KR102007078B1 - Anti-aging cosmetic composition containing novel heptapeptide monomer and dimer that promotes collagen synthesis - Google Patents

Abstract

The present invention relates to a cosmetic composition for preventing or improving skin aging containing a novel heptapeptide monomer and dimer represented by the following formulas 1 to 4 as an active ingredient, and more particularly, a novel heptapeptide monomer and dimer. It inhibits the expression of MMP-1, an important enzyme that causes skin aging, as an active ingredient, inhibits skin collagen degradation and increases the expression of procollagen, thereby preventing and improving skin aging. It relates to a cosmetic composition for. In particular, the heptapeptide monomers and dimers for preventing and improving skin aging can be added to various cosmetic compositions such as skins, lotions, creams, foundations, essences, gels, packs, foam cleansing or soap forms, and have no skin irritation. Skin stability is also excellent.
[Formula 1]
[Cys-Gly-Pro-Gln-X-Pro-Gln]
(2)
Gly-X-Gln-Gly-Pro-Gln-Cys
(3)
[Cys-Gly-Pro-Gln-X-Pro-Gln] ₂
[Chemical Formula 4]
Gly-X-Gln-Gly-Pro-Gln-Cys ₂
(In the formula 1 to 4, X is glutamic acid, aspartic acid, histidine, phenylalanine, alanine, cysteine, glycine, glutamine, asparagine, arginine, leucine, methionine, isoleucine, serine, tyrosine, threonine, lysine, tryptophan Selected from, proline and valine, and [] ₂ may represent a dimer of a thiol group (-SH), which is a cysteine residue, in a reduced form (-SS-).)

Description

Anti-aging cosmetic composition containing novel heptapeptide monomer and dimer that promotes collagen synthesis}

The present invention relates to a cosmetic composition comprising a skin functional heptapeptide monomer and a dimer as an active ingredient, and more particularly, to chemically stable, almost non-toxic and physiologically produced by dimerizing heptapeptide having an anti-aging function. It relates to a composition for preventing, improving or treating skin aging or skin wrinkles, which comprises a dimerized skin wrinkle improvement functional heptapeptide having excellent activity as an active ingredient as a cosmetic ingredient.

Skin aging is a product of a combination of hormones and external factors such as physical age, ultraviolet rays, smoking and pollution. As the standard of living gradually increases and the average life expectancy of humans increases, the tendency to manage external beauty increases, and the interest in the effect of aging on the skin is increasing. The skin is a body part that is in direct contact with the external environment and consists of various cells and structures that serve as an important barrier to protect the body from external factors including trauma, infection, and ultraviolet radiation. The skin is composed of epidermis, dermis and subcutaneous tissue. Epidermis is a thin layer of tissue composed mainly of keratinocytes and pigmented melanocytes and Langerhans cells. The dermis consists mainly of extracellular matrix produced by fibroblasts.

Type-I collagen, the most abundant protein in the extracellular matrix of skin dermis, contains other types of extracellular tissue proteins in addition to other types of collagen, elastin, proteoglycan, and fibronectin. Together they form a connective tissue. The newly produced type I procollagen is secreted into the extracellular space of the dermis and then undergoes fibrilogenesis to produce collagen, which contributes to the elasticity and strength of the skin.

In aging skin, keratinocyte cells and connective tissues are weakened, resulting in thinning of the epidermal and dermal layers. The connective tissue of the skin is basically composed of a bundle of fibrous collagen and elastic fibers. The denaturation of proteins such as collagen and elastin, which gives the skin durability and elasticity, leads to fragile skin and aging appearance.

From a histological point of view, skin aging is a change in the composition of extracellular matrix proteins that form a matrix in the dermis of the skin. Proteins in the dermal extracellular matrix impart strength and tension to the skin, thereby protecting the skin from external irritation or force and accounting for 90% of the dermal layer. It is closely related.

However, the precursors of collagen are significantly reduced in skin damaged by external factors such as photoaging, and the reduction of these proteins is known to be associated with clinical severity. Collagen decomposes as aging progresses and exposure to external stimuli such as ultraviolet light increases the biosynthesis of collagen degrading enzymes (MMPs), which reduces collagen synthesis and reduces the amount of collagen, which causes wrinkles on the skin.

Collagen degrading enzymes (MMPs) are zinc-dependent intracellular proteases that can degrade or restructure components of the extracellular structure of the dermis, of which MMP-1 acts on collagen in the dermis to synthesize collagen. Decreases. Therefore, the regulation of the amount and activity of the enzyme is an important factor for wrinkles.

To date, the most well-known substance that improves aging of skin by preventing wrinkles of skin is retinoid. Retinoids, derivatives of vitamin A, are known to inhibit collagen breakdown and improve skin wrinkles by inhibiting the production of collagen degrading enzyme MMP.

US Pat. No. 6,906,036 describes that healthy peptides can be maintained by inhibiting MMP, a major degrading enzyme of collagen, using peptides isolated from MMPs' proMMP. EP 2,510,919 discloses oleanoyl peptide peptides (Oleanoyl). It is described that pentapeptides are used to inhibit the action of MMPs, thereby improving the wrinkles of the skin.

This prevents the formation of wrinkles by preventing the breakdown of collagen through inhibition of MMP action, but it is difficult to expect a complete wrinkle improvement effect unless it promotes the further production of proteins such as collagen forming the matrix of the skin .

Patent US 6,620,419, filed by Sederma (France), describes the use of peptides at the carboxyl end of collagen to promote the production of collagen and is effective for aging. Patent US 6,974,799 uses a mixture of tripeptide and tetrapeptide. It has been described as showing an improved anti-aging effect by promoting the synthesis of collagen.

However, the above patents cannot expect the inhibitory effect of MMP, a collagen degrading enzyme, and can not expect complete anti-wrinkle effect because it only promotes the production of matrix protein.

Against this background, the inventors have developed heptapeptide monomers and dimers that can effectively inhibit the formation of collagen, which is a matrix protein of the skin, and at the same time, effectively inhibit MMP-1, which promotes the decomposition of the matrix protein. In order to confirm the anti-aging and anti-wrinkle effect of the heptapeptide monomer and dimer of the present invention, the in vitro assay using human dermal fibroblast was established to biosynthetic ability of collagen and collagen degrading enzymes (MMPs). The present invention was completed by confirming the anti-aging effect of the compound heptapeptide monomer and dimer.

