KR20040074795A - Cosmetic composition for preventing a wrinkles - Google Patents

Abstract

The present invention relates to a cosmetic composition having an anti-wrinkle effect and an antioxidant effect by containing a specific substance in a conventional cosmetic as a certain component, the anti-wrinkle cosmetic composition of the present invention having the ability to synthesize collagen and antioxidant effect

[Formula 1]

It is characterized by containing a decarboxy carnosine hydrochloride (Decarboxy Carnosine 2HCl; Carcinine).

As described above, the cosmetic composition containing dicarboxy carnosine hydrochloride according to the composition of the present invention increases the synthesis of collagen and has an antioxidant action, which not only prevents the formation of wrinkles in the human body, but also has excellent safety for the skin. The stability of the substance in the cosmetic composition is also excellent.

Description

Cosmetic composition for preventing wrinkles {Cosmetic composition for preventing a wrinkles}

The present invention relates to a cosmetic composition having an anti-wrinkle effect, and more particularly to a cosmetic composition having an anti-wrinkle effect and an antioxidant effect by containing a specific substance as a certain component in a conventional cosmetic.

Wrinkles are one of the natural aging phenomena of the skin that are usually formed by repeated muscle movements over long periods of time. Skin aging is largely divided into natural aging or endogenous aging and external aging (E.B. Doris, 1996). Because natural aging is caused by genetic causes, it is difficult to control, but external aging is caused by environmental causes. Therefore, studies to prevent external aging have been continued, and in particular, research on preventing wrinkle formation caused by exogenous photoaging due to prolonged ultraviolet exposure has been focused.

As described above, wrinkles formed by exogenous photoaging are involved with free radicals and various free radicals (Christina S et al, Photoaging is Associated with Protein Oxidation in Human Skin In Vivo, J Invest Dermatol 2002; 118: 618-625). The main causes are decreased collagen, increased elastin crosslinking, and decreased skin elasticity due to substrate degradation. In addition, decreased fibroblast proliferation decreases collagen synthesis, which leads to weakening of defense against external factors, thereby promoting wrinkle formation. Therefore, anti-wrinkle effect can be expected by promoting the generation of free radicals and collagen synthesis.

Retinoids, which have been used to improve wrinkles of the skin, are generically known as vitamin A and derivatives thereof such as retinoic acid, retinyl acetate, retinyl palmitate, and are known to prevent photoaging, inhibit collagen loss and promote collagen biosynthesis (Griffiths CE , The role of retinoids in the prevention and repair of aged and photoaged skin, Clin Exp Dermatol 2001 Oct; 26 (7): 613-8).

However, retinoic acid, which is well known as a collagen synthesis promoter, is unstable, and its usage is extremely limited due to safety problems such as irritation and redness when applying skin, and vitamin C, which is essential for antioxidant activity and collagen synthesis, is very unstable in heat, light, oxygen Therefore, there is a problem such as discoloration and odor caused by being exposed to the air or easily oxidized in an aqueous solution, and it is urgently required to develop a raw material that is safe for living organisms and has good collagen synthesis promoting effect.

Accordingly, it is an object of the present invention to provide a new material having collagen synthesis and antioxidant effects without the problems of instability and instability that were previously used for improving wrinkles of the skin.

Another object of the present invention is to provide an anti-wrinkle cosmetic composition which is excellent in safety and stability in use and contains a new substance having collagen synthesis and antioxidant effect.

Therefore, the present inventors have found that dicarboxy carnosine hydrochloride has a collagen production and antioxidant effect as a natural component that exists in mammalian tissue and includes an imidazole structure found in non-protein components. Reached.

1 is a graph showing the amount of TBARS produced over time of dicarboxy carnosine hydrochloride according to the present invention,

Figure 2 is a graph showing the degree of reduction of cytochrome c (Cytochrome c) over time of Example 4 containing 0.1% dicarboxy carnosine hydrochloride according to the present invention.

Anti-wrinkle cosmetic composition of the present invention for achieving the above object;

Formula 1 having collagen synthesis ability and antioxidant effect

[Formula 1]

It is characterized by containing a decarboxy carnosine hydrochloride (Decarboxy Carnosine 2HCl; Carcinine).

According to another configuration of the present invention, the dicarboxy carnosine hydrochloride is characterized in that it contains 0.1 to 2% by weight based on the total weight of the cosmetic composition.

