KR20150001370A - Composition for preventing and improving skin wrinkles - Google Patents

Abstract

The present invention relates to a composition for preventing and reducing skin wrinkles, which includes a human telomerase-derived peptide as an active component. The composition according to the present invention includes a human telomerase-derived peptide as an active component to produce excellent effects by increasing the biosynthesis of collagen. Therefore, the increased biosynthesis of collagen prevents or reduces wrinkles, and maintains or improves skin quality.

Description

Technical Field [0001] The present invention relates to a composition for preventing and improving skin wrinkles,

The present invention relates to a composition for preventing and improving skin wrinkles comprising a peptide derived from human telomerase as an active ingredient.

Skin aging is one of the most obvious evidences of aging, and skin aging processes are often caused by photo-aging due to persistent exposure to short wavelength UVs, often accompanied by wrinkles, facial, Such as solar lentigo and spotty pigmentation appearing in exposed areas of the skin, such as the eye.

The cause of wrinkles is largely divided into the generation of wrinkles by natural aging and the generation of wrinkles by exposure to ultraviolet rays (photoaging). However, the mechanism of wrinkling due to two causes has not been clarified yet. A common phenomenon found in association with skin changes due to natural aging and photoaging is the reduction and modification of collagen and elastin, the main constituent proteins in the dermis, and the reduction of skin elasticity due to the reduction of the hyaluronic acid, the substrate of the dermis. Particularly, according to the results thus far, reduction and modification of collagen and elastin, which are main proteins in the dermis, are directly related to the generation of wrinkles and the reduction of elasticity of skin.

Collagen is a major substrate protein produced in fibroblasts of the skin and exists in extracellular epilepsy. Its important functions are mechanical rigidity of skin, resistance of connective tissues and binding force of tissues, support of cell adhesion, division of cells and differentiation And the induction of wound healing). Such collagen is reduced by aging and photoaging by ultraviolet irradiation, which is known to be closely related to the wrinkling of the skin. In recent years, as a result of extensive research on skin aging, important functions of collagen in the skin have been revealed. Effective ingredients promoting collagen synthesis and exhibiting wrinkle-reducing effects are known.

Korean Patent Publication No. 1020060122844

Therefore, the inventors of the present invention have found that peptides derived from human telomerase can have a superior wrinkle-reducing effect as a result of efforts to develop a composition which is more safe for human body and excellent in the effect of improving wrinkles.

One aspect of the invention is to provide peptides useful for the skin.

One aspect of the present invention is to provide a composition comprising a peptide derived from human telomerase as an active ingredient.

One aspect of the present invention is to provide a peptide having skin wrinkle-improving effect.

One aspect of the present invention is to provide peptides useful for enhancing skin elasticity.

One aspect of the present invention is to provide a composition for external application for skin having an effect of preventing or improving skin wrinkles.

According to an aspect of the present invention, there is provided a peptide comprising an amino acid sequence of any one of SEQ ID NO: 1 to SEQ ID NO: 4 as an active ingredient, a peptide having 80% or more of sequence homology with the above peptide sequence, , Wherein the peptide or fragment thereof has collagen biosynthesis performance.

In the composition for preventing or improving skin wrinkles according to one aspect of the present invention, the fragment may be composed of three or more amino acids.

In the composition for preventing or improving skin wrinkles according to one aspect of the present invention, the peptide may be composed of an amino acid sequence of any one of SEQ ID NOS: 1 to 4.

In the composition for preventing or improving skin wrinkles according to one aspect of the present invention, the peptide may be derived from human telomerase.

In the composition for preventing or improving skin wrinkles according to one aspect of the present invention, the composition may be a composition for external application to the skin.

In the composition for preventing or improving skin wrinkles according to one aspect of the present invention, the composition may be a cosmetic composition.

In the composition for preventing or improving skin wrinkles according to one aspect of the present invention, the composition may be a pharmaceutical composition.

In the composition for preventing or improving skin wrinkles according to one aspect of the present invention, the composition may be a food composition.

