KR101204858B1 - Cosmetic Composition for for improving Skin-Wrinkle - Google Patents

Abstract

The present invention relates to a cosmetic composition for improving skin wrinkles containing sewage extract, coenzyme Q-10 and L-carnitine, wherein the cosmetic composition of the present invention is effective by simultaneously containing sewage extract, coenzyme Q-10 and L-carnitine. The present invention relates to a cosmetic composition for improving skin wrinkles, which effectively prevents skin aging by bringing synergistic effects on wrinkle improvement and skin elasticity improvement by ingredients.

Description

Cosmetic composition for improving skin wrinkles {Cosmetic Composition for for improving Skin-Wrinkle}

The present invention relates to a cosmetic composition for improving skin wrinkles containing sewage extract, coenzyme Q-10 and L-carnitine, wherein the cosmetic composition of the present invention is effective by simultaneously containing sewage extract, coenzyme Q-10 and L-carnitine. The present invention relates to a cosmetic composition for improving skin wrinkles, which effectively prevents skin aging by bringing synergistic effects on wrinkle improvement and skin elasticity improvement by ingredients.

Aging defines the limited replication life of normal somatic cells in humans with respect to cell replication, that is, the stage of stoppage of growth that is no longer able to divide after a defined cell division and eventually cannot be reversed (Tominaga et al. , Mech.Aging Dev. 123: 927-936 (2002)). Aging is associated with diseases such as neurodegenerative diseases, metabolic syndromes such as cancer or diabetes mellitus, and the causes of aging are genetic causes and accumulation of DNA damage (Sedelnikova et al., Nat. Cell Biol. 6: 168-). 170 (2004)), response to oxidative stress (Campisi et al., Exp. Gerontol. 36: 607-618 (2001)), damage to telomeres (Harrington and Robinson, Oncogene 21: 592-597 (2002)) Etc. are known.

In particular, O ₂ is constantly produced ^- an important role in, H ₂ O _2, the active oxygen species such as OH (reactive oxygen species, ROS) the aging process?. These ROS are produced from various enzymes such as oxidase, NADPH dehydrogenase, cyclooxygenase (COX), including mitochondria, peroxysomes and xanthine oxidase (XOD). In addition, reactive nitrogen species (RNS) such as NO, NO ₂ , HNO ₂ , and ONOO ^- in macrophage, neutrophils and other immune cells produce large amounts of RNS and ROS to attack foreign bodies during inflammatory reactions. . These free radicals damage cellular components, oxidize the phospholipids of biological membranes, and cause skin inflammatory reactions to promote skin aging (Clinics in dermatology, 14: 227-234 (1996)). These factors lead to rapid aging of the skin, resulting in rough wrinkles on the surface of the skin and a marked decrease in elasticity.

Recently, there is an increasing demand to develop substances that can regulate the expression of genes related to aging from natural products, and at the same time, they have antioxidant effects as part of suppressing skin wrinkles caused by the oxidation of free radicals (active oxygen, etc.). The development of materials that effectively prevent aging by applying materials is required.

The present inventors tried to find an effective substance for improving skin wrinkles, and as a result of applying the sewage extract, coenzyme Q-10, and L-carnitine to the skin, the present invention was confirmed by synergistically increasing the skin wrinkle improvement effect by each component. Completed.

Accordingly, an object of the present invention is to provide a cosmetic composition for improving skin wrinkles comprising sewage extract and coenzyme Q-10 and L-carnitine as an active ingredient.

In order to solve the above problems, the present invention may include the following means.

Cosmetic composition for improving skin wrinkles according to the present invention contains sewage extract, coenzyme Q-10 and L-carnitine as an active ingredient.

The sewage extract is 0.01 to 20.0% by weight based on the total weight of the composition, the coenzyme Q-10 is 0.01 to 10.0% by weight relative to the total weight of the composition, the L-carnitine contains 0.01 to 10.0% by weight relative to the total weight of the composition It is characterized by.

The cosmetic composition is characterized by having a formulation of a lotion, essence, lotion, cream, pack, gel, ointment, patch, or spray.

