KR20040040557A - Cosmetics composition having anti-wrinkle and anti-aging effect - Google Patents

Abstract

PURPOSE: Provided is a cosmetics composition having anti-wrinkle and anti-aging effects, which composition contains vitamin C or its derivatives and astaxanthin. Therefore, it has excellent effects on fibroblast proliferation, collagen synthesis, moisturization, skin elasticity and wrinkle removal without adverse side effect. CONSTITUTION: A cosmetics composition having anti-wrinkle and anti-aging effects is characterized by containing 0.01-10 wt.% of a mixture of vitamin C or its derivative and astaxanthin in a weight ratio of 1:0.001-1:20, based on the total weight of the composition.

Description

Wrinkle improvement and anti-aging cosmetic composition {COSMETICS COMPOSITION HAVING ANTI-WRINKLE AND ANTI-AGING EFFECT}

The present invention relates to a cosmetic composition having anti-wrinkle and anti-aging effects, and more particularly, to a cosmetic having anti-wrinkle and anti-aging effects by containing vitamin C or a derivative of vitamin C and astaxanthin.

In recent years, as women's desire for the function of cosmetics has increased, high-functional products have emerged that not only clean and aesthetic functions, which are the basic functions of cosmetics, but also whitening to bright and white skin, improving wrinkles and suppressing skin, etc. . In particular, wrinkle improvement products are of interest to many women who want to stay young, and thus efforts are being made to improve wrinkles on the skin.

In this regard, Japanese Patent Application Laid-open No. Hei 5-246838 discloses a method for smoothing collagen metabolism to improve skin wrinkles. When collagen biosynthesis is increased due to the loss of balance of collagen synthesis and degradation due to aging, or the activity of collagenase that degrades collagen metabolism is degraded, the production of cross-linked collagen is uncontrolled and aging of skin tissue Is promoted. Therefore, cosmetic compositions containing ammonium acetate, ammonium stannate and ammonium nitrate, which increase the activity of collagenase, revitalize collagen metabolism to prevent skin aging.

U.S. Patent No. 5,118,707 discloses a method for improving skin wrinkles containing benzofuran derivatives, including amiodarone. It is reported that applying the composition to the skin improves the skin wrinkles.

US Pat. No. 5,340,568 also contains a composition containing 2-Deoxy-2-halo-lysophosphatidic acid, which functions similar to growth factors and induces cell proliferation. It is described as inhibiting production.

In addition, WO 94/04184 describes a composition in which a protein growth factor is stabilized to prevent skin aging such as skin wrinkles and sagging skin. Protein growth factors used in this technique are epidermal growth factor (EGF), insulin-like growth factor (IGF), platelet-derived growth factor (PDGF), fibroblast growth factor (FGF), and the like.

However, the above described skin wrinkle improvement active ingredients cannot be used as some cosmetic ingredients or are very unstable, and it is not easy to transfer to the skin, so a special stabilization system and delivery system are needed, and the effect of improving wrinkles is not visible. There is a problem. Therefore, recently, attention has been gradually focused on skin protection agents containing retinoids. Currently, retinoids have been used as a means to solve photoaging phenomena such as wrinkles, skin thickening, sagging, and elasticity, which are a result of the accumulation of sunlight. However, these retinoids are very unstable compounds, and are sensitive to light (ultraviolet light), moisture, heat, and air (oxygen) and easily cause chemical changes, and researches for solving these problems continue.

Materials with high antiwrinkle effects, such as retinol and retinoic acid, are difficult to use as raw materials for cosmetics due to chemical instability, difficulty of use, complexity of formulation technology, and skin irritation as described above. Retinyl palmitate and retinyl ester, such as retinyl palmitate and retinyl acetate, which have high ease of use, and β-carotene, which is a precursor of vitamin A, are widely used as raw materials for cosmetics.

In addition, vitamin C is recognized as the most powerful antioxidant in relation to the aging phenomenon of the human body, but it is difficult to use in cosmetics because of stability problems. Accordingly, the present cosmetic industry has made a lot of efforts to use the vitamin C itself, but do not overcome the unstable properties, even if the effect is less stable than the vitamin C itself is using a more stable vitamin C derivative.

