KR20220085193A - PGC-1α-derived Peptide Derivative and Cosmetic Composition Comprising the Same - Google Patents

Abstract

The present invention provides a PGC-1α-derived peptide derivative and a cosmetic composition comprising the same. The peptide derivative according to the present invention exhibits increased PGC-1α expression, increased mitochondrial activity, increased collagen proliferation, antioxidant and anti-inflammatory effects, wrinkle improvement and pigmentation improvement effects, and thus can be effectively used in a cosmetic composition for preventing skin aging.

Description

PGC-1α-derived peptide derivative and cosmetic composition comprising the same {PGC-1α-derived Peptide Derivative and Cosmetic Composition Comprising the Same}

The present invention relates to a PGC-1α-derived peptide derivative and a cosmetic composition comprising the same. More specifically, the present invention relates to a peptide derivative that not only exhibits improvement of mitochondrial function through PGC-1α activity, but also has excellent collagen proliferation, antioxidant, and anti-inflammatory effects, and a cosmetic composition comprising the same.

Recently, with the aging of the population, the increase in social stress, and the increase in the amount of UV radiation due to the destruction of the ozone layer due to environmental pollution, the possibility of causing skin aging due to the active oxygen generated by UV light increases. Interest in materials that can be suppressed and social demand for cosmetics containing them are increasing. Accordingly, it is emerging as an important task to develop an anti-aging material that is biocompatible and excellent for maintaining cell homeostasis through intracellular antioxidant and anti-inflammatory activity.

Korean Patent Application Laid-Open No. 2018-0110636 discloses a natural activator conjugate and a cosmetic composition exhibiting free radical scavenging ability, fat autooxidation inhibitory ability, and anti-inflammatory effect, including a natural activator bound to a phytochemical fraction by a linker.

However, there is an urgent need for the development of anti-aging substances that exhibit excellent effects on wrinkle improvement and whitening as well as antioxidant and anti-inflammatory effects.

Republic of Korea Patent Publication No. 2018-0110636

The present inventors demonstrated PGC-1α activity by binding a specific peptide derived from peroxisome proliferator-activated receptor gamma coactivator-1α (PGC-1α) involved in mitochondrial synthesis and gallic acid, one of the phytochemicals. By promoting and improving mitochondrial function, it was discovered that it is possible to improve wrinkle improvement and whitening effects, as well as to increase antioxidant and anti-inflammatory effects, and completed the present invention.

Accordingly, it is an object of the present invention to provide a PGC-1α-derived peptide derivative that has antioxidative and anti-inflammatory, anti-wrinkle and pigmentation-improving effects, as well as excellent biocompatibility and safety without irritation to the skin.

Another object of the present invention is to provide a cosmetic composition for skin aging comprising the peptide derivative.

One embodiment of the present invention relates to a peptide derivative represented by the following formula (1) that can be used as a raw material for functional cosmetics.

[Formula 1]

In the above formula,

n is 1 or 2, preferably 2,

L is absent or a linking group, preferably Lys,

A is a PGC-1α-derived peptide comprising the amino acid residues of Thr-Pro-Pro-Thr-Thr-Pro.

The general rules for naming peptides herein are based on the application of the three-letter or one-letter amino acid code, unless specifically indicated otherwise. In other words, the center of the amino acid structure is denoted by a three-letter code (eg Thr, Lys) or a one-letter code (eg, T, K), and by placing “D-” in front of the three-letter code, D- The conformation (eg D-Thr, D-Lys) is assumed to be the L-conformation unless specifically indicated.

The amino acid residues constituting the peptide may be natural or non-natural amino acid residues.

The peptide derivative according to the present invention can be prepared by synthesizing a desired peptide and then reacting it with gallic acid for an amide bond.

The desired peptide may be obtained by extracting an in vivo protein and treating it with a proteolytic enzyme to reduce the molecular weight, or may be prepared using genetic recombination and a protein expression system, preferably by a chemical synthesis method using a peptide synthesizer, etc. have.

The peptide derivative according to the present invention is, for example,

(1) obtaining NH ₂ -protected peptide-resin by a conventional solid phase peptide synthesis (SPPS);

(2) reacting the obtained NH ₂ -protected peptide-resin with gallic acid; and

(3) It can be prepared including the step of removing the resin.

