KR20170049158A

KR20170049158A - Cosmetic composition for improving skin wrinkle comprising octapeptide

Info

Publication number: KR20170049158A
Application number: KR1020150150185A
Authority: KR
Inventors: 류남현; 김혜연; 이영수; 정제교; 김수아
Original assignee: 우리들제약(주)
Priority date: 2015-10-28
Filing date: 2015-10-28
Publication date: 2017-05-10

Abstract

The present invention relates to a cosmetic composition having octapeptide as a main component and having excellent skin wrinkle-reducing effect. The present invention relates to a cosmetic composition which is excellent in skin penetration and skin safety, has an effect of suppressing the formation of collagenase in the skin and improving the collagen biosynthesis Provides excellent cosmetics. In particular, the present invention relates to a cosmetic composition containing the octapeptide as a main component and having various properties such as liquid cosmetic lotion, emulsion, cream, essence, serum, pack, cleanser, ultraviolet screening cream, And can be applied to cosmetics.

Description

TECHNICAL FIELD The present invention relates to a cosmetic composition for improving skin wrinkles comprising an octapeptide as a main component,

Skin aging is affected by genetic, environmental exposure, hormonal changes, and metabolic processes. These factors lead to accumulated changes in the structure, function, and appearance of the skin. Human skin aging is largely classified into intrinsic aging that develops over time according to biological processes and external aging (photoaging) due to environmental conditions such as continuous light exposure, stress, and smoking. Clinically, the inner aged skin is characterized by thin, dry, pale, non-elastic and many fine wrinkles. On the other hand, skin aged by the external environment has deep wrinkles, spot coloration and coarse skin, It will have a greater impact. Although aged skin and photosensitized skin are morphologically different from each other in appearance, recent studies have shown that both aging promotes expression of collagenase, decreased synthesis of procollagen, and damage to connective tissues It is reported that they share important molecular features including signaling pathways. Environmental influences give more aesthetic qualities to skin and more obvious variations in tissue than internal effects, the most lethal external factor being UV radiation. Consistent with this molecular mechanism is believed to be that UV irradiation accelerates important aspects of aging due to the chronological age of human skin.

From a histological point of view, skin aging is caused by changes in the composition of extracellular matrix proteins that form the matrix in the dermis of the skin. Collagen proteins in the dermal extracellular tissues impart strength and tension to the skin, which protects the skin from external stimuli or forces. It accounts for 90% of the dermal layer. Collagen reduction is closely related to skin aging and wrinkle formation . The major cross-linking components of healthy dermis are type I collagen and type III collagen. However, in skin damaged by photoaging, the precursors of these two collagens are markedly reduced, and their reduction is associated with clinical severity. Collagen degradation may degrade as it ages, but when exposed to stimuli such as ultraviolet light, the biosynthesis of collagenase (MMPs) increases and collagen synthesis is reduced and wrinkles are formed. Collagen degrading enzymes (MMPs) are subtype-dependent intracellular proteases that are capable of degrading or restructuring the extracellular components of the dermis, among which MMP-1 acts on the collagen of the dermis . Therefore, regulation of the amount and activity of this enzyme is important for wrinkles.

The main ingredients for making wrinkle-improving functional cosmetics were retinol (2,500 IU / g), retinyl palmitate (10,000 IU / g), adenosine (0.04%), polyethoxylated retinamide (0.05-0.2 %). These products are also exempted from safety and efficacy screening data when they are used at the indicated concentrations. In addition, among the products approved as functional cosmetics by individual certification, hydroxyproproline, 7-dihydrocholesterol, carnitine, And bingo extract.

The wrinkle-reducing functional material is mainly composed of a component that regulates the differentiation and regeneration of epidermal cells, a substance that regulates an extracellular matrix (ECM), an antioxidant that cleans active oxygen, an anti-inflammatory component, a component that protects against damage by ultraviolet rays, The mainstream of similar Botox-like peptides that inhibit muscle movement is the mainstream. In addition to conventional retinoids and AHA, mevalonic acid has been developed and used as a regenerative substance for epidermal cell differentiation and regeneration. As ECM regulating component, paoniflorin extracted from peony or prangenidin extracted from white paper has been developed and used . In the case of prangenidin, PCIP enzyme immunoassay showed that the collagen biosynthesis was about 20 times better than vitamin C. On the other hand, a component that protects or regenerates DNA that is susceptible to damage by ultraviolet rays has been developed. A representative example is photolyase, which is extracted from phytoplankton and is known as an enzyme of DNA damage-healing function. In particular, retinol and retinoid derivatives are known to exhibit good effects in various fields such as psoriasis, aging, cancer, and acne. The retinoid is also known to inhibit collagenase (MMP-1) when it is applied to the skin to prevent collagen loss and to stimulate collagen formation, thereby preventing and restoring endogenous and photoaging. Despite these effects, however, it is known that when applied to the skin, it causes local skin irritation such as erythema, itching, psoriasis, and scaling. Therefore, it is required to develop a new wrinkle-reducing substance which is safe to the living body, inhibits collagenase (MMP-1) and has good collagen synthesis ability.