Astaxanthin, on the other hand, is a lipid-based substance that is a kind of carotenoid, and has the characteristics of a fat-soluble pigment. Contents of complex functions in metabolic aspects such as promotion have been studied. In addition, it is a substance that is increasingly used in various fields such as food and pharmaceuticals.

An object of the present invention is to obtain a cosmetic composition excellent in stability while excellent in anti-wrinkle effect by adding vitamin C or a derivative of vitamin C and astaxanthin.

1 is a graph comparing fibroblast proliferation ability,

2 is a collagen biosynthesis comparison graph.

In order to achieve the above object, according to the present invention, containing a mixture of vitamin C or a derivative of vitamin C and astaxanthin in a weight ratio of 1: 0.001 to 1:20, containing 0.01 to 10% by weight relative to the total weight of the composition There is provided a wrinkle improvement and anti-aging cosmetic composition.

Hereinafter, the present invention will be described in detail.

Vitamin C and vitamin C derivatives of the present invention can be prepared and used according to a known method or purchased commercially available. Preferred derivatives of vitamin C include tetrahexyldecyl ascorbate (VC-IP), magnesium ascorbyl phosphate (VC-PMG), ascorbyl glucoide (VC-G), and the like.

Astaxanthin may also be prepared and used according to a known method or commercially available one, but is preferably prepared as a fermentation metabolite using microorganisms because of good yield. In particular, it is preferably prepared using a strain of S. aphidomyces dendrose ATCC-96594.

The mixing ratio of vitamin C or derivatives of vitamin C to astaxanthin is 1: 0.001 to 1:20 by weight. If the weight ratio is less than 1: 0.001, there is almost no effect of improving wrinkles substantially, and if it exceeds 1:20, there is little effect increase due to the increase in the amount of astaxanthin added.

Vitamin C or derivatives of vitamin C and astaxanthin can be added to all cosmetics related to wrinkle improvement such as softening cream, nourishing cream, nourishing cream, massage cream, essence, pack, etc., and the amount added is 0.01 to the total weight of the cosmetic composition. It is preferable that it is -10 weight%, Preferably it is 0.1-10 weight%. When the concentration is less than 0.01% by weight, it is difficult to obtain a wrinkle improvement effect, and when it is blended at 10% by weight or more, the increase in the effect of wrinkle improvement is insignificant compared to the amount used, and it is uneconomical in terms of stability of the product.

Hereinafter, the present invention will be described in more detail with reference to Examples. These examples are merely illustrative of the present invention and the scope of the present invention is not limited to these examples.

Example

(Preparation of Vitamin C and Vitamin C Derivatives)

Vitamin C and vitamin C derivatives, tetrahexyldecyl ascorbate (VC-IP), magnesium ascorbyl phosphate (VC-PMG) and ascorbyl glucoide (VC-G), were supplied from cosmetic raw material suppliers.

(Production of Astaxanthin)

The culture method is described in detail below.

After incubating the strain of S. aphidomyces dendrose ATCC-96594 in malto liquid medium (0.3% malt extract, 0.5% peptone, 1.0% glucose), 1-methyl-3-nitro-1-nitrosoguanidine (NTG) Treatment killed about 98% of yeast. The surviving yeast was diluted and plated in malto-solid medium, and then cultured, and all of the resulting colonies were grown faster than the parent strain ATCC-96594 and were all glossy and dense red. The screening process was repeated four times in malto-solid medium to obtain pure colonies. The strains thus obtained were incubated for 5 days in malto medium. The culture was measured for absorbance at 660 nm and dry cells were measured to obtain the best yield. The culture solution was centrifuged at 10,000 x g for 30 minutes to remove the bacteria, 100 ml of propanol and 300 g of sodium chloride per liter were added to the remaining supernatant and allowed to stand at room temperature for 1 hour, followed by centrifugation to obtain a precipitate. The precipitate was dissolved in 20 mM sodium citrate buffer and then purified using a dialysis membrane.

