KR101030359B1 - Cosmetic compositions containing adenine for improving skin wrinkles and preventing the appearance thereof - Google Patents

Abstract

The present invention relates to a cosmetic composition for improving and inhibiting skin wrinkles comprising adenine.

The cosmetic composition of the present invention preferably comprises 0.001 to 1% by weight of adenine based on the total weight of the composition.

Cosmetic composition comprising adenine of the present invention provides excellent skin wrinkle improvement and production inhibitory effect.

Adenine, anti wrinkle, wrinkle improvement, collagen, collagenase

Description

Cosmetic compositions containing adenine for improving skin wrinkles and preventing the appearance according to the present invention.

Figure 1a is a photograph showing the result of measuring the collagen synthesis change by tissue staining method after applying to the animal the composition of the formulation of the formulation of Formulation 1 in order to confirm the effect of increasing the collagen synthesis by the composition of the present invention.

Figure 1b is a photograph showing the result of measuring the collagen synthesis change by tissue staining method after applying the composition of the cream formulation of Comparative Example 1 to the animal in order to confirm the effect of increasing the collagen synthesis by the composition of the present invention.

Figure 2a shows the result of measuring the collagenase synthesis change by tissue staining method after applying the composition of the formulation of the formulation of Formulation 1 to the animal, in order to confirm the inhibitory effect of collagenase synthesis by the composition of the present invention It is a photograph.

Figure 2b is to confirm the collagenase synthesis inhibitory effect by the composition of the present invention, after applying the composition of the cream formulation of Comparative Example 1 to the animal, the collagenase synthesis changes measured by tissue staining method The picture shown.

Figure 3a is a comparison of the improvement of the deep wrinkles (R1, R2, R3) measured by the image analysis in Test Example 4 when using the composition of the cream formulation of Formulation Example 1 and the composition of the cream formulation of Formulation Example 1 It is shown divided by when.

3B shows the improvement of the shallow wrinkles (R4, R5) measured by the image analysis in Test Example 4, when the cream formulation of Comparative Example 1 was used and the cream formulation of Formulation Example 1 was used. It is shown separately.

Figure 4a shows a photograph of a replica (Replca) before the use of the formulation 1 cream in Test Example 4.

Figure 4b shows a replica (Replca) photograph after 8 weeks of use of Formula 1 cream in Test Example 4.

Figure 5a shows a replica (Replca) before the use of Comparative Example 1 cream in Test Example 4.

Figure 5b shows a Replca picture after 8 weeks of use of Comparative Formula 1 Cream in Test Example 4.

The present invention relates to a cosmetic composition comprising adenine excellent in skin wrinkle improvement and suppression of production.

Human skin is changed by a number of internal and external factors as it ages. That is, internally, the secretion of various hormones that regulate metabolism decreases, the function of immune cells and the activity of cells decreases, thereby reducing the biosynthesis of immune proteins and constituent proteins necessary for the living body, and externally destroying the ozone layer. Due to the increase in the amount of ultraviolet rays in the sun's rays and the increase of environmental pollution, free radicals and free radicals increase, such as wrinkles of the skin is increased and the elasticity of the skin is reduced, blemishes, freckles and blotch There will be a number of changes, including an increase. Among the factors affecting skin aging, the most active research field is the study of changes in skin wrinkles.

Skin wrinkles are known to be produced by a number of factors, including skin moisture content, collagen content, and the ability to act on the environment. The greatest influence on the formation of double wrinkles is the amount of collagen produced and the amount and activity of collagenase, a collagen degrading enzyme.

Collagen is present in the dermal layer of the skin and accounts for about 70% of the total dry skin weight. The elasticity of the skin is known to be maintained by this collagen and elastin (elastic fiber). Collagen molecules are made in Fibroblasts and form triple helix structures after they are secreted out of the cell. The main functions of collagen are known to maintain the mechanical tightness of the skin, to strengthen the resistance of the connective tissue and the adhesion of the tissue, to sustain cell adhesion, induction of cell division and differentiation (Van der Rest et al., 1990). Such collagen is reduced due to the decrease of cellular activity due to natural aging and the increase of free radicals and free radicals caused by internal factors due to the activity of collagenase and external factors such as pollution, ultraviolet rays and stress (Ingrid Emerit et al., Free radical and aging, 1992).

