KR100553174B1 - The fusion peptide-ptd conjugates for improving skin-wrinkle and cosmetics containing the same - Google Patents

Abstract

The present invention relates to a fusion peptide for skin wrinkle improvement and a cosmetic for improving skin wrinkles containing the same. More specifically, the fusion peptide according to the present invention is a PTD peptide (Protein Transduction) to a peptide derived from the C-terminus of human SNAP-25. As a fusion peptide to which Domain) is bound, it has high in vivo permeability and stability, and suppresses the secretion of catecholamine, a neurotransmitter related to wrinkle formation, and has an excellent skin wrinkle improvement effect.

Potox, Wrinkle improvement, Anti-aging, Catecholamine, PTD

Description

The fusion peptide for skin wrinkle improvement and the cosmetic for improving skin wrinkles containing the same {THE FUSION PEPTIDE-PTD CONJUGATES FOR IMPROVING SKIN-WRINKLE AND COSMETICS CONTAINING THE SAME}

FIG. 1 shows the results of HPLC analysis of PTD-Potox (b) prepared using Faux Tox (a) and a solid phase automatic synthesizer.

2 is a genetic map of a recombinant vector for expressing PTD-potox according to the present invention.

FIG. 3 is a 15% SDS gel electrophoresis picture of the fusion peptide according to the present invention prepared and purified according to the method described in Example 2. FIG.

4 shows the results of in vivo experiments on the transdermal absorption efficiency of fusion peptides according to the present invention.

5 shows the catecholamine secretion inhibitory effect of the fusion peptide according to the present invention in PC12 cells.

The present invention relates to a fusion peptide for skin wrinkle improvement and a cosmetic for improving skin wrinkles containing the same, and more particularly, the fusion peptide according to the present invention improves wrinkles by inhibiting secretion of neurotransmitters, such as catecholamines, which are involved in skin wrinkle generation. Has an effect.

As you age, your skin ages and you get wrinkles. The causes of skin aging can be largely classified into internal factors such as aging and environmental factors. Depending on the area of occurrence, there are neck wrinkles, forehead wrinkles, brow wrinkles, fine lines under the eyes, fine lines around the lips, and fine wrinkles on the entire face. Skin wrinkles result from the reduction and degeneration of collagen fibers in the dermis. In addition, degeneration of the elastic fibers also degrades the elasticity of the skin, worsening wrinkles of the skin. Most of the fine wrinkles begin to develop in the late 20s and 30s, and become more pronounced when they are laughing.

As extensive research for the prevention and alleviation of the formation of wrinkles has been developed, important functions of collagen have been revealed in the skin, and to suppress skin wrinkles caused by aging by promoting collagen synthesis in the skin and preventing the collapse of collagen layer at its source. Research is underway. As a result, products that combine collagen with cosmetics have been released from collagen's skin moisturizing effect, but these cosmetics are applied to the surface of the skin, and it is said that it is essential to improve skin function because it is difficult to absorb the percutaneous absorption of collagen, which is a polymer. Can't. Retinoic acid, TGF-β (transforming growth factor), protein from animal placenta (JP8-231370), betulinic acid (JP8-208424), chlorella extract (JP10-36283) and the like, but in the case of retinoic acid is unstable and there is a limit to the amount of use due to stability problems such as irritation and redness when applying skin, and chlorella extract, etc., is insignificant and is substantially caused by the promotion of collagen synthesis of the skin. Can not expect improvement in skin function.

In addition to promoting collagen synthesis, new treatments for skin wrinkles include ultrasound therapy, laser dermabrasion, botulinum toxin injection, restilene injection, etc., but they do not show significant effects in terms of cost and duration.

