KR20180108970A - Cosmetic composition that prevents aging and improves wrinkles by containing novel heptapeptide monomer and dimer - Google Patents

Abstract

The present invention relates to a cosmetic composition for preventing and alleviating skin-aging or skin wrinkles, which comprises novel heptapeptide monomer and dimer of chemical formulas 1-4 as an active ingredient. In particular, the heptapeptide monomer and dimer inhibit the breakdown of skin collagens by suppressing the expression of MMP-1, and thus shows excellent effects in controlling skin-aging or reducing skin wrinkles. In particular, the heptapeptide monomer and dimer may be used by being added to various basic cosmetic compositions. [Chemical formula 1] Cys-Glu-Glu-Met-Gln-X-Arg [Chemical formula 2] Glu-X-Met-Gln-Arg-Arg-Cys [Chemical formula 3] Cys-Glu-Glu-Met-Gln-X-Arg [Chemical formula 4] Glu-X-Met-Gln-Arg-Arg-Cys

Description

TECHNICAL FIELD The present invention relates to a cosmetic composition containing a novel heptapeptide monomer and a dimer to prevent skin aging or skin wrinkles and to improve the cosmetic composition containing the novel heptapeptide monomer and dimer.

The present invention relates to a cosmetic composition for preventing skin aging and improving skin wrinkles containing heptapeptide monomers and dimers for preventing skin aging or skin wrinkles as an active ingredient. More particularly, the present invention relates to a cosmetic composition for preventing skin aging and improving skin wrinkles The present invention relates to novel heptapeptide monomers and dimers that promote the synthesis and expression of collagen, which is the most important factor in cosmetics, and a cosmetic composition containing the same as an active ingredient.

Skin changes through various aging factors. The skin in the aging process changes its ECM (Extracellular matrix) component, which is thinner in the epidermis, dermis and subcutaneous tissues, and gives elasticity to the skin. Collagen, which accounts for 70 ~ 80% of ECM, And the production thereof is rapidly lowered. Such decrease in collagen production is closely related to wrinkle formation. Collagen, elastin, proteoglycans, glucosaminoglycan, laminin, fibronectin, etc., which make up the skin connective tissue, By losing function, the skin loses its elasticity and wrinkles become excessively formed, and it becomes a senile skin.

The connective tissue of ECM (Extracellular Matrix) is composed of collagen fiber, collagen fiber, reticular fiber, elastic fiber and elastic fiber. Collagen, which accounts for about 1%, is mostly formed in fibroblasts of the skin. As the aging of the skin progresses with age, collagen content in the connective tissue of the skin decreases, which is a result of collagen synthesis degradation and accelerated degradation.

Collagen biosynthesis is regulated by a variety of factors involved in transcription and post-translational levels. Collagen degradation is caused by factors such as ultraviolet light and various degradation mechanisms in the body. Collagenase ) And matrix metalloproteases (MMPs), which promote the collagen degradation. Simultaneously with such degradation of synthesis and promotion of decomposition, collagen deformation is accelerated by the external environment such as UV, and the skin becomes wrinkled and deepened. As a result, skin aging phenomena such as skin sagging, loss of elasticity and wrinkles of skin are the result of non-homogenization of cells, disappearance of elastin, destruction of collagen, reduction of collagen synthesis and promotion of decomposition. Thus, skin aging occurs in the epidermis of the skin, but it can also be seen in the dermis.

Various cosmetic compositions have been studied in order to solve the problem of skin aging phenomenon, and in some cases, the wrinkle improvement effect of the skin is visible. Clinical results of skin wrinkle removal for retinol, which is widely used for improvement of skin wrinkles, spots and pigmentation, have been reported variously. The cosmetic compositions containing retinol have been reported to be effective for improving skin wrinkles and collagen synthesis and decomposition formed by natural light, such as those disclosed in U.S. Patent Nos. 4,603,146 and 4,877,805.

In addition, cosmetic composition containing L-ascorbic acid, which is an essential component in the collagen synthesis process, has been reported in USP 4,938,969 for improving skin wrinkles, ultraviolet light blocking, pigmentation such as spots / freckles / black spots. In addition, a cosmetic composition containing an α-hydroxy acid (AHA), which is effective in removing exfoliation, is also widely used for improving the skin by promoting cell activity along with exfoliation.

However, most of the active ingredients that help to improve the skin aging described above have many problems in safety and stability for use as a raw material of cosmetic materials, and examples of solving them are also reported. Retinol, which promotes collagen synthesis and improves skin wrinkles, is easily oxidized to light, moisture, air, and heat, making it very difficult to use as a raw material for cosmetics. USP 4,720,353 uses antioxidants such as BHA (Butylated Hydroxy Anisole) Propyl gallate, and tocopheryl linoleate. In EP0440,398B1, one or more water-soluble antioxidants and one or more oil-soluble antioxidants and chelating agents were added to the oil, ) -Type emulsion composition to stabilize retinol. In addition, various studies have been conducted to stabilize retinol, but even if the stability of cosmetics is solved, there is still a problem in use due to skin irritation.

USP 4,603,146 USP 4,877,805 USP 4,938,969 USP 4,720,353 EP0440,398B1

 Yaar M, Gilchrest BA. Aging versus photoaging: postulated mechanisms and effectors. J Invest Dermatol Symp Proc 1998: 3: 47-51.  Fisher GJ, Wang ZQ, Datta SC, Varani J, Kang S, Voorhees JJ. Pathophysiology of premature skin aging induced by ultraviolet light. N Engl J Med, 1997; 337: 1419-28.  Pilcher, BK, Sudbeck, BD, Dumin, JA, Welgus, HG, Parks, WC. Collagenase-1 and collagen in epidermal repair. Arch Dermatol Res, 1998; 290 (Suppl): S37-46. The present inventors have completed the present invention.

Under these circumstances, the present inventors have established an in vitro assay using a dermal fibroblast cell line to confirm the anti-aging effect and the anti-wrinkle effect of the compound heptapeptide monomers and dimers, and found that the biosynthetic ability of collagen and collagenase Confirming that the compound heptapeptide monomers and dimers have an effect of improving wrinkles, thereby completing the present invention.

Accordingly, an object of the present invention is to provide an external skin composition and a cosmetic composition which are harmless to human body, excellent in penetration of skin and excellent in stability, and which exhibit skin anti-aging and anti-wrinkle effect, However, these problems are illustrative, and the scope of the present invention is not limited thereto.

[Chemical Formula 1]

[Cys-Glu-Glu-Met-Gln-X-Arg]

(2)

[Glu-X-Met-Gln-Arg-Arg-Cys]

(3)

[Cys-Glu-Glu-Met-Gln-X-Arg] ₂

[Chemical Formula 4]

[Glu-X-Met-Gln-Arg-Arg-Cys] ₂

In the above Chemical Formulas 1 to 4, X represents an amino acid selected from the group consisting of glutamic acid, aspartic acid, histidine, phenylalanine, alanine, cysteine, glycine, glutamine, asparagine, arginine, leucine, methionine, isoleucine, serine, tyrosine, threonine, Selected from proline and valine,

[] 2 represents a dimer obtained by modifying a thiol group (-SH), which is a cysteine residue, with a reducing form (-S-S-).