Accordingly, the inventors of the present invention have made efforts to develop a dosage form containing octapeptide as a main ingredient capable of penetrating the skin and to confirm the effect on skin safety and wrinkle improvement. As a result, they have successfully produced octapeptide formulations and inhibited collagenase, Thereby improving the skin wrinkles, thereby completing the present invention.

KR 10-1355385 B

Accordingly, it is a technical object of the present invention to provide a cosmetic composition for improving wrinkles containing octapeptide as a main component which is safe for skin and permeable to the skin.

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

In the present invention, the octapeptide refers to a protein fragment or molecule composed of 8 amino acids forming a polypeptide chain. Preferably, the octapeptide has the following sequence: (a) a cosmetic composition for improving skin wrinkles,

DWYGFGDW-NH2

Here, D represents Asp, W represents Trp, Y represents Tyr, G represents Gly, and F represents Phe.

In the present invention, the octapeptide is contained in an amount of 0.001 to 0.1% by weight based on the total weight of the cosmetic composition.

In the present invention, the composition may further comprise ingredients such as water, oil, surfactant, moisturizing agent, thickening agent, lower alcohol, coloring agent, preservative, perfume, etc., A cosmetic composition for improving wrinkles is provided.

In the present invention, the formulation of the cosmetic composition may be in the form of a lotion, a gel, a water-soluble liquid, an emulsion, an essence, a serum, an ampoule, a cream, a pack, a cleanser, A basic cosmetic formulation comprising a water (W / O) type; A color cosmetic formulation consisting of a makeup base, a foundation, a skin cover, a lipstick, a lip gloss, a lip balm, a face powder, a two-way cake, an eye shadow, a teak collar and an eyebrow pencil; Hair cosmetic formulation consisting of hair shampoo, hair rinse, hair pack, hair treatment, scalp nutrition ampoule, hair essence, hair tonic, setting gel, setting spray, setting mousse, setting wax; Body cosmetic formulations comprising body cleansers, body lotions, body creams, body essences, and bar diesels; The present invention provides a cosmetic composition for improving skin wrinkles.

The present inventors tried to confirm the wrinkle-reducing effect by receiving octapeptide from "HYBRINE HYBRINE CO., LTD." Which jointly conducted the technical innovation task of the Small and Medium Business Administration. The octapeptide has the following sequence.

DWYGFGDW-NH2

Specifically, the present inventors established an in vitro assay system using a human fibroblast cell line and a UVA / UVB lamp to examine the ability of collagen and collagenase to synthesize biosynthesis in order to confirm the wrinkle-reducing effect of octapeptide , 20 healthy subjects were recruited and the wrinkle improvement effect was quantified by measuring the total wrinkle number, wrinkle length and wrinkle area through the replica analysis method. The evaluation was made through image analysis, and the present invention was completed.

According to the present invention, the octapeptide is excellent in skin safety and skin penetration, and is excellent in the effect of suppressing the formation of collagenase in skin and improving wrinkles due to the effect of promoting collagen biosynthesis. Accordingly, the cosmetics containing the above-mentioned octapeptide as a main component and containing cosmetics such as liquid lotion, emulsion, cream, essence, serum, pack, cleanser, ultraviolet screening cream and bibby cream, color cosmetics, hair cosmetics, It can be applied to various cosmetics.