The cosmetic composition according to the present invention may be composed of an aqueous phase or an oil phase, and the aqueous phase may further contain components such as a moisturizer, purified water, a divalent metal salt, a hydrophilic nonionic surfactant, or a water-soluble polymer, and the oil phase may be a higher alcohol. , Powder, oil-soluble polymer, ceramide or derivatives thereof, cholesterol or derivatives thereof, and the like.

As described above, the dicarboxy cannosine hydrochloride of the present invention having an antioxidant effect and the ability to inhibit collagen synthesis is a white powdery component stabilized by decarboxylation of cannosine. 2002-0044740), the treatment of renal failure (especially 1999-0076920), the prevention and treatment of inflammatory bowel disease (especially 0180556), etc. are known to have a photoprotective effect (Babizhayev MA et al. Biochemistry (Mosc) 1998 May; 63 (5): 523-8) Known.

Hereinafter, the present invention will be described in more detail with reference to Examples and Experimental Examples. However, these Examples and Experimental Examples are only for the purpose of understanding the present invention, and the scope of the present invention is not limited to the following Examples and Experimental Examples.

Example 1

To a flexible lotion containing dicarboxy carnosine hydrochloride in the configuration of Table 1 below.

ingredient Content (% by weight) Salicylic acid 0.5 1,3-butylene glycol 5.0 Monolauric acid polyoxyethylene sorbitan 1.3 Polyoxyethylene lauryl ether 0.7 ethanol 15.0 Spices 0.2 antiseptic Quantity Dicarboxy Carnosine Hydrochloride 0.1 Purified water Balance

Example 2

To prepare a lotion containing dicarboxy carnosine hydrochloride in the configuration of Table 2.

ingredient Content (% by weight) Stearic acid 0.5 ethanol 10.0 vaseline 1.0 Squalane 2.5 Liquid paraffin 6.0 Monooleic acid polyoxyethylene sorbitan 1.5 1,3-butylene glycol 8.0 Sodium Ethylenediaminetetraacetic Acid 0.05 Triethanolamine 0.8 Spices 0.2 antiseptic Quantity Antioxidant Quantity Dicarboxy Carnosine Hydrochloride 0.1 Purified water Balance

Example 3

To prepare an essence containing dicarboxy carnosine hydrochloride in the configuration of Table 3.

ingredient Content (% by weight) glycerin 5.0 1,3-butylene glycol 5.0 Sesquioleate sorbitan 0.4 Polylauric acid polyoxyethylene sorbitan 1.0 Paraoxybenzoic Acid Methyl 0.2 ethanol 2.0 Purified water Balance Vaseline 5.0 Cetanol 0.2 Dicarboxy Carnosine Hydrochloride 0.1 Tocopheryl Acetate 2.0

Example 4

To prepare a cream containing dicarboxy carnosine hydrochloride in the configuration of Table 4.

ingredient Content (% by weight) Stearic acid 0.5 ethanol 10.0 vaseline 1.0 Squalane 2.5 Liquid paraffin 6.0 Monooleic acid polyoxyethylene sorbitan 1.5 1,3-butylene glycol 8.0 Sodium Ethylenediaminetetraacetic Acid 0.05 Triethanolamine 0.8 Spices 0.2 antiseptic Quantity Antioxidant Quantity Dicarboxy Carnosine Hydrochloride 0.1 Purified water Balance

Experimental Example 1

Collagen Fiber Biosynthesis Effect

Fibroblasts (fibroblast W138) were incubated for 24 hours with 5 × 10 ⁴ cells / well in 48 well plates. After removal of the supernatant, dicarboxylic carnosine hydrochloride was treated by diluting the serum-free culture medium by concentration. After 24 hours, the supernatant of each well was collected and the amount of procollagen type IC-peptide (PICP) was measured using a collagen fiber biosynthesis kit (Procollagen Type IC-peptide EIA kit: Takara MK101). Absorbance was measured at 450 nm. TGF-β was used as a positive control and treated at 10 ng / ml.

Dicarboxy carnosine hydrochloride (ug / ml) O.D. Collagen (ng / mL) % of control 0 1.458 52.3 0 5 2.212 79.7 ** 52.7 10 2.357 84.6 ** 61.7 50 2.687 116.6 ** 122.9

As shown in Table 5, dicarboxy carnosine hydrochloride increased collagen synthesis in a concentration-dependent manner. The amount of collagen synthesis at 50 ug / ml of TGF-β used as a positive control was 118.9 ng / ml. In comparison, dicarboxy carnosine hydrochloride was found to have a significant collagen synthesis effect of about 98% at 50 ug / ml.

