KR101322353B1 - Composition for anti-wrinkling and whitening, comprising an extract of Meretrix as an effective component - Google Patents

Abstract

The present invention relates to an anti-wrinkle and whitening composition containing the anastomotic extract as an active ingredient, and specifically, an active ingredient is extracted from the anastomosis by heating with 10 to 100% by volume of a C 2 to C 5 alcohol solution as an extraction solvent. It relates to an anti-wrinkle and whitening composition containing the anastomosis extract as an active ingredient.
Since the composition according to the present invention is an extract extracted from natural substances, there are almost no side effects, and because it inhibits collagenase activity and tyrosinase activity of fibroblasts, it can be used as a medicine or cosmetics for treating or improving wrinkles and skin pigmentation, etc. In this case, anti-wrinkle and skin lightening effects can be expected.

Description

Composition for anti-wrinkling and whitening, comprising an extract of Meretrix as an effective component

The present invention relates to an anti-wrinkle and whitening composition containing the anastomotic extract as an active ingredient, and specifically, an active ingredient is extracted from the anastomosis by heating with 10 to 100% by volume of a C 2 to C 5 alcohol solution as an extraction solvent. It relates to an anti-wrinkle and whitening composition containing the anastomosis extract as an active ingredient.

Skin aging is one of the most evident evidence of aging, and the aging process of skin is usually caused by photo-aging due to continuous exposure to short-wave UV light (UVB), wrinkles, face, neck, Pigment changes such as solar lentigo and spot pigmentation appear on exposed areas such as forearms.

Photoaging is an activity that induces the expression and activation of matrix metalloproteinases (MMPs) in human skin by activating the mitogen-activated protein (MAP) kinase pathway due to UV. It has been reported to arise from the generation of reactive oxygen species (ROS). These MMPs contain collagenase and are considered a major factor in the photoaging process.

Melanogenesis associated with skin whitening is also induced by UV. The major regulator of melanogenesis is well known as tyrosinase, which is a copper-containing enzyme in animal tissues that catalyzes the production of melanin and plays an important role in the pigmentation process. It is one of the enzymes and affects L-DOPA oxidation.

Therefore, a substance that inhibits the expression or activity of collagen degrading enzymes such as MMPs or inhibits the production of reactive oxygen species may be usefully used for the development of medicines or cosmetics for anti-aging or anti-wrinkle, the expression of tyrosinase or Inhibitors can be developed as therapeutics or cosmetics for skin whitening.

Up to now, a lot of cosmetics and the like for the effect of anti-wrinkle and whitening has been developed, but most of it is to prevent the trouble of the skin by absorbing ultraviolet rays, side effects can also occur when sensitive skin needs to compensate for these problems.

Accordingly, the present inventors conducted various studies on anastomosis, which is a natural substance, in order to develop an anti-wrinkle and whitening composition having low side effects and excellent effects, and as a result, the anastomosis extract showed an excellent effect on anti-wrinkle and skin whitening. It confirmed and completed this invention.

Republic of Korea Patent No. 10-0758960. (September 14, 2007)

K. H. Leem. Effects of Meretrix Pharmacopuncture Extracts on the Elastase Activity and DPPH and NO Scavenging Activities. Journal of The Korean Acupuncture Society Vol. 28, No. 2, page 117-123. (2011. 4)

Therefore, the main object of the present invention is to provide an anti-wrinkle and whitening composition with less side effects and excellent effect.

According to one aspect of the present invention, the present invention provides a composition for anti-wrinkle and whitening containing the anastomosis extract as an active ingredient.

In the composition of the present invention, when preparing the anastomotic extract, it is preferable to prepare by using an alcohol solution of 2 to 5 carbon atoms of 10 to 100% by volume as the extraction solvent and heating, specifically 10 to 100 parts by weight of anastomosis. 100 to 10000 parts by weight of 100% by weight of ethanol solution is preferably prepared by heating for 10 minutes to 30 hours, more preferably, 1000 to 3000 parts by weight of 50 to 90% by weight of ethanol solution is added to 100 parts by weight of anastomosis, and 2 It is good to manufacture by heating for 4 hours. More preferably, it is preferable to concentrate the extracted extract, wherein the concentration method may be a reduced pressure concentration method and a freeze-drying method, it is preferable to freeze-dried after concentration under reduced pressure. The temperature at the time of heating is preferably 70 to 130 ° C, more preferably 90 to 110 ° C, and most preferably 100 ° C.