Recently, Fox-Tox, a peptide derived from the C-terminus of human SNAP-25 protein, has been marketed as a skin wrinkle improving agent. Potox is a fragment of the amino acid cut by botulinum neurotoxin at the C-terminal part of SNAP-25, a protein involved in the release of neurotransmitters from neurons in higher animals, resulting in hexapeptides. Inhibits the formation of the SNAPE complex, which leads to a decrease in the secretion of catecholamine (actecholamine), a neurotransmitter that causes muscle contraction and consequently prevents skin wrinkles. However, experts question the transdermal absorption of Potox, which is too large and hydrophilic.

As one method for increasing the skin absorption of the peptide anti-wrinkle agent, a method of grafting a long chain fatty acid such as palmitic acid at the end of the peptide to increase the fat solubility of the peptide (French Patent Application No. 2788058, PCT Publication WO 00/40611), but its absorption enhancement effect is not much improved.

Thus, there is a need for a general method for biologically effective delivery of biologically active macromolecules without damaging cells both inside and outside the body (LA Sternson, Ann. NY Acad. Sci., 57, 19-21 (1987). )). Examples of such methods include chemical addition of lipid peptides (P. Hoffmann et al. Immunobiol., 177, 158-170 (1998)) or methods using basic polymers such as polylysine or polyarginine (WC. Chen et al., Proc. Natl. Acad. Sci., USA, 75, 1872-1876 (1978)), but these methods have not yet been validated. Folic acid (CP Leamon and Low, Proc. Natl. Acd. Sci., USA, 88, 5572-5576 (1991)), used as a transporter, has been reported to be transported into the cell as a folate-salt conjugate, but to the cytoplasm. It is not confirmed yet. Pseudomonas Exotoxin is also used as a type of transporter (T. I. Prior et al., Cell, 64, 1017-1023 (1991)). However, even in these methods, the effect on the migration of biologically activated 'transferred' material into cells and its general applicability are not clear. Therefore, there is a continuous need for a method that can safely and effectively deliver biologically active substances into the cytoplasm or nucleus of living cells.

As a result of this study, PTD has been suggested, and the most studied is Tat protein, a transcription factor of human immunodeficiency virus-1 (HIV-1). It has been found that the protein is more effective in transcribing cell membranes when it is composed of parts of the 47th to 57th amino acids (YGRKKRRQRRR), which are concentrated in positively charged amino acids, rather than the complete form of 86 amino acids. (Fawell S. et al. Proc. Natl. Acad. Sci. USA 91, 664-668 (1994)). As another example of the effect as PTD, amino acids from 267 to 300 of VP22 protein of HSV-1 (Herpes Simplex Virus type 1) (Elliott G. et al. Cell, 88, 223-233 (1997)) And 339 to 355 amino acids (Schwarze SR et al. Trends Pharmacol Sci. 21, 45-48 (2000)) of the Drosophila ANTP (Antennapedia) protein, including artificial peptides that list electrically positive amino acids. The effect was confirmed (Laus R. et al. Nature Biotechnol. 18, 1269-1272 (2000)).

Accordingly, the present inventors have also intensively studied in order to further enhance the skin wrinkle improvement effect of the phototoxin by facilitating intracellular delivery, ie, skin absorption, of the phototoxin, a peptide having a skin wrinkle improvement effect. A peptide consisting of amino acids 858 to 868 of the murine Mph1 transcription factor was used as a peptide for intracellular material transfer, i.e., as PTD, to prepare a PTD-potox fusion peptide. As a result, the fusion peptide according to the present invention was prepared. When used to improve wrinkles, it has been found that there is no skin irritation, absorption from the skin is enhanced, stable, and the effect of improving wrinkles in vivo is maximized, thus completing the present invention.

Accordingly, an object of the present invention is to provide a fusion peptide having improved skin absorption efficiency as a peptide for improving skin wrinkles by inhibiting the secretion of neurons related to wrinkle formation.

In addition, an object of the present invention is to provide a cosmetic for improving skin wrinkles comprising the fusion peptide as an active ingredient.