When a functional group exists in the side chain of an amino acid residue constituting the desired peptide, a peptide can be synthesized using the amino acid in which the functional group is protected in step (1), and the protecting group bound to the functional group can be synthesized in step (3) is removed from

A specific example of a process for preparing a peptide derivative according to the present invention using an amino acid with a protected functional group is briefly shown in Scheme 1 below.

[Scheme 1]

In addition, another embodiment of the preparation process of the peptide derivative according to the present invention using an amino acid with a protected functional group is briefly shown in Scheme 2 below.

[Scheme 2]

One embodiment of the present invention relates to a cosmetic composition comprising the peptide derivative, in particular, a cosmetic composition for skin aging.

The peptide derivative according to the present invention exhibits a wrinkle improvement effect by promoting the production of PGC-1α inducing mitochondrial activity and collagen proliferation. In addition, the peptide derivative according to the present invention not only exhibits antioxidant and anti-inflammatory effects, but also exhibits a skin whitening effect, so that it can be effectively used in a cosmetic composition for preventing skin aging.

The cosmetic composition of the present invention may contain preferably 0.0001 to 10.0% by weight of the peptide derivative according to the present invention as an active ingredient. The content of the active ingredient may be appropriately determined according to the purpose of its use.

The cosmetic composition of the present invention may include components commonly used in cosmetic compositions, for example, conventional adjuvants such as antioxidants, stabilizers, preservatives and fragrances, and a carrier in addition to the peptide derivative according to the present invention as an active ingredient. have.

The cosmetic composition of the present invention may be prepared in any formulation commonly used in the art, for example, it may be formulated as a solution, suspension, emulsion, paste, gel, cream, powder, spray, etc.

When the formulation of the present invention is a paste, cream or gel, animal oil, vegetable oil, wax, paraffin, starch, tracanth, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc, zinc oxide, etc. may be used as a carrier component. can

When the formulation of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate, polyamide powder, etc. may be used as a carrier component. In particular, in the case of a spray, additional chlorofluorohydrocarbon, It may contain a propellant such as propane/butane or dimethyl ether.

When the formulation of the present invention is a solution or emulsion, a solvent, solubilizer or emulsifier as a carrier component, for example, water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, propylene glycol, glycerol fatty acid ester, sorbitan of fatty acid esters and the like may be used.

When the formulation of the present invention is a suspension, as a carrier component, a liquid diluent such as water, ethanol or propylene glycol, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester or polyoxyethylene sorbitan ester, microcrystals Adult cellulose, aluminum metahydroxide, bentonite, agar, tracanth and the like can be used.

The cosmetic composition of the present invention can be applied to cosmetics such as skin, lotion, cream, pack, color cosmetics, sun cream, face powder, compact, lipstick, and eye shadow.

The peptide derivative according to the present invention, in which gallic acid is bound to a PGC-1α-derived peptide, exhibits increased PGC-1α expression, increased mitochondrial activity, increased collagen proliferation, antioxidant and anti-inflammatory effects, wrinkle improvement and pigmentation improvement effects. In addition, the peptide derivative according to the present invention has excellent biocompatibility and no irritation to the skin, and thus has an excellent safety effect.

Therefore, the peptide derivative according to the present invention can be effectively used as a functional cosmetic material in a cosmetic composition, in particular, a cosmetic composition for skin aging.

1 is a graph showing the evaluation results of mitochondrial activity (mitochondrial membrane potential) of a peptide derivative according to the present invention.
2 is a graph showing the anti-inflammatory efficacy evaluation results of the peptide derivatives according to the present invention.
3 is a graph showing the antioxidant efficacy evaluation results of the peptide derivatives according to the present invention.
4 is a 3D image of the eye wrinkles after using the test group containing the peptide derivative according to the present invention and the control group not including the peptide derivative according to the present invention for 4 weeks and 8 weeks, respectively.
5 is a graph showing the amount of change in skin pigmentation ratio and pigmentation ratio after 4 weeks and 8 weeks, respectively, of a test group containing the peptide derivative according to the present invention and a control group not containing it.

Hereinafter, the present invention will be described in more detail by way of 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 limited to these examples.