In the composition of the present invention, the composition preferably contains 0.1 to 50% by weight of anastomotic extract, and more preferably 0.1 to 20% by weight. This is a content of the anastomotic extract necessary for the composition of the present invention to effectively exhibit the anti-wrinkle and whitening effect, and is determined based on the experimental results according to the concentration of the anastomotic extract.

Anastomosis is a clamshell of Meretrix meretrix or Cyclina sinensis , which is used to clean the lung heat and relieve large amounts of hard wall. It is prescribed for chest heat accumulation that causes chest compressions, wheezing, dyspnea, cough, and large amounts of sputum, softens and eliminates bumps. Clinically includes the treatment of scrofula and thyroid nodule. Neutralizes stomach acid to treat stomach pain, acid reflux and gastroduodenal ulcers, and also has a mild action to promote urination, which can cure edema and urination disorders. When used topically, it helps to heal wounds, burns and eczema. However, no studies have yet been conducted on the effects of such anastomosis on collagenase and tyrosinase.

In the present invention, in order to confirm the anti-wrinkle and whitening effect on anastomotic extract, HS68 human fibroblasts were used as test subjects, and type I procollagen production and collagenase activity according to UVB exposure when anastomotic extracts were treated by concentration The effect on and the effect on tyrosinase activity was confirmed.

According to the present invention, the anastomotic extract inhibits collagenase activity and tyrosinase activity in UVB-damaged cells, thereby exhibiting an excellent anti-wrinkle and skin lightening effect.

The composition of the present invention can be used for pharmaceuticals and cosmetics.

At this time, the composition according to the present invention can be formulated based on the formulation criteria of the Food and Drug Administration (KFDA) to a conventional pharmaceutical formulation or a dietary supplement of dietary supplements.

The composition of the present invention may be diluted in a conventional manner, mixed with a pharmaceutically acceptable carrier, or encapsulated in a container form according to the administration method, dosage form, and purpose.

When the carrier is used as a diluent, oral administration with at least one carrier selected from the group consisting of saline, buffer, dextrose, water, glycerol, Ringer's solution, lactose, sucrose, calcium silicate, methyl cellulose and ethanol For parenteral administration it is prepared in formulations such as powders, granules, injections, syrups, solutions, tablets, suppositories, pesaries, ointments, creams or aerosols. However, the carrier of the present invention is not limited to the above carrier. In this case, parenteral administration means administration of the active ingredient through rectal, intravenous, peritoneal, muscular, arterial, transdermal, nasal, inhalation, etc. in addition to orally.

The formulations may further comprise a filler, an anti-coagulant, a lubricant, a wetting agent, a flavoring agent, an emulsifier, an antiseptic, etc. to formulate the composition so as to provide rapid, sustained or delayed release of the active ingredient after administration to the mammal. And the dosage of the present invention can be adjusted according to the condition of the patient, the route of administration and the dosage form is not limited and those skilled in the art according to the symptoms can be obviously used within various ranges, In general, in the present invention, it is determined that the anastomotic extract according to the present invention in an experimentally effective amount may be continuously or intermittently administered 0.1 to 2000 mg per 1 kg of body weight per day.

On the basis of the effective amount, the present invention provides a cosmetic or cosmetic comprising a composition comprising a anastomosis extract itself or a cosmetically acceptable carrier as a functional ingredient, the cosmetic or cosmetics, such as lotion, nutrition lotion, nutrition cream, It can be prepared and used in a form selected from the group consisting of massage creams, packs and nutritional essences. At this time, the cosmetic preparation method and the carrier of the above-described form is obvious to those skilled in the art and description of the specific manufacturing method will be omitted.