Preferably, the peptides of the above formulas (1) to (4)

[Cys-Glu-Glu-Met-Gln-Leu-Arg] (Compound 1)

[Cys-Glu-Glu-Met-Gln-Phe-Arg] (Compound 2)

[Cys-Glu-Glu-Met-Gln-Ser-Arg] (Compound 3)

[Cys-Glu-Glu-Met-Gln-Tyr-Arg] (Compound 4)

[Cys-Glu-Glu-Met-Gln-Arg-Arg] (Compound 5)

[Cys-Glu-Glu-Met-Gln-Asp-Arg] (Compound 6)

[Cys-Glu-Glu-Met-Gln-Lys-Arg] (Compound 7)

[Cys-Glu-Glu-Met-Gln-Asn-Arg] (Compound 8)

[Glu-Gly-Met-Gln-Arg-Arg-Cys] (Compound 9)

[Glu-Leu-Met-Gln-Arg-Arg-Cys] (Compound 10)

[Glu-Ser-Met-Gln-Arg-Arg-Cys] (Compound 11)

[Glu-Tyr-Met-Gln-Arg-Arg-Cys] (Compound 12)

[Glu-Asp-Met-Gln-Arg-Arg-Cys] (Compound 13)

[Glu-Lys-Met-Gln-Arg-Arg-Cys] (Compound 14)

[Glu-Asn-Met-Gln-Arg-Arg-Cys] (Compound 15)

* Lipids (Stearic, Palmitic, arachidic, etc.)

[Cys-Glu-Glu-Met-Gln-Leu-Arg] (Compound 16)

[Cys-Glu-Glu-Met-Gln-Tyr-Arg] (Compound 17)

[Cys-Glu-Glu-Met-Gln-Arg-Arg] (Compound 18)

[Cys-Glu-Glu-Met-Gln-Lys-Arg] (Compound 19)

[Glu-Asp-Met-Gln-Arg-Arg-Cys] (Compound 20)

[Cys-Glu-Glu-Met-Gln-Leu-Arg] ₂ (Compound 21)

[Cys-Glu-Glu-Met-Gln-Phe-Arg] ₂ (Compound 22)

[Cys-Glu-Glu-Met-Gln-Ser-Arg] ₂ (Compound 23)

[Cys-Glu-Glu-Met-Gln-Tyr-Arg] ₂ (Compound 24)

[Cys-Glu-Glu-Met-Gln-Arg-Arg] ₂ (Compound 25)

[Cys-Glu-Glu-Met-Gln-Asp-Arg] ₂ (Compound 26)

[Cys-Glu-Glu-Met-Gln-Lys-Arg] ₂ (Compound 27)

[Cys-Glu-Glu-Met-Gln-Asn-Arg] ₂ (Compound 28)

[Glu-Gly-Met-Gln-Arg-Arg-Cys] ₂ (Compound 29)

[Glu-Leu-Met-Gln-Arg-Arg-Cys] ₂ (Compound 30)

[Glu-Ser-Met-Gln-Arg-Arg-Cys] ₂ (Compound 31)

[Glu-Tyr-Met-Gln-Arg-Arg-Cys] ₂ (Compound 32)

[Glu-Asp-Met-Gln-Arg-Arg-Cys] ₂ (Compound 33)

[Glu-Lys-Met-Gln-Arg-Arg-Cys] ₂ (Compound 34)

[Glu-Asn-Met-Gln-Arg-Arg-Cys] ₂ (Compound 35)

* Lipids (Stearic, Palmitic, arachidic, etc.)

[Cys-Glu-Glu-Met-Gln-Leu-Arg] ₂ (Compound 36)

[Cys-Glu-Glu-Met-Gln-Tyr-Arg] ₂ (Compound 37)

[Cys-Glu-Glu-Met-Gln-Arg-Arg] ₂ (Compound 38)

[Cys-Glu-Glu-Met-Gln-Lys-Arg] ₂ (Compound 39)

[Glu-Asp-Met-Gln-Arg-Arg-Cys] ₂ (Compound 40)

[2] represents a dimer obtained by modifying a thiol group (-SH), which is a cysteine residue, with a reducing form (-S-S-).

The present invention also provides a composition for improving wrinkles containing the peptide of the above formulas 1 to 4 or the peptide selected from the peptides of the above compounds 1 to 40.

Accordingly, the present invention can provide a pharmaceutical composition for improving wrinkles containing peptides of the above formulas (1) to (4) or peptides selected from one or more peptides of the above compounds 1 to 40. The peptide may be included in the pharmaceutical composition in an amount of 0.0001 to 1.0% by weight.

In another aspect, the present invention provides a cosmetic composition for improving wrinkles containing peptides of the above formulas (1) to (4) or peptides selected from one or more peptides of the above compounds 1 to 40. The peptide may be contained in the cosmetic composition in an amount of 0.0001 to 1.0% by weight. The cosmetics may be selected from lotions, lotions, gels, creams, essences, packs, ampoules, lotions, cleansing agents, soaps, body products, soaps, oils, lipsticks and foundations.

Hereinafter, the present invention will be described in detail.

Among the peptides of the present invention, the peptides of the compounds 21 to 40 are obtained by modifying the thiol group (-SH), which is a residue of cysteine (the cysteine located at the terminal of the peptide) of the peptide of the compounds 1 to 20, It is a dimer. For example, the compound obtained by modifying the thiol group, which is the cysteine residue of Compound 1, into a reducing form is Compound 21, and the compound obtained by modifying the thiol group, which is the cysteine residue of Compound 2, into a reducing form is Compound 22. Compounds 3 to 23 can be used to prepare the duplexes of compounds 1 to 20, respectively, in this manner.

The peptide of the present invention can be produced by, but not limited to, forming one or more amino acids or a suitably protected amino acid continuously in an amino acid skeleton constantly bonded to a solid phase. In addition, the peptide can be inserted, substituted or deleted with other amino acids within a range that does not significantly deteriorate the stability, and this also falls within the scope of the present invention.

In addition, the peptide of the present invention may further include a cell permeable peptide that promotes intracellular movement of the peptide, which is bound to the C-terminal or N-terminal of the peptide. Arg-Lys-Arg-Arg-Arg-Arg-Arg) and Tat-PTD peptide (Gly-Arg-Lys- Arg-Arg-Arg: Tat PTD). However, the present invention is not limited thereto. Any cell permeable peptide known in the art may be used as long as it does not inhibit the activity of the peptide according to the present invention. Do.

Meanwhile, the peptide of the present invention may exist in the form of a salt. The salt forms which can be used in the present invention may be those which are made during the final isolation and purification of the compound or by reacting the amino group with an appropriate acid. For example, an acid addition salt can be prepared by reacting an acid addition salt such as acetate, adipate, alginate, citrate, aspartate, benzoate, benzenesulfonate, bisulfate, butyrate, camphorate, camphorsulfonate, digluconate, glycerophosphate, Sulfates, heptanoates, hexanoates, formates, fumarates, hydrochlorides, hydrobromides, hydroiodides, 2-hydroxyethanesulfonates, lactates, maleates, mesitylenesulfonates, methanesulfonates, Naphthalene sulfonate, nicotinate, 2-naphthalene sulfonate, oxalate, pamoate, pectinate, persulfate, 3-phenylpropionate, picrate, pivalate, propionate, succinate, tartrate , Trichloroate, trifluoroacetate, phosphates, glutamate, bicarbonate, La-be-toluenesulfonate and undecanoate. However, the embodiment is not limited thereto. In addition, examples of acids that can be used to form acid addition salts include, but are not limited to, inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid and phosphoric acid, and organic acids such as oxalic acid, maleic acid, succinic acid and citric acid. At this time, a peptide form containing a triploloacetate salt or an acetate salt is most preferable.

The amino group or the carboxyl group of the amino acid used for producing the peptide of the present invention can be protected by a suitable protecting group. The protected amino acid can be reacted in solution by adding the following amino acid under conditions suitable for attachment to a solid support or to form an amide bond. In addition, the protecting group may be completely removed prior to the addition of the protected amino acid with a suitable protecting group. After all of the amino acids are linked as desired, the final desired peptide can be obtained by sequential or simultaneous separation from the free residual protecting group and the free solid support.

