KR102091587B1 - Composition for Improving Skin Wrinkle Containing gamma-Oligopeptide - Google Patents

Abstract

The present invention relates to a cosmetic composition for improving skin wrinkles containing a gamma oligopeptide or a salt thereof, and more specifically, a cosmetic that improves skin wrinkles by allowing skin penetration, inhibiting collagen degrading enzymes in the skin, and increasing collagen production. It relates to a composition.
The cosmetic composition for improving skin wrinkles according to the present invention inhibits collagen-degrading enzymes, enhances collagen synthesis, has little toxicity and side effects, and can be used for a long time, and is useful for improving skin wrinkles.

Description

Cosmetic composition for wrinkle improvement containing gamma oligopeptide or salt thereof {Composition for Improving Skin Wrinkle Containing gamma-Oligopeptide}

The present invention relates to a cosmetic composition for improving skin wrinkles containing a gamma oligopeptide or a salt thereof, more specifically, it is possible to penetrate the skin, inhibit collagen degrading enzymes in the skin, and improve collagen production to improve skin wrinkles The present invention relates to a cosmetic composition for improving wrinkles containing gamma oligopeptide or a salt thereof.

In the future, as the importance of functional cosmetics that emphasizes efficacy and effectiveness is emphasized in the development of cosmetics in the future, the expansion of the range of functional cosmetics in the future becomes a current task for the future development of the cosmetics industry. In other words, the concept of cosmetics is expected to expand from existing concepts such as skin care, anti-aging and cell revitalization, whitening and UV protection, to products of the current quasi-drugs and medicinal cosmetics such as wound healing, anti-atopy, and body improvement. With the implementation of the Cosmetics Act and the expansion of the range of functional cosmetics, spurs are being spurred on the development of cosmetics research and development and manufacturing technology, and patent applications and approvals for functional cosmetics are expected to gradually increase as a cornerstone to preempt the era of functional cosmetics. There is a growing tendency to be recognized for technology through domestic and foreign patents.

Functional cosmetics, which mainly focused on product development for the purpose of skin protection, such as UV protection in the early days, have been developing products for cosmetic purposes such as wrinkle improvement in the past 10 years, and multiple products capable of whitening, wrinkle improvement, and color simultaneously The demand for functional cosmetics, aka BB cream, has also increased.

Unlike whitening, the exact mechanism for wrinkle improvement is relatively unclear. The main cause of wrinkles is skin aging, and aging can be roughly divided into two cases: natural aging and photoaging. That is, wrinkles on the skin are caused by reduction of elasticity fibers such as collagen or elastin due to a decrease in cell regeneration ability, lipid peroxide formation by free radicals, degeneration due to oxidation of biomaterials, and fatigue due to excessive muscle exercise. It is known as. Therefore, in order to prevent aging or improve wrinkles, functions such as promoting collagen synthesis, antioxidant activity, and preventing muscle fatigue are required.

Retinol (2500 IU / g), retinyl palmitate (10000 IU / g), adenosine (0.04%), polyethoxylated retinamide (0.05∼0.2) %). In the case of using the published concentration, they are also exempt from the submission of safety validation data, and in addition, among the products approved as functional cosmetics through individual certification, hydroxyproline, 7-dihydrocholesterol, kineticin, peony extract, betel nut Self-extract

Wrinkle-improving functional materials are mainly components that control the differentiation and regeneration of epidermal cells, substances that regulate extracellular matrix (ECM), antioxidants that scavenge free radicals, anti-inflammatory components, components that protect against damage by UV rays, and others. Peptides of the similar botox concept, which degrade muscle movement, are the mainstream. In addition to conventional retinoids and AHAs, mevalonic acid has been developed and used as a substance that controls the differentiation and regeneration of epidermal cells, and components such as paoniflorin extracted from peony or prangenidin extracted from white paper have been developed and used as an ECM control component. . In the case of Prangenidin, the collagen biosynthesis ability through PCIP enzyme immunoassay was found to be about 20 times superior to vitamin C. On the other hand, a component that protects or regenerates DNA that is susceptible to damage by ultraviolet rays has been developed, and a representative example is photolyase, which is extracted from phytoplankton and known as an enzyme for DNA damage-healing function.

Polygammaglutamic acid (γ-PGA) is a mucolytic material produced by microorganisms. Poly gamma glutamic acid is produced from Bacillus isolated from Cheonggukjang, a traditional Korean fermented food using rice straw. Poly-gamma-glutamic acid is an edible, water-soluble, anionic, and biodegradable polymer material that can be used as a raw material and functional food material for cosmetics such as hygroscopic and moisturizing agents.