As described above, the present invention relates to a cosmetic composition for improving skin wrinkles containing sewage extract, coenzyme Q-10, and L-carnitine, wherein the cosmetic composition of the present invention comprises sewage extract and coenzyme Q-10 and L-carnitine. Containing at the same time brings a synergistic effect on wrinkle improvement and skin elasticity improvement by the active ingredient.

In particular, as can be seen in the experimental example below, the sewage extract exhibits the action of i) promoting the proliferation of skin cells, ii) promoting the expression of sirt-1 protein, iii) antioxidant activity, and iv) a combination thereof, and coenzyme Q-10 It was confirmed that i) promotes tropoelastin and fibrin production, ii) antioxidant activity, and iii) a combination thereof, and L-carnitine showed i) promoting the proliferation of skin cells.

The cosmetic composition for improving skin wrinkles according to the present invention contains sewage extract, coenzyme Q-10 and L-carnitine as active ingredients.

Hereinafter, the present invention will be described in detail.

Sewage ( Polygonum multiflorum ) used in the present invention It is known to come from China and has been cultivated for a long time. The height is 3 ~ 4m and the roots are extended to the side, often forming tubers. Heart-shaped leaves are about 3 ~ 6㎝ long and 2.5 ~ 4.5㎝ wide, with petioles and chin leaves, but without sawtooth at edges. Flowers bloom in August-September in conical inflorescences. There are no petals and 5 calyxes. Fruits are ovate, glossy, 2.5mm long. The reddish brown tuber is called Shouo in one shot. Shouo is famous as a herb that strengthens kidney function, enhances energy, darkens hair and makes people live longer without disease. The function of the liver to get rid of tiredness, multiply the skin, strengthen the bones and muscles, and strengthen the heart is also effective in nervous breakdowns and insomnia. Its excellent hematopoietic effect makes it an excellent treatment for anemia and is widely used in women's menstrual disorders, uterus and chronic constipation. Sewage is also a good medicine to boost the energy of the elderly. Improves kidney function so that hair does not whiten, hair does not fall out, and eating longer prevents aging. Chinese people consider HaoSuo to be the three greatest medicines with ginseng and wolfberry.

Sewage extract according to the present invention can be prepared as follows.

The sewage is washed thoroughly with water and dried in the shade. Dry sewage is placed in an extraction container and extracted for a predetermined time at an appropriate temperature in an appropriate amount of extraction solvent. The extract may be filtered and concentrated using a vacuum rotary evaporator or the like. The amount of the extraction solvent is 2 to 10 times the dry weight of the herbal medicine. The extraction method may be an extraction method such as hot water extraction, reflux cooling extraction, ultrasonic extraction, etc., and may be extracted once to five times, preferably as an extraction method of hot water extraction. The immersion temperature is preferably 30 to 70 ° C, more preferably 40 to 60 ° C, and most preferably 50 ° C or so, which is most effective in preventing decay, discoloration and detachment. The immersion time is 5 to 20 hours, preferably 10 hours. After the sewage extract is obtained, solids are removed using a filter paper, the suspension is centrifuged, and the supernatant can be filtered under reduced pressure. The filtered extract may be concentrated under reduced pressure at 20 ~ 100 ℃, preferably 50 ~ 70 ℃ with a vacuum rotary concentrator.

Various active ingredients in sewage, provided by the present invention, can be obtained using various extraction methods known in the art, and preferably, (a) water, (b) anhydrous or hydrous lower alcohol having 1 to 4 carbon atoms. (Methanol, ethanol, propanol, butanol, etc.), (c) a mixed solvent of the lower alcohol with water, (d) acetone, (e) ethyl acetate, (f) chloroform or (g) 1,3-butylene glycol Can be obtained as an extraction solvent.

On the other hand, it will be apparent to those skilled in the art that the extract of the present invention can be obtained not only the above-described extraction solvent but also herbal extracts having substantially the same effect using other extraction solvents.

In addition, the extract of the present invention includes not only the extract by the above-described extraction solvent, but also the extract that has undergone a conventional purification process. Obtained by various additional purification methods, such as, for example, separation using ultrafiltration membranes having a constant molecular weight cut-off value, separation by various chromatography (manufactured for separation according to size, charge, hydrophobicity or affinity). The fraction is also included in the extract of the present invention.