US 6,906,036 EP 2,510,919 US 6,620,419 US 6,974,799

[Non-Patent Document 1] Pilcher BK, Sudbeck BD, Dumin JA, Welgus HG, Parks WC, Collagenase-1 and collagen in epidermal repair: Arch. Dermatol. Res., 290 (Suppl.): S37-S46 (1998) [Non-Patent Document 2] Griffiths CEM, Russman AN, Majmudar G, Singer RS, Hamilton TA, Voorhees JJ. Restoration of collagen formation in photodamaged human skin by tretinoin (retinoic acid): N Engl J Med, 329: 530-5.8 (1993) [Non-Patent Document 3] Talwar HS, Griffiths CEM, Fisher GJ, Hamilton TA, Voorhees JJ. Reduced type I and type III procollagens in adult human skin: J Invest Dermatol, 105: 285-90 (1995) [Non-Patent Document 4] Rittie R, Fisher GJ, UV-light-induced signal cascades and skin aging; Age Res Rev, 1: 705-720 (2002) By confirming the sound, the invention was completed.

In order to solve the above problems of the prior art, the present invention promotes collagen formation, which is a major protein of the skin matrix, and at the same time, heptapeptide which can effectively inhibit the expression of MMP-1, an enzyme that degrades collagen, which is a matrix protein. An object of the present invention is to provide a composition for preventing skin aging through preventing, improving or treating skin aging or wrinkles including monomers and dimers as an active ingredient.

In addition, the present invention has excellent skin aging prevention and improvement effect due to heptapeptide monomer and dimer that inhibits expression of MMP-1 and procollagen type II protein expression, which is an important enzyme causing skin aging, and prevents skin aging without cytotoxicity. An object of the present invention is to provide a composition for improvement or treatment.

[Formula 1]

[Cys-Gly-Pro-Gln-X-Pro-Gln]

[Formula 2]

Gly-X-Gln-Gly-Pro-Gln-Cys

(3)

[Cys-Gly-Pro-Gln-X-Pro-Gln] ₂

[Formula 4]

Gly-X-Gln-Gly-Pro-Gln-Cys ₂

In Formulas 1 to 4, X is glutamic acid, aspartic acid, histidine, phenylalanine, alanine, cysteine, glycine, glutamine, asparagine, arginine, leucine, methionine, isoleucine, serine, tyrosine, threonine, lysine, tryptophan, Selected from proline and valine, [] 2 represents a dimer of a cysteine residue, a thiol group (-SH), in a reduced form (-SS-).

Preferably, the peptide of Formula 1 to 4,

Gly-Pro-Gln-Pro-Pro-Gln-Cys (Compound 1)

Gly-Pro-Gln-Asp-Pro-Gln-Cys (Compound 2)

Gly-Pro-Gln-Asn-Pro-Gln-Cys (Compound 3)

Gly-Pro-Gln-Tyr-Pro-Gln-Cys (Compound 4)

Gly-Pro-Gln-Leu-Pro-Gln-Cys (Compound 5)

[Cys-Gly-Leu-Gln-Gly-Pro-Gln] (Compound 6)

[Cys-Gly-Pro-Gln-Gly-Pro-Gln] (Compound 7)

[Cys-Gly-Asp-Gln-Gly-Pro-Gln] (Compound 8)

[Cys-Gly-Lys-Gln-Gly-Pro-Gln] (Compound 9)

[Cys-Gly-Ser-Gln-Gly-Pro-Gln] (Compound 10)

Geological (Stearic, Palmitic, arachidic, etc.)

Gly-Pro-Gln-Asn-Pro-Gln-Cys (Compound 11)

[Cys-Gly-Pro-Gln-Gly-Pro-Gln] (Compound 12)

[Cys-Gly-Asp-Gln-Gly-Pro-Gln] (Compound 13)

*

Gly-Pro-Gln-Pro-Pro-Gln-Cys ₂ (compound 14)

Gly-Pro-Gln-Asp-Pro-Gln-Cys ₂ (Compound 15)

Gly-Pro-Gln-Asn-Pro-Gln-Cys ₂ (Compound 16)

Gly-Pro-Gln-Tyr-Pro-Gln-Cys ₂ (Compound 17)

Gly-Pro-Gln-Leu-Pro-Gln-Cys ₂ (Compound 18)

[Cys-Gly-Leu-Gln-Gly-Pro-Gln] ₂ (Compound 19)

[Cys-Gly-Pro-Gln-Gly-Pro-Gln] ₂ (Compound 20)

[Cys-Gly-Asp-Gln-Gly-Pro-Gln] ₂ (Compound 21)

[Cys-Gly-Lys-Gln-Gly-Pro-Gln] ₂ (Compound 22)

[Cys-Gly-Ser-Gln-Gly-Pro-Gln] ₂ (Compound 23)

Geological (Stearic, Palmitic, arachidic, etc.)

Gly-Pro-Gln-Asn-Pro-Gln-Cys ₂ (Compound 24)

[Cys-Gly-Pro-Gln-Gly-Pro-Gln] ₂ (Compound 25)

[Cys-Gly-Asp-Gln-Gly-Pro-Gln] ₂ (Compound 26)

In the compound, [] 2 represents a dimer of a cysteine residue, a thiol group (-SH), in a reduced form (-S-S-).

In another aspect, the present invention provides a composition for improving wrinkles containing at least one peptide selected from the peptides of Formulas 1 to 4, or the compounds 1 to 26.

Thus, the present invention can provide a pharmaceutical composition for improving wrinkles containing at least one peptide selected from the peptides of Formulas 1 to 4, or the compounds 1 to 26. The peptide may be included in the pharmaceutical composition at 0.0001 to 1.0% by weight.

*

In still another aspect, the present invention provides a cosmetic composition for improving wrinkles containing at least one peptide selected from the peptides of Formulas 1 to 4 or the peptides of Compounds 1 to 26. The peptide may be included in the cosmetic composition in an amount of 0.0001 to 1.0% by weight. The cosmetics may be selected from lotion, milky lotion, gel, cream, essence, pack, ampoule, lotion, cleaning agent, soap, body products, soap, oil, lipstick and foundation.

Hereinafter, the present invention will be described in detail.

Among the peptides of the present invention, the peptides of the compounds 14 to 26 are obtained by modifying the thiol group (-SH), which is a cysteine (cysteine located at the terminal of the peptide) residues of the peptides of the compounds 1 to 13, respectively, into a reduced form (-SS-). It is a dimer. For example, a duplex in which a thiol group, which is a cysteine residue of Compound 1, is modified, is Compound 14, and a duplex, in which a thiol group, which is a cysteine residue of Compound 2, is reduced, is Compound 15. In this manner, the duplexes of Compounds 1 to 13 may be prepared using Compounds 3 to 16, respectively.

Peptides of the invention can be prepared in such a way that one or more amino acids or suitably protected amino acids are continuously formed in the amino acid backbone, which are constantly bound to a solid phase, to form amide bonds. In addition, the peptide can be inserted, substituted, deleted other amino acids in a range that does not significantly reduce the stability, which also belongs to the scope of the present invention.