As described above, since the composition according to the present invention is an extract extracted from natural substances, there are almost no side effects, and because it inhibits collagenase activity and tyrosinase activity of fibroblasts, it is necessary to treat or improve wrinkles and skin pigmentation, etc. When used as a medicine or cosmetics, excellent anti-wrinkle and skin whitening effect can be expected.

1 is a graph illustrating the cytotoxicity of the anastomotic extract in HS68 human fibroblasts. B is a control group treated with distilled water instead of anastomotic extract and not UVB treated, and C is a control group treated with distilled water and UVB treated instead of anastomotic extract. 10, 30, 100 are the experimental groups treated with anastomotic extract of 10, 30, 100 ㎍ / ㎖ concentration and UVB treatment, respectively. Each result is expressed as the mean ± standard error (SEM) for the results of three replicates.
Figure 3 is a graph showing the experiment of the effect of anastomotic extract on collagenase activity of HS68 human fibroblasts. B is a control group treated with distilled water instead of anastomotic extract and not UVB treated, and C is a control group treated with distilled water and UVB treated instead of anastomotic extract. 10, 30, 100 are the experimental groups treated with anastomotic extract of 10, 30, 100 ㎍ / ㎖ concentration and UVB treatment, respectively. Each result is expressed as the mean ± standard error (SEM) for the results of three replicates. * Indicates significant difference compared to the control (p <0.05).
Figure 4 is a graph showing the experimental experiment of the effect of anastomotic extract on tyrosinase activity. C is a control group treated with distilled water instead of the anastomotic extract, and 0.1, 1, and 10 are experimental groups treated with anastomotic extracts at concentrations of 0.1, 1, and 10 mg / ml. Each result is expressed as the mean ± standard error (SEM) for the results of three replicates. * Indicates significant difference compared to the control (p <0.05).

Hereinafter, the present invention will be described in more detail with reference to Examples. Since these examples are only for illustrating the present invention, the scope of the present invention is not to be construed as being limited by these examples.

Example 1. Anastomosis Extract Preparation

Anastomosis was purchased from Omniherb, Korea.

2,000 g of 70% ethanol was added to 100 g of anastomosis and heated to 100 ° C. in a heat extractor for 3 hours to obtain an extract.

In addition, the obtained extract was lyophilized using a lyophilizer to obtain 0.09 g of a lyophilisate, and then the lyophilisate was dissolved in water or a buffer solution at each concentration, and the microfilter paper (Whatman no. 2, 0.45 ~ 0.2 μm) was filtered three times to prepare a sample for the experiment. Samples were stored in sterile glass bottles sealed.

Experimental Example 1

The effect of the anastomotic extract of Example 1 on HS68 human fibroblasts was confirmed.

Reagents used in this experiment were purchased from Sigma-Aldrich, St. Louis, Mo., USA, with some exceptions.

The experimental results are expressed as standard error of mean ± mean. The significance of the change was determined using one-way ANOVA as Dunnett's post hoc test. p <0.05 values indicate statistically significant.

1-1. Cell culture

HS68 human fibroblasts (Health Protection Agency Culture Collections, UK) were tested at 37 ° C in Dulbecco's Modified Eagle's medium (Gibco, USA) containing 10% fetal bovine serum and 1% antibiotic. Incubated in humidified conditions of 5% CO ₂ .

1-2. UVB treatment

In order to treat UV in the cells of the subject, a UVB lamp (Vilber Lourmat, France) was used as a UVB source. HS68 cells were washed twice with phosphate-buffered saline (PBS) and all UVB treatments were performed under a layer of thin PBS (200 μl / well). Immediately after UVB treatment, fresh medium without serum was added to the cells and after a 24 hour incubation period, the reaction was measured. The control group followed the same manner as above without performing only UVB treatment.

1-3. Cytotoxicity test

Cell viability was determined using MTT [3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyltetrazolium bromide] (reduced with formazan).