In general, at 80 years of age, collagen is reduced by 65% compared to 20 years, resulting in thinner skin, which is known to be closely related to wrinkle formation of the skin (Arthur K. Balin et al., Aging and the skin, 1989). In recent years, extensive research on skin aging has emphasized the importance of collagen in the skin. When collagen metabolism is activated by the synthesis of collagen in the skin, the components of the dermal matrix are increased, and thus, the effects of wrinkle improvement, elasticity enhancement, skin strengthening and wound healing can be expected.

In order to obtain such an effect, conventionally, collagen was formulated into a cosmetic product. However, these cosmetics are to apply collagen to the surface of the skin, it is difficult to absorb the percutaneous absorption of collagen, a polymer, its function can not be expected. Recently, collagen is also injected directly into the dermis of the skin. However, this method is also difficult to expect correct skin wrinkles due to side effects.

In recent years, interest in substances that promote collagen synthesis has been increased to solve this problem, and collagen synthesis promoters include retinoic acid and proteins derived from animal placenta (Japanese Patent Laid-Open No. 8-231370). Became known. However, retinoic acid is unstable and complex formulation technology, and its use is limited due to safety problems such as irritation and redness on the skin, and since the placenta-derived protein is prohibited from using bovine extracts from mad cow disease, substantial skin collagen It is difficult to expect a skin wrinkle improvement effect by promoting the synthesis of.                         

Therefore, there is an urgent need for the development of new collagen production promoters, which are safe for the living body, are easy to use, have high formulation stability, and have excellent collagen production promoting effects.

In this context, the present inventors have also extensively studied and examined how to effectively control various factors that have a great influence on the formation of wrinkles, thereby improving wrinkles in the skin, or preventing or delaying the formation of wrinkles. As a result, adenine was found to be excellent in improving wrinkles and suppressing the formation of wrinkles, thereby completing the present invention.

That is, it is an object of the present invention to provide an adenine composition having an excellent effect of improving wrinkles of the skin or preventing or delaying the formation of wrinkles on the skin.

The present invention relates to a cosmetic composition for improving and inhibiting skin wrinkles comprising adenine represented by the following formula (1).

The cosmetic composition of the present invention preferably comprises 0.001 to 1% by weight of adenine based on the total weight of the composition.                     

Adenine is well known as a DNA component that stores and transmits biogenetic information. DNA consists of four bases that carry genetic information and sugar and phosphate groups that perform structural functions. DNA bases are further divided into purine bases and pyrimidine bases, and adenine belongs to the double purine bases. That is, adenine may be said to be the most basic component of DNA that stores and transmits genetic information of a living body. Adenine is also an essential ingredient in making ATP, a bioenergy. The living body needs free energy to generate biomolecules such as muscle relaxation, cell movement, active transport of various molecules and ions, and proteins. It is ATP that transports this free energy. ATP consists of one adenine, one sugar and three phosphate groups. In addition, adenine is an important component in the components of NAD and NADH, which perform important functions as electron acceptors of molecules that generate energy in vivo such as ATP. In addition, adenine is also known as an important ingredient in NADPH, which functions to transfer electrons.

In the present invention, adenine is a commercially available product of SAMLONG CHEMICALS, China, and has a white particle form.

The cosmetic composition of the present invention is used for the purpose of improving the wrinkles of the skin, preventing the formation of wrinkles, and delaying them, and is not particularly limited in the formulation thereof. For example, supple cosmetics, nourishing cosmetics, massage It may be a cosmetic composition having a formulation of a cream, a nourishing cream, a pack, an essence, a gel or a skin adhesive cosmetic, and may also be a transdermal formulation such as a lotion, an ointment, a cream, a patch or a spray.

Hereinafter, the configuration and effect of the present invention will be described in more detail with reference to Examples and Test Examples. However, the present invention is not limited by the following examples and test examples.

Prescription Example 1.

The composition of the cream formulation of the cosmetic composition containing adenine of the present invention is as follows.

ingredient Content (% by weight) glycerin 5.0 Carbomer 0.4 Stearic acid 3.0 Cetyl alcohol 2.0 Squalane 3.0 Dimethicone 1.0 Triethanolamine 0.1 Adenine 1.0 antiseptic 0.2 incense 0.1 Purified water  Balance system 100

Prescription Example 2.