In addition, the cell permeable peptide (cell permeable peptide) for promoting intracellular migration of the peptide of the present invention may be further included by binding to the peptide C-terminal or N-terminal. For example, the cell permeable peptide includes TAT peptide (Arg-Lys-Lys -Arg-Arg-Tyr-Arg-Arg-Arg) and Tat-PTD peptide (Gly-Arg-Lys-Lys-Arg -Arg-Gln- Arg-Arg-Arg: Tat PTD), but the present invention is not limited thereto, and any cell-penetrating 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.

On the other hand, the peptide of the present invention may exist in the form of a salt. Forms of salts usable in the present invention may be made during the final separation and purification of the compound or by reacting an amino group with a suitable acid. For example, as acid addition salts, acetates, adipates, alginates, citrate, aspartates, benzoates, benzenesulfonates, bisulfates, butyrates, camphorates, camphorsulfonates, digluconates, glycerophosphates, hemi Sulfate, heptanoate, hexanoate, formate, fumarate, hydrochloride, hydrobromide, hydroyodide, 2-hydroxyethane sulfonate, lactate, maleate, mesitylenesulfonate, methanesulfonate, Naphthylenesulfonate, nicotinate, 2-naphthalenesulfonate, oxalate, pamoate, pectinate, persulfate, 3-phenylpropionate, picrate, pivalate, propionate, succinate, tartrate Trichloroacetate, trifluoroacetate, phosphate, glutamate, bicarbonate, La-be-toluenesulfonate and undecanoate. However, the embodiment is not limited thereto. In addition, examples of acids that may be used to form acid addition salts may 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, the peptide form containing the trifluoroacetate salt or the acetate salt is most preferred.

The amino groups or carboxyl groups of the amino acids used for preparing the peptides of the invention may be protected by suitable protecting groups. Protected amino acids can be reacted in solution by adding the following amino acids under conditions suitable to attach to a solid support or to form an amide bond. In addition, the protecting group can be removed completely before adding the amino acid protected with a suitable protecting group. After all amino acids have been linked as desired, the final desired peptide can be obtained by successive or simultaneous separation from the free residual protecting group and free solid support.

In the present invention, a condensation condensation obtained by synthesizing a desired hexapeptide by condensation of an appropriately protected tetrapeptide with another appropriately protected dipeptide to form an amide bond under the condition that the chiral center is not racemized. Peptides can be prepared using reaction techniques.

The most preferable synthetic method for preparing the peptide compound of the present invention may be a solid phase peptide synthesis method synthesized using a solid phase polymer retardation, wherein the α-amino group of the peptide prepared by the above method is acid or base sensitive. It can be protected by a functional group. The protecting group of the amino acid at this time must have a stable property in the conditions of peptide condensation reaction, and must be easily removable without breaking the extended peptide chain or without racemicization of any chiral center contained therein. Thus, suitable protecting groups include 9-fluorenylmethyloxycarbonyl (Fmoc), t-butoxycarbonyl (Boc), benzyloxycarbonyl (Cbz), biphenylisopropyl-oxycarbonyl, t-amyloxycarbonyl , Isobornyloxycarbonyl, (α, α) -dimethyl-3,5-dimethoxybenzyloxycarbonyl, O-nitrophenylsulfenyl, 2-cyano-t-butyloxycarbonyl, and the like, and such Other suitable protecting groups known in the art for the purpose are also usable within the scope of the present invention.

As the most preferable protecting group of the amino acid used in the peptide synthesis of the present invention, a 9-flu orenylmethyloxycarbonyl (Fmoc) protecting group can be used.

In particular, protecting groups for amino acid residues used in the peptide synthesis of the present invention are, for N-methyl glutamic acid, t-butyl (t-Bu); For lysine, t-butoxycarbonyl (Boc); For serine, 7t-butyl (t-Bu); For threonine and allothreonine, it is t-butyl (t-Bu); In the case of cysteine, it is preferable that it is trityl (Trt), but this invention is not limited to this.

In solid phase peptide synthesis methods, the C-terminal amino acid may be attached to a suitable solid support or resin. Suitable solid supports useful for such synthesis are preferably reagents of the staged condensation-deprotection reaction and materials which are inert to the reaction conditions and are insoluble in the medium used, for example rink amid or rink amide 4- It may be a methyl benzyl hydryl amine resin (rink amid MBHA resin).

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

C-terminal amide is a solvent such as dichloromethane, N-methylpyridone (NMP) or DMF at a temperature of 10 ℃ to 50 ℃, preferably at a temperature of 30 ℃ for 1 to 24 hours 4-dimethylaminopyridine (DMAP), 1-hydroxybenzotriazole (HOBt), N-methylmorpholine (NMM), benzotriazol-1-yloxy-tris (dimethylamino) phosphonium-hexafluorophosphate N, N'-dicyclohexylcarbodiimide (DCC), 'N, N'-diisoxo in the presence or absence of (BOP) or bis (2-oxo-3-oxazolidinyl) phosphine chloride (BOP CI) Propylcarbodiimide (DIC), [O- (7-azabenzotriazol-1-yl) -1,1,3,3-tetramethylurojunium hexaflurophosphate] (HATU) or O-benzotriazole Activated carboxylic acid in -1-yl-N, N, N ', N'-tetra-methyluronium hexafluorophosphate (HBTU) to be condensed (bonded, coupled) to a resin or solid support via condensation There.

If the solid support is a link amide 4-methylbenzylhydrylamine resin, the preferred moiety as a protecting group is that the Fmoc functional group is excess of a secondary amine solution, preferably 20% piperidine DMF solution before condensing to the C-terminal amino acid. Use to cut. Preferred reagents used for condensation of the desired amithenoic acid in the deprotected '4- (2' ', 4'-dimethoxyphenyl-Fmoc-aminomethyl) phenoxyacetamidoethyl resin are DMF for suitably protected amino acids. N-methylmorpholine (NMM), 1-hydroxybenzotriazole (H OBt) and O- (7-azabenzotriazol-1-yl) -1,1,3,3-tetramethyluronium in a solvent Hexafluorophosphonate] (HATU), O-benzotriazol-1-yl-N, N, N ', N'-tetramethyluronium hexafluorophosphate (HBTU), N, N'-dicyclohex Condensation reaction reagents such as silk carbodiimide (DCC) or N, N'-diisopropenylcarbodiimide (DIC).

Condensation of consecutive amino acids carried out in the present invention can be carried out manually or manually using an automated peptide synthesizer well known in the art. Preferred conditions for the synthesis reaction include deprotection of the α-amino acid protected with Fmoc group with a secondary amine solution, preferably piperidine, followed by washing with a sufficient excess of solvent and another protection desired for condensation. 3 to 7-fold molar excess of the amino acid can then be added, and the reaction can be preferably carried out in a DMF solvent.