Experimental Example 2

Antioxidative effect

1) MDA measurement

The experiment was performed by Novak (1991), and the reaction was caused by the addition of 0.01 U / ml of Xanthine-Oxidase. The reaction was terminated by adding 1 ml of trichloroacetic acid (60 g / l), and 0.5 ml of thiobarbituric acid (10 g / l) was added thereto and boiled for 15 minutes to obtain an appropriate color for analysis. The treatment concentration was 10 mM and the absorbance at 532 nm was determined using hydrogen peroxide as a control and the results are shown in Table 6.

Control Dicarboxy Carnosine Hydrochloride Carnosine MDA generation amount (nM / mn) 280 ± 8 94 ± 5 134 ± 9

As shown in Table 6, it can be seen that dicarboxy carnosine hydrochloride has an inhibitory effect of MDA production of about 66% based on the control group.

2) TBARS measurement (Thiobarbituric acid-reactive substances assay)

This experiment used a method devised by Babizhayev (1989). The peroxidation of liposomes was started by adding 2.5 μl of FeSO ₄ and 200 uM of ascorbate and ended by adding 2 ml of 0.25N HCl containing 15% of trichloroacetic acid. 0.125% of chiovabituric acid was added, boiled for 15 minutes, cooled and centrifuged at 3,000 rpm for 15 minutes, and the absorbance of the supernatant was measured at 532 nm. Standard curves of malondialdehyde (MDA) were obtained from 1,1,3,3 and tetra ethoxypropane. Inhibition of production of TBARS by 10 mM dicarboxy carnosine hydrochloride was measured.

As shown in Figure 1 showed the highest oxidation between 10-20 seconds and dicarboxy carnosine hydrochloride reduced TBARS production compared to the control.

3) SOD-like activity measurement

This experiment used tissues obtained by biopsy from pig ears instead of human skin (P. Dick and al. 1991). The tissues were heated at 70 ° C. for 70 seconds before physically separating the epidermis-dermis. Example 4 was treated with separated skin sections for 5 minutes and washed with 1% TRITON X 100 solution. After UVA-UVB irradiation (0.8 J / cm 2), it was crushed in buffer and diluted in 1% TRITON X 100 and stored at 0 ° C. for 2 hours. The supernatant was then obtained by centrifugation and similar SOD activity was measured.

SOD activity was initiated by adding 10 μl skin / ml tissue solution to 50 mM buffer containing 1.25 mM DTPA and adding 100 μl of Xanthine oxidase (0.06 U / ml). The reduction rate of cytochrome c on a spectrometer maintained at 25 ° C. was measured by absorbance at 550 nm. The protective effect on skin was calculated | required based on the following formula.

Protective Effect (%) = (Irradiation 0%-Irradiation 0.1%) / (Irradiation 0%-No Irradiation) × 100

* No irradiation: Absorption change in non-UV treated skin tissue, SOD-like activity in natural state of skin tissue.

* Irradiation 0%: Change of absorbance of cream containing 0% of dicarboxy carnosine hydrochloride, showing the best effect of ultraviolet rays on SOD-like activity of skin tissue.

Irradiation 0.1%: Dicarboxy carnosine hydrochlory 0.1%, i.e. after treatment with Example 4 shows SOD-like activity against ultraviolet irradiation.

As shown in FIG. 2, when the UV irradiation was not performed, 0.1 OD / min was 0.17 OD / min in 0% of dicarboxy carnosine hydrochlorine and 0.14 OD / min in 0.1%. It showed a protective effect.

Experimental Example 3

Wrinkle improvement human efficacy test

1) Image analysis by replica

The study was conducted for 12 weeks with 25 women (mean age 42.3 years) aged 32-55 years. The skin was used only once a day for a week and then twice a day (morning and evening). The dermal wrinkle replica was made using Silflo (manufactured by Flexico, Inc., UK) to obtain a wrinkle pattern on the tail of the subject's eye area, which was stabilized in a constant temperature and humidity room (22 ± 2 ° C, 50 ± 5% humidity). For wrinkle replica image analysis, the replica was irradiated with light of about 38 degrees to the replica after 6 weeks and 12 weeks of use at room temperature, and the shadow of wrinkles was taken with a CCD camera to measure the depth of wrinkles instead. A computer imaging system for measuring wrinkle depth was analyzed using C + K's Skin-Visiometer SV 600 software. R1, R2, and R3 of arbitrary units, R1, R2, and R3, are values for deep wrinkles. When two or more variables among three variables are significant, they are interpreted as effective against deep wrinkles. R4 and R5 are shallow. When any one of the R4 or R5 value was found to be significant for wrinkles, it was interpreted as showing efficacy against shallow wrinkles.

