WO2014058084A1 - Cosmetic composition containing gamma-oligopeptide or salts thereof for improving wrinkles - Google Patents

Abstract

The present invention relates to a cosmetic composition containing gamma-oligopeptide or salts thereof for improving wrinkles on the skin, and more specifically, to a cosmetic composition which can penetrate the skin, inhibit collagen-dissolving enzymes in the skin, and increase collagen generation to improve wrinkles on the skin. The cosmetic composition for improving wrinkles on the skin according to the present invention inhibits collagen-dissolving enzymes, increases collagen synthesis, is not toxic and has few side effects and thus can be used for a long time, and is useful in improving wrinkles on the skin.

Description

Cosmetic composition for anti-wrinkle containing gamma oligopeptide or salt thereof

The present invention relates to a cosmetic composition for improving skin wrinkles containing gamma oligopeptides or salts thereof, and more particularly to skin permeation, inhibiting collagen degrading enzymes in the skin, and increasing collagen production to improve skin wrinkles. It relates to a cosmetic composition for improving wrinkles containing gamma oligopeptide or a salt thereof.

The future development of cosmetics is emphasized the importance of functional cosmetics that have enhanced efficacy and effectiveness compared to general cosmetics that emphasize stability, and the expansion of the range of functional cosmetics in the future is becoming an issue for the future development of the cosmetics industry. In other words, the concept of cosmetics will be expanded from existing concepts such as skin care, anti-aging and cell regeneration, whitening and UV protection to products of current quasi-drugs and medicinal cosmetics such as wound healing, anti-atopy, and body improvement. The enforcement of the Cosmetic Act and the expansion of the range of functional cosmetics are accelerating the development of R & D manufacturing technology of cosmetics, and the patent application and approval of functional cosmetics is expected to gradually increase as a cornerstone for preoccupation of the era of functional cosmetics. There is a growing tendency to be recognized for technology through domestic and foreign patent acquisition.

Functional cosmetics, which were mainly focused on product development for the purpose of skin protection, such as UV protection in the early years, have been developing products for cosmetic purposes such as wrinkle improvement in the past 10 years, and they can simultaneously whiten, wrinkle and color. The market demand for functional cosmetics, also known as BB creams, has also increased.

Unlike whitening, the exact mechanism for wrinkle improvement is relatively unclear. The main cause of wrinkles is skin aging. Aging can be classified into two types: natural aging and photo aging. In other words, the wrinkles of the skin may be caused by a decrease in elastic fibers such as collagen or elastin due to the deterioration of cell regeneration ability, the generation of lipid peroxide by active oxygen, degeneration due to oxidation of biological components, and fatigue caused by excessive muscle movement. Known as Therefore, in order to prevent aging or to 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%), Polyethoxylatediretinamide (0.05-0.2) 4 types). They are also exempted from submitting safety validation data when using the indicated concentrations.In addition, products approved as functional cosmetics through individual certification are hydroxyproline, 7-dihydrocholesterol, kainetin, peony extract, and betel nut. There are self extracts

Anti-wrinkle functional materials are mainly used to control the differentiation and regeneration of epidermal cells, substances to regulate extracellular matrix (ECM), antioxidants to eliminate free radicals, anti-inflammatory components, components to protect against UV damage Peptides with a similar Botox concept that slow muscle movement are the mainstream. In addition to conventional retinoids and AHAs, mevalonic acid has been developed and used as an agent for controlling the differentiation and regeneration of epidermal cells, and ingredients such as paoniflorin extracted from peony or prangenidin extracted from white paper have been developed and used as ECM control ingredients. . In the case of Prangenidin, collagen biosynthesis through PCIP enzyme immunoassay was found to be about 20 times better than vitamin C. On the other hand, a component that protects or regenerates the DNA susceptible to UV damage has been developed, a typical example is a photolyase extracted from phytoplankton, known as an enzyme of DNA damage healing function.

Polygamma glutamic acid ([gamma] -PGA) is a mucous substance produced by microorganisms. Poly-gamma-glutamic acid is produced from Bacillus isolates from Cheonggukjang, a traditional Korean fermented soybean meal using rice straw. Polygamma glutamic acid is an edible, water-soluble, anionic and biodegradable polymer that can be used as a raw material and functional food material for cosmetics such as moisture absorbents and moisturizers.