1 is a graph showing the toxicity of octapeptide according to the present invention to HDF-N cells.
FIG. 2 is a graph showing the evaluation of the wrinkle-reducing effect using the Elastase inhibition test (* P <0.05, ** P <0.01, *** P <0.005)
FIG. 3 is a graph showing the wrinkle-reducing effect using the Procollagen combination performance test (* P <0.05, ** P <0.01, *** P <0.005)
FIG. 4 is a graph showing the evaluation of the wrinkle-reducing effect using the MMP-1 inhibitory activity test (* P <0.05, ** P <0.01, *** P <0.005)
FIG. 5 is a photograph showing the skin permeability of the octapeptide according to the present invention. FIG.
FIG. 6 is a graph showing changes in visual evaluation of eye wrinkles at a time point.
7 is a photograph showing changes in the degree of eye wrinkles of a subject according to the use of the product according to the present invention.
8 is a graph showing changes in eye wrinkles (replicas) at a time point.
FIG. 9 is a photograph showing the change of the eye wrinkle replica of the subject according to the use of the product according to the present invention.

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

&Lt; Example 1 > Evaluation of cytotoxicity of octapeptide

(1) Cell line and culture

The cell lines used in this test were normal human primary dermal fibroblasts-Neonatal (HDF-N) derived from human neonatal foreskin and were inoculated on the bottom of the culture dish and then treated with 0.1% insulin, 0.1% rhFGF, 0.1% gentamicin, Fibroblast Basal Medium (FBM, lonza, CC-3131) medium was added and incubated at 37 ° C in an incubator containing 5% carbon dioxide.

(2) Of the test solution pharmacy

The test solution used in this test was provided as a powder with octapeptide provided by Hewlett-Packard, Inc. It was dissolved in DMSO and prepared at 1000x and diluted to the appropriate concentration in cell culture medium. In addition, preliminary experiments were carried out using the WST-1 assay, and the concentrations that did not show cytotoxicity were selected. The test substance was diluted with the cell culture medium. After preliminary testing, three concentrations of the test substance were determined and the skin-related efficacy of the test solution was evaluated. Positive controls were also diluted with cell culture medium.

(3) Cytotoxicity test

To confirm the toxicity of the test solution, cytotoxicity in HDF-N cells was measured. HDF-N cells were seeded at 6 × 10 ³ cells / well in a 96-well plate and cultured for 24 hours under cell culture conditions. After incubation, starvation was maintained for 12 hours, and the test solution and fresh medium (except media supplement) were added and cultured for 24 hours. After 24 hours, the WST-1 reaction solution was diluted 1/10 in the supplemented medium to measure the cell survival rate. The cells were treated for 100 hours in each well, and the absorbance was measured at 450 nm .

The in vitro cytotoxicity of octapeptide was measured by WST-1 reaction solution, which is dependent on the mitochondrial activity of the cell and can determine cell survival. Cell viability was assessed in HDF-N cells and expressed as a percentage of control (control) except for the sample. As a result, as shown in FIG. 1, the sample octapeptide exhibited cell viability of 80% or more at a concentration of 1 mM or less in HDF-N cells. Therefore, the highest concentration of the sample used in the subsequent test using HDF-N cells was selected to be 1 mM.

[Table 1: Toxicity of octapeptide to HDF-N cells (n = 4)]

< Example 2> Octapeptide Evaluation of wrinkle-improving efficacy

(One) Elastase inhibition assay

In this test, skin fibroblast HDF-N cell was used as an enzyme elastase to measure the elastase activity of the sample. To this end, 0.2 M Tris-HCl (pH 8.0) containing 0.1% triton X-100 was added to the cultured HDF-N cells, and the resultant was dissolved in an ultrasonic mill and centrifuged at 3,000 rpm for 20 minutes to obtain supernatant , And the activity of enzyme was measured by total protein amount.

To measure the elastase activity of the sample, 200 μl of homogenized fibroblast elastase, 0.2 M Tris-HCl buffer and sample were added in concentration, and 50 ml STANA specific for elastase was added and cultured at 37 Absorbance was measured at 405 nm. The efficacy was calculated for inhibition of elastase activity versus control without test substance. Phosphoramidon was used as a positive control.

Octapeptide on the activity of elastase. For this purpose, HDF-N cells were used as an enzyme in the skin fibroblast, and each sample was treated with 5 uM, 10 uM, 50 uM, and 100 uM of the cytotoxicity-free concentrations and then the activity was measured and compared with the untreated control Respectively.

[Table 2: Evaluation of wrinkle-improving efficacy (Elastase inhibition test) (n = 3)]

As a result, as shown in FIG. 2, the activity of the octapeptide was 103.9%, 82.3%, 21.6% and 16.3%, respectively, as compared with the control group, and all of the test concentrations inhibited the activity of the elastase enzyme in a concentration-dependent manner .