In the final step of the synthesis of the peptide using the solid resin of the present invention, the peptide to be obtained from the resin can be removed in a continuous or single operation, and the protecting groups protecting the residues of the amino acids can be deprotected. Removal conditions of the peptide from the resin and deprotection conditions of the protecting groups present at the residues generally include cleavage reagent cocktails that cleave the bond between the resin-peptide, for example trifluoroacetic acid (TFA), triisopropylsilane. It can be obtained by treating dichloromethane mixed cocktail solution consisting of (TIS), thioanisol, water or ethanedithiol (EDT). The mixed solution thus obtained can produce a precipitate by excessively treating the refrigerated diethyl ether solvent. The precipitate obtained as described above was centrifuged to completely settle the precipitate, and the precipitate was solidified by first removing the excessive amounts of trifluoroacetic acid, triisopropylsilane, thioanizol, water and ethanedithiol, and repeating the above procedure two or more times. Can be obtained. At this time, the fully deprotected peptide salt can be separated and purified using a mixed solvent consisting of water and acetnitrile solvent and reversed phase high performance liquid chromatography (HPLC). The separated and purified peptide solution can be completely concentrated and dried using lyophilization to obtain a solid peptide.

Peptides of the present invention has a collagen synthesis effect.

According to a preferred embodiment of the present invention, the composition of the present invention may be provided as a pharmaceutical composition or cosmetic composition for wrinkle improvement.

When the composition of the present invention is prepared as a pharmaceutical composition, the pharmaceutical composition of the present invention includes a pharmaceutically acceptable carrier. The pharmaceutically acceptable carrier is conventionally used in the preparation, lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia rubber, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose , Polyvinylpyrrolidone, cellulose, purified water, syrup, methyl cellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, mineral oil, and the like. In addition to the above components, the pharmaceutical composition of the present invention may further include conventionally used additives such as lubricants, wetting agents, sweeteners, flavoring agents, emulsifiers, suspending agents, preservatives and the like. The pharmaceutical composition of the present invention is preferably parenteral administration, more preferably applied by topical administration by application.

Suitable dosages of the pharmaceutical compositions of the present invention may be prescribed in various ways depending on factors such as formulation method, mode of administration, age, weight, sex, morbidity, condition of food, time of administration, route of administration, rate of excretion and response to response of the patient. Can be.

The dosage of the peptide, which is an active ingredient included in the pharmaceutical composition of the present invention, is 0.001 to 100 mg / kg, preferably 0.1 to 100 mg / kg, more preferably 1 to 50 mg / kg, based on an adult. Dosages may be administered once or several times a day. In addition, the peptide of the present invention may preferably contain 0.0001 to 1.0% by weight, more preferably 0.001 to 1.0% by weight, most preferably 0.001 to 0.01% by weight relative to the total weight of the pharmaceutical composition. It is not limited to this.

The cosmetic composition of the present invention includes not only peptides, which are effective ingredients thereof, but also components commonly used in cosmetic compositions, and include, for example, conventional auxiliaries such as antioxidants, stabilizers, solubilizers, qubitatamines, pigments, and perfumes, and carriers. It includes. In addition, as the carrier, purified water, monohydric alcohols (ethanol or propyl alcohol), polyhydric alcohols (glycerol, 1,3-butyrene glycol or propylene glycol), higher fatty acids (palmitylic acid or quilizonolenic acid), Oils and fats such as wheat germ oil, camellia oil, jojoba oil, olive oil, squalene, sunflower oil, bovine macadamia peanut oil, avocado oil, or fatty acid glycerides may be used, but are not limited thereto. If necessary, a surfactant, a humectant, a preservative, an antioxidant and the like can be added.

Surfactants that can be used in the cosmetic composition of the present invention, as anionic surfactant 알킬, alkylbenzene sulfonate, polyoxyalkylene alkyl sulfate ester salt, alkyl sulfate ester salt, olefin sulfonate, alkyl phosphate, polyoxyalkylene Alkyl ether phosphate, dialkyl sulfosuccinate, fatty acid salt, etc. are mentioned, As a nonionic surfactant, polyoxyethylene alkyl ether, polyoxyethylene fatty acid ester, polyhydric alcohol fatty acid partial ester, polyoxyethylene polyhydric alcohol fatty acid partial Ester, polyglycerol fatty acid ester, polyoxyethylene cured castor oil derivative, fatty acid diethanolamide, and the like. Examples of the cationic surfactant include tertiary aliphatic amine salts, alkyltrimethylammonium halides, and dialkyldimethylammonium halides. Examples of the amphoteric surfactants include amide betaine type, imidazolinium betaine type, and sulfobeta. Like dolls. Examples of the moisturizing agent include glycerin, propylene propylene glycol, 1,3-butylene glycol, dipropylene glycol, sorbitol, and the like. Examples of the preservative include benzoic acid, dehydroacetic acid, paraoxybenzoic acid esters (methyl paraoxybenzoate and butyl paraoxybenzoate), phenoxyethanol and the like. As the antioxidant, ascorbic acid, BHA, and the like can be cited. In addition, ultraviolet absorbers, anti-inflammatory agents, and refreshing agents can be added.

Peptide of the present invention may preferably contain 0.0001 to 1.0% by weight, more preferably 0.001 to 1.0% by weight, most preferably 0.001 to 0.01% by weight relative to the total weight of the cosmetic composition, but is not limited thereto. .

Moreover, the kind of cosmetics is not specifically limited, For example, skin care cosmetics, lipsticks, such as a lotion, an emulsion, a gel, cream, an essence, a pack, an ampoule, a lotion, a cleaning agent, a soap, body products, soap, oil, Makeup cosmetics, such as a foundation, etc. are mentioned, The formulation is not specifically limited.

The cosmetic composition of the present invention may be used daily and may also be used for an indefinite period of time. Preferably, the amount of use, frequency of use and duration may 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, heptapeptide monomers and dimers, which increase the collagen formation, a major protein of the skin matrix, and at the same time, inhibit the expression of MMP-1, an enzyme that degrades collagen and cause skin aging, are included as an active ingredient. It is excellent in preventing and improving effect, and it is not toxic to skin cells, so it is suitable for use as a cosmetic, pharmaceutical and food composition for skin aging, prevention of skin wrinkles, improvement or treatment.

1 graphically depicts the rate of procollagen type II production of the compounds of the present invention.
Figure 2 graphically depicts the inhibition of MMP-1 production of the compounds of the present invention.
Figure 3 shows the cell viability of the compounds of the present invention.
Figure 4 is a graphical representation of the eye wrinkle improvement effect of the composition containing a compound of the present invention.

Hereinafter, a preferred embodiment of the present invention will be described in detail. However, the present invention is not limited to the embodiments described herein and may be embodied in other forms. Rather, the information introduced herein is intended to be thorough and complete, and to fully convey the spirit of the present invention to those skilled in the art.