In order to achieve the above object, the fusion peptide according to the present invention is characterized in that a peptide derived from the C-terminus of human SNAP-25, that is, a PTD peptide (Protein Transduction Domain) having excellent cell permeability, is bound to Potox. In addition, the peptides according to the present invention are non-irritating and easily and safely penetrate into the outer skin and the endothelium, without causing problems of skin diseases, and have a continuous effect of inhibiting the secretion of catecholamines, which cause the collapse of the collagen layer to produce skin wrinkles.

The fusion peptide according to the invention relates to, for example, a fusion peptide (also referred to herein as 'PTD-potox') in which a PTD peptide is bound to a peptide derived from the C-terminus of human SNAP-25. The peptide sequence thereof may be represented by the following Chemical Formula 1:

[Formula 1]

_Wherein [] _PTD is an autologous cell penetrating PTD peptide containing at least 30% of arginine, and R ₁ is at least one selected from the group consisting of tyrosine, alanine, arginine, valine, glycine and proline side chains. L is an integer from 5 to 30;

[] _gly connects autologous cell penetrating PTD peptide with human SNAP-25 C-terminal peptide and enhances the fluidity between PTD peptide and Potox, enhancing the binding efficiency between the infiltrated PTD-Potox and SNAPE complexes. A peptide maximizing, wherein m is an integer from 0 to 3;

Faux Tox is a peptide derived from the C-terminus of human SNAP-25, consisting of amino acids 186 to 206].

Examples of Faux Tox that can be used in the present invention include the following peptide sequences consisting of amino acids 186 to 206 of human SNAP-25.

N-Ile-Met-Glu-Lys-Ala-Asp-Ser-Asn-Lys-Thr-Arg-Ile-Asp-Glu-Ala-Asn-Gln-Arg-Ala-Thr-Lys-Met-Leu-Gly- Ser-Gly-C (SEQ ID NO: 1)

However, if the effect of the original Potox, which can suppress the secretion of neurotransmitters, such as catecholamines involved in the production of wrinkles, and can exhibit a wrinkle improvement effect, any modification, ie substitution, insertion and It will be understood by those skilled in the art that any deletions or the like can also be used as the Potox portion of the fusion peptide according to the invention.

Examples of [] _{PTDs that} can be used in the present invention include peptides having the following sequences consisting of amino acids 858 to 868 of the murine Mph1 transcription factor.

N-Thr-Ala-Arg-Val-Arg-Arg-Arg-Gly-Asp-Arg-Arg-C (SEQ ID NO: 2)

However, if the effect of increasing the skin absorption rate of the protein can be substantially maintained or elevated, any modifications, ie, substitutions, insertions, deletions, etc., to the sequence of SEQ ID NO. It will be understood by those skilled in the art that they can be used as part.

According to the present invention, by inserting about 0 to 3 glycine residues between the peptide sequence of Potox and PTD, it is possible to increase the efficiency of delivery and absorption of Potox into the cells. However, it has been found by the present inventors that when using other peptide residues known in the art other than glycine, the yield of reaction tends to be poor in obtaining a linked fusion peptide between Potox and PTD.

Meanwhile, the method for preparing the fusion peptide according to the present invention is not particularly limited as long as the fusion peptide as the final product prepared can exhibit an improved effect on skin absorption efficiency as well as skin wrinkle improvement. In addition, methods well known in the art, such as a solid phase synthesis method or a synthesis method using a recombinant expression system may be used. However, the method for producing the fusion peptide according to the present invention is not limited only by this method.

The present invention provides a method for preparing a fusion peptide (PTD-potox) in which a peptide derived from the C-terminus of human SNAP-25, ie, a potox, is bound to a self-cell permeable PTD peptide. For example, the PTD-potox fusion peptide of the present invention can be prepared by the following method.

First, the synthesis of pure PTD-potox fusion peptides can be performed using an organic synthesis device for peptide synthesis. This method is Merrifield's solid phase peptide synthesis ( J. Am. Chem. Soc . 85, 2149-21, 54 (1963)) and PTD using a peptide semi-automatic synthesizer (Peti-Syzer Model PSS-510). A fusion peptide is synthesized by condensation reaction of a monomer having a C-terminus and an N-terminus reactivity with an amino acid of Potox.