In the present invention, the chiral center is condensed with another appropriately protected dipeptide to form an amide bond and then deprotected to obtain the desired hexapeptide, which is obtained by synthesizing the desired hexapeptide The peptide can be prepared using reaction techniques.

As the most preferable synthesis method for producing the peptide compound of the present invention, a solid phase peptide synthesis method of synthesizing by using a solid phase polymer latex can be used. The α-amino group of the peptide prepared by the above method is acid or base sensitive Can be protected by a functional group. The protecting group of the amino acid at this time should have a stable property in the peptide condensation reaction condition and should have a property that is easily removable without destroying the extended peptide chain or without any racemization of any chiral center contained therein. Accordingly, suitable protecting groups include 9-fluorenylmethyloxycarbonyl (Fmoc), t-butoxycarbonyl (Boc), benzyloxycarbonyl (Cbz), biphenylisopropyl- oxycarbonyl, t- amyloxycarbonyl , Isobornyloxycarbonyl, (α, α) -dimethyl-3,5-dimethoxybenzyloxycarbonyl, O-nitrophenylsulfanyl, 2-cyano-t-butyloxycarbonyl and the like, Other suitable protecting groups known in the art for the purpose are also within the scope of the present invention.

The 9-fluorenylmethyloxycarbonyl (Fmoc) protecting group can be used as the most preferable protecting group of the amino acid used in the peptide synthesis of the present invention.

In particular, the protecting group of the amino acid residue used in the peptide synthesis of the present invention is t-butyl (t-Bu) in the case of N-methylglutamic acid; For lysine, t-butoxycarbonyl (Boc); In the case of serine, 7t-butyl (t-Bu); For threonine and allotreonine, t-butyl (t-Bu); In the case of cysteine, trityl (Trt) is preferable, but the present invention is not limited thereto.

In the solid phase peptide synthesis method, the C-terminal amino acid can be attached to a suitable solid support or resin. Suitable solid supports useful for this synthesis include reagents of the staged condensation-deprotection reaction and those which are inert to the reaction conditions and which are insoluble in the medium used, for example rink amid or link amide 4- Methylbenzylhydryl amine resin (rink amid MBHA resin).

In particular, the preferred solid support for the C-terminal amide peptide may be the linkamide 4-methylbenzylhydrylamine resin available from Novabiochem Cor poration.

The C-terminal amide is reacted in a solvent such as dichloromethane, N-methylpyridone (NMP) or DMF at a temperature of 10 ° C to 50 ° C, preferably at a temperature of 30 ° C, for 1 to 24 hours (DMAP), 1-hydroxybenzotriazole (HOBt), N-methylmorpholine (NMM), benzotriazol-1-yloxy-tris (dimethylamino) phosphonium-hexafluorophosphate N'-dicyclohexylcarbodiimide (DCC), N, N'-diisobutyl ketone (BOC) in the presence or absence of boron trichloride (BOP) or bis (2-oxo-3-oxazolidinyl) phosphine chloride Propyl carbodiimide (DIC), [O- (7-azabenzotriazol-1-yl) -1,1,3,3-tetramethyluronium hexafluorophosphate] (HATU) or O-benzotriazole (Coupling, coupling) of the carboxylic acid to the resin or solid support via condensation by activating the carboxylic acid on N, N, N ', N'-tetramethyluronium hexafluorophosphate (HBTU) There.

When the solid support is a linkamide 4-methylbenzylhydrylamine resin, as a preferred protecting group, the Fmoc functional group is excessively dissolved in a secondary amine solution, preferably a 20% piperidine DMF solution, before condensation with the C-terminal amino acid . Preferred reagents used to condense the desired amino acid with the deprotected 4- (2 ', 4'-dimethoxyphenyl-Fmoc-aminomethyl) phenoxyacetamidoethyl resin include, but are not limited to, DMF (NMM), 1-hydroxybenzotriazole (HOBt) and O- (7-azabenzotriazol-1-yl) -1,1,3,3-tetramethyluronium (HATU), O-benzotriazol-1-yl-N, N, N ', N'-tetramethyluronium hexafluorophosphate (HBTU), N, N'-dicyclohexyl (DCC) or N, N'-diisopropanol carbodiimide (DIC).

Condensation of consecutive amino acids performed in the present invention can be carried out directly or manually using an automatic peptide synthesizer as is well known in the relevant art. Preferred synthetic reaction conditions include deprotection of the α-amino acid protected with the Fmoc group by treatment with a secondary amine solution, preferably piperidine, followed by washing with a sufficient excess of the solvent, Followed by 3 to 7-fold molar excess of the amino acid, preferably in a DMF solvent.

Synthesis of peptides using the solid phase resin of the present invention In the final step, the peptides to be obtained from the resin can be removed successively or in a single operation to deprotect the protective groups protecting the amino acid residues, respectively. The removal of the peptide from the resin and the deprotection conditions of the protecting groups present in the residue generally include a cleavage reagent cocktail that cleaves the bond between the resin and the peptide, such as trifluoroacetic acid (TFA), triisopropylsilane (TIS), thioanisole, water or ethanedithiol (EDT), or the like. The resulting mixed solution can be precipitated by treating excess refrigerated diethyl ether solvent. The precipitate thus obtained was centrifuged to complete precipitation, excess trifluoroacetic acid, triisopropylsilane, thioanisole, water and ethanedithiol were firstly removed and the above procedure was repeated twice or more to obtain a solidified precipitate Can be obtained. At this time, the completely deprotected peptide salt can be separated and purified using a mixed solvent composed of water and an acetonitrile solvent and reverse phase high performance liquid chromatography (HPLC). The purified and purified peptide solution can be completely concentrated and dried using lyophilization to obtain a solid peptide.

The peptides of the present invention have collagen synthesis effect.

According to a preferred embodiment of the present invention, the composition of the present invention can be provided as a pharmaceutical composition for improving wrinkles or a cosmetic composition.

When the composition of the present invention is made from a pharmaceutical composition, the pharmaceutical composition of the present invention includes a pharmaceutically acceptable carrier. Such pharmaceutically acceptable carriers are those conventionally used in the field of the present invention and include lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia rubber, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose But are not limited to, polyvinylpyrrolidone, cellulose, purified water, syrup, methylcellulose, methylhydroxybenzoate, propylhydroxybenzoate, calcium stearate, mineral oil and the like. The pharmaceutical composition of the present invention may further contain additives commonly used such as lubricants, wetting agents, sweeteners, flavors, emulsifying agents, suspending agents, preservatives and the like in addition to the above components. The pharmaceutical composition of the present invention is preferably parenterally administered, and more preferably, is applied by a local administration method by application.

The appropriate dosage of the pharmaceutical composition of the present invention may vary depending on such factors as formulation method, administration method, age, body weight, sex, pathological condition, food, administration time, route of administration, excretion rate, .

The dose of the peptide as an active ingredient contained in the pharmaceutical composition of the present invention is 0.001 to 100 mgkg, preferably 0.1 to 100 mg / kg, more preferably 1 to 50 mg / kg on an adult basis, May be administered once or several times a day. The peptides of the present invention may be contained in an amount of preferably 0.0001 to 1.0% by weight, more preferably 0.001 to 1.0% by weight, and most preferably 0.001 to 0.01% by weight, based on the total weight of the pharmaceutical composition But is not limited thereto.

