing 16-hydroxy-octadeca-9,17-diene-12,14-diynyl ester acetic acid of the present invention is very effective in improving skin elasticity.

[Test Example 5] Confirmation of skin wrinkle improvement efficacy

Formulation Example 1 and Comparative Formulation Example 1 were used to confirm the effect of improving wrinkles in humans by the composition of the present invention.

In order to confirm the wrinkle improvement effect of Formulation Example 1 and Comparative Formulation Example 1, the following evaluation was performed. 20 healthy women in their 40s were divided into 2 groups of 10 people for each of the 2 groups of Formulation Example 1 and Comparative Formulation Example 1, and a nourishing cream was applied to the face once a day for 12 weeks. The condition of the wrinkles was measured with a skin meter (visiometer, SV600, Courage+Khazaka electronic GmbH, Germany) and image analysis was performed. The results are shown in Table 6 below. The values in Table 6 below represent the average of the values obtained by subtracting the value of the variable before application from the value of each parameter 12 weeks after application.

8 weeks after use
Clinical results R1 R2 R3 R4 R5 Formulation Example 1 0.20 0.19 0.15 0.02 0.02 Comparative Formulation Example 1 0.27 0.26 0.21 0.03 0.03

R1: The difference between the maximum and minimum values of the wrinkle contour line

R2: After randomly dividing the wrinkle contour line by 5 spaces, the average of R1 values among them

R3: The highest value among R1 values divided by 5

R4: Average value obtained by subtracting each peak and valley value from the baseline of the wrinkle contour line

R5: The difference value of the value obtained by subtracting each wrinkle contour from the baseline of the wrinkle contour line

As shown in Table 6, it was found that the composition for external application of Formulation Example 1 was very excellent in improving skin wrinkles.

[Test Example 6] Measurement of biosynthesis inhibitory efficacy of gelatinase A (MMP-9) and gelatinase B (MMP-2) by ultraviolet rays

Human keratinocytes (cell line: HaCaT; source: ATCC) were cultured in a 24 hole plate incubator at a concentration of 1x10 ⁴ , and after 24 hours, UV B was irradiated with 30 mJ/cm ² , and then 16-hydroxy-octane Deca-9,17-diene-12,14-diynyl ester was replaced with a medium containing 10 ppm of acetic acid. On the second day of culture, the supernatant was harvested, zymography was performed using gelatin gel, and the amounts of MMP-2 and MMP-9 produced were measured with a densitometer and quantified. At this time, the experimental group irradiated with UVB and replaced with a medium that did not contain a separate sample was used as a control group, and the comparison values were shown in Table 7 with the control group as 100.

Biosynthesis rate of gelatinase A (MMP-9) and gelatinase B (MMP-2) ingredient MMP-2(%) MMP-9(%) 16-hydroxy-octadeca-9,17-diene-12,14-diynyl ester acetic acid 73 74 Control 100 100

As shown in Table 7 above, 16-hydroxy-octadeca-9,17-diene-12,14-diynyl ester acetic acid is used as MMP-2, an enzyme that degrades type 4 collagen and type 7 collagen, which are skin substrates. It can be seen that it is an ingredient that can protect the skin by reducing the biosynthesis of MMP-9, preventing the decomposition of the boundary between the epidermis and the dermis.

Claims (5)

A composition for external application for skin for anti-aging containing 16-hydroxy-octadeca-9,17-diene-12,14-diynyl ester acetic acid represented by the following formula (1) as an active ingredient.
[Formula 1]

The composition for external application for skin according to claim 1, wherein the active ingredient is contained in an amount of 0.001 to 50% by weight based on the total weight of the composition. The composition for external application for skin according to claim 1, wherein the composition is for enhancing skin elasticity. The composition for external application for skin according to claim 1, wherein the composition is for improving skin wrinkles. The composition for external application for skin according to claim 1, wherein the composition is for suppressing photoaging.