Example One: degaloyle - heptapeptide (Lysil- threonyl - prolyl - prolyl - threonyl - threonyl -proline) production

1.1 Synthesis of NH ₂ -protected heptapeptide-resin

The peptide was synthesized by a conventional solid phase peptide synthesis (SPPS) using 9-fluorenylmethoxycarbonyl (Fmoc) as a protecting group for the amino group, and N-hydroxybenzotriazole ( Amino acid residues were extended using N-Hydroxybenzotriazole (HOBt) and N,N'-Dicyclohexylcarbodiimide: DCC) as activators [References: Wang C. Chan, Perter D. White, 'Fmoc solid phase peptide synthesis', Oxford] .

Specifically, in a glass reactor, 0.07 g (0.1 mmole) of Trityl Resin to which proline is protected with Fmoc in an amino group is swelled in 6 ml of N,N'-Dimethylformamide (DMF) as a solvent for 30 minutes. , the solvent was removed and treated twice with 6ml of a 20% piperidine solution to completely remove Fmoc.

The Fmoc-removed proline resin was treated with 6 ml of DMF 5 times to wash the resin, and then reacted with a threonine solution activated with HOBt-DCC at room temperature for about 2 hours. In this process, NH ₂ -protected heptapeptide (lysyl-threonyl-prolyl-prolyl-threonyl-threonyl-proline)- obtained by sequentially reacting threonine, proline, proline, threonine, and lysine The resin was washed 5 times with DMF.

1.2 Synthesis of digalloyl heptapeptide

The NH ₂ -protected peptide (lysyl-threonyl-prolyl-prolyl-threonyl-threonyl-proline)-resin synthesized above was treated twice with 6ml of a 20% piperidine solution to Fmoc After removing the resin, the resin was washed 5 times with 6ml of DMF to completely remove the remaining piperidine. Each of 10 equivalents of gallic acid, HOBt, and DCC was added to the resin, and the reaction was terminated by stirring at room temperature overnight. After the reaction was completed, the resin was washed 3 times with DMF and 3 times with DCM, and then the resin was completely dried.

The dried digalloyl-heptapeptide resin was reacted with a mixture of trifluoroacetic acid: water: thioanisole: ethanedithiol (87.5: 5: 5: 2.5, v/v) at room temperature for 2-3 hours to react the peptide. After removing the t-butyl (t-Bu) group and t-butoxycarbonyl (Boc) group, which are protecting groups of the amino acid residues constituting the Royl-heptapeptide was precipitated.

The obtained digalloyl-heptapeptide was subjected to reverse phase high performance liquid chromatography using water containing 0.05% trifluoroacetic acid and acetonitrile as solvents (Reverse phase high performance liquid chromatography; column: vydac, C18, 110A, 250*20mm ), and then freeze-dried to obtain a white powder.

Yield: 48%

Example 2: galloyle - Hexapeptide ( threonyl - prolyl - prolyl - threonyl - threonyl -proline) production

2.1 Synthesis of NH ₂ -protected hexapeptide-resin

In the same manner as in Example 1, NH ₂ -protected hexapeptide (threonyl-prolyl-prolyl-threonyl-threonyl-proline)-resin was synthesized.

2.2 Synthesis of galloyl-hexapeptide

NH ₂ -protected peptide (threonyl-prolyl-prolyl-threonyl-threonyl-proline)-resin synthesized above was treated twice with 6ml of 20% piperidine solution to remove Fmoc, and then , The resin was washed 5 times with 6ml of DMF to completely remove the remaining piperidine. Each of 10 equivalents of gallic acid, HOBt, and DCC was added to the resin, and the reaction was terminated by stirring at room temperature overnight. After the reaction was completed, the resin was washed 3 times with DMF and 3 times with DCM, and then the resin was completely dried.

The dried galloyl-peptide resin is reacted with a mixture of trifluoroacetic acid: water: thioanisole: ethanedithiol (87.5: 5: 5: 2.5, v/v) at room temperature for 2-3 hours to form a peptide. After removing the t-butyl (t-Bu) group, which is a protecting group of the amino acid residue, and separating the galloyl-hexapeptide from the resin, the galloyl-hexapeptide was precipitated with cold diethyl ether.

The obtained galloyl-hexapeptide was subjected to reverse phase high performance liquid chromatography using water containing 0.05% trifluoroacetic acid and acetonitrile as solvents (Reverse phase high performance liquid chromatography; column: vydac, C18, 110A, 250*20mm) After purification by lyophilization, white powder was obtained.