The composition according to the present invention exhibits an excellent collagen biosynthesis-increasing effect by containing a human telomerase-derived peptide as an active ingredient. Therefore, by promoting collagen biosynthesis of the skin, it is possible to prevent or prevent wrinkles of the skin, or to maintain or enhance a healthy skin condition by improving the wrinkles.

FIG. 1 is a graph showing the results of measurement of collagen synthesis (%) of peptides according to one aspect of the present invention.
FIG. 2 is a graph showing cell viability (%) for confirming toxicity to cells when the peptides according to the present invention are treated at a maximum concentration value.

The present invention can be variously modified and can have various embodiments, and the present invention will be described in more detail as follows. It is to be understood, however, that the invention is not to be limited to the specific embodiments, but includes all modifications, equivalents, and alternatives falling within the spirit and scope of the invention. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS Hereinafter, the present invention will be described in detail with reference to the accompanying drawings.

The peptides shown in SEQ ID NO: 1 to SEQ ID NO: 4 are shown in Table 1 below. SEQ ID NO: 5 is the sequence of the human telomerase full-length protein. The "name" in Table 1 below is what the peptides are named for. In another aspect of the present invention, at least one of the peptides of SEQ ID NOS: 1 to 4 includes a "synthetic peptide" synthesized by selecting peptides at corresponding positions among the peptides contained in telomerase. As used herein, the term "pep" is a generic term for a peptide having any one of SEQ ID NOS: 1 to 4, or a peptide having 80% or more sequence homology with the sequence, or a fragment thereof.

SEQ ID NO: name Telomerase phase location order Length One. pep-RIA-109 [617-620] LTSR 4 aa 2. pep-RIA-129 [613-615] RPA 3 aa 3. pep-RIA-134 [614-620] PALLTSR 7 aa 4. pep-RIA-136 [614-618] PALLT 5 aa 5. Telomerase [1-1132] MPRAPRCRAVRSLLRSHYREVLPLATFVRR LGPQGWRLVQRGDPAAFRALVAQCLVCVPWDARPPPAAPSFRQVSCLKELVARVLQRLCERGAKNVLAFGFALLDGARGGPPEAFTTSVRSYLPNTVTDALRGSGAWGLLLRRVGDDVLVHLLARCALFVLVAPSCAYQVCGPPLYQLGAATQARPPPHASGPRRRLGCERAWNHSVREAGVPLGLPAPGARRRGGSASRSLPLPKRPRRGAAPEPERTPVGQGSWAHPGRTRGPSDRGFCVVSPARPAEEATSLEGALSGTRHSHPSVGRQHHAGPPSTSRPPRPWDTPCPPVYAETKHFLYSSGDKEQLRPSFLLSSLRPSLTGARRLVETIFLGSRPWMPGTPRRLPRLPQRYWQMRPLFLELLGNHAQCPYGVLLKTHCPLRAAVTPAAGVCAREKPQGSVAAPEEEDTDPRRLVQLLRQHSSPWQVYGFVRACLRRLVPPGLWGSRHNERRFLRNTKKFISLGKHAKLSLQELTW KMSVRDCAWLRRSPGVGCVPAAEHRLREEILAKFLHWLMSVYVVELLRSFFYVTETTFQKNRLFFYRKSVWSKLQSIGIRQHLKRVQLRELSEAEVRQHREARPALLTSRLRFIPKPDGLRPIVNMDYVVGARTFRREKRAERLTSRVKALFSVLNYERARRPGLLGASVLGLDDIHRAWRTFVLRVRAQDPPPELYFVKVDVTGAYDTIPQDRLTEVIASIIKPQNTYCVRRYAVVQKAAHGHVRKAFKSHVSTLTDLQPYMRQFVAHLQETSPLRDAVVIEQSSSLNEASSGLFDVFLRFMCHHAVRIRGKSYVQCQGIPQGSILSTLLCSLCYGDMENKLFAGIRRDGLLLRLVDDFLLVTPHLTHAKTFLRTLVRGVPEYGCVVNLRKTVVNFPVEDEALGGTAFVQMPAHGLFPWCGLLLDTRTLEVQSDYSSYARTSIRASLTFNRGFKAGRNMRRKLFGVLRLKCHSLFLDLQ VNSLQTV CTNIYKILLLQAYRFHACVLQLPFHQQVWKNPTFFLRVISDTASLCYSILKAKNAGMSLGAKGAAGPLPSEAVQWLCHQAFLLKLTRHRVTYVPLLGSLRTAQTQLSRKLPGTTLTALEAAANPALPSDFKTILD 1132 aa

