KR102051998B1 - Cosmetic composition for anti-wrinkle and anti-aging comprising protease-activated recepter2(par2) agonist - Google Patents

Abstract

The present invention relates to a wrinkle improvement, anti-aging functional cosmetic composition comprising a PAR2 receptor agonist having a type I collagen synthesis inducing function. The cosmetic composition of the present invention can improve skin wrinkles and maintain skin elasticity by selecting the PAR2 agent having the best type I collagen synthesis effect among the known PAR2 receptor agonists. Can be used.

Description

Cosmetic composition for skin wrinkle improvement or anti-aging comprising an agent of protease activating receptor (PAR2) {COSMETIC COMPOSITION FOR ANTI-WRINKLE AND ANTI-AGING COMPRISING PROTEASE-ACTIVATED RECEPTER2 (PAR2) AGONIST}

The present invention relates to a cosmetic composition for skin wrinkle improvement or anti-aging comprising an agent of a protease activating receptor (PAR2) having a type I collagen synthesis inducing function.

Collagen is a major matrix protein constituting skin and bone. It is present in extracellular epilepsy, accounts for 30% of the total weight of biological proteins, has a solid triple helix structure, and a total of 28 are known. It is known to induce mechanical firmness of skin, resistance of connective and connective tissues, support of cell adhesion, cell division and differentiation.

In the case of skin, in particular, 80% of type I collagen and 15% of type II collagen are reported, and the proportion of type I collagen decreases relatively as aging progresses (Brain Behav Immun. 2009 Nov; 23 (8): 1089-95). As the skin ages, Type I collagen decreases, resulting in a relatively different type of collagen, resulting in thickening and loss of elasticity. It is also known that the amount of type I collagen in the skin decreases due to stress and smoking. Therefore, in order to slow down the aging of the skin, it is necessary to develop a technology that can selectively induce or preserve the synthesis of type I collagen. In the wound healing process, if collagen metabolism is activated by promoting the synthesis of collagen in the skin, the dermal matrix component is increased, which is very important for rapid healing. In addition, expression of various related genes such as Smad2 and TGF-β is also known to be involved in collagen synthesis (Arch Dermatol Res. 2008 Apr; 300 Suppl 1: S57-64). If the expression of these genes can be raised, the synthesis of collagen in the tissue can be accelerated. Synthesis of type I collagen in tissues is also known to occur via PAR2, a protease activating receptor (Hypertension 2011 Aug; 58: 264-270). In this pathway, the activation of the PAR-2 receptor by the tryptase activates the MEK 1/2 pathway, which in turn activates the ERK 1/2 pathway, leading to the formation of procollagen (procollagen I & III). It is known that the synthesis of type I collagen is regulated through this pathway.

Cosmetics and medicines to improve skin wrinkles have been continuously developed so far, and recently, cosmetics such as sunscreens, moisturizers and antioxidants are widely used to prevent wrinkles caused by external factors. The effect of substantially improving wrinkles is still marginal. Therefore, substances that promote collagen synthesis have been developed as skin wrinkle improving substances. Retinoic acid, Retinol (vitamin A) and vitamin C are typical examples. In addition to the retinoic acid, retinol, betulinic acid (betulinic acid; US Patent Publication US 5,529,769), Foskolin (Hakeda, Y. et al, J. Biochem, 101, 1163-14699 (1987)), Betoleti ( Bertholletia Extract (US Pat. No. 6,004,568), Landing (Radix Phytolacca esculenta) or Semen Cuscutae Extract (Korean Patent Publication No. 2000-13588), Agastache rugosa, Lagenaria siceria var.clavata Ser And Aquilaria agallocha Roxb extract (Korean Patent Publication No. 2000-184670) and the like are those developed as antiwrinkle cosmetics that promote collagen synthesis.

However, the development of such raw materials is time-consuming and expensive, which may impair product productivity in the field of wrinkle relief cosmetics or medical product development.

The present invention has been made to overcome the limitations of the prior art as described above, the problem to be solved in the present invention is to find a PAR2 agonist excellent in collagen synthesis inducing ability of the known PAR2 receptor agonist (agonist) and using it By increasing the synthesis of type I collagen in the skin tissue, the company aims to secure leading technologies by creating functional products that help to improve wrinkles and prevent skin aging.

Accordingly, an object of the present invention to provide a functional cosmetic composition for skin wrinkle improvement or anti-aging comprising the peptide of SEQ ID NO: 1.

In order to achieve the above object, the present invention provides a functional cosmetic composition for skin wrinkle improvement or anti-aging comprising a peptide of SEQ ID NO: 1.