Yield: 52%

Example 3: galloyle - Heptapeptide (lysyl-threonyl) - prolyl - prolyl - threonyl - threonyl -proline) production

3.1 Synthesis of NH ₂ -protected heptapeptide-resin

In the same manner as in Example 1, NH ₂ -protected heptapeptide (lysyl-threonyl-prolyl-prolyl-threonyl-threonyl-proline)-resin was synthesized.

3.2 Synthesis of Galloyl Heptapeptide

The NH ₂ -protected heptapeptide (lysyl-threonyl-prolyl-prolyl-threonyl-threonyl-proline)-resin synthesized above was treated twice with 6ml of a 20% piperidine solution. After removing Fmoc, the resin was washed 5 times with 6ml of DMF to completely remove the remaining piperidine. Each of 10 equivalents of gallic acid, HOBt, and DCC was added to the resin, and the reaction was terminated by stirring at room temperature overnight. After the reaction was completed, the resin was washed 3 times with DMF and 3 times with DCM, and then the resin was completely dried.

The dried galloyl-heptapeptide resin was reacted with a mixture of trifluoroacetic acid: water: thioanisole: ethanedithiol (87.5: 5: 5: 2.5, v/v) at room temperature for 2-3 hours to obtain the peptide. After removing the t-butyl (t-Bu) group and t-butoxycarbonyl (Boc) group, which are protecting groups of the amino acid residues constituting the amino acid residue, the galloyl-heptapeptide was separated from the resin, and then the galloyl- Heptapeptide was precipitated.

Reverse phase high performance liquid chromatography (column: vydac, C18, 110A, 250*20mm) using the obtained galloyl-heptapeptide in water containing 0.05% trifluoroacetic acid and acetonitrile as solvents After purification by lyophilization, white powder was obtained.

Yield: 51.3%

comparative example One: heptapeptide (Lysil- threonyl - prolyl - prolyl - threonyl - threonyl -proline) production

4.1 Synthesis of NH ₂ -protected heptapeptide-resin

In the same manner as in Example 1, NH ₂ -protected heptapeptide (lysyl-threonyl-prolyl-prolyl-threonyl-threonyl-proline)-resin was synthesized.

4.2 Synthesis of heptapeptide

The NH ₂ -protected heptapeptide (lysyl-threonyl-prolyl-prolyl-threonyl-threonyl-proline)-resin synthesized above was treated twice with 6ml of a 20% piperidine solution. After removing Fmoc, the resin was washed 5 times with 6ml of DMF to completely remove the remaining piperidine. The resin was washed 3 times with DMF and 3 times with DCM, and then the resin was completely dried.

The dried heptapeptide resin is reacted with a mixture of trifluoroacetic acid: water: thioanisole: ethanedithiol (87.5: 5: 5: 2.5, v/v) at room temperature for 2 to 3 hours to form the peptide. After removing the t-butyl (t-Bu) group and t-butoxycarbonyl (Boc) group protecting the amino acid residue, the heptapeptide was isolated from the resin, and then the peptide was precipitated with cold diethyl ether.

The obtained heptapeptide was purified by reverse phase high performance liquid chromatography (column: vydac, C18, 110A, 250*20mm) using water containing 0.05% trifluoroacetic acid and acetonitrile as solvents. It was freeze-dried to obtain a white powder.

Yield: 56.2%

comparative example 2: Hexapeptide ( threonyl - prolyl - prolyl - threonyl - threonyl -proline) production

5.1 Synthesis of NH ₂ -protected hexapeptide-resin

In the same manner as in Example 1, NH ₂ -protected hexapeptide (threonyl-prolyl-prolyl-threonyl-threonyl-proline)-resin was synthesized.

5.2 Synthesis of hexapeptide

The NH ₂ -protected hexapeptide (threonyl-prolyl-prolyl-threonyl-threonyl-proline)-resin synthesized above was treated twice with 6ml of a 20% piperidine solution to remove Fmoc. Then, the resin was washed 5 times with 6ml of DMF to completely remove the remaining piperidine. The resin was washed 3 times with DMF and 3 times with DCM, and then the resin was completely dried.