<Example 1-1. Synthesis of Compound 1>

Names and abbreviations of amino acids used in the present invention are indicated as follows.

Ala: Alanine / Cys: Cysteine / Gly: Glycine / Val: Valine / Pro: Proline / Phe: Phenylalanine / Met: Metheonine / Trp: Tryptophan / Glu: Glutamine /

71.4 mg (0.10 mmol) of 2-chlorotrityl chloride resin (1.4 mmol loaded per gram) purchased from Novabiochem corporation was weighed into a reaction vessel. The resin was solvated with 3 ml of DMF and allowed to sweep for 5 minutes, then 3 ml of 20% (w / v) piperidine DMF solution was added and shaken for 20 minutes and the piperidine DMF solution After removal, the mixture was washed 5 times with 10 ml of DMF solvent (5 times with 10 ml each). * 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. Add 3 ml of 20% (w / v) piperidine DMF solution, shake for 10 minutes, remove the piperidine solution, and then add 20% (w / v) piperidine DMF solution for 20 minutes. After the reaction, the Fmoc protecting group protected in the resin was completely removed and washed 5 times with 10 ml of DMF solvent (5 times with 10 ml each). The deprotection of the Fmoc protecting group at this stage was determined by 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 condensed (coupled) continuously according to the same synthesis cycle as described below.

(1) 5 washes with DMF solvent (10 mL);

(2) deprotection twice for 10 min using 20% (w / v) piperidine DMF solution (3 mL);

(3) five washes with DMF solvent (10 mL);

(4) addition of Fmoc-amino acid;

(5) addition of a condensation reagent to activate amino acids and condensation for 2 hours;

(6) washing 5 times with DMF solvent (10 mL);

(1) to (6) were repeated repeatedly, at which time Fmoc-protected amino acid (0.80 mmol) after Fmoc-Cys (Trt) -OH was added to the resin reaction vessel and condensed in the order described below. .

(i) Fmoc-Gln-OH;

(ii) Fmoc-Pro-OH;

(iii) Fmoc-Pro-OH;

(iv) Fmoc-Gln-OH;

(v) Fmoc-Pro-OH;

(vi) Fmoc-Gly-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 the end of the synthesis, a mixture of trifluoroacetic acid / thioanizol / ethanedithiol / triisopropylsilane / water (95: 5: 2.5: 2.5: 2.5) Using (10 mL), the peptide was cleaved from the resin. The precipitate was produced by treating 100 ml of the diethyl ether solvent refrigerated and stored in the thus obtained mixed solution. The precipitate obtained was centrifuged to completely settle and the first removal of trifluoroacetic acid, thioanisol and ethanedithiol was followed by the above procedure (100 ml of diethyl ether solvent was added to wash the precipitate and centrifugation-first removal). Trifluoroacetic acid, thioaniazole and ethanedithiol) were tried twice to obtain a precipitate which solidified. 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 fractions were lyophilized to give collagen synthesis promoter H ₂ N- [Gly-Pro-Gln-Pro-Pro-Gln-Cys] -CO ₂ H (100 mg) as a trifluoroacetate salt in white powder form. Got it.

Compound 1: H ₂ N- [Gly-Pro-Gln-Pro-Pro-Gln-Cys] -CO ₂ H

MS (ESI) m / e, [M + H] ⁺ = 725.92; (100 mg)

<Example 1-2. Synthesis of Compounds 2-10>

Using the same production process as in Example 1-1, but by changing the order of amino acids protected with Fmoc to prepare a peptide of the compound 2-10.

Compound 2: H ₂ N- [Gly-Pro-Gln-Asp-Pro-Gln-Cys] -CO ₂ H

MS (ESI) m / e, [M + H] ⁺ = 743.89; (108mg)

Compound 3: H ₂ N- [Gly-Pro-Gln-Asn-Pro-Gln-Cys] -CO ₂ H

MS (ESI) m / e, [M + H] ⁺ = 742.91; (105 mg)

Compound 4: H ₂ N- [Gly-Pro-Gln-Tyr-Pro-Gln-Cys] -CO ₂ H

MS (ESI) m / e, [M + H] ⁺ = 791.98; (104 mg)

Compound 5: H ₂ N- [Gly-Pro-Gln-Leu-Pro-Gln-Cys] -CO ₂ H

MS (ESI) m / e, [M + H] ⁺ = 741.95; (110 mg)

Compound 6: H ₂ N- [Cys-Gly-Leu-Gln-Gly-Pro-Gln] -CO ₂ H

MS (ESI) m / e, [M + H] ⁺ = 701.89; (103 mg)

Compound 7: H ₂ N- [Cys-Gly-Pro-Gln-Gly-Pro-Gln] -CO ₂ H

MS (ESI) m / e, [M + H] ⁺ = 685.86; (101mg)

Compound 8: H ₂ N- [Cys-Gly-Asp-Gln-Gly-Pro-Gln] -CO ₂ H

MS (ESI) m / e, [M + H] ⁺ = 703.83; (120 mg)

Compound 9: H ₂ N- [Cys-Gly-Lys-Gln-Gly-Pro-Gln] -CO ₂ H

MS (ESI) m / e, [M + H] ⁺ = 716.92; (101mg)

Compound 10: H ₂ N- [Cys-Gly-Ser-Gln-Gly-Pro-Gln] -CO ₂ H

MS (ESI) m / e, [M + H] ⁺ = 675.82; (110 mg)

<Example 1-3. Synthesis of Compound 14>

The collagen synthesizing peptide H ₂ N- [Cys-Gly-Pro-Gln-Pro-Pro-Gln] -CO ₂ H (50 mg) was dissolved in 2 ml of dimethyl sulfoxide (DMSO) and 10 ml of water was added thereto. Then, the mixture was stirred at room temperature for 3 days to obtain a final collagen synthesizing peptide (50 mg) in the form of a dimer in which a thiol group (-SH), which is a cysteine residue, was converted into a reduced form (-SS-).

Compound 14: H ₂ N- [Gly-Pro-Gln-Pro-Pro-Gln-Cys] ₂ -CO ₂ H

MS (ESI) m / e, [M + H] ⁺ = 1451.84; (50 mg)

<Example 1-4. Synthesis of Compounds 15-23>

Using the same production process as in Example 1-3, but by changing the order of amino acids protected with Fmoc to prepare the peptide of the compound 15 ~ 23.