Extracts of the present invention may be prepared in powder form by additional processes such as distillation under reduced pressure and freeze drying or spray drying.

Coenzyme Q-10, another active ingredient of the cosmetic composition of the present invention, is biosynthesized in all cells, but its amount is very small, and in normal cases, a sufficient amount is produced and consumed in the human body, Insufficient physical labor, various stresses, disturbances of the immune system, and mitochondrial diseases can cause shortages in the body.

L-carnitine, another active ingredient of the cosmetic composition of the present invention, is an amino acid-like substance synthesized from lysine and methionine, which are amino acids in the liver. Carnitine helps the body convert fat into energy. To metabolize fatty acids to produce energy, the fatty acids in the cytoplasm must be transported to the mitochondria, where carnitine aids in the transport of fatty acids and removes them when excess amounts of organic acids, the metabolites of mitochondria, are produced. L-carnitine is used as a composition for controlling cellulite in cosmetics (Korean Patent Publication No. 2005-67837).

The cosmetic composition having an anti-aging effect according to the present invention is not particularly limited in its formulation, but may be prepared, for example, in a formulation such as lotion, essence, cream, pack, gel, lotion, ointment, patch or spray. have. The content of the extract in the formulation may be contained in 0.01 to 20% by weight, preferably 0.1 to 10% by weight relative to the total weight of the composition. If the content of the extract is less than 0.01% by weight based on the total weight, it is difficult to expect the desired effect in the present invention, when more than 20% by weight may cause problems in formulation stability. The content of the coenzyme Q-10 and L-carnitine is included in 0.01-10.0% by weight, preferably 0.1-5.0% by weight relative to the total weight of the cosmetic composition. At this time, when the total weight of coenzyme Q-10 and L-carnitine is less than 0.01%, the effect is less likely to occur, and when it is 10.0% or more, it is highly likely to cause irritation on the skin and may have a great influence on stabilization of the formulation.

Hereinafter, the present invention will be described in more detail with reference to Examples. These examples are only for illustrating the present invention in more detail, it is to those skilled in the art that the scope of the present invention is not limited by these examples in accordance with the gist of the present invention. Will be self-evident.

Preparation Example 1 Preparation of Sewage Extract

The sewage of this example used what was collected in Korea.

50 g of the collected plant was immersed in 200 ml of 100% ethanol and warmed for 10 hours in a 50 ℃ water bath. The obtained extract was cooled to room temperature and filtered with filter paper to obtain a filtrate. Ethanol was evaporated from the filtrate using nitrogen, concentrated, and then dissolved in 10% ethanol to prepare an extract.

Experimental Example 1 Fibroblast Proliferation Effect

In order to examine the effects of sewage extract and L-carnitine on fibroblast proliferation, human normal fibroblasts (Korea Cell Line Bank) were inoculated to 1 x 10 ⁴ cells in each well of a 96-well microplate. And incubated at 37 ° C. for 24 hours in DMEM medium (Dulbecco's modified eagle medium, Sigma). Subsequently, the experimental group replaced with the serum-free DMEM medium containing the sewage extract and L-carnitine 50-500 µg / ml and the control group replaced with the serum-free DMEM medium containing no sample for 24 hours. It was. Then, 10 μl each of MTT solution (3- (4,5-dimethylthiazol-2-yl) 2,5-diphenyltetrazolium bromide: 5 mg / ml) was added thereto, followed by incubation for 4 hours. The medium was removed, shaken for 20 minutes by addition of 100 μl of DMSO solution per well, and the absorbance of the supernatant was measured at 570 nm using a microplate reader (Molecular Devices, USA). The measured values were substituted into the following Equation 1 to calculate the cell proliferation effect, and the results are shown in Table 1 below.

Sample (ug / ml)
Cell proliferation effect (%) Sewao Extract L-carnitine Control group - - 50 17.2 25.1 100 31.5 42.8 200 50.8 59.4 300 62.8 70.9 500 69.0 80.2

As can be seen in Table 1, compared to the control group not treated with the sample, the experimental group treated with sewage extract and L-carnitine showed a very high fibroblast proliferation effect, especially L-carnitine is very effective in improving skin wrinkles It could be assumed to be effective.