The present inventors produce poly-gamma-glutamic acid using a high molecular weight poly-gamma-glutamic acid and a material patent for a method of using the same (Korea Registered Patent No. 399091), a salt-resistant strain of Bacillus subtilis cheonggukjang strain, which produces a high-molecular weight poly-gamma-glutamic acid Patent on how to do this (Korea Registered Patent No. 500796), in addition, anti-cancer composition containing polygamma glutamic acid, adjuvant, immune enhancer and patent for the inhibition of viral infection (Republic of Korea Registered Patent No. 496606, Republic of Korea registered Patent No. 517114 and Korea Patent No. 475406, Korea Patent No. 0873179) have been obtained. In addition, by revealing the anti-cancer function through immune enhancement of polygamma glutamic acid [Poo, HR et al., Journal of Immunology , 178: 775, 2007, Poo, HR et al., Cancer Immunol Immunother , 59:11, 2010] In addition, research on the pharmaceutical use of polygamma glutamic acid has been conducted, and various effects have been studied while developing continuous use for polygamma glutamic acid.

Conventional cosmetic compositions containing polygamma glutamic acid provide application aspects as moisturizing (Korea Patent Publication No. 2010-0101328), skin whitening (Korea Patent Publication No. 2010-0086728) or mineral skin absorption enhancement (2009-0132372) However, it has not been applied for skin wrinkle improvement.

Accordingly, the present inventors developed a gamma oligopeptide which is a poly-gamma-glutamic acid formulation capable of skin permeation, and as a result of diligent efforts to confirm the effect on wrinkle improvement, as a result of successfully preparing a skin-permeable poly-gamma-glutamic acid formulation, this material is a collagen degrading enzyme And inhibited and improved collagen synthesis, thereby improving skin wrinkles, and completed the present invention.

An object of the present invention is to provide a cosmetic composition for improving skin wrinkles containing a gamma oligopeptide or a salt thereof having an effect on improving skin wrinkles.

In order to achieve the above object, the present invention provides a cosmetic composition for improving skin wrinkles containing gamma oligopeptide or a salt thereof as an active ingredient.

The cosmetic composition for improving skin wrinkles according to the present invention inhibits collagen-degrading enzymes, enhances collagen synthesis, has little toxicity and side effects, and can be used for a long time, and is useful for improving skin wrinkles.

1 shows the ability of gamma oligopeptide to inhibit collagen degrading enzyme activity.
Figure 2 shows the ability of gamma oligopeptide to synthesize procollagen.

In the present invention, it was intended to confirm whether the gamma oligopeptide having a low molecular weight of polygamma glutamic acid or a salt thereof is effective in improving skin wrinkles.

In the embodiment of the present invention, a gamma oligopeptide having a low molecular weight of polygamma glutamic acid or a salt thereof was prepared. The molecular weight of the gamma oligopeptide or a salt thereof prepared from the low molecular weight polygamma glutamic acid used in the present invention is characterized in that it is 600 to 5,000 Da, and the gamma oligopeptide is characterized in that it is a peptide composed of 5 to 15 glutamic acid. , The salt was selected from the group consisting of sodium salt, potassium salt, calcium salt, magnesium salt, ammonium salt and zinc salt.

In another embodiment of the present invention, UVA is irradiated to HDF-N cells to confirm the inhibitory ability of collagen degrading enzyme, and then the activity of collagen degrading enzyme is analyzed after culturing by adding gamma oligopeptide (0.05, 0.1%, 0.5%). As a result, the activity of the collagen degrading enzyme in the control group and the activity of the degrading enzyme in the gamma oligopeptide treatment group were measured to be 177%, 154%, and 133%, respectively, compared to the control group not irradiated with UVA, and thus the gamma oligopeptide was not treated. It was confirmed that the activity of the collagen degrading enzyme was significantly reduced compared to the value of the non-experimental group of 183% (FIG. 1).

In another embodiment of the present invention, as a result of analyzing the concentration of procollagen after culturing by adding gamma oligopeptide (0.01, 0.05%, 0.1%) to HDF-N cells to confirm collagen production capacity, compared with the control group It increased to 129%, 168% and 199%, respectively (FIG. 2). Through this, gamma oligopeptide was confirmed to show efficacy in wrinkle improvement by increasing the concentration of procollagen synthesis in a concentration-dependent manner.

The cosmetic composition for improving skin wrinkles of the present invention may be added to a functional cosmetic product for improving wrinkles. In the cosmetic composition for improving wrinkles of the present invention, the content of gamma oligopeptide is preferably 0.005% to 10%, and if it is less than 0.005%, the effect of improving skin wrinkles cannot be expected, and if it is more than 10%, the effect of improving wrinkles according to the increase in content Not only is it unpredictable, it is expensive, and it is not economical.