The present inventors produce polygamma glutamic acid using a high-molecular weight polygamma glutamic acid and a material patent for the method of using the same (Korean Patent No. 399091), a salt-tolerant strain Bacillus subtilis Chungkukjang strain producing high molecular weight poly gamma glutamic acid. Acquired a patent for a method (Korean Patent No. 500796), and a patent for anti-cancer composition containing polygamma glutamic acid, an immunoadjuvant, an immune enhancer, and a virus infection inhibition (Korean Patent No. 496606, Republic of Korea) Patent No. 517114, Republic of Korea Patent No. 475406, Republic of Korea Patent No. 0873179). In addition, by revealing the anti-cancer function through immuno-enhancing 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 medical use of polygamma glutamic acid has been carried out, and various effects have been studied through continuous development of polygamma glutamic acid.

The cosmetic composition containing the conventional polygamma glutamic acid provides an application aspect as a moisturizing (Korean Patent Publication No. 2010-0101328), skin whitening (Korean Patent Publication No. 2010-0086728) or for enhancing the skin absorption of minerals (2009-0132372). However, it has not been applied for skin wrinkle improvement.

Therefore, the present inventors have made efforts to confirm the effect on wrinkle improvement by developing gamma oligopeptide, a polygamma glutamic acid formulation capable of permeation of the skin, and thus successfully prepared a skin permeable poly gamma glutamic acid formulation to produce a collagen degrading enzyme. It was confirmed that to improve the skin wrinkles by inhibiting and promoting collagen synthesis, to complete the present invention.

Summary of the Invention

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

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.

Figure 1 shows the collagenase enzyme inhibitory activity of the gamma oligopeptide.

Figure 2 shows the procollagen synthesis of gamma oligopeptides.

Detailed Description of the Invention and Specific Embodiments

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

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

In another embodiment of the present invention, to investigate the inhibitory ability of the collagenase, UVA was irradiated to HDF-N cells, and then cultured by adding gamma oligopeptide (0.05, 0.1%, 0.5%) and analyzing the activity of collagenase. As a result, the activity of collagenase and gamma oligopeptide in the control group were measured to be 177%, 154% and 133%, respectively, compared to the control group without UVA irradiation. Compared with the 183% of the experimental group did not significantly reduce the activity of collagenase enzymes (Fig. 1).

In another embodiment of the present invention, the concentration of procollagen was analyzed by adding gamma oligopeptide (0.01, 0.05%, 0.1%) to HDF-N cells to confirm collagen production ability, and compared with the control group. Increased to 129%, 168%, and 199%, respectively (FIG. 2). Through this, gamma oligopeptide was shown to increase the procollagen synthesis capacity in a concentration-dependent manner to show the effect on wrinkle improvement.

The cosmetic composition for improving skin wrinkles of the present invention may be added to wrinkle improvement functional cosmetics. The content of gamma oligopeptide in the cosmetic composition for wrinkle improvement of the present invention is preferably 0.005% to 10%, when the content is less than 0.005%, anti-wrinkle effect cannot be expected, and when the content is more than 10%, the wrinkle improvement effect is increased. Not only can it not be expected, it is expensive, and it is not economical.

Cosmetics prepared from the compositions of the present invention can be prepared in the form of common emulsifying and solubilizing formulations. Cosmetics of the emulsified formulations include nutrient cosmetics, creams, essences, etc., and cosmetics of the solubilized formulations are flexible cosmetics. In addition, the compositions of the present invention may be prepared in adjuvant form for topical or systemic application commonly used in the field of dermatology by containing a dermatologically acceptable medium or base in addition to cosmetics.

Formulations of suitable cosmetics include, for example, emulsions, suspensions, emulsions, pastes, microemulsions, microcapsules, microgranules or ionic (liposomes) obtained by dispersing an oil phase in a solution, gel, solid or pasty anhydrous product, aqueous phase. , In the form of non-ionic vesicle dispersants, creams, skins, lotions, powders, soaps, ointments, sprays, surfactant-containing cleansing, oils, powder foundations, emulsion foundations, wax foundations or conceal sticks Can be provided. It may also be prepared in the form of a foam or in the form of an aerosol composition further containing a compressed propellant.