Experimental Example 2 Investigation of Sirt-1 Protein Expression of Sewage Extract

For the sewage extract, Sirt-1 protein expression was measured by Western blot according to the following method. Normal human fibroblasts of 4-8 passages and fibroblasts of 20-30 passages at 37 ° C. in DMEM medium (Sigma) containing 10% bovine serum (Gibco / BRL) at a density of 1 × 10 ⁶ in a 60 mm culture dish. Incubated for 24 hours in a 5% CO ₂ incubator. Thereafter, the cells were exchanged with a serum-free medium, and the samples were added at a concentration of 0.5 and 1.0% and incubated at 37 ° C. for 24 hours. Thereafter, the cell cultures were concentrated by using a concentrated filter Millipore (Millipore), and the protein was quantified according to the BCA method (Sigma) and used for western blotting. The same amount of protein was electrophoresed in a SDS-PAGE gel consisting of a 7% separating gel and a 5% staking gel and a semi-dry transfer device (Biorad). Transfer the electrophoretic protein to PVDF membrane, process the primary antibody Anti-SIRT-1 antibody, and after the reaction, process the secondary antibody conjugated with HRP and enhance the ECL (enhanced chemiluminescence (Intron) reaction). It was then confirmed by photosensitive to an X-ray (X-ray) film. The expression level of the Sirt-1 protein was quantified by the image analyzer (Image analyzer, Biorad), and the experimental results are shown in Table 2, and 1.2mM CaCl ₂ was used as a control.

sample sirt-1 protein synthesis amount (%) Young fibroblasts Aged fibroblasts Untreated group 100 76 1.2 mM CaCl ₂ 141 111 0.5% Sewage Extract 112 95 1.0% Sewage Extract 135 107

As shown in the results of Experimental Example 2, sewage extract was found to increase sirt-1 protein synthesis. The percentage of sirt-1 protein synthesis increased by 35% and 41% in young fibroblasts and aged fibroblasts, respectively. Therefore, the sewage extract of the present invention was shown to effectively activate sirt-1 protein expression.

Experimental Example 3 Tropoelastin and Fibrillin Formation Effects in Human Skin Fibroblasts

Coenzyme Q-10 increased the production of tropoelastin and fibrinine, which play an important role in elasticity in human skin. Human normal fibroblasts were seeded to 5 × 10 ⁴ cells in each well of a 24-well microplate and allowed to attach for 24 hours. After attaching the test material to the concentrations of the following Table 3, the cells were harvested after 2 days. At this time, the test material was not treated as a control. Enzyme immunoassay (ELISA) was used to quantify tropoelastin and fibrin in harvested media. The results are shown in Table 3 below, and the control group was reported as 100 and expressed as a relative ratio.

Test substance Concentration (ppm) Tropoelastin production (%) Fibrillin production (%) Coenzyme Q-10
10 120 114 100 131 129 Vitamin E
10 122 121 100 135 134

As can be seen in Table 3 above, the production of tropoelastin and fibrillin increased depending on the concentration of coenzyme Q-10.

Experimental Example 4 Antioxidative effect

Free Radical Scavenging Activity Test was conducted to investigate the antioxidant effects of sewage extract and coenzyme Q-10 obtained according to the preparation of the present invention. Free radical scavenging activity was modified from the method of Kim et al. (Kor. J. Pharmacogn., 24 (4), 299-303 (1993)), which is a stable free radical DPPH (1,1-diphenyl-2- picryhydrazyl, Sigma) reagent was used. 150 μl of a 0.2mM DPPH solution (ethanol in the case of Blanc) was prepared at various concentrations, and 150 μl of these samples were added and mixed, and allowed to stand at room temperature for 30 minutes, and then an experimental group was measured for absorbance at 517 nm. Vitamin E acetate was used as a comparison group, and ethanol was used as a control group. After absorbance measurement for the experimental group and the control group, the free radical scavenging activity was obtained using Equation 2 below and the results are shown in Table 4 below.

Test substance Concentration (ug / ml) Free radical scavenging effect (%) Sewage Extract
100 50.8 200 82.1 Coenzyme Q-10
10 70.6 20 85.4 Vitamin E Acetate
10 40.1 20 52.3

As can be seen through the results shown in Table 4, both of the sewage extract and coenzyme Q-10 of the present invention was confirmed that the free radical scavenging ability is very excellent.