Cultures were 0.5% nitrogen, 5-15 g / l glucose, 8-11 g / l CSL, 2-4 g / l yeast extract, 0.02 g / l potassium phosphate, 0.03 g / l magnesium sulfate, 0.02 copper sulfate In a liquid fermentation broth containing g / l and 0.05 g / l of calcium chloride, air and oxygen were mixed at a ratio of 10: 1, and fermented and cultured in a batch manner while being sucked at 4 to 6 vvm aeration rate.

The prepared raw materials were prepared as follows alone or by mixing two or more.

Treatment Raw material mixing One Vitamin c 2 VC-IP 3 VC-PMG 4 VC-G 5 Astaxanthin 6 Vitamin C + VC-IP (1: 0.5) 7 Vitamin C + VC-PMG (1: 0.5) 8 Vitamin C + VC-G (1: 0.5) 9 Vitamin C + Astaxanthin (1: 0.5) 10 VC-IP + Astaxanthin (1: 0.5) 11 VC-PMG + Astaxanthin (1: 0.5) 12 VC-G + Astaxanthin (1: 0.5)

(Test Example 1)

Fibroblast Proliferation Capacity Comparison Experiment

Cell proliferation was tested using human skin derived fibroblasts. Fibroblasts were used by proliferating cells stored in -70 ° C or liquid nitrogen. Fibroblasts cultured in T-75 flasks were washed 2-3 times with PBS, and the adherent cells were dropped using trypsin, followed by centrifugation at 1500 rpm for 5 minutes (Heraeus, Labofuge 400R) to collect the cells. After centrifugation, the supernatant was discarded, and 10 ml of DMEM (10% FBS) was added to the precipitated cells and mixed evenly. By measuring the number of cells using a blood cell counter ⁴ to 10 ea / well degree of cell to each well of a 96 well it was dispensed by 200㎕. When cultured at 37 ° C., 5% CO ₂ and about 50% of the well area grew, the medium was exchanged with fresh DMEM (FBS10%), and each concentration (0.001%, 0.01%, 0.1%, 1%, 10%, 15%), cells were incubated for 24 hours at 37 ℃, 5% CO ₂ .

TT {(3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyl tetrazolium bromide (Sigma)} was dissolved in PBS to 2 mg / ml and stored in the refrigerator, Each well was treated with 20ul of MTT reagent (5mg / ml in PBS), incubated for 2 more hours, and then discarded the supernatant and mixed with 150µl of DMSO or 100µl of 0.04N-HCl. After the step, the absorbance was measured at 560 nm using an ELISA reader (μQuant, Bio-Tek instrument INC.) (Reference: 650 nm).

Cell proliferation was calculated according to the following equation, and the results are shown in FIG. 1.

According to Figure 1, compared to vitamin C alone or vitamin C derivatives alone, when astaxanthin was added to vitamin C or derivatives of vitamin C showed an enhancement effect.

(Test Example 2)

Collagen Biosynthesis Effect

Fibroblasts incubated in T-75 flasks were washed 2-3 times with PBS, the adhered cells were dropped using trypsin, and the cells were collected by centrifugation (Heraeus; Labofuge 400R) at 1500 rpm for 5 minutes. After centrifugation, the supernatant was discarded, and 10 ml of DMEM (10% FBS) was added to the collected cells and mixed evenly. By measuring the number of cells using a hemocytometer, cells of about 10 ⁴ ea / well were dispensed into each 200-well well of 96 wells. When cells at about 50% of the well area were grown while incubating at 37 ° C and 5% CO ₂ , the cells were exchanged with fresh DMEM (FBS10%), and each concentration (0.001%, 0.01%, 0.1%) was used for treatments 1 to 12. , 1%, 10%, 15%), the cells were incubated for 24 hours at 37 ℃, 5% CO ₂ .