The cosmetic composition of the present invention contains not only the peptide as an active ingredient but also the components commonly used in cosmetic compositions, and includes conventional additives such as antioxidants, stabilizers, solubilizers, vitamins, pigments and perfumes, . As the carrier, there may be mentioned water, purified water, monohydric alcohols (ethanol or propyl alcohol), polyhydric alcohols (glycerol, 1,3-butylene glycol or propylene glycol), higher fatty acids (palmitic acid or linoleic acid) But are not limited to, fats and oils (such as wheat germ oil, camellia oil, jojoba oil, olive oil, squalane, sunflower oil, macadamia peanut oil, avocado oil or fatty acid glycerides). If necessary, a surfactant, a moisturizer, a preservative, an antioxidant and the like may be added.

Examples of the surfactant that can be used in the cosmetic composition of the present invention include anionic surfactants such as alkylbenzenesulfonates, polyoxyalkylene alkylsulfate ester salts, alkylsulfate ester salts, olefin sulfonates, alkyl phosphates, polyoxyalkylene Alkyl ether phosphates, dialkyl sulfosuccinates, fatty acid salts, and the like. Nonionic surfactants include polyoxyethylene alkyl ethers, polyoxyethylene fatty acid esters, polyhydric alcohol fatty acid partial esters, polyoxyethylene polyhydric alcohol fatty acid moieties Esters, polyglycerin fatty acid esters, polyoxyethylene hydrogenated castor oil derivatives, and fatty acid diethanolamides. Examples of the cationic surfactant include tertiary aliphatic amine salts, alkyltrimethylammonium halides, and dialkyldimethylammonium halides. Examples of amphoteric surfactants include amide betaine, imidazolinium betaine, sulfobetaine, Dolls, and the like. Examples of the moisturizing agent include glycerin, propylene glycol, 1,3-butylene glycol, dipropylene glycol, and sorbitol. Examples of the preservative include benzoic acid, dehydroacetic acid, paraoxybenzoic acid esters (such as methyl parahydroxybenzoate, butyl parahydroxybenzoate), and phenoxyethanol. Examples of the antioxidant include ascorbic acid and BHA. In addition, an ultraviolet absorber, an anti-inflammatory agent and a refreshing agent may be added.

The peptide of the present invention may be contained in an amount of preferably 0.0001 to 1.0% by weight, more preferably 0.001 to 1.0% by weight, and most preferably 0.001 to 0.01% by weight, based on the total weight of the cosmetic composition, but is not limited thereto .

The kind of the cosmetic material is not particularly limited, and examples thereof include skin care cosmetic materials such as lotion, milky lotion, gel, cream, essence, pack, ampoule, lotion, cleanser, soap, body products, soap, oil, Makeup cosmetics such as foundation, and the like, and the formulations thereof are not particularly limited.

The cosmetic composition of the present invention can be used everyday or can be used for an unspecified period. Preferably, the amount of usage, the number of times of use, and the period of time can be adjusted according to the age, skin condition or skin type of the user, and the concentration of the peptide.

According to the present invention, it has been confirmed that a composition containing as an active ingredient a heptapeptide monomer and a dimer excellent in skin stability without skin irritation exhibits an effect of improving collagen biosynthesis and a wrinkle-reducing effect due to inhibition of collagenase-producing MMP-1 production And thus it is used as a skin pharmaceutical composition and a cosmetic composition having excellent efficacy for preventing skin aging and improving wrinkles.

1 is a graphical representation of the effect of the compounds of the present invention on increasing collagen synthesis.
2 is a graphical representation of the inhibitory effect of the compounds of the present invention on MMP-1 production.
Figure 3 illustrates the cell viability of the compounds of the present invention.
FIG. 4 is a graphical representation of the effect of improving the wrinkles of the eyes of a composition containing the compound of the present invention.

Hereinafter, preferred embodiments of the present invention will be described in detail. However, the present invention is not limited to the embodiments described herein but may be embodied in other forms. Rather, the teachings herein are thorough and complete, and are provided to enable those skilled in the art to convey the ideas of the invention in sufficient detail.

≪ Example 1-1. Synthesis of Compound 1 >

Nomenclature and abbreviations of amino acids used in the present invention are expressed as follows.

Ala: alanine / Cys: cysteine / Gly: glycine / Val: valine / Pro: proline / Phe: phenylalanine / Met: methonein / Trp: tryptophan / Glu: glutamine /

71.4 mg (0.10 mmol) of 2-chlorotrityl chloride resin (resin loaded with 1.4 mmol per gram) purchased from Novabiochem corporation was weighed into a reaction vessel. The resin was solubilized with 3 ml of DMF and sufficiently swelled for 5 minutes, then 3 ml of a 20% (w / v) piperidine DMF solution was added, shaking for 20 minutes, and a solution of piperidine DMF After removing, it was washed 5 times with 10 ml of DMF solvent (washing 5 times in 10 ml). * DMF: Dimethylformamide.

Fmoc-Gly (Trt) -OH (468.6 mg, 0.80 mmol), HOBt (108.1 mg, 0.80 mmol) and DIC (0.124 mL, 0.80 mmol) were completely dissolved in 2 mL of DMF solvent and then added to the resin. The reaction solution was shaken at room temperature for 8 hours and then washed 5 times with 10 ml of DMF solvent. 3 ml of a 20% (w / v) piperidine DMF solution was added, shaking the mixture for 10 minutes, removing the piperidine solution, adding 20% (w / v) piperidine DMF solution, After the reaction, the Fmoc protecting group protected by the resin was completely removed and washed 5 times with 10 ml of DMF solvent (5 times in 10 ml portions). At this stage, the deprotection of the Fmoc protecting group was carried out using the Kaiser test [E. Kaiser et al. Anal. Biochem., 1970, 34 (2), 595-598). * Fmoc: 9-fluorenylmethyloxycarbonyl / HOBt: 1-hydroxybenzotriazole / DIC: N, N'-diisopropylcarbodiimide.

Next, the peptides were continuously condensed (coupled) according to the same synthesis cycle as described below.

(1) washing 5 times with DMF solvent (10 ml);

(2) deprotection twice with 10% (w / v) piperidine DMF solution (3 ml) for 10 minutes;

(3) washing 5 times with DMF solvent (10 ml);

(4) Fmoc-amino acid addition;

(5) Amino acid activation and 2 hour condensation by addition of condensation reagent;

(6) washed 5 times with DMF solvent (10 ml);

The above steps (1) to (6) were repeated, wherein the Fmoc-protected amino acid (0.80 mmol) after Fmoc-Cys (Trt) -OH was added to the resin reaction vessel in the sequence described below and condensed .

(i) Fmoc-Arg-OH;

(ii) Fmoc-Leu-OH;

(iii) Fmoc-Gln-OH;

(iv) Fmoc-Met-OH;

(v) Fmoc-Glu-OH;

(VI) Fmoc-Glu-OH;

(VII) Fmoc-Cys (Trt) -OH;

After (7) after Fmoc-Cys (Trt) -OH condensation, finally, 20% piperidine DMF solution (3 ml) was treated.

Immediately after such synthesis termination, the peptide condensed resin was treated with a mixture of trifluoroacetic acid / thioanisole / ethanediol / triisopropylsilane / water (95: 5: 2.5: 2.5: 2.5) (10 ml), and the peptide was cleaved from the resin. The thus obtained mixed solution was treated with 100 ml of refrigerated diethyl ether solvent to produce a precipitate. The obtained precipitate was centrifuged to completely precipitate, and then trifluoroacetic acid, thioanisole and ethanedithiol were firstly removed, and 100 ml of a diethyl ether solvent was added to wash the precipitate, followed by centrifugation, , A process for removing trifluoroacetic acid, thioanisole and ethanedithiol) was repeated twice to obtain a solidified precipitate. The precipitate (peptide) was purified by HPLC using a 5% to 100% acetonitrile / water gradient solvent system containing 0.01% trifluoroacetic acid over 50 minutes using a C-18 column. The pure purified fraction was lyophilized to obtain collagen synthesis promoting peptide H ₂ N- [Cys-Glu-Glu-Met-Gln-Leu-Arg] -CO ₂ H (100 mg) as a white powder type trifluoroacetate salt .