The cosmetic composition of the present invention can improve skin wrinkles and maintain skin elasticity by selecting the PAR2 agent having the best type I collagen synthesis effect among the known PAR2 receptor agonists. Can be used.

1 is a structural formula of four candidate groups among known PAR2 receptor agonists. In particular, in the present invention, in order to increase the ease of absorption in the skin, a small molecular weight PAR2 agent was selected as a candidate group.
FIG. 2 shows cytotoxicity experiments of PAR2 receptor agonist candidates PCIF-1, 2, 3, and 4. FIG.
3 is a graph comparing the expression level of the type I collagen gene at the RNA level after PAR2 receptor agonist candidate PCIF-1, 2, 3 and 4 were treated to human skin fibroblasts by concentration (1 uM, 5 uM, 10 uM). ^* p <0.05; ^** p <0.01; ^*** p <0.001; ^**** ｐ <0.0001; Significant value was measured by One-way ANOVA test).
4 to 6 shows the expression level of the TGF-βI gene (FIG. 4), Smad2 (FIG. 5) and Smad3 (FIG. 6) after PCIF-2 was treated to human skin fibroblasts at different concentrations (1 uM, 5 uM, and 10 uM). A graph comparing at the RNA level ( ^* p <0.05; ^** p <0.01; ^*** p <0.001; ^**** ｐ <0.0001; Significant value was determined by One-way ANOVA test).
7 is a diagram showing the results of confirming the expression level of type I collagen by Western blot after PCIF-2 was treated with human fibroblasts (Human Dermal Fibroblast, HDFn) 1, 5 and 10 uM, respectively.

Hereinafter, the present invention will be described in detail.

In the present invention, the term "skin wrinkle" refers to wrinkle formation on skin.

In addition, the term "skin aging" refers to aging of the skin caused by intrinsic and external factors, preferably may be skin photoaging accompanied by the formation of skin wrinkles, more preferably skin wrinkles Skin photoaging by ultraviolet stimulation accompanied by formation.

The present invention provides a functional cosmetic composition for improving skin wrinkles or preventing aging comprising the peptide of SEQ ID NO: 1.

The inventors have found that some of the known PAR2 agonists induce collagen synthesis and completed the present invention. Specifically, among several known PAR2 agonists, a substance capable of activating a signaling pathway associated with type I collagen synthesis was selected, and among them, a small molecular weight PAR2 agonist easily absorbed into the skin was selected as a candidate. In one embodiment of the present invention by treating a number of candidate substances to human skin fibroblasts to confirm the expression of type I collagen, as a result, the peptide PCIF-2 of SEQ ID NO: 1 (SLIGKV-NH ₂ ) is the best type I collagen The present invention was completed by confirming that it has the expression ability.

Therefore, the functional cosmetic composition for skin wrinkle improvement or anti-aging of the present invention may be preferably characterized in that it comprises a peptide of SEQ ID NO: 1.

In the present invention, the peptide may be composed of the amino acid sequence of SEQ ID NO: 1 or may include the amino acid sequence as part. In addition, the peptide may comprise a functional equivalent of the peptide comprising the amino acid sequence. The term "functional equivalent" means at least 40%, preferably at least 80%, more preferably at least 90%, more preferably at least 95% sequence with the amino acid sequence as a result of the addition, substitution or deletion of an amino acid. By homology, it means a protein having substantially the same biological activity as the amino acid sequence.

The term "substantially homogeneous physiological activity" refers to skin condition improving activity, such as anti-wrinkle or anti-aging due to the structural and functional homology of the peptide. The sequence homology is determined relative to the two optimally arranged sequences and the comparison region, and some nucleic acid sequences in the comparison region may be added or deleted.

The content of the peptide included in the cosmetic composition of the present invention is not particularly limited as long as it does not exhibit cytotoxicity and exhibits sufficient PAR-2 activity, but may be contained in 0.00006 to 5.0% by weight relative to the total weight of the composition. And preferably 0.0006 to 1.0% by weight, more preferably 0.0001 to 0.001% by weight, and most preferably 0.00015 to 0.0003% by weight. If the total content of the peptide is less than 0.00006% by weight it is difficult to expect a substantial wrinkle improvement effect, if it exceeds 5.0% by weight may cause irritation to the skin or affect the stabilization of the formulation.

In addition, the peptide contained in the cosmetic composition may be characterized in that the concentration of 1uM to 10uM, more preferably may be a concentration of 3uM to 8uM, most preferably may be a concentration of 4uM to 6uM. In one embodiment of the invention, the peptide may be a concentration of 1uM to 10uM.