In the cosmetic composition containing adenine of the present invention, the composition of nutrient longevity (milk lotion) is as follows.

ingredient Content (% by weight) Adenine 1.0 Stearic acid 1.0 Cetyl alcohol 1.0 Glyceryl Monostearate 1.0 Polyoxyethylene sorbitan monostearate 1.0 Sorbitan sesquioleate 1.0 Liquid paraffin 4.0 Squalane 3.0 Carlylic / Capric Triglycerides 4.0 Carboxy Vinyl Polymer 0.1 Butylene glycol 5.0 Triethanolamine 0.2 antiseptic 0.2 incense 0.1 Purified water Balance system 100.0

Comparative Prescription 1.

Cream formulations were prepared in compositions of Formulation Example 1, except for adenine.

ingredient Content (% by weight) glycerin 5.0 Carbomer 0.4 Stearic acid 3.0 Cetyl alcohol 2.0 Squalane 3.0 Dimethicone 1.0 Triethanolamine 0.1 Adenine - antiseptic 0.2 incense 0.1 Purified water Balance system 100

Experimental Example 1 Measurement of Collagen Synthetic Efficacy

Human fibroblasts were placed in a 96-well microtiter plate containing Dulbecco's Modified Eagle's Media (DMEM) medium containing 2.5% fetal bovine serum at 5,000 cells / well. Incubate until 80% growth. Then, adenine was treated at a concentration of 0.001% for 1 day, and then the cell culture was collected. The collected cell culture medium was measured for collagen production using a commercially available collagen protein measuring instrument (Catalog #: MK101 manufactured by Takara Shuzo, Japan). First, the collected cell culture solution was placed in a 96-well plate uniformly coated with primary collagen antibodies, and the antigen-antibody reaction was performed in a thermostat for 3 hours. After 3 hours, the chromophore-bound secondary collagen antibody was placed in a 96-well plate and reacted again for 15 minutes. After 15 minutes, a color-causing substance is added to induce color development at room temperature for 15 minutes. Then, 1M sulfuric acid is added to stop the color reaction. A yellowish 96-well plate was measured at 450 nm using an absorbance meter, and the degree of synthesis of collagen was calculated by Equation 1 below. At this time, the reaction absorbance of the cell culture liquid collected from the group not treated with adenine was used as a control. The measurement results are shown in Table 1.

matter Collagen Synthesis (%) Adenine treatment group 45 Control 0

From Table 1, it can be seen that the group treated with adenine is superior in promoting collagen production than the control group not treated with adenine.

Test Example 2 Measurement of Collagenase Inhibition Effect

Human fibroblasts were placed in a 96-well microtiter plate containing Dulbecco's Modified Eagle's Media (DMEM) medium containing 2.5% fetal bovine serum at 5,000 cells / well and 70-80%. Incubate until grown. Then, after treatment with adenine at a concentration of 0.001% for 24 hours, the cell culture was collected.

The collected cell culture solution was measured by using a commercially available collagenase measuring instrument (Amersham Pharmacia's Catalog #: RPN 2610, USA). First, the collected cell culture solution was put into a 96-well plate uniformly coated with primary collagenase antibody, and the antigen-antibody reaction was performed in a thermostat for 3 hours. After 3 hours, the chromophore-conjugated secondary collagen antibody was placed in a 96-well plate and reacted again for 15 minutes. After 15 minutes, a color-causing substance is added to cause color development at room temperature for 15 minutes, and when 1M sulfuric acid is added again to stop the color reaction, the color of the reaction solution is yellow and the degree of yellow color varies depending on the degree of reaction. The yellowish 96-well plate was measured at 405 nm using an absorbance meter, and the degree of synthesis of collagenase was calculated by the following Equation 2. At this time, the reaction absorbance of the cell culture liquid collected from the group not treated with adenine was used as a control. The measurement results are shown in Table 2.

matter Collagenase Expression (%) Adenine treatment group -20 Control 0

As a result of measuring the expression inhibition of collagenase in the cell from Table 2, it was confirmed that the group treated with adenine inhibited the expression of collagenase more effectively than the control group not treated with adenine.

Experimental Example 3. Wrinkle removal effect through animal experiment

(1) test method                     

40 hairless mice were irradiated with UV rays once daily for 8 weeks (UVB 70mJ / cm 2), and then divided into 2 groups of 20 animals in each group. Next, the cream-like composition of Formulation Example 1 (including adenine) and the cream-like composition of Comparative Formulation Example 1 (including adenine ratio) were each applied to the back area for 100 days each day for 1 month, followed by biopsy and histologically. Collagen formation and the content of collagenase, a collagen degrading enzyme, were measured.