Cosmetics prepared from the compositions of the present invention can be prepared in the form of general emulsifying and solubilizing formulations. Cosmetics in emulsified formulations include nutrient makeup, cream, essence, etc., and cosmetics in solubilized formulations include softened makeup. In addition, the composition of the present invention may be prepared in the form of an adjuvant for topical application or systemic application commonly used in the field of dermatology by containing a dermatologically acceptable medium or base in addition to cosmetics. Specifically, the cosmetic composition may be characterized in that it is any one formulation selected from the group consisting of cosmetics, essences, skins, lotions, creams, packs, foundations, and makeup bases.

Suitable cosmetic formulations include, for example, solutions, gels, solid or dough anhydrous products, emulsions, suspensions, emulsions, pastes, microemulsions, microcapsules, microgranules or ionic forms (liposomes) obtained by dispersing an oil phase in an aqueous phase. In the form of nonionic vesicle dispersants, in the form of creams, skins, lotions, powders, soaps, ointments, sprays, surfactant-containing cleansing, oil, powder foundations, emulsion foundations, wax foundations or conceal sticks Can be provided. In addition, it may be prepared in the form of a foam or aerosol composition further containing a compressed propellant.

The "dermatologically acceptable carrier" that can be used depending on the desired formulation is purified water, oil, wax, fatty acid, fatty acid alcohol, fatty acid ester, surfactant, humectant, thickening agent, antioxidant, viscosity stability It includes, but is not limited to, topical agents (viscosity stabilizers), chelating agents, buffering agents, preservatives, lower alcohols, etc., the types and concentrations of which are varied, and those skilled in the art are modified without departing from the spirit and scope of the present invention. And parts that can be changed. Whitening agents, moisturizers, anti-inflammatory agents, anti-bacterial agents, anti-fungal agents, vitamins, sunscreens, antibiotics, anti-acne agents, perfumes, and dyes may be included, if necessary. Can be included. 0.001 to 50% by weight per total weight of the composition is suitable.

Specifically, as the oil, hydrogenated vegetable oil, perm oil, cottonseed oil, olive oil, palm oil, jojoba oil, avocado oil may be used, and wax includes beeswax, sperm, carnauba, candelilla, montan, ceresin, liquid paraffin , Lanolin can be used. As fatty acids, stearic acid, linoleic acid, linolenic acid, and oleic acid may be used, and cetyl alcohol, octyl dodecanol, oleyl alcohol, panthenol, lanolin alcohol, stearyl alcohol, and hexadecanol may be used as fatty acid alcohol. , As a fatty acid ester, isopropyl myristate, isopropyl palmitate, butyl stearate, etc. may be used, but are not limited thereto.

Examples of surfactants include anionic surfactants such as sodium stearate, sodium cetyl sulfate, polyoxyethylene lauryl ether phosphate, and sodium N-acyl glutamate; Cationic surfactants such as stearyldimethylbenzylammonium chloride and stearyltrimethylammonium chloride; Amphoteric surfactants such as alkylaminoethylglycine hydrochloride and lecithin; Glycerin monostearate, sorbitan monostearate, propylene glycol monostearate, polyoxyethylene oleyl ether, polyethylene glycol monostearate, polyoxyethylene sorbitan monopalmitate, polyoxyethylene coconut fatty acid monoethanol arniide (monoethaolarnide) ), Polyoxypropylene glycol, polyoxyethylene castor oil, and nonionic surfactants such as polyoxyethylene lanolin.

Glycerin, 1,3-butylene glycol and propylene glycol may be used as the absorbent, and ethanol and isopropanol may be used as the lower alcohol. Examples of thickeners include sodium alginate, sodium caseinate, gelatin agar, xanthan rubber, starch, cellulose ethers (e.g., hydroxyethyl cellulose, methyl cellulose, carboxymethyl cellulose, hydroxypropylmethyl cellulose), polyvinyl pyrrolidone, Polyvinyl alcohol, polyethylene glycol, sodium carboxymethyl cellulose, and the like, but is not limited thereto. Butylated hydroxytoluene, butylated hydroxyanisole, propyl gallate, citric acid, and ethoxyquin are available as antioxidants, and disodium edetate and ethanehydroxy diphosphate are used as chelating agents. Available, citric acid, sodium citrate, boric acid, borax, disodium hydrogen phosphate can be used as a buffer, methyl parahydroxybenzoate, ethyl parahydroxybenzoate, dihydro as a preservative Acetic acid, salicylic acid, and benzoic acid are available, but are not limited thereto.