Human fibroblasts (HS68) were seeded in 24-well plates to a density of 2 × 10 ⁵ cells / ml per well and incubated at 37 ° C. with 5% CO ₂ . Prior to UVB treatment, the cells were each pretreated for 24 hours with the sample of Example 1 above. After UVB treatment, the cells were retreated with each corresponding sample and further incubated for 24 hours before treatment with MTT at 0.05 mg / ml (final concentration). Untreated and control groups were incubated without treatment of the sample of Example 1. Cells were then further incubated at 37 ° C. for 4 hours. The medium containing MTT was removed, the resulting MTT formazan was extracted with 200 μl of DMSO, and the absorbance was measured at 595 nm with reference to a wavelength of 690 nm. Cell viability was calculated by the following formula.

[formula]

% Cell viability = [(OD595 of sample) / (OD595 of control)] × 100

The results are shown in FIG. 1, and cell viability was recalculated using the control as 100%. Cell viability was found to be 103.1 ± 3.1%, 100.4 ± 1.1% and 100.1 ± 0.5% at concentrations of 10, 30 and 100 μg / ml, respectively. Thus, up to 100 μg / ml, no cytotoxicity was observed.

1-4.

H68 human fibroblasts were seeded (2 × 10 ⁵ cell / well) in 24-well plates and incubated at 5% CO ₂ conditions and 37 ° C. The cells were pretreated for 24 hours with the samples of Example 1 (10, 30, 100 μg / ml concentrations) before UVB treatment, respectively. After UVB treatment, cells were retreated with the sample of each Example 1 and further incubated for 24 hours. Untreated and control groups were incubated without the sample treatment of Example 1. After incubation, supernatants were collected from each well.

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1-5. Collagenase Inhibition Assay

In order to evaluate collagenase activity, matrix metalloproteinase-1 (MMP-1) activity was measured, and matrix metalloproteinase-1 (MMP-1) human biotrak ELISA system for the supernatant of 1-4. It was measured using (Amersham life science, USA).

MMP-1 activity was recalculated with the control as 100% (see Figure 3). As a result, MMP-1 activity was significantly decreased (33.4 ± 0.6%, 20.3 ± 0.6%, 25.0 ± 0.3%) at all concentrations of 10, 30 and 100 µg / ml.

1-6. Tyrosinase Inhibition Assay

Tyrosinase activity was determined according to existing methods. To 20 μl of the sample of Example 1 dissolved in 0.1 M sodium phosphate buffer (pH 6.5) (0.1, 1 and 10 mg / ml) was added 40 U of mushroom tyrosinase and 40 μl of 1.5 mM L-tyrosine. 220 μl of 0.1 M sodium phosphate buffer was added to the reaction mixture. The mixture (300 µl) was reacted at 37 ° C for 10 minutes, and then the absorbance was measured at 490 nm. The same but untreated sample of Example 1 was used as a control.

The control was recalculated to 100% (see Figure 4). As a result, tyrosinase activity was significantly decreased (58.1 ± 5.6%, p <0.05) at a concentration of 10 mg / ml, and tyrosinase activity of the groups treated with 0.1 and 1 mg / ml was not significant (90.1 ± 0.3). %, 80.7 ± 10.9%).

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Claims (5)

Anti-wrinkle and whitening cosmetic composition comprising the anastomosis extract as an active ingredient to inhibit collagenase activity and tyrosinase activity. According to claim 1, wherein the anastomosis extract
A cosmetic composition for anti-wrinkle and whitening, characterized in that the extract using the alcohol solution of 2 to 5 carbon atoms of 10 to 100% by volume as an extraction solvent and extracted from the anastomosis by heating. According to claim 1, wherein the anastomosis extract
A cosmetic composition for anti-wrinkle and whitening, which is an extract obtained by adding 100 to 10000 parts by weight of an ethanol solution of 10 to 100% by volume to anastomosis and heating for 10 minutes to 30 hours. According to claim 1, wherein the anastomosis extract
A cosmetic composition for anti-wrinkle and whitening, wherein 100 to 10000 parts by weight of an ethanol solution of 10 to 100% by weight of anastomosis is added and the extract obtained by heating for 10 minutes to 30 hours is lyophilized. According to claim 1, wherein the composition is anti-wrinkle and whitening cosmetic composition, characterized in that it contains 0.1 to 50% by weight anastomosis extract.

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