Solid phase synthesis begins by coupling an amino acid having an α-amino group protected to a suitable resin from the amino acid at the carboxyl end. At this time, the amino acid in which the α-amino group is protected is attached to the hydroxy methyl resin or the chloromethylated resin as an ester bond. As a group protecting the α-amino group, Fmoc (9-fluorenyl methoxycarbonyl) is used. (Fmoc-protected amino acids can be purchased from Bidtech Co., Ltd.). For example, the amino group of Arg is reactive. Reactive moieties of Gly, Lys, Gln are Fmoc-amino acids protected with appropriate groups such as tert-butyl (t-Bu), Pmc (2,2,5,7,8-pentamethylchroman-6-sulfonyl) Use Peptide synthesis proceeds by sequentially removing protecting groups with reagents such as trifluoroacetic acid (TFA), phenol, on the amino terminus of the peptide chain attached to a solid support resin. Peptides can be isolated from TFA solution by extraction with diethyl ether and purified by high performance liquid chromatography (HPLC).

Second, the method for producing a PTD-potox fusion peptide using a biological method, (a) can be expressed in the form of a fusion peptide by cloning the PTD-potox gene in the expression vector pPEPTIDE capable of expressing a specific peptide. Preparing a recombinant vector, and (b) mass expression in the form of a recombinant transporter-PTD-potox peptide in E. coli using the pPEPTIDE-PTD-potox expression vector, followed by pure separation and purification. It includes. At this time, in order to prepare a recombinant vector, the base sequence encoding the fusion peptide can be easily inferred from the amino acid sequence disclosed by those skilled in the art, a cloning vector for inserting the base sequence, prepared using a cloning technique A recombinant expression vector, a host cell for transforming with the recombinant vector to express the desired fusion peptide, a method for transforming the recombinant vector into a host, a method for expressing the desired fusion peptide from the transformed host cell, and a target from the final product Selection and general technical matters, such as how to obtain fusion peptides, will be readily appreciated by those skilled in the art.

Meanwhile, the present invention provides a skin comprising a fusion peptide in which the autologous cell penetrating PTD peptide is bound to a peptide derived from the C-terminus of human SNAP-25, which has excellent cell permeability and excellent wrinkle improvement effect and persistence of the effect. Provide cosmetics for wrinkle improvement. Cosmetics for improving skin wrinkles according to the present invention may contain a fusion peptide according to the present invention at a concentration of 0.01 mg / ml to 100 mg / ml, but exhibits an effect of improving skin wrinkles according to the administration method and the skin condition of the subject It may also contain the fusion peptide according to the present invention in an amount outside the above range, and the concentration of such a suitable fusion peptide can be easily recognized by those skilled in the art.

In addition, to prepare cosmetics for improving skin wrinkles, other ingredients commonly known in the art may be appropriately selected according to cosmetic formulations, and the amount of fusion peptides according to the present invention for exhibiting skin wrinkle improvement is difficult. Easy to choose without In addition, it will be understood by those skilled in the art that the fusion peptide according to the present invention may be used in combination with various other anti-wrinkle ingredients known in the art, unless deteriorating the effect of the fusion peptide according to the present invention.

Hereinafter, a method for preparing PTD-potox fusion peptide according to the present invention represented by Chemical Formula 1, its chemical stability, skin wrinkle improvement effect and its persistence, and safety excellent skin wrinkle improver through inhibition of excellent neurotransmitter The usefulness as will be described in more detail through Examples and Experimental Examples. However, it will be understood by those skilled in the art that the present invention is not limited by these examples.