Collagen synthesis performance measurement kit (Procollagen Type-I C-peptide EIA kit (Takara, MK 101)) was used. For measurement using immunological methods, 100 μl of Ab-POD conjugate solution was transferred to one well and aliquoted, and 20 μl of cell culture or standard solution was added. The 96 well plate was wrapped with foil etc., and it incubated for 3 hours at 37 degreeC. After 3 hours, all the solutions in each well were removed and washed 4 times with PBS. After the washing step, 100 μl of substrate solution was added to each well and incubated at 20-30 ° C. for 15 minutes. In the final step, 100 µl of 1N H ₂ SO ₄ , the reaction termination solution, was added and mixed gently. The absorbance was measured with an ELISA reader and the result was used. After the reaction solution was added, the plate was stored at room temperature for about 1 hour and absorbance was measured at 450 nm.

Synthesis (%) was calculated according to the following equation, and the results are shown in FIG. 2.

In FIG. 2, as shown in the results of cell proliferation ability, when astaxanthin was added to vitamin C or a derivative of vitamin C, an enhancement effect was observed as compared to the treatment using vitamin C alone or a derivative thereof alone.

(Examples 1 to 4 and Comparative Examples 1 to 3)

According to the conventional cosmetic preparation method, the components of the following Table 1 (unit:%) were mixed to prepare a homogeneously mixed essence cosmetic through a heating and cooling process after stirring with a homomixer.

(Test Example 3)

Moisturizing and elasticity test

Moisture was measured by the high frequency impedance method using SKICON-200 (skin surface moisture content meter, IBS).

Twenty healthy men and women were banned from using moisturizers or lotions on their legs two weeks before the start of the trial. Examples 1-4 and Comparative Examples 1-3 were applied daily for 15 days. The test subjects were treated with the composition twice a day for three weeks, once in the morning and then in the evening. The subjects were evaluated and averaged according to the following criteria before and after 21 days of treatment.

0 points = flaking, peeling (<10μΩ)

1 point = slightly dry skin (10 to 30 µΩ)

2 points = lightly dry skin (30-50μΩ)

3 points = smooth, no evidence of dryness (> 50μΩ)

In order to measure the elasticity, 140 women in their 30s and 40s were divided into seven groups, with 20 as one group. In each group, Examples 1-4 and Comparative Examples 1-3 were applied to the face once daily for 1 month, and then a cutometer (SEM575, Courage + Khazaka electronic GmbH) capable of measuring skin elasticity after 1 month. Germany), the elasticity of the facial area was measured and compared with before application.

Skin elasticity was calculated according to the following formula.

The results are shown in Table 2 below.

Effect Production Example Moisturizing Skin elasticity improvement (%) Before treatment After treatment Comparative Production Example 1 1.2 points 2.3 points 48 Preparation Example 1 1.2 points 4.3 points 72 Preparation Example 2 1.2 points 4.0 points 70 Preparation Example 3 1.2 points 3.9 points 69 Preparation Example 4 1.2 points 3.6 points 67 Comparative Production Example 2 1.2 points 2.5 points 52 Comparative Production Example 3 1.2 points 1.4 points 14

(Test Example 4)

Wrinkle Relaxation Test

Twenty healthy women, 20-40 years old, were applied twice a day to Example 1-4 and Comparative Example 1-3 at the corners of the eyes, and then the tail of the eyes was measured. A transparent replica was produced and image analysis was performed using a Skin Visiometer (C + K company) on the day of 0, 30 and 60 days. The evaluation method was based on three wrinkle measurements: wrinkles clearly improved (1), slight wrinkles improved (2) and wrinkles not improved (3).

The results are shown in Table 3.

Wrinkle Relief (1) Slight Wrinkle Relief (2) No effect (3) Comparative Production Example 1 17 people 2 people 1 person Preparation Example 1 20 people 0 people 0 people Preparation Example 2 19 people 1 person 0 people Preparation Example 3 19 people 1 person 0 people Preparation Example 4 19 people 1 person 0 people Comparative Production Example 2 18 people 2 people 0 people Comparative Production Example 3 0 people 2 people 18 people

In Table 3, in the case of the present invention showed more than 90% wrinkle improvement compared to Comparative Example 1.

(Test Example 5)

Human patch experiment

In order to confirm that the cosmetic composition of the present invention is free of skin irritation and other side effects, a patch test was performed on the human body of the compositions of Examples 1-4 and Comparative Examples 1-3.