The dried hexapeptide resin is reacted with a mixture of trifluoroacetic acid: water: thioanisole: ethanedithiol (87.5: 5: 5: 2.5, v/v) at room temperature for 2 to 3 hours to compose the peptide. After removing the t-butyl (t-Bu) group, which is a protecting group of the amino acid residue, and separating the hexapeptide from the resin, the hexapeptide was precipitated with cold diethyl ether.

The obtained hexapeptide was purified by reverse phase high performance liquid chromatography (column: vydac, C18, 110A, 250*20mm) using water containing 0.05% trifluoroacetic acid and acetonitrile as solvents. It was freeze-dried to obtain a white powder.

Yield: 55.8%

Experimental example 1: Assess mitochondrial activity

In order to confirm the activity of mitochondria, the mitochondria membrane potential was measured.

Epithelial cells were aliquoted into a 96-well plate for fluorescence measurement, 2.0×10 ⁴ cells per well, and DMEM medium supplemented with penicillin/streptomycin 1% and FBS 10% was used at 37°C, 5% CO ₂ condition for 1 day. After incubation, the untreated group, the material-treated group (Examples 1, 2, 3, Comparative Examples 1 and 2 each at 100 μM concentration) and the UV-treated group were washed with DPBS, respectively, and then JC-1 dye prepared at 10ug/ml was added and incubated for about 20 minutes.

Wash it again with DPBS to remove excess dye, add an appropriate amount of DPBS, and measure the fluorescence intensity with a fluorescence plate reader (Green: Ex=485nm, Em=530nm / Red: Ex=530nm, Em=590 nm). The ratio of Green was expressed as a relative value compared to that of the UV irradiation control group.

The results are shown in FIG. 1 .

1, it was confirmed that the digalloyl heptapeptide prepared in Example 1 and the galloyl heptapeptide prepared in Example 3 showed excellent mitochondrial activity (mitochondrial membrane potential).

Experimental example 2: PGC - 1α activity evaluation

Real time quantitative polymerase chain reaction (RT-qPCR) was used to confirm the expression level of PGC-1α. Human fibroblasts were treated with DMEM (Dulbecco's Modified Eagle's Medium, Gibco, USA) containing 10% FBS, Penicillin (50U/mL) and Streptomycin (50/mL) while maintaining constant humidity in an incubator at 37 °C and 5% CO ₂ 500 μL of 1×10 ⁵ cells/mL was dispensed into a 24 well plate, followed by incubation for 24 hours.

After that, each sample was treated at 100 μM, washed twice with an appropriate amount of DPBS at 2 hours, 8 hours, and 24 hours, RNA was recovered using TRIzol reagent, and then iScript™ cDNA Synthesis Kit (Biorad, Cat no 1708891) to synthesize cDNA. The synthesized cDNA was amplified using SYBR Green Supermix (Biorad, Cat no. 1708882) and CFX Connect™ Real-Time PCR Detection System (Biorad, Cat no. 1855201) to measure the expression level. The results are shown in Table 1 below.

sample name order treatment concentration PGC-1α expression rate
(% of control) 2h 8h 24h Control 1 (untreated group, H ₂ O) - - 100 Example 1 (GA) ₂ -KTPPTTP 100 uM 122 210 123 Example 2 GA-TPPTTP 75 102 95 Example 3 GA-KTPPTTP 90 115 107 Comparative Example 1 KTPPTTP 78 139 80 Comparative Example 2 TPPTTP 85 140 87

From Table 1, it was confirmed that the digalloyl heptapeptide prepared in Example 1 exhibited a remarkably high expression rate of PGC-1α.

Experimental example 3: procollagen ( Procollgen ) synthesis evaluation

DMEM (Dulbecco's Modified Eagle's Medium, containing ₁₀ % FBS, Penicillin (50U/ml), Streptomycin (50/ml), Gibco, USA), 1×10 ⁵ pieces/ml, and 500 μl of each in a 24 well plate, followed by incubation for 24 hours. The untreated group, Examples 1, 2, 3, and Comparative Examples 1 and 2 at a concentration of 100uM were added and cultured at 37°C, 5% CO ₂ in an incubator for 48 hours. After 24 hours, 20 μl of each supernatant of the medium was taken and the amount of newly synthesized collagen was measured using Procollagen Type I C-Peptide EIA Kit (PICP, Takara, Cat No. MK101). The amount of PICP was converted to ng/1×10 ⁵ cells, calculated according to the following equation, and the results are shown in Table 2 below.