“Dermatologically acceptable carriers” that can be used according to the desired formulation include purified water, oils, waxes, fatty acids, fatty alcohols, fatty acid esters, surfactants, humectants, thickening agents, antioxidants, and viscosity stabilizers. Examples include, but are not limited to, topical stabilizers, chelating agents, buffers, preservatives, lower alcohols, and the like, and their types and concentrations are varied and modifications may be made by those skilled in the art without departing from the spirit and scope of the present invention. And parts that can be changed. If necessary, a whitening agent, a moisturizer, an anti-inflammatory agent, an antibacterial agent, an antifungal agent, vitamins, sunscreens, antibiotics, anti-acne agents, perfumes, dyes may be included, and these may be included in the cosmetic composition according to the present invention in an amount commonly used in the cosmetic field. May be included. Suitable from 0.001 to 50% by weight per total weight of the composition.

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

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

Glycerin, 1,3-butylene glycol, propylene glycol may be used as the moisture absorbent, and ethanol or isopropanol may be used as the lower alcohol. Examples of thickeners include sodium alginate, sodium caseate, gelatin agar, xanthan gum, starch, cellulose ethers (e.g., hydroxyethyl cellulose, methyl cellulose, carboxymethyl cellulose, hydroxy propylmethyl cellulose), polyvinyl pyrrolidone, Polyvinyl alcohol, polyethylene glycol, sodium carboxymethyl cellulose, and the like, but are not limited thereto. As antioxidants, butylated hydroxytoluene, butylated hydroxyanisole, propyl gallate, citric acid, ethoxyquin can be used, and chelating agents include disodium edetate and ethane hydroxy diphosphate. Citric acid, sodium citrate, boric acid, borax, disodium hydrogen phosphate are available as buffers, and methyl parahydroxybenzoate, ethyl parahydroxybenzoate, dihydro as preservatives. Acetic acid, salicylic acid, benzoic acid are available, but are not limited to these.

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

Example 1 Preparation of Gamma Oligopeptide and Its Salts

Basic medium for the production of polygamma glutamic acid (medium with 3% L-glutamic acid; 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) to sterilize, 5L fermenter ( Jar fermentor, working volume was fermented by inoculating 4% seed culture (LB medium) of Bacillus subtilis var chungkookjang (KCTC 0697BP) in 3L). The stirring speed was 500rpm, the air injection speed was 1.0vvm, and fermented at 37 ° C. for 48 hours to remove the cells using a filter press (containing diatomaceous earth) to obtain a polygamma glutamic acid-containing sample liquid.

The polygamma glutamic acid-containing sample solution was adjusted to pH 2.0 using 2N sulfuric acid solution and then left at 10 ° C. for 15 hours to obtain a polygamma glutamic acid precipitate. The obtained polygamma glutamic acid precipitate was washed with a sufficient amount of cold distilled water of 10 ° C. or less (pH 3.5 or more), and then obtained polygamma glutamic acid precipitate using a Licech filter, followed by lyophilization to prepare a high molecular weight polygamma glutamic acid.

Example 2 Collagen Degrading Enzyme Inhibition Function

Among various mechanisms in which wrinkles are formed due to natural aging and photo aging, this test was intended to evaluate the wrinkle improvement effect of the sample by measuring the activity of collagen degrading enzyme MMP-1 (Matrix metalloproteinases) by immuno ELISA assay (FIG. One).

Cell lines were 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 Was added and maintained at 37 ° C. and incubated 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 24 well plates and incubated for 48 hours under cell culture conditions. After 48 hours, the medium was discarded and washed with DPBS, and then UV-A 5J / cm 2 was irradiated by adding 200 µl of DPBS. Samples were diluted in medium at appropriate concentrations and then treated with cells and incubated in cell culture conditions for 24 hours. Retinoic acid (RA) was used as a positive control and the concentration was 1 uM. The culture medium was taken and the amount of MMP-1 was measured using a human total MMP-1 ELISA kit (R & D system, DY901). The amount of MMP-1 measured was corrected to the total protein amount.