The test consisted of 30 healthy men and women who used a Finn chamber on the back to put cosmetic products directly into aluminum discs or put them on paper discs and treated them with 40 μl and applied them to the skin with scanpore tape. It fixed, the patch was removed after 24 hours, and the result was determined after 4 hours.

Results are expressed as levels depending on the degree of erythema and edema, and the criteria the International Contact Dermatitis Research Council; it depends on the basis of the (International Contact Dermatitis Research Group ICDRG) (Wooding et al, 1967; Rietschell, 1982; Fischer & Maibach, 1984; Aberer et al , 1993).

The criteria for evaluation are as follows.

0: no redness

1: erythema

2: redness and papules

3: redness, papules and vesicles

4: severe edema and vesicles

The stimulus degree was calculated according to the following formula based on the score determined according to the above criterion.

The results are shown in Table 4 below.

Comparative Production Example 1 Preparation Example 1 Preparation Example 2 Preparation Example 3 Preparation Example 4 Comparative Production Example 2 Comparative Production Example 3 Irritation degree (%) 5.0 0.5 0.01 0.01 0.1 0.1 1.0

As shown in Table 4, the cosmetic composition according to the present invention showed little skin inflammation or irritation as a result of skin primary irritation test.

(Manufacture example 5)

The essence was prepared by mixing the components of the following table (unit:%).

(Comparative Production Example 4)

Essence was prepared in the same manner as in Preparation Example 5, except that only vitamin C was used instead of vitamin C and astaxanthin.

(Comparative Production Example 5)

Essence was prepared in the same manner as in Preparation Example 5, except that only vitamin C derivatives (magnesium ascorbyl phosphate) were used instead of vitamin C and astaxanthin.

(Test Example 6)

Stability experiment

Preparation Example 5, Comparative Preparation Example 4 and Comparative Preparation Example 5 were stored in an opaque glass container in a constant temperature chamber maintained at 45 ℃ constant stored for 12 weeks, opaque in a completely shaded refrigerator maintained at 4 ℃ constant For samples stored in a vial container for 12 weeks and samples stored for 12 weeks in a circulation chamber (once / day) circulating at 37 ° C. at −5 ° C., the degree of separation, discoloration and precipitation was measured.

The degree of product separation and discoloration was evaluated by classifying into the following six grades.

Product discoloration evaluation criteria:

0: no change 1: very little separation (discoloration)

2: Slightly separated (discolored) 3: Slightly separated (discolored)

4: Extremely separated (discolored) 5: Extremely separated (discolored)

The results are shown in the table below.

Temperature Preparation Example 5 Comparative Production Example 4 Comparative Production Example 5 45 ℃ 0 3 3 4 ℃ 0 3 2 cycle 0 3 2

Through the above table, astaxanthin and vitamin C-containing cosmetics according to the present invention was stable without discoloration or separation symptoms at 4 ℃, 45 ℃ and circulation tests, but in the case of cosmetics containing only vitamin C or vitamin derivatives without astaxanthin Separation (discoloration) was found to be unstable.

In addition, the degree of product precipitation was evaluated by classifying into the following six grades.

Product discoloration evaluation criteria:

0: no change 1: very slight precipitation

2: little precipitate 3: little precipitation

4: settling heavily 5: settling extremely severe

The results are shown in the table below.

Temperature Preparation Example 5 Comparative Production Example 4 Comparative Production Example 5 45 ℃ 0 3 3 4 ℃ 0 3 2 cycle 0 3 2

Through the table, it can be seen that the astaxanthin and vitamin C-containing cosmetics according to the present invention do not precipitate and stabilize.

Therefore, according to the present invention, a wrinkle containing vitamin C or a derivative of vitamin C and astaxanthin, which is not only excellent in fibroblast proliferation, collagen biosynthesis, moisturizing, skin elasticity and wrinkle alleviating effect, but also has little side effects and irritation. Improved and anti-aging cosmetic compositions can be obtained.

Claims (1)

Wrinkle improvement and anti-aging cosmetic composition comprising a mixture of vitamin C or a derivative of vitamin C and astaxanthin in a weight ratio of 1: 0.001 to 1:20, 0.01 to 10% by weight based on the total weight of the composition.