Preparation Example 2 Examples and Comparative Examples

According to the configuration of the present invention was prepared a cosmetic of the composition shown in the following table.

ingredient
Content (% by weight) Example Comparative Example 1 Comparative Example 2 Comparative Example 3 Sashou extract 2.0 2.0 - - Coenzyme Q-10 1.0 - 1.0 - L-carnitine 1.0 - 1.0 - Lipophilic monostearate 2.0 2.0 2.0 2.0 Cetearyl alcohol 2.2 2.2 2.2 2.2 Stearic acid 1.5 1.5 1.5 1.5 Wax 1.0 1.0 1.0 1.0 Polysorbate 60 1.5 1.5 1.5 1.5 Sorbitan stearate 0.6 0.6 0.6 0.6 Hardened vegetable oil 1.0 1.0 1.0 1.0 Squalane 3.0 3.0 3.0 3.0 Mineral oil 5.0 5.0 5.0 5.0 Trioctanoine 5.0 5.0 5.0 5.0 Dimethicone 1.0 1.0 1.0 1.0 Sodium magnesium silicate 0.1 0.1 0.1 0.1 glycerin 5.0 5.0 5.0 5.0 Betaine 3.0 3.0 3.0 3.0 Triethanolamine 1.0 1.0 1.0 1.0 Sodium hyaluronate 4.0 4.0 4.0 4.0 Incense, preservative a very small amount a very small amount a very small amount a very small amount Distilled water To 100 To 100 To 100 To 100

Experimental Example 5 Skin Wrinkle Improvement Effect

The skin wrinkle improvement effect of the cosmetic composition containing the sewage extract of this invention, coenzyme Q-10, and L-carnitine was measured in 40 healthy women aged 35 to 50 years old. First divided into four groups, Example A in Group A, Comparative Example 1 in Group B, Comparative Example 2 in Group C, and the formulation of Comparative Example 3 in Group D twice daily (morning and evening) Application was made for 3 months. After 3 months, the degree of wrinkle improvement was evaluated by subject's questionnaire and image analysis of wrinkles.

The subject's questionnaire was judged as four stages of no improvement, slight improvement, moderate improvement, and significant improvement compared to before use, and the results are shown in Table 6.

Image analysis of wrinkles was performed by taking a replica of the eye under the eye before the experiment was started, and replicas immediately after the experiment was completed at the same area under the eye. Was measured. The measurement results of wrinkle density by image analysis are shown in Table 7 as a reduction ratio against wrinkle density before use.

sample No improvement Minor improvements Moderate improvement A significant improvement Example - 2 4 4 Comparative Example 1 One 3 4 2 Comparative Example 2 One 2 5 2 Comparative Example 3 9 One - -

sample Wrinkle Density Reduction (%) Example 50.3 Comparative Example 1 19.7 Comparative Example 2 21.3 Comparative Example 3 1.5

As in the results of Tables 6 and 7, the sewage extract, coenzyme Q-10, and L-carnitine were simultaneously compared with Comparative Example 1, which was treated with sewage extract alone, and Comparative Example 2, which simultaneously contained coenzyme Q-10 and L-carnitine. The containing example showed the better skin wrinkle improvement effect.

Experimental Example 6 Skin Elasticity Effect

The skin elasticity enhancing effect of the cosmetic composition containing the sewage extract of this invention, coenzyme Q-10, and L-carnitine was measured. 40 healthy women aged 20 years or older (mean age 37 years) were divided into four groups, with Examples in Group A, Comparative Example 1 in Group B, Comparative Example 2 in Group C and Comparative Example 3 in Group D. After applying twice a day (morning and evening) around the eyes for 12 weeks, use a skin elasticity tester (Cutometer MPA580, Courage + Khazaka, Germany) to give skin elasticity at a temperature of 24-26 ° C and a humidity of 75. Was measured. The test results are described as R5 (R5 (12 weeks) -R5 (0 weeks)) of the cutometer MPA580. R5 values indicate the properties of net elasticity, and the closer to 1, the better the elasticity. The results are shown in Table 8 below.