An aspect of the present invention provides a peptide comprising SEQ ID NOS: 1 to 4, or a peptide having 80% or more sequence homology with the peptide sequence, or a polynucleotide encoding the fragment. Peptides can be mass produced using the polynucleotides. For example, a peptide containing a polynucleotide encoding a peptide can be cultured in a host cell to produce a large amount of peptide.

The peptides disclosed herein may comprise peptides having 80% or more, 85% or more, 90% or more, 95% or more, or 96% or more of the sequence homology with the peptides of SEQ ID NOS: 1-4. In addition, the peptide disclosed in the present specification may be a peptide comprising SEQ ID NOS: 1 to 4, or a peptide having 80% or more of sequence homology with the peptide sequence, or a fragment thereof and one or more amino acids, two or more amino acids, An amino acid, at least four amino acids, at least five amino acids, at least six amino acids, or at least seven amino acid altered peptides.

In one aspect of the invention, the amino acid change is of a property that causes the physicochemical properties of the peptide to change. For example, amino acid changes such as improving the thermal stability of the peptide, altering the substrate specificity, changing the optimum pH, etc. can be performed.

As used herein, the term "amino acid" includes D-isomers and modified amino acids as well as the 22 standard amino acids that are naturally incorporated into the peptide. Accordingly, in one aspect of the present invention, the peptide may be a peptide comprising a D-amino acid. In another aspect of the present invention, the peptide may include post-translationally modified non-standard amino acids and the like. Examples of post-translational modifications include phosphorylation, glycosylation, acylation (including, for example, acetylation, myristoylation and palmitoylation), alkylation ), Carboxylation, hydroxylation, glycation, biotinylation, ubiquitinylation, changes in chemical properties (such as, for example, beta-depleted deamidation , Deamidation) and structural changes (e.g., formation of a disulfide bridge). Also included are amino acid changes such as changes in amino acids, such as changes in amino groups, carboxy groups or side chains, caused by chemical reactions that take place during binding with crosslinkers to form peptide conjugates.

The peptides disclosed herein may be wild type peptides identified and isolated from natural sources. Meanwhile, the peptides disclosed in the present specification may include a peptide comprising SEQ ID NOS: 1 to 4, or a peptide having 80% or more sequence homology with the peptide sequence, or a peptide having one or more amino acids substituted, deleted and / Or an inserted amino acid sequence. Amino acid changes in wild-type polypeptides as well as in artificial variants include conservative amino acid substitutions that do not significantly affect folding and / or activity of the protein. Examples of conservative substitutions include, but are not limited to, basic amino acids (arginine, lysine and histidine), acidic amino acids (glutamic acid and aspartic acid), polar amino acids (glutamine and asparagine), hydrophobic amino acids (leucine, isoleucine, valine and methionine) Tryptophan and tyrosine), and small amino acids (glycine, alanine, serine and threonine). In general, amino acid substitutions that do not alter specific activity are known in the art. The most commonly occurring interactions are Ala / Ser, Val / Ile, Asp / Glu, Thr / Ser, Ala / Gly, Ala / Thr, Ser / Asn, Ala / Val, Ser / Gly, Tyr / Lys / Arg, Asp / Asn, Leu / Ile, Leu / Val, Ala / Glu, and Asp / Gly, and vice versa.

According to another aspect of the present invention, there is provided a composition comprising the peptides derived from human proteins as described above, or peptides or fragments thereof having 80% or more sequence homology with the above peptide sequences as an active ingredient.