The cosmetic composition of the present invention may further include, as an active ingredient, a known substance having a skin wrinkle improvement or an anti-aging effect, without stimulating the skin, in addition to the above-described peptide.

In addition, the cosmetic composition of the present invention may include, in addition to the peptides described above, conventional adjuvants and carriers such as antioxidants, stabilizers, solubilizers, vitamins, pigments and flavors, and may be formulated with the carrier.

Cosmetic compositions of the present invention may be prepared in any formulation conventionally prepared in the art and include, for example, solutions, suspensions, emulsions, pastes, gels, creams, lotions, powders, soaps, surfactant-containing cleansing , Oils, powder foundations, emulsion foundations, wax foundations and sprays, and the like, but are not limited thereto. More specifically, flexible lotion (skin lotion), nourishing lotion (milk lotion), nourishing cream, massage cream, essence, eye cream, hair tonic, shampoo, rinse, cleansing cream, cleansing foam, cleansing water, pack, spray and It may be prepared in the form of a powder.

When the formulation of the present invention is a paste, cream or gel, animal oils, vegetable oils, waxes, paraffins, starches, trachants, cellulose derivatives, polyethylene glycols, silicones, bentonites, silicas, talc or zinc oxide may be used as carrier components. Can be.

When the formulation of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used, in particular in the case of a spray, additionally chlorofluorohydrocarbon, propane Propellant such as butane or dimethyl ether.

When the formulation of the present invention is a solution or emulsion, a solvent, solubilizer or emulsifier is used as the carrier component, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1 Fatty acid esters of, 3-butylene glycol, glycerol aliphatic ester, polyethylene glycol or sorbitan.

When the formulation of the present invention is a suspension, liquid carrier diluents such as water, ethanol or propylene glycol, suspending agents such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol esters and polyoxyethylene sorbitan esters, and microcrystals are used as carrier components. Soluble cellulose, aluminum metahydroxy, bentonite, agar or tracant and the like can be used.

When the formulation of the present invention is a surfactant-containing cleansing, the carrier component is aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyltaurate, sarcosinate, fatty acid amide. Ether sulfates, alkylamidobetaines, aliphatic alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, lanolin derivatives or ethoxylated glycerol fatty acid esters and the like can be used.

Hereinafter, the present invention will be described in detail by way of examples. However, the following examples are merely to illustrate the present invention is not limited to the contents of the present invention. The embodiments of the present invention are provided to more completely explain the present invention to those skilled in the art.

Example  1. Candidate agonist ( agonist ) Set

Among the known PAR2 receptor agonists, substances were determined to be related to type I collagen synthesis first, and among them, a small molecular weight agent that was easily absorbed into the skin was selected as a candidate group. Selected candidate agents are shown in FIG. 1.

Example  2. Securing the safety of candidate agents

In order to ensure the safety of the candidate PAR2 receptor agonist PCIF-1, 2, 3, 4 prepared in Example 2, after treatment with cultured human dermal fibroblasts PCIF-1, 2, 3, 4 and measuring the cell survival rate It was. Specifically, each agent candidate was treated with 100 nM, 500 nM and 1 mM for 24 hours and cytotoxicity was confirmed using MTT reagent. Positive control was treated with DMSO. The results are shown in FIG.

As shown in FIG. 2, the majority of cells were killed in the positive control group (DMSO), while the cells treated with PCIF-1, 2, 3, and 4 showed similar survival of the cells regardless of the concentration. It was.

Example  3. Increased Type I Collagen Expression in Human Dermal Fibroblasts by Candidate Agent Treatment

PCIF-1, 2, 3, and 4 were treated on human skin fibroblasts to purify total RNA through RNA separation, and then synthesized cDNA using type I collagen (collagen I, col I) and TGF. The degree of expression of -βI, Smad2 and Smad3 genes was compared with the untreated group. The results are shown in FIG.

As shown in FIG. 3, it was confirmed that the type I collagen gene was highestly expressed when PCIF-2 was treated among four candidate agents. In particular, when PCIF-2 was treated at a concentration of 5 μM, it was confirmed that the expression of type I collagen gene was significantly increased. Thus, the following experiment was conducted using the PCIF-2.

Example  4. PCIF Of Gene Expression Related to Type I Collagen Synthesis in Human Dermal Fibroblasts

PCIF-2, which confirmed the highest type I collagen synthesis ability in Example 3, was treated to human skin fibroblasts by concentration (1 uM, 5 uM, 10 uM) to confirm the type I collagen synthesis-related gene expression. The results are shown in FIGS. 4 to 6.