(2) Test result

1) collagen production rate

The back of the hairless mice was biopsied and immunostained using collagen antibody. Next, the collagen synthesis change was measured using an optical microscope. As a result, as shown in FIGS. 1A and 1B, the collagen production rate (FIG. 1A) of the group to which the composition of the cream formulation of Formulation Example 1 (including adenine) was applied was shown to the composition (including the adenine ratio) of Comparative Formula 1. It was confirmed that it was significantly higher than the collagen production rate (Fig. 1b) of the applied group.

2) collagenase production inhibition rate

The dorsal area of hairless mice was biopsied and immunostained using collagenase antibodies. Next, the collagen synthesis change was measured using an optical microscope. As a result, as shown in FIGS. 2A and 2B, the collagenase production rate (FIG. 2A) of the group to which the composition of the cream formulation of Formulation Example 1 (including adenine) was applied was compared to that of the cream formulation of Comparative Formula 1 (including adenine ratio). It was confirmed that significantly lower than the collagenase production rate (Fig. 2b) of the group coated with.

Test Example 4. Measurement of wrinkle improvement effect through human experiment

The skin wrinkle improvement effects of the creams prepared in Formulation Example 1 and Comparative Formulation Example 1 were compared. In other words, the wrinkle improvement effect of the skin was divided into the left and right face for 32 women in their 30s and 40s, giving the products of Prescription Example 1 and Comparative Prescription Example 1 and applied twice a day for 8 weeks . After 8 weeks, the replica was floated using silicone and the depth of the wrinkles was measured by image analysis using a visiometer (SV500, Courage + Khazaka electronic, Germany). The result is shown in Table 3 by the value of R1-R5. In addition, a questionnaire was asked about whether the effect of wrinkle improvement after 8 weeks of use is improved. The results are shown in Table 4.

In Table 3 below, R1 to R3 represent the degree of deep wrinkles, and R4 and R5 represent the degree of shallow wrinkles.

Result of wrinkle depth measurement through image analysis 32 persons Comparative Prescription 1 (without adenine) Prescription Example 1 (Including Adenine) medium R1 R2 R3 R4 R5 R1 R2 R3 R4 R5 Before using the product 1.355 1.301 0.764 0.132 0.299 1.612 1.566 0.861 0.151 0.322 8 weeks after use 1.516 1.469 0.854 0.149 0.310 1.349 1.304 0.758 0.133 0.295

The wrinkle change rate was calculated by substituting the values of R1 to R5 measured in the following equations, and the results are shown in FIGS. 3A and 3B.

Wrinkle change rate (%) = [(measured value before use-measured value after 8 weeks of application) / measured value before use] × 100

In addition, before the use of Formula 1 cream (Fig. 4a) and after 8 weeks of use (Fig. 4b) and replica (Replca) photograph and the use of prescription Example 1 cream (Fig. 5a) and after 8 weeks of use (Fig. 5b) Replica pictures of are shown in Figures 4a, 4b and 5a, 5b.

Table 3 above; 3A, 3B; 4A, 4B; 5A and 5B, the cosmetic composition of Formulation Example 1 containing adenine is statistically compared with the cosmetic composition of Comparative Formulation 1, which does not contain adenine, as a result of using the Independent T-TEST statistical analysis method. As it has a significant level (P <0.05) wrinkle improvement effect was confirmed that the improvement effect of deep wrinkles (R1 ~ R3) and shallow wrinkles (R4, R5) is excellent.

Survey Results on Wrinkle Improvement Wrinkle improvement effect Number of people ratio(%) no effect 2 6.3 Prescription Example 1 is excellent 20 62.5 Comparative Prescription 1 is excellent 5 15.6 Both are effective 5 15.6

As a result of conducting the questionnaire survey as described above, it was confirmed that the composition of the present invention prepared in Formulation Example 1 containing adenine exhibited superior effects.

As described above, the cosmetic composition comprising adenine provided by the present invention can provide a cosmetic composition exhibiting excellent skin wrinkle improvement and production inhibitory effect.

Claims (3)

A cosmetic composition for improving and inhibiting skin wrinkles comprising adenine represented by Formula 1 below: [ Formula 1 ]

. The cosmetic composition for improving and inhibiting skin wrinkles according to claim 1, comprising adenine in an amount of 0.001 to 1% by weight based on the total weight of the composition. The method of claim 1, wherein the skin wrinkles characterized in that the formulation of the softening cream, nourishing cream, massage cream, nourishing cream, pack, essence, gel, skin adhesive type, lotion, ointment, cream, patch, or spray Cosmetic composition for improvement and suppression of production.

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