Compound 1: H ₂ N- [Cys-Glu-Glu-Met-Gln-Leu-Arg] -CO ₂ H

MS (ESI) m / e, [M + H] < ⁺ > = 1039.35; (100 mg)

<Examples 1-2. Synthesis of Compounds 2 to 15>

Using the same manufacturing procedure as in Example 1-1, peptides of the following compounds 2 to 15 were prepared by changing the order of amino acids protected with Fmoc.

Compound 2: H ₂ N- [Cys-Glu-Glu-Met-Gln-Phe-Arg] -CO ₂ H

MS (ESI) m / e, [M + H] &lt; ⁺ &gt; = 1073.38; (110 mg)

Compound 3: H ₂ N- [Cys-Glu-Glu-Met-Gln-Ser-Arg] -CO ₂ H

MS (ESI) m / e, [M + H] &lt; ⁺ &gt; = 1013.28; (105 mg)

Compound 4: H ₂ N- [Cys-Glu-Glu-Met-Gln-Tyr-Arg] -CO ₂ H

MS (ESI) m / e, [M + H] &lt; ⁺ &gt; = 1089.38; (107 mg)

Compound 5: H ₂ N- [Cys-Glu-Glu-Met-Gln-Arg-Arg] -CO ₂ H

MS (ESI) m / e, [M + H] &lt; ⁺ &gt; = 1082.39; (102 mg)

Compound 6: H ₂ N- [Cys-Glu-Glu-Met-Gln-Asp-Arg] -CO ₂ H

MS (ESI) m / e, [M + H] &lt; ⁺ &gt; = 1041.29; (101 mg)

Compound 7: H ₂ N- [Cys-Glu-Glu-Met-Gln-Lys-Arg] -CO ₂ H

MS (ESI) m / e, [M + H] &lt; ⁺ &gt; = 1054.38; (105 mg)

Compound 8: H ₂ N- [Cys-Glu-Glu-Met-Gln-Asn-Arg] -CO ₂ H

MS (ESI) m / e, [M + H] &lt; ⁺ &gt; = 1040.31; (180 mg)

Compound 9: H ₂ N- [Glu-Gly-Met-Gln-Arg-Arg-Cys] -CO ₂ H

MS (ESI) m / e, [M + H] &lt; ⁺ &gt; = 1010.33; (101 mg)

Compound 10: H ₂ N- [Glu-Leu-Met-Gln-Arg-Arg-Cys] -CO ₂ H

MS (ESI) m / e, [M + H] &lt; ⁺ &gt; = 1066.42; (110 mg)

Compound 11: H ₂ N- [Glu-Ser-Met-Gln-Arg-Arg-Cys] -CO ₂ H

MS (ESI) m / e, [M + H] &lt; ⁺ &gt; = 1040.35; (106 mg)

Compound 12: H ₂ N- [Glu-Tyr-Met-Gln-Arg-Arg-Cys] -CO ₂ H

MS (ESI) m / e, [M + H] &lt; ⁺ &gt; = 1116.45; (107 mg)

Compound 13: H ₂ N- [Glu-Asp-Met-Gln-Arg-Arg-Cys] -CO ₂ H

MS (ESI) m / e, [M + H] &lt; ⁺ &gt; = 1068.36; (106 mg)

Compound 14: H ₂ N- [Glu-Lys-Met-Gln-Arg-Arg-Cys] -CO ₂ H

MS (ESI) m / e, [M + H] &lt; ⁺ &gt; = 1081.45; (102 mg)

Compound 15: H ₂ N- [Glu-Asn-Met-Gln-Arg-Arg-Cys] -CO ₂ H

MS (ESI) m / e, [M + H] &lt; ⁺ &gt; = 1067.38; (103 mg)

<Examples 1-3. Synthesis of Compound 21 >

The collagen synthesis promoting peptide H ₂ N- [Cys-Glu-Glu-Met-Gln-Leu-Arg] -CO ₂ H (50 mg) was dissolved in 2 ml of dimethylsulfoxide (DMSO) And stirred at room temperature for 3 days to obtain a final collagen synthesis promoting peptide (50 mg) in the form of a dimer in which the cysteine residue thiol group (-SH) was modified to a reduced form (-SS-).

Compound 21: H ₂ N- [Cys-Glu-Glu-Met-Gln-Leu-Arg] ₂ -CO ₂ H

MS (ESI) m / e, [M + H] &lt; ⁺ &gt; = 2078.7; (50 mg)

<Examples 1-4. Synthesis of Compounds 22 to 35 >

Using the same procedure as in Example 1-3, peptides of the following compounds 22 to 35 were prepared by changing the order of amino acids protected by Fmoc.

Compound 22: H ₂ N- [Cys-Glu-Glu-Met-Gln-Phe-Arg] ₂ -CO ₂ H

MS (ESI) m / e, [M + H] &lt; ⁺ &gt; = 2146.76; (47 mg)

Compound 23: H ₂ N-Cys-Glu-Glu-Met-Gln-Ser-Arg] ₂ -CO ₂ H

MS (ESI) m / e, [M + H] &lt; ⁺ &gt; = 2178.76; (48 mg)

Compound 24: H ₂ N- [Cys-Glu-Glu-Met-Gln-Tyr-Arg] ₂ -CO ₂ H

MS (ESI) m / e, [M + H] &lt; ⁺ &gt; = 2178.76; (44 mg)

Compound 25: H ₂ N- [Cys-Glu-Glu-Met-Gln-Arg-Arg] ₂ -CO ₂ H

MS (ESI) m / e, [M + H] &lt; ⁺ &gt; = 2164.78; (44 mg)

Compound 26: H ₂ N- [Cys-Glu-Glu-Met-Gln-Asp-Arg] ₂ -CO ₂ H

MS (ESI) m / e, [M + H] &lt; ⁺ &gt; = 2082.58; (47 mg)

Compound 27: H ₂ N- [Cys-Glu-Glu-Met-Gln-Lys-Arg] ₂ -CO ₂ H

MS (ESI) m / e, [M + H] &lt; ⁺ &gt; = 2108.76; (46 mg)

Compound 28: H ₂ N- [Cys-Glu-Glu-Met-Gln-Asn-Arg] ₂ -CO ₂ H

MS (ESI) m / e, [M + H] &lt; ⁺ &gt; = 2080.62; (48 mg)

Compound 29: H ₂ N- [Glu-Gly-Met-Gln-Arg-Arg-Cys] ₂ -CO ₂ H

MS (ESI) m / e, [M + H] &lt; ⁺ &gt; = 2020.66; (50 mg)

Compound 30: H ₂ N- [Glu-Leu-Met-Gln-Arg-Arg-Cys] ₂ -CO ₂ H

MS (ESI) m / e, [M + H] &lt; ⁺ &gt; = 2132.84; (52 mg)

Compound 31: H ₂ N- [Glu-Ser-Met-Gln-Arg-Arg-Cys] ₂ -CO ₂ H

MS (ESI) m / e, [M + H] &lt; ⁺ &gt; = 2080.7; (51 mg)

Compound 32: H ₂ N- [Glu-Tyr-Met-Gln-Arg-Arg-Cys] ₂ -CO ₂ H

MS (ESI) m / e, [M + H] &lt; ⁺ &gt; = 2232.9; (50 mg)

Compound 33: H ₂ N- [Glu-Asp-Met-Gln-Arg-Arg-Cys] ₂ -CO ₂ H

MS (ESI) m / e, [M + H] &lt; ⁺ &gt; = 2136.72; (56 mg)