R1: difference between the highest and lowest of the wrinkle contour

R2: Average of R1 values after randomly dividing 5 lines of wrinkles

R3: Highest value of R1 divided by 5

R4 is the mean value of the baseline of the wrinkle contour, minus the value of each top and valley.

The smaller the value of the variable, the smaller the wrinkles.

Table 7 shows the value of △ after subtracting the wrinkle replica value from the wrinkle replica value analyzed based on the horizontal lines after using Example 3 for 12 weeks, and the significance between the family and the control group through an independent t-test. Whether or not was confirmed by biological statistical criteria (p <0.05).

Replica Measurement Variables R1 R2 R3 R4 R5 Control 0.423 0.432 0.101 0.012 -0.011 product family -0.219 -0.214 -0.103 -0.011 -0.04 p-value 0.0010 0.010 0.027 0.011 0.036

From the above results, it can be seen that Example 3 exhibits a significant level of wrinkle improvement in deep and shallow wrinkles when compared to the control group.

Table 8 below was a circle (circle) as a criterion analysis parameter and used as shown in Table 7 for Δ value measured after 12 weeks.

Replica Measurement Variables R1 R2 R3 R4 R5 Control 0.200 0.210 0.054 0.000 -0.009 product family -0.182 -0.188 -0.101 -0.007 -0.003 p-value 0.050 0.044 0.065 0.460 0.727

From the above results it can be seen that Example 3 shows a significant level of wrinkle improvement effect in the deep wrinkles and shallow wrinkles compared to the control group.

2) Evaluation of wrinkle improvement through clinical photography and visual judgment

Twenty-five subjects were visually evaluated by two dermatologists. Wrinkle values were recorded as integers ranging from '1' without wrinkles to '13' with many wrinkles. Visual wrinkle evaluation was measured before use, after 6 weeks of use, and after 12 weeks of use according to Example 3, and the difference between after use and before use was expressed as Δ value. Independent t-test confirmed the improvement of wrinkles at the significance level p <0.05.

Visual Evaluator 1 Visual evaluator 2 term 6 weeks-0 weeks 12 weeks-0 weeks 6 weeks-0 weeks 12 weeks-0 weeks division product family Control product family Control product family Control product family Control Average -0.32 0.00 -0.48 -0.08 -0.36 -0.04 -0.68 -0.28 p-value 0.052 0.019 0.061 0.027

According to the above results, after 6 weeks of product use according to Example 3, there was no statistically significant level of wrinkle improvement, but after 12 weeks of use of the product, it showed statistically significant wrinkle improvement effect when compared to the control group. .

Experimental Example 4

Product safety assessment

1) Evaluation of subject safety through questionnaire

For 25 subjects as in Experimental Example 3, the safety evaluation of the product according to Example 3 by questionnaires and questionnaires was carried out after 6 and 12 weeks.

Skin safety Subject Survey Assessment Dermatologist Evaluation Week 6 Week 12 Week 6 Week 12 stimulus 0 0 0 0 Non-irritating 25 25 25 25

As can be seen in Table 10, all 25 people did not experience skin irritation due to the continuous use of the product according to Example 3 of the present invention in the questionnaire and showed the same result in the safety evaluation of dermatologists.

As described above, the cosmetic composition containing dicarboxy carnosine hydrochloride according to the composition of the present invention increases the synthesis of collagen and has an antioxidant action, which not only prevents the formation of wrinkles in the human body, but also has excellent safety for the skin. The stability of the substance in the cosmetic composition is also excellent.

Claims (2)

Formula 1 [Formula 1] A cosmetic composition for preventing wrinkles, comprising dicarboxy carnosine hydrochloride (Decarboxy Carnosine 2HCl; Carcinine) and additionally containing a conventional cosmetic composition component. The anti-wrinkle cosmetic composition of claim 1, wherein the dicarboxy carnosine hydrochloride is contained in an amount of 0.1 to 2 wt% based on the total weight of the cosmetic composition.