Compound 15: H ₂ N- [Gly-Pro-Gln-Asp-Pro-Gln-Cys] ₂ -CO ₂ H

* MS (ESI) m / e, [M + H] ⁺ = 1487.78; (45 mg)

Compound _{16: H 2 N- [Gly-} Pro-Gln-Asn-Pro-Gln-Cys] 2 -CO 2 H

MS (ESI) m / e, [M + H] ⁺ = 1485.82; (46 mg)

Compound 17: H ₂ N- [Gly-Pro-Gln-Tyr-Pro-Gln-Cys] ₂ -CO ₂ H

MS (ESI) m / e, [M + H] ⁺ = 1583.96; (49 mg)

Compound 18: H ₂ N- [Gly-Pro-Gln-Leu-Pro-Gln-Cys] ₂ -CO ₂ H

MS (ESI) m / e, [M + H] ⁺ = 1483.9; (42 mg)

Compound 19: H ₂ N- [Cys-Gly-Leu-Gln-Gly-Pro-Gln] ₂ -CO ₂ H

MS (ESI) m / e, [M + H] ⁺ = 1403.78; (47 mg)

Compound 20: H ₂ N- [Cys-Gly-Pro-Gln-Gly-Pro-Gln] ₂ -CO ₂ H

MS (ESI) m / e, [M + H] ⁺ = 1371.72; (48 mg)

Compound 21: H ₂ N- [Cys-Gly-Asp-Gln-Gly-Pro-Gln] ₂ -CO ₂ H

MS (ESI) m / e, [M + H] ⁺ = 1407.66; (49 mg)

Compound 22: H ₂ N- [Cys-Gly-Lys-Gln-Gly-Pro-Gln] ₂ -CO ₂ H

MS (ESI) m / e, [M + H] ⁺ = 1433.84; (47 mg)

Compound 23: H ₂ N- [Cys-Gly-Ser-Gln-Gly-Pro-Gln] ₂ -CO ₂ H

MS (ESI) m / e, [M + H] ⁺ = 1351.64; (54 mg)

<Example 1-5. Synthesis of Compound 11>

Of the three peptides selected from the 10 peptides, DMF (4ML), 1.06 mL (10 eq) and DIPEA 1.96 mL (10 eq) were added to the compound 3 peptide before the cleavage of the peptide from the resin. 5 washes with 10 mL of DMF solvent (5 washes with 10 mL each) * DMF: Dimethylformamide reaction was confirmed by Kaiser test, and then trifluoroacetic acid / thio Peptides were cleaved from the resin using a mixture of anisol / ethanedithiol / triisopropylsilane / water (95: 5: 2.5: 2.5: 2.5) (10 mL). The precipitate was produced by treating 100 ml of the diethyl ether solvent refrigerated and stored in the thus obtained mixed solution. The precipitate obtained was centrifuged to completely settle and the first removal of trifluoroacetic acid, thioanisol and ethanedithiol was followed by the above procedure (100 ml of diethyl ether solvent was added to wash the precipitate and centrifugation-first removal). Trifluoroacetic acid, thioaniazole and ethanedithiol) were tried twice to obtain a precipitate which solidified. 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 fractions were lyophilized to give the collagen synthesizing peptide Stearyl- [Gly-Pro-Gln-Asn-Pro-Gln-Cys] -CO ₂ H (102 mg) as a trifluoroacetate salt in white powder form. In this manner, compounds 11 to 13 were synthesized.

* Type of Geology

Ste) Stearic acid

Ii) Palmitic acid

Arachidic acid

O) Oleic acid

Erucic acid

Lin) Linoleic acid

Iii) Linolenic acid

Through experiments of collagen synthesis and mmp-1 synthesis of the ten compounds of Examples 1-1 and 1-2 as described above, the experiment was performed by selectively attaching the top three high collagen synthesis rates.

Compound 11: Stearyl- [Gly-Pro-Gln-Asn-Pro-Gln-Cys] -CO ₂ H

Compound 12: Palmitoyl- [Cys-Gly-Pro-Gln-Gly-Pro-Gln] -CO ₂ H

Compound 13: Erucyl- [Cys-Gly-Asp-Gln-Gly-Pro-Gln] -CO ₂ H

<Example 1-6. Synthesis of Compound 24>

The collagen synthetase-promoting peptide Stearyl- [Gly-Pro-Gln-Asn-Pro-Gln-Cys] -CO ₂ H (53 mg) methyl sulfoxide (DMSO) obtained in Example 1-5 was dissolved in 10 ml. After adding water and stirring for 3 days at room temperature, the final collagen synthesizing peptide Stearyl- [Gly in the form of a dimer in which the thiol group (-SH), a cysteine residue, was transformed into a reduced form (-S-S-). -Pro-Gln-Asn-Pro-Gln-Cys] ₂ -CO ₂ H (50 mg). Compounds 24-26 were synthesized in this manner.

Compound 24: Stearyl- [Cys-Gly-Pro-Gln-Asn-Pro-Gln] ₂ -CO ₂ H

Compound 25: Palmitoyl- [Cys-Gly-Pro-Gln-Gly-Pro-Gln] ₂ -CO ₂ H

Compound 26: Erucyl- [Cys-Gly-Asp-Gln-Gly-Pro-Gln] ₂ -CO ₂ H

Examples 1-3, 1-4 as described above through the experiment of collagen synthesis ability and mmp-1 synthesis inhibitory activity, the experiment was carried out by selectively attaching the top three high collagen synthesis rate lipids, the same method as the monomer Was selected.

Experimental Example 1 Determination of Procollagen II Synthesis Promoting Effect of Peptides ( in vitro )

0.5 ml of human dermal fibroblasts (HDF: Human dermal fibroblast) were dispensed on a 24 well plate at a concentration of 0.2 ⁴ cells / ml, and cultured at 37 ° C., 5% CO ₂ , and humidified conditions for 1 day. As the culture medium, 0.5 ml of each medium containing FBS (manufactured by Gibco) at 5% in DMEM medium (Dulbecco's Modified Eagle's Medium, manufactured by Gibco) was used.

Subsequently, the cell culture solution was washed with PBS, exchanged with DMEM (Dulbecco's Modified Eagle's Medium, manufactured by Sigma) without FBS (Gibco), and cultured for 12 hours at 37 ° C., 5% CO ₂ , and humidified conditions. Proceeded. After incubation, the cells were washed with 0.5 ml PBS (Phosphate Buffered Saline, manufactured by Sigma), and then treated with 100 uM of each compound of the present invention. 100 μl of PBS that did not dissolve the compound was used as a control.

In the procollagen II production promotion test, two days after the addition of the compound of the present invention, the culture medium was collected, and the concentration of type II pro collagen secreted in the culture medium was measured by enzyme-linked immunoassay (ELSIA, Human Procollagen type II ELISA Kit). (Prepared by R & D system).

Based on the quantitative results, the amount of synthesized type II pro collagen (pg / ml) was measured, and the type II pro collagen production rate was calculated according to Equation 1 below, and the results are shown in Table 1 and FIG. 1.