Compound 34: H ₂ N- [Glu-Lys-Met-Gln-Arg-Arg-Cys] ₂ -CO ₂ H

MS (ESI) m / e, [M + H] &lt; ⁺ &gt; = 2162.9; (59 mg)

Compound 35: H ₂ N- [Glu-Asn-Met-Gln-Arg-Arg-Cys] ₂ -CO ₂ H

MS (ESI) m / e, [M + H] &lt; ⁺ &gt; = 2134.76; (52 mg)

&Lt; Examples 1-5. Synthesis of Compound 16 >

DMF (4 ML), 1.06 mL (10 eq) of stearyl chloride and 1.96 mL (10 eq) of DIPEA were added to 1 peptide of 5 compounds selected from the above 15 peptides before the peptide was cleaved from the resin. DMF: Dimethylformamide The reaction was completed by Kaiser test. The lipid and peptide-condensed resin was dissolved in trifluoroacetic acid / thio (3-hydroxyphenyl) thiourea for 3 hours. The peptide was cleaved from the resin by using (10 ml) a mixture of anisole / ethanedithiol / triisopropylsilane / water (95: 5: 2.5: 2.5: 2.5). The thus obtained mixed solution was treated with 100 ml of refrigerated diethyl ether solvent to produce a precipitate. The obtained precipitate was centrifuged to completely precipitate, and then trifluoroacetic acid, thioanisole and ethanedithiol were firstly removed, and 100 ml of a diethyl ether solvent was added to wash the precipitate, followed by centrifugation, , A process for removing trifluoroacetic acid, thioanisole and ethanedithiol) was repeated twice to obtain a solidified precipitate. The precipitate (peptide) was purified by HPLC using a 5% to 100% acetonitrile / water gradient solvent system containing 0.01% trifluoroacetic acid over 50 minutes using a C-18 column. By freeze-drying the purified fractions of collagen synthesis as acetate, trifluoroacetate type of white powder peptide promotes Stearyl- [Cys-Glu-Glu- Met-Gln-Leu-Arg] -CO 2 H (102㎎) was obtained. Compounds 16 to 20 were synthesized in this manner.

* Types of lipids

I) Stearic acid

Ii) Palmitic acid

Iii) arachidic acid

Iv) Oleic acid

V) Erucic acid

Vi) Linoleic acid

Ⅶ) Linolenic acid

The collagen synthesis performance and the mmp-1 synthesis inhibitory activity of the 15 compounds of Examples 1-1 and 1-2 were examined, and the top five collagen synthesis rates were selectively applied with lipid.

Compound 16: Stearyl- [Cys-Glu- Glu-Met-Gln-Leu-Arg] -CO 2 H

Compound 17: Erucyl- [Cys-Glu- Glu-Met-Gln-Tyr-Arg] -CO 2 H

Compound 18: Palmitoyl- [Cys-Glu- Glu-Met-Gln-Arg-Arg] -CO 2 H

Compound 19: Linolenyl- [Cys-Glu- Glu-Met-Gln-Lys-Arg] -CO 2 H

Compound 20: Linoleyl- [Glu-Asp- Met-Gln-Arg-Arg-Cys] -CO 2 H

<Examples 1-6. Synthesis of Compound 36 >

Embodiment the collagen synthesis promotion obtained in Example 1-5 peptide Stearyl- of [Cys-Glu-Glu-Met -Gln-Leu-Arg] -CO 2 H (53mg) was dissolved in methyl sulfoxide (DMSO) 2㎖ 10㎖ Water was added and stirred at room temperature for 3 days to obtain a final collagen synthesis promoter peptide Stearyl- [Cys (-S-S-)] in which the cysteine residue thiol group (-SH) -Glu-Glu-Met-Gln- Leu-Arg] 2 -CO 2 to obtain H (50㎎). Compounds 36 to 40 were synthesized in this manner.

Compound 36: Stearyl- [Cys-Glu- Glu-Met-Gln-Leu-Arg] 2 -CO 2 H

Compound 37: Erucyl- [Cys-Glu- Glu-Met-Gln-Tyr-Arg] 2 -CO 2 H

Compound 38: Palmitoyl- [Cys-Glu- Glu-Met-Gln-Arg-Arg] 2 -CO 2 H

Compound 39: Linolenyl- [Cys-Glu- Glu-Met-Gln-Lys-Arg] 2 -CO 2 H

Compound 40: Linoleyl- [Glu-Asp- Met-Gln-Arg-Arg-Cys] 2 -CO 2 H

Through experiments of collagen synthesis and mmp-1 synthesis inhibition of 15 compounds of Examples 1-3 and 1-4, experiments were carried out by selectively attaching lipids to the top five collagen synthesis ratios. Respectively.

Experimental Example  One : Of peptide  Measurement of promoting collagen synthesis in vitro )

0.5 ml of human dermal fibroblast (HDF: Human dermal fibroblast) was dispensed into a 24-well plate at a concentration of 1.0 ⁴ cells / ml and cultured for 1 day at 37 ° C, 5% CO ₂ , and humidified conditions. The culture medium used was a medium containing 10% FBS (Gibco) in DMEM medium (Dulbecco's Modified Eagle's Medium, manufactured by Gibco) in an amount of 0.5 ml per well.

Subsequently, the cells were exchanged with DMEM (Dulbecco's Modified Eagle's Medium, Sigma) not containing FBS (Gibco), washed again with 0.5 ml of PBS (Phosphate Buffered Saline, Sigma) uM treatment. 100 占 퐇 of PBS in which the compound was not dissolved was used as a control.

In the collagen production acceleration test, the culture solution was collected for 2 days, and the concentration of type I procollagen secreted in the culture solution was measured by enzyme-linked immunosorbent assay (ELSIA, Procollagen type I c-peptide EIA kit, manufactured by R & D system) Respectively.

Based on the quantitative results, the amount (pg / ml) of type I procollagen synthesized was measured and the type I procollagen production rate was calculated according to the following equation (1). The results are shown in Table 1 and FIG.

[Equation 1]

Type I procollagen production rate (%) = (amount of test type I procollagen / amount of control type I procollagen) * 100

Comparison of collagen production rate of each compound division Skin fibroblast
Collagen production (%) division Skin fibroblast
Collagen production (%) Compound 1 159.8 Compound 21 186.3 Compound 2 158.4 Compound 22 180.2 Compound 3 157.7 Compound 23 183.7 Compound 4 169.9 Compound 24 192.8 Compound 5 162.4 Compound 25 189.2 Compound 6 157.3 Compound 26 179.6 Compound 7 160.1 Compound 27 188.6 Compound 8 159.1 Compound 28 180.1 Compound 9 143.2 Compound 29 185.9 Compound 10 141.6 Compound 30 179.9 Compound 11 141.6 Compound 31 178.5 Compound 12 148.8 Compound 32 182.3 Compound 13 167.5 Compound 33 195.4 Compound 14 149.7 Compound 34 185.8 Compound 15 148.3 Compound 35 183.0 Compound 16 130.2 Compound 36 172.3 Compound 17 131.4 Compound 37 172.1 Compound 18 127.6 Compound 38 168.9 Compound 19 128.9 Compound 39 166.6 Compound 20 130.1 Compound 40 162.7

Referring to Table 1 and FIG. 1, it was confirmed that the compounds 1 to 40 of the present invention all exhibited an effect of promoting collagen production, and exhibited sufficient effects for preventing skin wrinkles.