Experimental Formulation Skin elasticity effect Example 0.27 Comparative Example 1 0.09 Comparative Example 2 0.12 Comparative Example 3 0.01

As shown in the results of Table 8, it was confirmed that the skin elasticity effect of the Example containing the sewage extract, coenzyme Q-10, L-carnitine at the same time than the Comparative Examples 1 and 2.

Formulation Example 1: Softener (Skin Lotion)

The number of softening times was determined according to the usual method as shown in the following table.

Compounding ingredient Content (% by weight) Sewao Extract 0.5 Coenzyme Q-10 0.2 L-carnitine 0.2 1.3-butylene glycol 6.0 glycerin 4.0 Oleyl alcohol 0.1 Polysorbate 20 0.5 ethanol 15.0 Benzophenone-9 0.05 Incense, preservative a very small amount Purified water to 100

Formulation Example 2. Nutrients (Milk Lotion)

Nutrients were prepared according to a conventional method as shown in the table below.

Compounding ingredient Content (% by weight) Sewao Extract 1.0 Coenzyme Q-10 0.5 L-carnitine 0.5 Propylene glycol 6.0 glycerin 4.0 Triethanolamine 1.2 Tocophenylacetate 3.0 Liquid paraffin 5.0 Squalane 3.0 Macadamia Nut Oil 2.0 Polysorbate 60 1.5 Sorbitan sesquioleate 1.0 Carboxyvinyl Polymer 1.0 Viechti 0.01 EDTA-2Na 0.01 Incense, preservative a very small amount Purified water to 100

Formulation Example 3. Nutrition Cream

Nutritive creams were prepared according to conventional methods as shown in the following table.

Compounding ingredient Content (% by weight) Sewao Extract 2.0 Coenzyme Q-10 1.0 L-carnitine 1.0 Cetostearyl alcohol 2.0 Glyceryl stearate 1.5 Trioctanoine 5.0 Polysorbate 60 1.2 Sorbitan stearate 0.5 Squalane 5.0 Liquid paraffin 3.0 Cyclomethicone 3.0 BH 0.05 Delta-tocopherol 0.2 Concentrated glycerin 4.0 1,3-butylene glycol 2.0 Xanthan gum 0.1 EDTA-2Na 0.05 Incense, preservative a very small amount Purified water to 100

Formulation Example 4 Massage Cream

Massage cream was prepared according to a conventional method as shown in the table below.

Compounding ingredient Content (% by weight) Sewao Extract 0.5 Coenzyme Q-10 0.2 L-carnitine 0.2 Propylene glycol 6.0 glycerin 4.0 Triethanolamine 0.5 Wax 2.0 Tocopheryl acetate 0.1 Polysorbate 60 3.0 Sorbitan sesquioleate 2.5 Cetearyl Alcohol 2.0 Liquid paraffin 30.0 Carboxyvinyl polymer 0.5 Incense, preservative a very small amount Purified water to 100

Formulation Example 5 Pack

Packs were prepared according to conventional methods as shown in the table below.

Compounding ingredient Content (% by weight) Sewao Extract 1.0 Coenzyme Q-10 0.5 L-carnitine 0.5 Propylene glycol 2.0 glycerin 4.0 Carboxyvinyl Polymer 0.3 ethanol 7.0 Fiji-40 Hydrogenated Castor Oil 0.8 Triethanolamine 0.3 Viechti 0.01 EDTA-2Na 0.01 Incense, preservative a very small amount Purified water to 100

Claims (3)

Cosmetic composition for improving skin wrinkles containing sewage extract, coenzyme Q-10 and L-carnitine as an active ingredient. According to claim 1, wherein the sewage extract is 0.01 to 20.0% by weight relative to the total weight of the composition, the coenzyme Q-10 is 0.01 to 10.0% by weight relative to the total weight of the composition, the L-carnitine is 0.01 to about the total weight of the composition A cosmetic composition for improving skin wrinkles, comprising 10.0% by weight. The cosmetic composition of claim 1, wherein the cosmetic composition has a formulation of a lotion, an essence, a lotion, a cream, a pack, a gel, an ointment, a patch, or a spray.