[Equation 1]

Collagen biosynthesis increase rate (%) = [Test group collagen amount / Control collagen amount]×100

sample name order treatment concentration Collagen biosynthesis increase rate
(% of control) Control group (untreated group) - - 100 Example 1 (GA) ₂ -KTPPTTP 100 uM 123 Example 2 GA-TPPTTP 122 Example 3 GA-KTPPTTP 118 Comparative Example 1 KTPPTTP 105 Comparative Example 2 TPPTTP 100

From Table 2, it was confirmed that the digalloyl heptapeptide prepared in Example 1, the galloyl hexapeptide prepared in Example 2, and the galloyl heptapeptide prepared in Example 3 showed an excellent increase rate of collagen biosynthesis.

Experimental example 4: Anti-inflammation efficacy evaluation

In order to confirm the anti-inflammatory effect, the degree of inhibition of iNOS activity was confirmed. Specifically, after activation of a HaCaT (keratinocyte) cell line with 1 g of LPS, treatment with L-NMMA (positive control), TPPPTTP, GA-TPPTTP, KTPPTTP, GA-KTPPTTP, (GA) ₂ -KTPPTTP 24 incubated for hours. After incubation, 100 μl of the cell culture supernatant and 100 μl of Griess reagent (1% sulfanilamide in 5% phosphoric acid and 1% naphthylamide in H ₂ O) were mixed, reacted in a 96-well-plate for 10 minutes, and absorbance was measured at 540 nm. By doing so, the amount of NO (NO ₂ form) was measured. The concentration of NO ₂ was obtained by measuring the absorbance by diluting sodium nitrate. The results are shown in Figure 2,

As shown in FIG. 2 , it was found that the anti-inflammatory effect of digalloyl heptapeptide prepared in Example 1 was the best.

Experimental example 5: DCF -Evaluation of anti-oxidation efficacy in cells using DA Assay

DCF-DA assay was used to measure intracellular antioxidant capacity. Skin fibroblasts were seeded in a 6-well plate with cover glass at 1 × 10 ⁶ cells/well, cultured for 24 hours, and adhered. It was treated at a concentration of 100 uM and incubated for 24 hours. Thereafter, oxidative stress was induced by treatment with 1 mM hydrogen peroxide for 1.5 hours, washed twice with sterile PBS (pH 7.4), and then 1 mL of 3.7% paraformaldehyde solution was added for cell fixation for 15 minutes. After standing at room temperature, it was removed. After treatment with 10 μM DCF-DA, the reaction was carried out at 37° C. for 2 hours, taking care not to enter light, and then mounted using Gel/Mount and observed under a microscope. In addition, intracellular fluorescence values were measured by DCF (EX = 485 nm; Em = 525 nm) using a fluorescent microplate reader (Sunrise, Tecan Austria Gmbh, Austria), and the results are shown in FIG. 3 .

3, it was confirmed that the digalloyl heptapeptide prepared in Example 1 showed the best intracellular antioxidant efficacy.

Experimental example 6: Human application evaluation test for the improvement of wrinkles around the eyes

A cream containing the digalloyl heptapeptide prepared in Example 1 was prepared, and a human application test was performed on the effect of improving the wrinkles around the eyes when used in parallel.

6.1 Test Evaluation Methods

(1) Subjects for test evaluation: 23 (43-60 years old female, average age 53.7 years old)

The age ratio of the test subjects is shown in Table 3 below.

age ratio (%) 40's 21.74 50's 73.91 60's 4.35

(2) Test evaluation period: 8 weeks (before use, after 4 weeks, after use 8 weeks)

(3) The evaluation method after 4 weeks and 8 weeks after use was as follows.

- PRIMOS ^R Evaluation of eye wrinkles using lite (Canfield, USA)

- Subjective questionnaire evaluation by test subjects

- Visual evaluation by a dermatologist

(4) How to use the sample: Take an appropriate amount of the product twice a day (morning and evening), spread it evenly over the entire face, and tap it lightly for absorption.

(5) Sample information:

- Test group: Cream containing (Galloyl) ₂ -KTPPTTP peptide

- Control: Cream without active ingredient

6.2 Test Evaluation Results

(1) PRIMOS ^R Statistical analysis results were calculated for the average wrinkle depth (Rz) and average roughness (Ra) of the eye area using lite.