EXAMPLE

Example 1 Preparation of PTD-Potox Conjugate (YARVRRRGPRRGGGIMEKADSNKTRIDEANQRATKMLGSG)

(1) Preparation of PTD-Potox Conjugate

PTD-Potox conjugates according to the invention having the amino acid sequence of YARVRRRGPRRGGGIMEKADSNKTRIDEANQRATKMLGSG were synthesized by solid phase peptide synthesis using a peptide semi-auto synthesizer (Peti-Syzer Model PSS-510). 0.1 mmol of Rink Amide MBHA resin was placed in a standard reaction vessel, and the synthesis was started by adding 0.5 mmol of Fmoc-G, the first amino acid of C-terminal of the peptide to be synthesized. From the C-terminal amino acid to the N-terminal amino acid, the corresponding amino acid residues were reacted three times with 0.5 mmol each.

 Deprotection of Fmoc was performed three times for 10 minutes using 20% piperidine diluted with dimethyl formamide (DMF), washed for 10 minutes with DMF and coupling for 100 minutes. Coupling used N-hydroxybenzotriazole (HOBT) and 1,3-diisopropylcarbodiimide (DIC), and the system which wash | cleaned with DMF for 10 minutes was used.

(2) Isolation and Purification of PTD-Potox Conjugates

After the synthesis was completed, the PTD-Potox conjugate was placed in a 50 mL corning tube, cooled to 0 ° C., 0.75 g of phenol, 0.25 mL of 1,2-ethanedithiol (EDT), 0.25 mL of tianisol, A cutting solvent having a mixing ratio of 0.5 mL of distilled water and 8.25 mL of TFA was added thereto, followed by reaction at room temperature for 4 hours. After the reaction was completed, the resin separated from the peptide was filtered and the remaining solution was added with 30 ml of diethyl ether below 50 ° C. to precipitate the peptide, followed by washing with diethyl ether added twice more. The precipitate thus obtained was dried and then dissolved in MeOH and purified by HPLC. The column used for purification used the C18 analytical column 218TP54 (VyDac) and the C18 preparative column 218TP1022 (VyDac). Purification conditions were 1 mL / min and 25 mL / min, respectively. The solvent was eluted with A: 100% H ₂ O + 0.01% TFA and B: 100% acetonitrile + 0.01% TFA. Acetonitrile of the eluted peptide was removed and lyophilized to obtain a high purity peptide.

HPLC analysis was performed on the peptides obtained in this example to obtain the results of FIG. 1.

Example 2. Preparation and Purification of Fusion Peptide (PTD-Potox) Expression Vectors Using a Microbial Expression System

(1) Preparation of PTD- Potox Expression Vector

To prepare a nucleotide sequence encoding the PTD-potox fusion peptide, Hind III, PshB I at the N-terminus, and BamH I at the C-terminus were put in pUC 19 to prepare a template. The PTD-Potox template was isolated by restriction enzymes PshB I and BamH I to bind the poly His tag region for high expression and easy purification of PTD-potox and the nucleotide sequence encoding the high expression protein. Purification was made using an Auquiquick tablet kit.

The expression vector, pPEPTIDE vector, was processed using restriction enzymes PshB I, BamH I, purified using a Quiaquick purification kit, and then cloned into a nucleotide sequence encoding the prepared PTD-Potox recombinant expression vector. Was prepared. The gene map of the schematic expression vector vector is shown in Figure 2, and the protein expressed from the expression vector was named PTD-Potox-carrier.

(2) Preparation of E. coli transformants and expression and purification of fusion peptides

E. coli BL21 was transformed by the heat shock transformation method using the expression vector PTD-Potox prepared above, and the transformed E. coli was inoculated in 100% LB medium in an amount of 1% by 1%. Pre-incubated with stirring at 37 ° C. for 12 hours. It was then inoculated again in 1000 mL of LB medium and incubated for 4 hours at 37 ° C., followed by addition of 1 mM IPTG (isopropyl β-D-thiogalactopyranoside, GibcoBRL cat. # 15529-019). Expression of lac operon was induced and cultured for 8 hours to induce the expression of the fusion peptide.