[Equation 1]

Type II Pro Collagen Production Rate (%) = (Experiment Type II Pro Collagen Amount / Control Type II Pro Collagen Amount) * 100

Comparison of Procollagen II Production Rate of Each Compound division Skin fibroblasts
Procollagen II production rate (%) division Skin fibroblasts
Procollagen II production rate (%) Compound 1 140.2 Compound 14 152.2 Compound 2 158.1 Compound 15 159.4 Compound 3 173.9 Compound 16 186.7 Compound 4 160.1 Compound 17 161.5 Compound 5 164.2 Compound 18 170.3 Compound 6 165.5 Compound 19 165.9 Compound 7 174.2 Compound 20 189.9 Compound 8 178.6 Compound 21 191.1 Compound 9 168.1 Compound 22 182.2 Compound 10 170.2 Compound 23 180.0 Compound 11 180.2 Compound 24 193.2 Compound 12 175.1 Compound 25 190.7 Compound 13 174.0 Compound 26 189.8

Referring to Table 1 and FIG. 1, all of the compounds 1 to 26 of the present invention show a procollagen II production promoting effect and have a sufficient effect on skin aging prevention and skin wrinkle prevention.

On the other hand, it is very important that the peptide is rapidly decomposed by the hydrolases present in the body after permeation in the body, so that its capacity decreases rapidly, thereby slowing down the degradation rate of the peptide. In general, hydrolases decompose the amide bond (-C (= O) NH-) portion of the peptide structure with carboxylic acid (-CO2H) and amine (-NH2). It has its own cysteine dimer (-SS-) backbone, which lowers or disrupts the degradation of common hydrolytic enzymes. That is, among the peptide compounds, compounds 14 to 26 in dimer form are slower to be degraded by hydrolase in vivo than compounds 1 to 13 in monomer form. Therefore, among the peptide compounds of the present invention, in the case of compounds 14 to 26 having cysteine dimers, it is determined that the stability in the body is very excellent. Furthermore, the lipidated peptides 11-13 and 24-26 had similar or higher levels of collagen production promotion than peptides that did not.

Experimental Example 2 Measurement of Inhibitory Effect of Peptide MMP-1 Production ( in vitro )

Human dermal fibroblasts (HDF: Human dermal fibroblast) (HDF) were dispensed in 0.5 ml aliquots in 24 well plates at a concentration of 0.2 ⁴ cells / ml, and cultured daily at 37 ° C., 5% CO ₂ , and humidified conditions. As the culture medium, 0.5 ml of each well of a medium containing 10% of FBS (manufactured by Gibco) in DMEM medium (Dulbecco's Modified Eagle's Medium, manufactured by Gibco) was used.

Subsequently, the cell culture solution was washed with PBS, exchanged with DMEM (Dulbecco's Modified Eagle's Medium, manufactured by Sigma) without FBS (Gibco), and cultured for 12 hours at 37 ° C., 5% CO ₂ , and humidified conditions. Proceeded. After incubation, washed again with 0.5 ml of PBS (Phosphate Buffered Saline, manufactured by Sigma) and then treated with 100 uM of each compound of the present invention. 100 μl of PBS that did not dissolve the compound was used as a control.

In the MMP-1 production inhibition test, after culturing for 2 days, the culture solution was taken and the concentration of MMP-1 secreted in the culture was quantified by enzyme-linked immunoassay (ELSIA, MMP-1 EIA Kit; manufactured by R & D system).

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

[Equation 2]

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

Comparison of the inhibition rate of MMP-1 production of each compound division Skin fibroblasts
% Inhibition of MMP-1 production division Skin fibroblasts
% Inhibition of MMP-1 production Compound 1 73.4 Compound 14 65.5 Compound 2 70.2 Compound 15 65.0 Compound 3 63.1 Compound 16 50.7 Compound 4 67.3 Compound 17 58.2 Compound 5 69.4 Compound 18 55.4 Compound 6 69.9 Compound 19 56.1 Compound 7 62.4 Compound 20 54.3 Compound 8 57.2 Compound 21 49.8 Compound 9 70.7 Compound 22 58.0 Compound 10 75.5 Compound 23 57.9 Compound 11 54.2 Compound 24 47.7 Compound 12 61.9 Compound 25 50.2 Compound 13 63.0 Compound 26 53.6

Referring to Table 2 and Figure 2, all of the compounds 1 to 26 of the present invention was confirmed that the MMP-1 production inhibitory effect is excellent, based on this it is confirmed that exhibits a sufficient effect on the prevention of skin wrinkles. Among the plurality of peptide compounds of the present invention, the lipidated peptides 11-13 and 24-26 had similar or higher inhibitory effects on MMP-1 production than other peptides.

Experimental Example 3: Confirmation of the safety of heptapeptide monomer and dimer ( in vitro )

Human dermal fibroblasts (HDF: Human dermal fibroblast, manufactured by Thermo) were dispensed at a concentration of 0.75 ³ cells / ml in 96-well plates at 100 ul, and cultured at 37 ° C., 5% CO ₂ , and humidified conditions for 2 days. . As the culture solution, a medium containing 10% of FBS (manufactured by Gibco) in 10% DME medium (Dulbecco's Modified Eagle's Medium, manufactured by Gibco) was used for each well.

Subsequently, it was exchanged with DMEM (Dulbecco's Modified Eagle's Medium, manufactured by Sigma) without FBS (Gibco), washed again with 100 ul of PBS (Phosphate Buffered Saline, manufactured by Sigma), and then the compound of the present invention was 200 uM. Treated and incubated. 100 μl of PBS that did not dissolve the compound was used as a control.

The test sample was added and cultured for 72 hours, then treated with MTT and incubated for 2 hours. Thereafter, the culture solution was removed, 200 μl of DMSO was added, and the resulting formazan was sufficiently dissolved, and the absorbance was measured at 540 nm. Based on the measured values, cell viability was calculated according to the following Equation 3, and the results are shown in Table 3 and FIG. 3.

&Quot; (3) &quot;

% Cell viability = (Experimental group 540 nm absorbance / control group 540 nm absorbance) * 100

Cell viability comparison of each compound division Cell survival rate (%) division Cell survival rate (%) Control 100 Control 100 Compound 1 103.2 Compound 14 101.2 Compound 2 102.4 Compound 15 100.4 Compound 3 100.4 Compound 16 100.4 Compound 4 100.5 Compound 17 102.1 Compound 5 100.9 Compound 18 101.9 Compound 6 101.1 Compound 19 102.0 Compound 7 102.6 Compound 20 100.7 Compound 8 99.9 Compound 21 99.9 Compound 9 98.5 Compound 22 99.5 Compound 10 100.4 Compound 23 101.3 Compound 11 98.2 Compound 24 98.7 Compound 12 98.1 Compound 25 98.6 Compound 13 99.3 Compound 26 98.6

Referring to Table 3 and FIG. 3, it was confirmed that the heptapeptide monomer and the dimer compound of the present invention show cell viability of 98% or more when the cell viability is compared with the control group. Through the results of this experiment, all compounds of the present invention was confirmed that the safety (Safety) in animal cells including human skin fibroblasts (Safety) is a safe raw material.