Hereinafter, the present invention will be described in more detail through examples. These examples are only for illustrating the present invention, it will be apparent to those skilled in the art that the scope of the present invention is not to be construed as limited by these examples.

Example 1: Preparation of gamma oligopeptide and salts thereof

Basic medium for polygamma glutamic acid production (3% L-glutamic acid added medium; glucose 3%, (NH ₄ ) ₂ SO ₄ 1%, KH ₂ PO ₄ 0.27%, Na ₂ HPO ₄ / 12H ₂ O 0.17% , NaCl 0.1%, sodium citrate 0.5%, Soypeptone 0.02%, MgSO ₄ / 7H ₂ O 0.7%, Vitamin solution 10ml / ℓ, pH 6.8) prepared by sterilization, 5L fermenter ( Jar fermentor, working volume 3ℓ) was inoculated with 4% seed culture (LB medium) of Bacillus subtilis var chungkookjang (KCTC 0697BP). The stirring speed was 500 rpm, and the air injection speed was 1.0 vvm. After fermentation at 37 ° C. for 48 hours, the cells were removed using a filter press (containing diatomaceous earth) to obtain a sample solution containing polygamma glutamic acid.

After adjusting the pH to 2.0 using a 2N sulfuric acid solution in the sample solution containing polygamma glutamic acid, the polygamma glutamic acid precipitate was obtained by standing at 10 ° C for 15 hours. After washing the obtained poly-gamma-glutamic acid precipitate with a sufficient amount of cold distilled water at a temperature of 10 ° C. or less (pH 3.5 or higher), a polygamma-glutamic acid precipitate was obtained using a Nutsche filter and lyophilized to prepare a high molecular weight poly-gamma-glutamic acid.

After adding the prepared poly-gamma-glutamic acid to distilled water and warming it at 121 ° C for 2 hours, sodium hydroxide, potassium hydroxide, calcium hydroxide, and magnesium hydroxide were added to dissolve them at a neutral pH to freeze-dried the gamma oligopeptide salt solution. Powders were prepared. The average molecular weight of the prepared gamma oligopeptide is 1,300 Da and the number of glutamic acid oligomers is 10.

Example 2: collagen degrading enzyme inhibitory function

Among several mechanisms in which wrinkles are generated due to natural aging and photoaging, this test was intended to evaluate the wrinkle improvement efficacy of a sample by measuring the activity level of collagen degrading enzyme MMP-1 (Matrix metalloproteinases) using an immuno ELISA assay (Fig. One).

Cell line used was Normal Human Primary Dermal fibroblasts-Neonatal (HDF-N) derived from Human neonatal foreskin. Fibroblast Basal Medium (FBM, lonza, CC-3131) medium containing 0.1% insulin, 0.1% rhFGF, 0.1% gentamicin, and 2% FBS after inoculating HDF-N (ATCC PCS-201-010) on the bottom of the culture dish And maintained at 37 ° C. and cultured in an incubator containing 5% carbon dioxide.

In order to evaluate the ability of gamma oligopeptide to inhibit collagen degrading enzyme (MMP-1), HDF-N cells were dispensed at 2.0 × 10 ⁴ cells / well in a 24 well plate, and then cultured for 48 hours under cell culture conditions. After 48 hours, the medium was discarded, washed with DPBS, and 200 μl of DPBS was added to irradiate UV-A 5J / cm 2. After diluting the sample in a medium at an appropriate concentration, the cells were treated and cultured for 24 hours under cell culture conditions. Retinoic acid (RA) was used as a positive control, and the concentration was treated with 1 uM. The culture was taken and the amount of MMP-1 was measured using a Human total MMP-1 ELISA kit (R & D system, DY901). The measured MMP-1 amount was corrected by the total protein amount.

First, as a result of irradiating UVA wavelength 5J / cm ² to HDF-N cells, the experimental group irradiated with UVA increased MMP-1 production by 183.03% compared to the control group not irradiated. Thus, it was found that irradiation of UVA 5J / cm ² effectively induced MMP-1 production in HDF-N cells. On the other hand, after treating gamma oligopeptide with 0.05, 0.1, and 0.5% concentrations, it was measured with Human total MMP-1 ELISA kit (R & D system, DY901), and showed 177.34%, 154.43%, and 133.34%, respectively, compared to the control group. . Through this, it was confirmed that gamma oligopeptide reduces MMP-1 in a concentration-dependent manner.