The culture solution was centrifuged at 5,000 rpm at 20 ° C. for 20 minutes to remove the supernatant, leaving only the pellet. Then, 10 mL of buffer solution (50 mM NaH ₂ PO) containing 1 mg / mL lysozyme (Sigma, cat, # L-7651) ₄ , 300mM NaCl, 10mM imidazole, pH 8.0), the pellet was left for 30 minutes on ice, and then ultrasonically infused for 10 seconds at an intensity of 300 W using an ultrasonic grinder (Heat systems, ultrasonic processor XL). The cooling process was repeated for 10 seconds to obtain a cumulative ultrasonic injection time of 3 minutes. The eluate was centrifuged at 12,000 rpm for 20 minutes at 4 ° C. to remove the lysate of E. coli, and only pure eluate was separated.

2.5 mL of 50% Ni ²⁺ -NTA agarose slurry (Qiagen, cat # 30230) was added to the separated eluate, and stirred at 200 rpm for 4 hours at 4 ° C to combine the fusion protein with Ni ²⁺ -NTA agarose. The mixture was poured into a 0.8 × 4 cm column for chromatography (BioRad, cat # 731-1550) and flowed.

After washing twice with 4 mL of Buffer 2 (50 mM NaH ₂ PO ₄ , 300 mM NaCl, 20 mM imidazole, pH 8.0), 0.5 mL of Buffer 3 (50 mM NaH ₂ PO ₄ , 300 mM NaCl, 250 mM The fusion protein was divided into four times with dozol, pH 8.0), and subjected to SDS-PAGE, followed by Cowmash blue staining, and the results are shown in FIG. 3. In Figure 3, lane 1 is the standard molecular weight protein, lane 2 is the fusion protein PTD-potox-transporter.

2mL ice-cold 4M NH ₂ OH, 0.4MK ₂ CO ₃ , 6M GuHCl, pH 9.0 was added to the fusion protein until the final buffer concentration reached 2M NH ₂ OH, 0.2MK ₂ CO ₃ , 3M GuHCl, pH 9.0. After stirring for 5 hours at 225rpm to separate the fusion protein into the fusion peptide and the transporter protein, 12N HCl was added to adjust the pH between 2-3 and stirred for 10 minutes to complete the reaction.

12.5N NaOH was added to neutralize the pH to 6.5, and the separated transport protein was separated by filling in a packed Ni _²⁺ -NTA agarose column, and then gel-filtered to remove salts from the buffer solution and freeze-dried. Pure fusion peptides were obtained.

Example 3 Percutaneous Absorption (Skin Cell Penetration) Experiment

Hereinafter, penetration into the skin cells of the compound represented by the formula (1) is referred to as transdermal absorption. Transdermal absorption experiment was conducted using the compound represented by General formula (1).

In detail, the fusion peptide was applied to the skin of the pig's back 2 days after birth by about 5 mmole, and after 2 hours, the skin was cut with a 1 cm punch, and frozen sectioned to measure the level of transdermal absorption with a confocal lens. It was. The results are shown in FIG. As can be seen from Figure 4, it can be seen that the percutaneous absorption of the compound represented by the formula (1) used in the present invention is excellent.

Example 4. Skin wrinkle improvement effect test by PTD-Potox in vitro.

In order to measure the usefulness of the PTD-potox fusion peptide prepared in Example 2 as a skin wrinkle improver, it was compared with Potox, which is known as a substance having an excellent inhibitory effect of catecholamine.

PC12 cells ( Peochromocytoma , ATCC CRL-1721) were thawed in T75 (Nunc) in 15 mL of F-12K medium (Gibco) in 2 mM L-glutamine, 1.5 g / L sodium bicarbonate, 15% horse serum, 2.5% FBS, After culturing for 2 days, the cell density was 625,000 cells / cm 2, the supernatant was removed by centrifugation, and cultured for one day by adding F-12K medium, Potox and PTD-Potox of the same volume, respectively, and secreted catecholamines. The amount of was analyzed via fluorescence HPLC.