Experimental Example 4 Preparation of Cosmetic Formulation

In order to evaluate the effect of the cosmetic composition including the new heptapeptide monomer and dimer, which is a peptide having collagen synthesis promotion and collagenase inhibitory ability, a cosmetic formulation in the form of a cream was prepared by combining the components shown in Table 4 below.

Cream Formulation to Assess Wrinkle Improvement ingredient Experimental Example 1 (% by weight) Comparative Example 1 (% by weight) Heptapeptide Monosaccharide 0.01 - Heptapeptide dimer 0.01 - Argan oil 5 5 Shea Butter 5 5 Green Tea Seed Oil 3 3 Avocado Oil One One Isoami Laurate 5 5 Cetiaryl Alcohol 3 3 Hyaluronic acid 10 10 Beta Glucan Solution 10 10 Spices Quantity Quantity Purified water Remaining amount Remaining amount Sum 100.0 100.0

Experimental Example 5: Evaluation of the wrinkle improvement effect around the eyes (visual evaluation)

In order to confirm the wrinkle improvement effect of the cosmetic composition according to the present invention, 23 Korean women aged 40 to 55 years were evaluated in the same manner as the daily use of the cream for each 4 weeks.

The cosmetic preparations prepared according to Experimental Example 1 and Comparative Example 1 were used separately on each side of the face, and the wrinkle improvement effect of the subject was evaluated through visual observation by an experienced inspector.

Eye wrinkle improvement effect evaluation result division Wrinkle improvement effect Great slightly none Effective rate (%) Experimental Example 1 12 8 3 69.5% Comparative Example 1 3 10 10 36.8%

Referring to Table 5, the cosmetic prepared by including the heptapeptide monomer and dimer having the collagen synthesis promoting and collagen degrading enzyme inhibitory effect as in Experimental Example 1 was prepared without containing the components as in Comparative Example 1 It was confirmed that the wrinkle improvement effect was significantly improved than cosmetics.

Experimental Example 6: application of the human eye wrinkle improvement effect (PRIMOS High Resolution)

In order to confirm the anti-wrinkle effect of the cosmetic composition according to the present invention, 18 Korean women aged 35 to 55 years were evaluated in the same manner as the daily use of cream for 6 weeks. The evaluation was performed by using PRIMOS High Resolution (Phaseshift Rapid In vivo Measurement Of Skin high resolution, manufactured by GFMesstechinik GmbH, Germany). The value of the eye wrinkle variable through roughness analysis is as follows.

Ra (Average roughness): The average of the absolute values of the length from the centerline to the cross-sectional curve of the surface when the centerline is drawn at the roughness height of the cross section.

Root mean square roughness (Rq): The root mean square root of the length from the center line to the cross-sectional curve of the surface when the center line is drawn at the roughness height of the cross section.

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

The smaller the value of the variable analyzed by the roughness measurement, the better the wrinkles. The decrease rate of the roughness parameter due to PRIMOS High Resolution was calculated according to Equation 4. The results of the calculated values for each variable are shown in Table 5 and FIG. 4.

&Quot; (4) &quot;

% Reduction = (Measurement before application-Measurement after application / Measurement before application) * 100

Eye wrinkle improvement human application test results Roughness Variable 3 weeks after application 6 weeks after application Ra 3.996 6.251 Rq 3.020 5.438 Rmax 1.899 4.577

As a result of analyzing the measured values of the roughness parameters of the wrinkles around the eye area using PRIMOS High Resolution, the Ra value after the use of Experiment 1 cream was applied at a statistically significant level (p <0.05) after 3 weeks of application. After 6 weeks, 3.996% and 6.251% decreased, respectively, and the Rq values were statistically significant (p <0.05) compared to before sample application. In addition, the Rmax value decreased 4.577% after 6 weeks of application to a statistically significant level (p <0.05) compared to the sample application. Therefore, the cream of Experimental Example 1 was found to help improve eye wrinkles after 6 weeks of application.

Although the excellent wrinkle improvement effect and safety of the present invention are described in detail in the above-described embodiments and experimental examples, it will be apparent to those skilled in the art that various modifications and modifications can be made within the technical spirit of the present invention. And modifications belong to the appended claims. Therefore, it is to be understood that the embodiments and the experimental examples described above are exemplary in all respects and not restrictive.

Claims (7)

Collagen synthesis promoting peptides corresponding to the following compounds 14 to 23

Gly-Pro-Gln-Pro-Pro-Gln-Cys ₂ (compound 14),
Gly-Pro-Gln-Asp-Pro-Gln-Cys ₂ (compound 15),
Gly-Pro-Gln-Asn-Pro-Gln-Cys ₂ (compound 16),
Gly-Pro-Gln-Tyr-Pro-Gln-Cys ₂ (compound 17),
Gly-Pro-Gln-Leu-Pro-Gln-Cys ₂ (compound 18),
Cys-Gly-Leu-Gln-Gly-Pro-Gln ₂ (compound 19),
Cys-Gly-Pro-Gln-Gly-Pro-Gln ₂ (compound 20),
[Cys-Gly-Asp-Gln-Gly-Pro-Gln] ₂ (Compound 21),
Cys-Gly-Lys-Gln-Gly-Pro-Gln ₂ (compound 22), and
Cys-Gly-Ser-Gln-Gly-Pro-Gln ₂ (compound 23),
And [] ₂ to compounds 14 to 23 are peptides for promoting collagen synthesis, wherein the thiol group (-SH), which is a cysteine residue, is converted into a reduced form (-SS-).

delete Composition for preventing and improving skin wrinkles, characterized in that it contains at least one peptide selected from the peptide of claim 1.

delete The method of claim 3, wherein
The peptide is a pharmaceutical composition for preventing and treating skin wrinkles, characterized in that it comprises 0.0001 to 1.0% by weight in the pharmaceutical composition.
Cosmetic composition for preventing and improving skin wrinkles, characterized in that it contains at least one peptide selected from the peptide of claim 1.
The method of claim 6,
The peptide is a cosmetic composition for improving wrinkles, characterized in that contained in the cosmetic composition 0.0001 ~ 1.0% by weight, lotion, milk, gel, cream, essence, pack, ampoule, lotion, cleaning agent, soap, body products, soap, Cosmetic composition for preventing and improving skin wrinkles, characterized in that selected from oils, lipsticks and foundations.

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