Example 3: Procollagen synthesis function

To evaluate the collagen precursor procollagen (procollagen) synthesis ability, HDF-N cells were dispensed into a 48 well plate at 1.0 × 10 ⁴ cells / well and cultured for 24 hours under cell culture conditions. Thereafter, the medium was removed, and the starvation state of the cells was maintained for 24 hours, and then, the test substance was diluted in cell culture medium FBM (excluding media supplement), treated to cells by concentration, and then cultured for 24 hours. After 24 hours, the medium of the cultured cells was collected and the amount of procollagen was measured using a procollagen type I c-peptide (PIP) EIA kit (TAKARA, MK101) (FIG. 2). On the other hand, the cells attached to the bottom were washed with DPBS, lysis with 1N NaOH, and the total protein amount was measured to obtain the amount of procollagen synthesis per protein. The amount of each procollagen and the amount of protein was calculated according to the calibration curve.

As a result, when treated with 10 ng / ml of TGF-β used as a positive control, procollagen synthesis increased by 195.23% compared to the control group. On the other hand, BLS-PGA increased to 129.34%, 168.74%, and 199.60%, respectively, compared to the control group when the concentrations of 0.01%, 0.05%, and 0.10% were treated with 0.10% as the highest concentration. Through this, procollagen synthesis ability was increased in a concentration-dependent manner, and all three concentrations showed statistically significant results. Accordingly, gamma oligopeptide is thought to show efficacy in improving wrinkles by increasing the concentration of procollagen synthesis in a concentration-dependent manner at 0.01%, 0.05%, and 0.10%.

Hereinafter, a formulation example for the composition of the present invention is illustrated. However, the formulation comprising the composition of the present invention is not limited now.

Formulation Example 1: Flexible lotion (skin)

Gamma Oligopeptide 5.00 (%)

1,3-butylene glycol 1.00 (%)

Disodium IDT 0.05 (%)

Allantoin 0.10 (%)

Dipotassium glycyrrhide 0.05 (%)

Citric Acid 0.01 (%)

Sodium citrate 0.02 (%)

Glyceres-26 1.00 (%)

Arbutin 2.00 (%)

Hydrogenated castor oil 1.00 (%)

Ethanol 30.00 (%)

Preservative trace

Colorant trace

Flavoring agent trace

Purified water level

Formulation Example 2: Nutrition Cream

Gamma Oligopeptide 5.00 (%)

1,3-butylene glycol 7.0 (%)

Glycerin 1.0 (%)

D-panthenol 0.1 (%)

Plant extract 3.2 (%)

Magnesium aluminum silicate 0.3 (%)

PEG-40 stearate 1.2 (%)

Stearic Acid 2.0 (%)

Polysorbate 60 1.5 (%)

Lipophilic glyceryl stearate 2.0 (%)

Sorbitan sesquioleate 1.5 (%)

Cetearyl alcohol 3.0 (%)

Mineral oil 4.0 (%)

Squalane 3.8 (%)

Carlylic / Capric Triglyceride 2.8 (%)

Vegetable oil 1.8 (%)

Dimethicone 0.4 (%)

Dipotassium glyceride trace amount

Allantoin trace

Sodium hyaluronate trace

Tocopheryl Acetate

Triethanolamine

Preservative dosage

Flavoring amount

Purified water level

As the specific parts of the present invention have been described in detail above, it will be apparent to those of ordinary skill in the art that these specific techniques are only preferred embodiments, and the scope of the present invention is not limited thereby. will be. Accordingly, the substantial scope of the present invention will be defined by the appended claims and their equivalents.

Claims (7)

As a cosmetic composition for the improvement of skin wrinkles containing gamma oligopeptide or a salt thereof as an active ingredient,
The total content of the gamma oligopeptide or salt thereof is a cosmetic composition for improving skin wrinkles, characterized in that 0.01 to 0.5% by weight relative to the total weight of the cosmetic composition.
The method of claim 1, wherein the gamma oligopeptide or a salt thereof is a cosmetic composition for improving skin wrinkles, characterized in that it is prepared by reducing the molecular weight of polygamma glutamic acid.
[6] The cosmetic composition for improving skin wrinkles according to claim 1, wherein the gamma oligopeptide or a salt thereof has a molecular weight of 600 to 5,000 Da.
According to claim 1, The gamma oligopeptide cosmetic composition for improving skin wrinkles, characterized in that the peptide consisting of 5 to 15 glutamic acid.
According to claim 1, The salt is sodium salt, potassium salt, calcium salt, magnesium salt, ammonium salt and zinc salt cosmetic composition for improving wrinkles, characterized in that selected from the group consisting of salts.
delete The cosmetic composition of claim 1, wherein the cosmetic composition is any one selected from the group consisting of cosmetic products, essences, skins, lotions, creams, packs, foundations, and makeup bases. Cosmetic composition.

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