The concentration of Potox and PTD-Potox was added at a concentration (M) of 10 ^-3 to 10 ^-13 . The results are shown in FIG. As can be seen in Figure 5 it can be seen that the compound represented by the formula (1) used in the present invention has an excellent wrinkle improvement effect.

Example 5. Wrinkle improvement effect measurement

In order to determine the anti-wrinkle effect of the PTD-potox fusion peptide prepared in Example 1, 10 females in their 30s do not include the experimental group including the compound represented by the formula (1) of the wrinkles around the eyes of the present invention The controls were applied once a day for 3 months each. Then, the wrinkle improvement state was visually observed and the improvement state of wrinkles was determined.

As a result, the compound represented by the formula (1) used in the present invention showed a wrinkle improving effect in the wrinkled skin around the eyes, from which the compound represented by the formula (1) of the present invention to reduce the wrinkles when used as an external application to the skin, Improvement could be predicted.

 As described above, the PTD-potox fusion peptide according to the present invention, in which a specific autologous cell penetrating PTD peptide having a self-penetrating signal is bound to a peptide derived from the C-terminus of human SNAP-25, has an absorption ability to skin. It is excellent, non-irritating, and has an excellent secretion effect of catecholamines, through which it can be seen that it has an anti-wrinkle effect and persistence of the effect.

<110> FORHUMAN TECH CO.LTD. <120> THE FUSION PEPTIDE-PTD CONJUGATES FOR IMPROVING SKIN-WRINKLE AND          COSMETICS CONTAINING THE SAME <130> P04-0656 <160> 2 <170> KopatentIn 1.71 <210> 1 <211> 26 <212> PRT <213> Artificial Sequence <220> <223> Human SNAP-25 <400> 1 Ile Met Glu Lys Ala Asp Ser Asn Lys Thr Arg Ile Asp Glu Ala Asn   1 5 10 15 Gln Arg Ala Thr Lys Met Leu Gly Ser Gly              20 25 <210> 2 <211> 11 <212> PRT <213> Artificial Sequence <220> <223> Murine Transcription factor Mph1 <400> 2 Thr Ala Arg Val Arg Arg Arg Gly Asp Arg Arg   1 5 10

Claims (6)

A fusion peptide in which a PTD peptide is bound to a peptide derived from the C-terminus of SNAP-25 represented by Formula 1 below: [Formula 1]

_Wherein [] _PTD is an autologous cell penetrating PTD peptide containing at least 30% of arginine, and R ₁ is at least one selected from the group consisting of tyrosine, alanine, arginine, valine, glycine and proline side chains. L is an integer from 5 to 30; [] _gly connects autologous cell penetrating PTD peptide with human SNAP-25 C-terminal peptide and enhances the fluidity between PTD peptide and Potox, enhancing the binding efficiency between the infiltrated PTD-Potox and SNAPE complexes. A peptide maximizing, wherein m is an integer from 0 to 3; Faux Tox is a peptide derived from the C-terminus of human SNAP-25, consisting of amino acids 186 to 206]. The fusion peptide of claim 1, wherein [] _PTD has a sequence of SEQ ID NO: 2 and Faux Tox has a sequence of SEQ ID NO: 1. 8. The fusion peptide of claim 1, wherein the fusion peptide is used to improve skin wrinkles. 4. The fusion peptide according to claim 3, wherein the improvement of skin wrinkles is shown by improving the skin permeation and absorption efficiency of FAUX TOX domain by [] _PTD domain. The fusion peptide according to claim 3, wherein the skin wrinkle improvement is obtained by inhibiting secretion of catecholamine, a neurotransmitter. A cosmetic composition for improving skin wrinkles, comprising the fusion peptide according to any one of claims 1 to 5 as an active ingredient.

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