On the other hand, it is very important to slow down the degradation rate of the peptide, since the peptides are rapidly degraded by various hydrolytic enzymes when they enter the living body and its ability is rapidly reduced. In general, various hydrolytic enzymes decompose an amide bond (-C (= O) NH-) site in a peptide structure into a carboxylic acid (-CO2H) and an amine (-NH2) The dimer structure has a (-SS-) backbone that functions to lower or disturb the degrading ability of common hydrolytic enzymes. That is, among the above peptide compounds, the compound 21 to 40 in the form of a dendritic form is less decomposed by hydrolytic enzymes in vivo than the compounds 1 to 20 in the monomer form. Therefore, among the peptide compounds of the present invention, compounds 21 to 40 having cysteine dimers are considered to have excellent stability in the body. Furthermore, lipidated peptides 16-20 and 36-40 were similar in promoting collagen production to peptides that were not.

Experimental Example  2 : Of peptide MMP -1 Production inhibition effect measurement ( in vitro )

0.5 ml of human dermal fibroblast (HDF: Human dermal fibroblast) was dispensed into a 24-well plate at a concentration of 1.0 ⁴ cells / ml and cultured for 1 day at 37 ° C, 5% CO ₂ , and humidified conditions. The culture medium used was a medium containing 10% FBS (Gibco) in DMEM medium (Dulbecco's Modified Eagle's Medium, manufactured by Gibco) in an amount of 0.5 ml per well.

Subsequently, the cells were exchanged with DMEM (Dulbecco's Modified Eagle's Medium, Sigma) not containing FBS (Gibco), washed again with 0.5 ml of PBS (Phosphate Buffered Saline, Sigma) uM treatment. 100 占 퐇 of PBS in which the compound was not dissolved was used as a control.

The concentration of MMP-1 secreted in the culture medium was quantitated by enzyme-linked immunosorbent assay (ELSIA, MMP-1 EIA Kit; manufactured by R & D system) after culturing for 3 days in the inhibition rate of MMP- Respectively.

Based on the quantitative results, the amount of synthesized MMP-1 (pg / ml) was measured and the inhibition rate of MMP-1 production was calculated according to the following formula (2). The results are shown in Table 2 and FIG.

&Quot; (2) &quot;

Inhibition rate of MMP-1 production (%) = (amount of MMP-1 in the experimental group / amount of MMP-1 in the control) * 100

Comparison of MMP-1 inhibition rate of each compound division Skin fibroblast
Inhibition rate of MMP-1 production (%) division Skin fibroblast
Inhibition rate of MMP-1 production (%) Compound 1 69.8 Compound 21 52.8 Compound 2 70.0 Compound 22 54.3 Compound 3 71.4 Compound 23 54.3 Compound 4 64.0 Compound 24 49.7 Compound 5 68.2 Compound 25 52.1 Compound 6 69.9 Compound 26 55.8 Compound 7 67.7 Compound 27 54.0 Compound 8 70.3 Compound 28 60.2 Compound 9 70.3 Compound 29 60.7 Compound 10 73.5 Compound 30 62.4 Compound 11 73.8 Compound 31 62.6 Compound 12 73.7 Compound 32 64.0 Compound 13 61.9 Compound 33 50.5 Compound 14 70.1 Compound 34 54.9 Compound 15 74.3 Compound 35 58.9 Compound 16 67.6 Compound 36 62.1 Compound 17 65.8 Compound 37 60.0 Compound 18 70.4 Compound 38 60.3 Compound 19 73.0 Compound 39 55.8 Compound 20 72.1 Compound 40 57.2

Referring to Table 2 and FIG. 2, it was confirmed that the compounds 1 to 40 of the present invention all had an excellent inhibitory effect on MMP-1 production, and it was confirmed that the compounds 1 to 40 exhibited sufficient effect against the skin wrinkle on the basis thereof. Of the many peptide compounds, the lipidated peptides 16-20 and 36-40 were less effective at inhibiting MMP-1 production than the peptides.

Experimental Example  3: Collagen production Of peptide  Safety check ( in vitro )

100 μl of human dermal fibroblast (HDF: Human dermal fibroblast, manufactured by Thermo) was dispensed into a 96-well plate at a concentration of 1.0 ³ cells / ml and cultured for 1 day at 37 ° C and 5% CO ₂ under humidified conditions . The culture medium used was 100 μl of a medium containing 10% of FBS (manufactured by Gibco) in DMEM medium (Dulbecco's Modified Eagle's Medium, manufactured by Gibco).

Subsequently, the cells were exchanged with DMEM (Dulbecco's Modified Eagle's Medium, Sigma) containing no FBS (Gibco), and further washed with 100 μl of PBS (Phosphate Buffered Saline, Sigma) And cultured. 100 占 퐇 of PBS in which the compound was not dissolved was used as a control.

After the test sample was added, it was cultured for 24 hours, treated with MTT and cultured for 3 hours. The culture was then removed and the absorbance was measured at 540 nm with DMSO. Based on the measured values, the cell viability was calculated according to Equation 3. The results are shown in Table 3 and Fig.

&Quot; (3) &quot;

Cell survival rate (%) = (absorbance at 540 nm in the experimental group / absorbance at 540 nm in the control) * 100

Cytotoxicity of each compound division Cell survival rate (%) division Cell survival rate (%) Control group 100 Control group 100 Compound 1 98.2 Compound 21 99.2 Compound 2 99.1 Compound 22 97.6 Compound 3 98.8 Compound 23 97.7 Compound 4 99.0 Compound 24 100.0 Compound 5 98.4 Compound 25 96.9 Compound 6 98.2 Compound 26 96.2 Compound 7 98.2 Compound 27 100.7 Compound 8 97.7 Compound 28 98.4 Compound 9 97.4 Compound 29 99.6 Compound 10 97.0 Compound 30 99.7 Compound 11 99.2 Compound 31 99.3 Compound 12 98.1 Compound 32 99.0 Compound 13 96.9 Compound 33 99.0 Compound 14 99.2 Compound 34 98.4 Compound 15 100.4 Compound 35 97.7 Compound 16 95.8 Compound 36 100.3 Compound 17 97.3 Compound 37 100.1 Compound 18 97.1 Compound 38 99.2 Compound 19 97.7 Compound 39 98.5 Compound 20 98.3 Compound 40 95.9

Referring to Table 3 and FIG. 3, all of the compounds of the present invention showed cell viability of 95% or more when compared with the control group. From the results of this experiment, it was confirmed that the compounds of the present invention were safe and confirmed as safe materials in animal cells containing human dermal fibroblasts.

Experimental Example  4: Manufacture of Cosmetic Formulation

In order to evaluate the effects of the new heptapeptide monosaccharide and the cosmetic comprising the dimer, which are the collagen synthesis promoting peptide, the ingredients as shown in Table 4 below were blended to prepare a cream form cosmetic formulation.

Formulation of cream to evaluate wrinkle improvement effect ingredient Experimental Example 1 (% by weight) Comparative Example 1 (% by weight) Heptapeptide monosaccharide 0.01 - Heptapeptide dimer 0.01 - glycerin 5.0 5.0 Capric glyceride 4.5 4.5 Mineral oil 3.5 3.5 Cetyl alcohol 2.0 2.0 vaseline 2.0 2.0 Stearic acid 1.5 1.5 Polysorbate 60 1.5 1.5 FAGE -100 stearate 1.0 1.0 Wax 1.0 1.0 Glyceryl monostearate 0.5 0.5 Triethanolamine 0.2 0.2 Preservative, coloring, fragrance Suitable amount Suitable amount Purified water All remaining capacity All remaining capacity Sum 100.0 100.0

Experimental Example  5: Eye wrinkle improvement effect Human body application test ( PRIMOS  High Resolution)

In order to confirm the wrinkle-reducing effect of the cosmetic composition according to the present invention, 22 Korean women aged 35 to 55 years were evaluated by the same method as routine cream use for each 4 weeks. The evaluation was performed by using the PRIMOS High Resolution (Phaseshift Rapid In Vivo Measurement of Skin high resolution, GFMesstechinik GmbH, Germany). The values of the eye wrinkle variables through the roughness analysis are as follows.