On the other hand, the phosphoramidon used as the positive control group showed a statistically significant difference at 10 uM when the ELASase activity was 51.2% as compared with the control group.

(2) Procollagen increased test

HDF-N cells were plated in 48 well plates at 1 × 10 ⁴ cells / well and cultured for 24 h in cell culture conditions. Thereafter, the medium was removed, and the starvation state of the cells was maintained for 24 hours. After that, the test substance was diluted in the cell culture medium, FBM (without media supplement) Then, the culture medium was harvested and procollagen type I c-peptide (PIP) EIA kit (TAKARA, MK101) was used to measure the amount of procollagen. Cells attached to the bottom were washed with PBS, lysed with 1N NaOH, and the total amount of protein was measured to calculate the amount of procollagen synthesis per protein.

In order to investigate the procollagen aggregation performance of the samples, the toxicity was determined by the previously conducted cytotoxicity test, and the amount of procollagen was calibrated by the total protein amount.

[Table 3: Evaluation of wrinkle-improving efficacy (n = 4) using Procollagen combination performance test]

As a result, as shown in FIG. 3, treatment of TGF-β 10 ng / ml used as a positive control group resulted in 169.5% procollagen amount compared to the control group, and thus HDF-N cells promoted procollagen biosynthesis by TGF-β Could know.

On the other hand, when the sample octapeptide was treated with 0.1, 0.5, and 1 mM, respectively, procollagen production performance was evaluated. As a result, octapeptide produced 107.7%, 128.2%, and 135.9% of procollagen, respectively. This was increased in a concentration-dependent manner, especially at 0.5 mM and 1 mM concentrations, respectively.

(3) Ultraviolet MMP -1 expression inhibition test

The effect of the test substance on the expression of MMP-1 by ultraviolet light was evaluated through this test. HDF-N cells were seeded at 2 × 10 ⁴ cells / well in a 24-well plate and cultured for 24 hours under cell culture conditions. After 24 hours, the medium was discarded and washed with DPBS, followed by addition of DPBS 200 and UVA 5J / irradiation. The sample was diluted to the appropriate concentration in the medium, treated with the cells, and cultured for 24 hours under cell culture conditions. After that, the amount of MMP-1 was measured by using a human total MMP-1 ELISA kit (R & D system, DY901), and the measured amount of MMP-1 was corrected to the total protein amount.

As a result, the amount of MMP-1 was 147.3% as compared with that of the non-irradiated group, and it was found that HDF-N cells induced MMP-1 expression by UVA 5J /.

On the other hand, HDF-N cells were treated with 1 μM, 5 μM, and 10 μM of sample octapeptide and then measured with a human total MMP-1 ELISA kit (R & D system, DY901).

As a result, the amounts of octapeptide produced 141.5%, 124.4%, and 111.2% of MMP-1, respectively, as compared with the UV-untreated group, which showed a tendency to decrease in a concentration-dependent manner. Especially, at 5uM and 10uM concentration, statistically significant difference was shown.

On the other hand, retinoic acid, a positive control used in this test, showed 107.8% of MMP-1 production as compared with UV-untreated group, and it was statistically decreased compared to UV irradiation group. (Fig. 4 and Table 4)

[Table 4: Wrinkle-improving efficacy evaluation using MMP-1 inhibitory test (n = 4)]

<Example 3> Skin penetration test of octapeptide

FIG. 5 shows the result of photographing the octapeptide with an in vivo imaging system (Maestro 2) after applying an octapeptide having a fluorescent substance (FITC) attached to the skin of a pig to measure skin permeability.

Fig. 5 shows the results of comparing immediately after applying octapeptide and after 40 minutes. The distribution of the luminescent portion after 40 minutes was widened as compared with immediately after the application of the octapeptide, and it was confirmed that the octapeptide penetrated.

Examples of formulations for the cosmetic composition of the present invention are illustrated below. As an example of the formulation, an essence was prepared to evaluate the effect of skin irritation and wrinkle improvement in clinical practice. However, the formulation containing the composition of the present invention is not limited to the following formulation examples.