As shown in Figures 4 to 6, it was confirmed that the expression of TGF-βI, Smad2 and Smad3 genes related to type I collagen synthesis were also increased during PCIF-2 treatment. In particular, it was confirmed that the expression of TGF-βI, Smad2 and Smad3 genes was significantly increased at 5 uM concentration.

In addition, the PCIF-2 was treated with human fibroblasts (Human Dermal Fibroblast, HDFn) by concentration (1, 5, 10 uM) and subjected to Western blot, and the results are shown in FIG. 7. As shown in Figure 7, it was confirmed that the amount of type I collagen protein significantly increased at a concentration of 5 uM.

Through the above results, it was confirmed that the type I collagen synthesis ability of PCIF-2 is excellent. Therefore, the present invention has been confirmed that by selecting the PAR2 agonist having the best type I collagen synthesis effect among the known PAR2 receptor agonists can effectively improve skin wrinkles and skin elasticity.

Production Example

Functional cosmetic composition for skin wrinkle improvement or anti-aging according to the present invention can be prepared in various forms of cosmetics according to the purpose. The following are some examples of the preparation of functional cosmetics including the peptide of SEQ ID NO: 1 according to the present invention, but the present invention is not limited thereto.

Production Example  1. Preparation of flexible lotion (skin lotion)

A flexible lotion was prepared according to a conventional method as follows.

1.00 parts by weight of peptide of SEQ ID NO: 1

1.00 parts by weight of 1,3-butylene glycol

0.05 parts by weight of disodium ethane

0.05 parts by weight of dipotassium glycylizate

0.01 part by weight of citric acid

Sodium citrate 0.02 parts by weight

Glyceres-26 1.00 parts by weight

Arbutin 2.00 parts by weight

Polysorbate 20 0.50 parts by weight

Ethanol 30.0 parts by weight

Trace amount of colorant

Trace amount of flavor

Purified water level

Production Example  2. Manufacturing of Nutritional Cream

A nutritious cream was prepared according to a conventional method as follows.

1.00 parts by weight of peptide of SEQ ID NO: 1

1,3-butylene glycol 7.00 parts by weight

1.00 parts by weight of glycerin

0.10 part by weight of D-panthenol

0.30 parts by weight of magnesium aluminum silicate

PEG-40 stearate 1.20 parts by weight

2.00 parts by weight of stearic acid

Polysorbate 60 1.50 parts by weight

Lipid glyceryl stearate 2.00 parts by weight

1.50 parts by weight of sorbitan sesquioleate

Cetearyl Alcohol 3.00 parts by weight

4.00 parts by weight of mineral oil

3.80 parts by weight of squalane

Carlylic / capric triglycerides 2.80 parts by weight

1.80 parts by weight of vegetable oil

Dimethicone 0.40 part by weight

Diphosphium glycylizate traces

Allantoin Trace

Sodium Hyaluronate Trace

Tocopheryl acetate appropriate amount

Triethanolamine appropriate amount

Flavoring Agent

Purified water level

So far, the present invention has been described with reference to the preferred embodiments or preparations. Those skilled in the art will appreciate that the present invention can be implemented in a modified form without departing from the essential features of the present invention. Therefore, the disclosed embodiments or preparations should be considered in descriptive sense only and not for purposes of limitation. The scope of the present invention is shown in the claims rather than the foregoing description, and all differences within the scope will be construed as being included in the present invention.

<110> Pusan National University Industry-University Cooperation Foundation <120> COSMETIC COMPOSITION FOR ANTI-WRINKLE AND ANTI-AGING COMPRISING          PROTEASE-ACTIVATED RECEPTER2 (PAR2) AGONIST <130> PNU1-363P-1 <150> KR 10-2017-0160222 <151> 2017-11-28 <160> 1 <170> KoPatentIn 3.0 <210> 1 <211> 6 <212> PRT <213> Artificial Sequence <220> <223> PCIF-2 <400> 1 Ser Leu Ile Gly Lys Val   1 5

Claims (5)

Functional cosmetic composition for improving skin wrinkles comprising a peptide of SEQ ID NO: 1 in a concentration of 3 to 8μM.
delete The composition of claim 1 wherein the cosmetic composition consists of a solution, suspension, emulsion, paste, gel, cream, lotion, powder, soap, surfactant-containing cleansing, oil, powder foundation, emulsion foundation, wax foundation and spray A functional cosmetic composition for improving skin wrinkles, comprising a formulation selected from the group.
The functional cosmetic composition according to claim 1, wherein the skin wrinkles are due to skin aging.
The functional cosmetic composition according to claim 4, wherein the skin aging is skin photoaging due to UV stimulation.

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