- Ra (Average roughness): Average of the absolute value of the length from the center line to the section curve of the surface when drawing the center line at the roughness height of the section.

- Root mean square roughness (Rq): Square root mean square of the length from the centerline to the surface curvature of the surface when drawing the center line at the roughness height of the section.

- Rmax (Maximum roughness depth): The difference between the highest acid and lowest bone within a single measurement range.

The decrease in the value of the parameter analyzed by the roughness measurement means that the wrinkles in the eye are improved, and the rate of decrease in the value of the roughness parameter due to the PRIMOS High Resolution is calculated according to Equation (4). The calculated values for each variable are shown in Table 5 and FIG.

&Quot; (4) &quot;

Decrease rate (%) = (Measured value before applying sample - Measured value after applying sample / Measured value before applying sample) * 100

Test results for human eye wrinkles Roughness variable After 2 weeks of application After 4 weeks of application Ra 4.103 6.847 Rq 2.986 5.871 Rmax 1.395 4.624

As a result of analyzing the measured value of the roughness of the eye wrinkle area using PRIMOS High Resolution, the Ra value after using the cream of Experimental Example 1 was statistically significant (p <0.05) (P <0.05), which was 2.986% and 5.871%, respectively, after 2 weeks and 4 weeks after application, respectively. In addition, Rmax value was statistically significant (p <0.05) compared to before application of sample, and decreased by 4.624% after 4 weeks of application. Therefore, the cream of Experimental Example 1 was found to be effective for improving the wrinkles of the eye after 2 weeks of application and after 4 weeks of application.

It will be apparent to those skilled in the art that various modifications and variations can be made in the present invention without departing from the spirit and scope of the invention as set forth in the appended claims. And modifications are within the scope of the appended claims. It is therefore to be understood that the above-described embodiments and examples are illustrative in all respects and not restrictive.

Claims (7)

The collagen synthesis promoting peptide corresponding to the following compounds 1 to 15 and 21 to 35

[Cys-Glu-Glu-Met-Gln-Leu-Arg] (Compound 1)
[Cys-Glu-Glu-Met-Gln-Phe-Arg] (Compound 2)
[Cys-Glu-Glu-Met-Gln-Ser-Arg] (Compound 3)
[Cys-Glu-Glu-Met-Gln-Tyr-Arg] (Compound 4)
[Cys-Glu-Glu-Met-Gln-Arg-Arg] (Compound 5)
[Cys-Glu-Glu-Met-Gln-Asp-Arg] (Compound 6)
[Cys-Glu-Glu-Met-Gln-Lys-Arg] (Compound 7)
[Cys-Glu-Glu-Met-Gln-Asn-Arg] (Compound 8)
[Glu-Gly-Met-Gln-Arg-Arg-Cys] (Compound 9)
[Glu-Leu-Met-Gln-Arg-Arg-Cys] (Compound 10)
[Glu-Ser-Met-Gln-Arg-Arg-Cys] (Compound 11)
[Glu-Tyr-Met-Gln-Arg-Arg-Cys] (Compound 12)
[Glu-Asp-Met-Gln-Arg-Arg-Cys] (Compound 13)
[Glu-Lys-Met-Gln-Arg-Arg-Cys] (Compound 14)
[Glu-Asn-Met-Gln-Arg-Arg-Cys] (Compound 15)
[Cys-Glu-Glu-Met-Gln-Leu-Arg] ₂ (Compound 21)
[Cys-Glu-Glu-Met-Gln-Phe-Arg] ₂ (Compound 22)
[Cys-Glu-Glu-Met-Gln-Ser-Arg] ₂ (Compound 23)
[Cys-Glu-Glu-Met-Gln-Tyr-Arg] ₂ (Compound 24)
[Cys-Glu-Glu-Met-Gln-Arg-Arg] ₂ (Compound 25)
[Cys-Glu-Glu-Met-Gln-Asp-Arg] ₂ (Compound 26)
[Cys-Glu-Glu-Met-Gln-Lys-Arg] ₂ (Compound 27)
[Cys-Glu-Glu-Met-Gln-Asn-Arg] ₂ (Compound 28)
[Glu-Gly-Met-Gln-Arg-Arg-Cys] ₂ (Compound 29)
[Glu-Leu-Met-Gln-Arg-Arg-Cys] ₂ (Compound 30)
[Glu-Ser-Met-Gln-Arg-Arg-Cys] ₂ (Compound 31)
[Glu-Tyr-Met-Gln-Arg-Arg-Cys] ₂ (Compound 32)
[Glu-Asp-Met-Gln-Arg-Arg-Cys] ₂ (Compound 33)
[Glu-Lys-Met-Gln-Arg-Arg-Cys] ₂ (Compound 34)
[Glu-Asn-Met-Gln-Arg-Arg-Cys] ₂ (Compound 35)
, And [] ₂ in compounds 21 to 35 represents a duplex in which a thiol group (-SH) as a cysteine residue of compounds 1 to 15 is modified to a reduced form (-SS-). .
The method according to claim 1,
Peptides corresponding to each of the 10 lipidated compounds 16 to 20 and 36 to 40, each having excellent collagen aggregation performance and MMP-1 inhibitory ability among the compounds 1 to 15 and 21 to 35 peptides,
Stearyl- [Cys-Glu-Glu- Met-Gln-Leu-Arg] -CO 2 H ( compound 16),
Erucyl- [Cys-Glu-Glu- Met-Gln-Tyr-Arg] -CO 2 H ( compound 17),
Gly-Arg-Arg-CO ₂ H (Compound 18), &lt; RTI ID = 0.0 &
Gly-Lys-Arg-CO ₂ H (Compound 19), Linolenyl- [Cys-Glu-
Linoleyl- [Glu-Asp-Met- Gln-Arg-Arg-Cys] -CO 2 H ( compound 20),
Stearyl- [Cys-Glu-Glu- Met-Gln-Leu-Arg] 2 -CO 2 H ( compound 36),
Erucyl- [Cys-Glu-Glu- Met-Gln-Tyr-Arg] 2 -CO 2 H (Compound 37),
Palmitoyl- [Cys-Glu-Glu- Met-Gln-Arg-Arg] 2 -CO 2 H ( compound 38),
Lys-Arg- _2- CO ₂ H (Compound 39), and Linolenyl- [Cys-Glu-Glu-Met-
Linoleyl- [Glu-Asp-Met- Gln-Arg-Arg-Cys] 2 -CO 2 H ( compound 40)
In which collagen aggregation performance and MMP-1 inhibitory ability are similar to those of compounds 1 to 15 and 21 to 35, respectively.
A composition for preventing or improving skin aging or skin wrinkles, which comprises a peptide selected from one or more peptides according to any one of claims 1 to 2.
A pharmaceutical composition comprising at least one peptide selected from the peptides according to any one of claims 1 to 2.
5. The method of claim 4,
Wherein the peptide is contained in the pharmaceutical composition in an amount of 0.0001 to 1.0% by weight.
A cosmetic composition for improving skin aging or skin wrinkle prevention, characterized by containing at least one peptide selected from the peptides according to any one of claims 1 to 2.
The method according to claim 6,
Wherein the peptide is contained in the cosmetic composition in an amount of 0.0001 to 1.0% by weight. The cosmetic composition for improving wrinkles is characterized in that it contains a lotion, a lotion, a gel, a cream, Wherein the composition is selected from oil, lipstick, and foundation.

Images