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

Example 3 Procollagen Synthesis Function

In order to evaluate the procollagen (procollagen) synthesis ability of the collagen precursor HDF-N cells were dispensed at 1.0 × 10 ⁴ cells / well in 48 well plate and incubated for 24 hours in the cell culture conditions. Thereafter, the medium was removed, and the cells were starved for 24 hours, after which the test substance was diluted in FBM (except for media supplement), which is a cell culture medium, and treated with the cells by concentration, followed by incubation for 24 hours. After 24 hours, the cultured cells were harvested and the amount of procollagen was measured using procollagen type I c-peptide (PIP) EIA kit (TAKARA, MK101) (FIG. 2). On the other hand, the cells attached to the bottom was washed with DPBS and lysed with 1N NaOH to measure the total protein amount to determine the amount of procollagen synthesis per constant protein. Procollagen and protein amounts were calculated according to the calibration curve.

As a result, the procollagen synthesis increased by 195.23% when 10 ng / ml of TGF-β was used as a positive control. On the other hand, BLS-PGA increased to 129.34%, 168.74%, and 199.60%, respectively, when treated with 0.01%, 0.05%, and 0.10% concentrations with 0.10% as the highest concentration. This increased the procollagen synthesis capacity in a concentration-dependent manner, and the three concentrations showed statistically significant results. Accordingly, gamma oligopeptides are thought to show efficacy in wrinkle improvement by increasing procollagen synthesis ability in a concentration-dependent manner at 0.01%, 0.05%, and 0.10%.

Hereinafter, the formulation example for the composition of this invention is illustrated. However, formulations comprising the compositions of the present invention are not limited in any way.

Formulation Example 1 Flexible Lotion (Skin)

Gamma Oligopeptide 5.00 (%)

1,3-butylene glycol 1.00 (%)

Disodium ID 0.05 (%)

Allantoin 0.10 (%)

Dipotassium glycylizate 0.05 (%)

Citrix Acid 0.01 (%)

Sodium Citrate 0.02 (%)

Glyceres-26 1.00 (%)

Arbutin 2.00 (%)

Hydrogenated Castor Oil 1.00 (%)

Ethanol 30.00 (%)

Preservative Trace

Trace amount of colorant

Trace amount of flavor

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 (%)

Botanical Extracts 3.2 (%)

Magnesium Aluminum Silicate 0.3 (%)

PEG-40 Stearate 1.2 (%)

Stearic Acid 2.0 (%)

Polysorbate 60 1.5 (%)

Lipid Glyceryl Stearate 2.0 (%)

Sorbitan Sesquioleate 1.5 (%)

Cetearyl Alcohol 3.0 (%)

Mineral oil 4.0 (%)

Squalane 3.8 (%)

Carlylic / Capric Triglycerides 2.8 (%)

Vegetable Oil 1.8 (%)

Dimethicone 0.4 (%)

Dipotassium glycylizate traces

Allantoin Trace

Sodium Hyaluronate Trace

Tocopheryl acetate appropriate amount

Triethanolamine appropriate amount

Preservative

Flavoring Agent

Purified water level

As described above in detail specific parts of the present invention, it will be apparent to those skilled in the art that these specific descriptions are merely preferred embodiments, and thus the scope of the present invention is not limited thereto. will be. Thus, the substantial scope of the present invention will be defined by the appended claims and their equivalents.

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, which is useful for improving skin wrinkles.

Claims (7)

1. A cosmetic composition for improving skin wrinkles comprising gamma oligopeptide or a salt thereof as an active ingredient.
2. The cosmetic composition for improving skin wrinkles according to claim 1, wherein the gamma oligopeptide or a salt thereof is prepared by lowering polygamma glutamic acid.
3. The cosmetic composition for improving skin wrinkles according to claim 1, wherein the gamma oligopeptide or its salt has a molecular weight of 600 to 5,000 Da.
4. The cosmetic composition for improving skin wrinkles according to claim 1, wherein the gamma oligopeptide is a peptide consisting of 5 to 15 glutamic acids.
5. The cosmetic composition for improving skin wrinkles according to claim 1, wherein the salt is selected from the group consisting of sodium salt, potassium salt, calcium salt, magnesium salt, ammonium salt and zinc salt.
6. The cosmetic composition for improving skin wrinkles according to claim 1, wherein the total content of the gamma oligopeptide or its salt is 0.001 to 10% by weight based on the total weight of the cosmetic composition.
7. According to claim 1, wherein the cosmetic composition is a skin wrinkle improvement, characterized in that any one selected from the group consisting of cosmetics, essences, skins, lotions, creams, packs, foundations and makeup bases Cosmetic composition.

Images