The results are shown in Table 4 below.

parameter
(Parameter) measurement group
(Group) measurement point
(Time point) Average
(Mean, μm) Standard Deviation
(SD) Significance Probability
( p -value) average rate of change
(%) Rz test group before product use 60.83 11.27 - - after 4 weeks of use 59.17 11.94 0.169 (-) 2.45 after 8 weeks of use 54.52 10.49 0.000 (-) 10.15 control before product use 60.34 8.10 - - after 4 weeks of use 60.15 7.50 0.896 (-) 0.48 after 8 weeks of use 60.10 7.77 0.836 (-) 0.01 Ra test group before product use 12.99 2.55 - - after 4 weeks of use 12.70 2.68 0.275 (-) 1.87 after 8 weeks of use 11.65 2.34 0.000 (-) 10.05 control before product use 12.89 1.80 - - after 4 weeks of use 12.84 1.81 0.866 (-) 0.31 after 8 weeks of use 12.84 1.68 0.817 (-) 0.02

From Table 4, it can be confirmed that the average wrinkle depth (Rz) and the average roughness (Ra) of the eye area wrinkle area decreased by 10.15% and 10.05%, respectively, to a statistically significant level ( p < 0.05) in the test group after 8 weeks of use. could

(2) After 4 weeks and 8 weeks of use, 3D images of wrinkles around the eyes according to the use of the product (test group, control group) are shown in FIG. 4 .

From FIG. 4, it was confirmed that wrinkles around the eyes were improved after 8 weeks of use in the test group using the cream containing digalloyl heptapeptide.

Experimental example 7: Human application evaluation test for skin pigmentation improvement effect

A cream containing the digalloyl heptapeptide prepared in Example 1 was prepared, and a human application test was conducted for the effect of improving skin pigmentation of concurrent use.

7-1. Assessment Methods

The degree of skin pigmentation was evaluated using a Spectrophotometer ^® CM2600d (Minolta, Japan), and pictures were taken using a Janus Pro (PIE, Korea). This device measures the spectral reflectance of an object's color and calculates the tristimulus values using CIELAB's colorimetric system L*, a*, b*. In the L*a*b* color space, brightness is indicated by L*, and chromaticity indicating hue and saturation is indicated by a* and b*.

In this test, the L* value of the cheekbone protrusion (exposed area) without pigment disease and the upper arm (unexposed area) not exposed to light on the inside of the arm (unexposed area) was measured 3 times before using the product, after 4 weeks of use, and after 8 weeks of use to obtain the average value, and the ratio of the L* value of the exposed and unexposed areas was calculated and analyzed. The closer the pigmentation ratio to 1, the smaller the difference in skin color due to photoaging.

Sample information:

Test group: Cream containing (Galloyl) ₂ -KTPPTTP peptide

Control group: Cream without active ingredient

7-2 Evaluation Results

Fig. 5 shows a graph of skin pigmentation ratio analysis results according to product use (mean±deviation, probability of significance p<0.05*, probability of significance between groups p<0.05†).

As shown in FIG. 5 , compared with before product use, the skin pigmentation ratio of the test group significantly increased by 0.24% after 4 weeks of use and 0.50% after 8 weeks of use (p<0.05). In addition, there was a significant difference between the two groups in the amount of change in the skin pigmentation ratio after 8 weeks of use (p<0.05), confirming that the test group improved skin pigmentation compared to the control group.

Claims (7)

A peptide derivative represented by the following formula (1):
[Formula 1]

In the above formula,
n is 1 or 2,
L is absent or is a linking group,
A is a PGC-1α-derived peptide comprising the amino acid residues of Thr-Pro-Pro-Thr-Thr-Pro. The peptide derivative according to claim 1, wherein n is 2. The peptide derivative according to claim 1, wherein L is Lys. The peptide derivative according to claim 1, wherein n is 2 and L is Lys. A cosmetic composition comprising the peptide derivative according to any one of claims 1 to 4. [Claim 6] The cosmetic composition according to claim 5, wherein the composition is for skin aging prevention. [Claim 6] The cosmetic composition according to claim 5, wherein it is used for wrinkle improvement, whitening, antioxidant or anti-inflammatory.

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