Ingredients Table 1: Peptide Repair Skin Essence

< Example 4> Formulation example Human skin primary stimulation test

Twenty of 20 women between the ages of 50 and 50 were injected 20 times into the IQ chamber of the formulation prepared with different octapeptide contents and added to the test site. After 24 hours, the test area was marked with a skin marker. After 30 minutes and 24 hours, the skin reaction of the test area was observed under the magnifying glass.

[Table 5] Skin Response Results after Removal of Patches by Formulation (n = 32)]

* Mean: 24 hour response + 48 hour response

As shown in Table 5, the 1 + grade skin reaction was observed in 1 patient in the control group and in the preparation example 1 24 hours after removal of the patch. None of the other three species showed any skin reaction.

Example 5 Effect of Formulation Example 3 (test group) on wrinkles on human skin

Twenty women in their 30s and 60s were randomly selected to use the control and formulation 3 (test group) products continuously for 12 weeks, twice a day, morning and evening, respectively. Before and after 4 weeks, 8 weeks, and 12 weeks after the use, the wrinkles of the right and left eyes of the subjects were visually evaluated and evaluated. The visual evaluation was evaluated by the two researchers according to the visual evaluation criteria (Guideline for evaluating the efficacy of the functional food cosmetics and the Food and Drug Administration, 2005.07), and the results were shown in FIGS. 6 and 7 Was significantly reduced (improved) in Formulation Example 3 (test group) compared to the control group at the end of 12 weeks.

For the evaluation of the device, transparancy profilometry analysis was carried out on the skin patch (replica) produced by the control and preparation example 3 (test group) using a visiometer (SV600, Courage + Khazaka electronic GmbH, Germany) (R1: skin roughness, R2: maximum roughness, R3: average roughness, R4: smoothness depth, R5: arithmetic average roughness) were analyzed for the wrinkle parameters R1, R2, R3, R4 and R5. The results are shown in Figs. 8 and 9.

Compared with before use, formulation 3 (test group) showed statistically significant decrease (improvement) in all parameters at 4 weeks, 8 weeks, and 12 weeks after use.

While the foregoing embodiments of the present invention have been described in detail with reference to the preferred embodiments thereof, it will be understood by those skilled in the art that various changes and modifications may be made without departing from the spirit and scope of the invention as defined in the appended claims. It is natural that the claims fall within the scope of the claims.

INDUSTRIAL APPLICABILITY As described above, the octapeptide according to the present invention is excellent in skin penetration and skin safety, and is excellent in the effect of suppressing the formation of collagenase in skin and improving wrinkles due to the effect of promoting collagen biosynthesis. Accordingly, the cosmetics containing the above-mentioned octapeptide as a main component and containing cosmetics such as liquid lotion, emulsion, cream, essence, serum, pack, cleanser, ultraviolet screening cream and bibby cream, color cosmetics, hair cosmetics, It can be applied to various cosmetics.

Claims

A cosmetic composition for improving skin wrinkles containing octapeptide as an active ingredient.

The cosmetic composition for improving skin wrinkles according to claim 1, wherein the octapeptide has the following sequence:
DWYGFGDW-NH2
Here, D represents Asp, W represents Trp, Y represents Tyr, G represents Gly, and F represents Phe.

The cosmetic composition for improving skin wrinkles according to claim 1, wherein the octapeptide is contained in an amount of 0.001 to 0.1% by weight based on the total weight of the cosmetic composition.

The composition of claim 1, wherein the composition further comprises ingredients such as water, oil, surfactant, moisturizer, thickeners, lower alcohols, pigments, preservatives, perfumes, A cosmetic composition for improving skin wrinkles.

The cosmetic composition according to claim 1, wherein the cosmetic composition is at least one selected from the group consisting of lotion, gel, aqueous liquid, emulsion, essence, serum, ampoule, cream, pack, cleanser, ultraviolet screening cream, A basic cosmetic formulation comprising a w / o type; A color cosmetic formulation consisting of a makeup base, a foundation, a skin cover, a lipstick, a lip gloss, a lip balm, a face powder, a two-way cake, an eye shadow, a teak collar and an eyebrow pencil; Hair cosmetic formulation consisting of hair shampoo, hair rinse, hair pack, hair treatment, scalp nutrition ampoule, hair essence, hair tonic, setting gel, setting spray, setting mousse, setting wax; Body cosmetic formulations comprising body cleansers, body lotions, body creams, body essences, and bar diesels; Wherein the cosmetic composition is a cosmetic composition for improving skin wrinkles.