As the skin gets older, the secretion of various hormones that regulate metabolism is reduced internally, and the function of immune cells and the activity of cells are lowered, and the biosynthesis of immune proteins and biologic proteins necessary for the living body is reduced. In addition, externally, ozone layer breakage increases the amount of ultraviolet rays reaching the surface of the sunlight and increases the environmental pollution, thereby increasing free radicals and active noxious oxygen, thereby reducing skin thickness and wrinkling , And the elasticity is decreased. Therefore, as the aging progresses, the content and arrangement of collagen, elastin, hyaluronic acid, and glycoprotein, which constitute the skin, change or decrease. In other words, decrease of cellular activity due to natural endogenous aging and decrease of biosynthesis of substrate material by microinflammation, increase of stress due to various harmful environment, increase of active oxygen species by sunlight, As the degradation and denaturation are accelerated, the skin substrate is destroyed and thinned, and various symptoms of skin aging appear.

Telomere is a genetic material that is repeatedly present at the end of a chromosome, and is known to prevent damage to the chromosome or its binding to other chromosomes. As the cell divides, the length of the telomere is getting shorter. When there is more than a certain number of cell divisions, the telomere becomes very short, and the cell stops dividing and dies. On the other hand, it is known that lengthening the telomeres prolongs the life of the cells. For example, it is known that the cancer cells secrete an enzyme called telomerase and prevent the shortening of the telomeres.

Accordingly, one aspect of the present invention provides a peptide comprising the peptide of SEQ ID NOS: 1 to 4, a peptide having 80% or more sequence homology with the peptide sequence, or a fragment thereof as an active ingredient, When applied or administered to the skin, the biosynthesis of collagen in the skin reduced by aging can be promoted. Thus, the composition according to the present invention can prevent or improve wrinkles of the skin, maintain or improve skin elasticity, and prevent or improve the aging of the skin.

The composition according to one aspect of the present invention comprises the peptide of SEQ ID Nos: 1 to 4 or a peptide having the sequence homology of 80% or more with the peptide sequence, or a fragment thereof as an active ingredient in an amount of 0.1 μg / mg to 1 mg / Mg / mg to 0.5 mg / mg, more specifically from 10 mu g / mg to 0.1 mg / mg. When it is included in the above range, it is not only suitable for exhibiting the intended effect of the present invention but also can satisfy both the stability and safety of the composition, and may be suitably included in the above range in terms of cost effectiveness.

The composition according to one aspect of the present invention can be applied to all animals including humans, dogs, chickens, pigs, cattle, sheep, guinea pigs or monkeys.

The pharmaceutical composition according to one aspect of the present invention can be administered orally, rectally, transdermally, intravenously, intramuscularly, intraperitoneally, intramuscularly, intradermally or subcutaneously.

Formulations for oral administration may be, but are not limited to, tablets, pills, soft or hard capsules, granules, powders, solutions or emulsions. Formulations for parenteral administration may be, but are not limited to, injections, drops, lozenges, ointments, gels, creams, suspensions, emulsions, suppositories, patches or spraying agents.

The pharmaceutical composition according to one aspect of the present invention may contain additives such as a diluent, an excipient, a lubricant, a binder, a disintegrant, a buffer, a dispersant, a surfactant, a colorant, a fragrance or a sweetener as necessary. The pharmaceutical composition according to one aspect of the present invention can be prepared by a conventional method in the art.

The effective ingredients of the pharmaceutical composition according to one aspect of the present invention will vary depending on the age, sex, weight, pathological condition and severity of the subject to be administered, route of administration, or judgment of the prescriber. Determination of the amount of application based on these factors is within the level of ordinary skill in the art and its daily dose is, for example, from 0.1 [mu] g / kg / day to 1 g / kg / day, Kg / day, more specifically, 10 μg / kg / day to 1 mg / kg / day, and more particularly 50 μg / kg / day to 100 μg / kg / day. The pharmaceutical composition according to one aspect of the present invention may be administered once to three times a day, but is not limited thereto.

The composition for external application for skin according to one aspect of the present invention may be provided in all formulations suitable for topical application. For example, it may be provided as a solution, an emulsion obtained by dispersing an oil phase in an aqueous phase, an emulsion obtained by dispersing an oil phase in water, a suspension, a solid, a gel, a powder, a paste, a foam or an aerosol. Such formulations may be prepared according to conventional methods in the art.

In addition, the cosmetic composition according to one aspect of the present invention may be provided as an injection preparation. The cosmetic composition of the present invention can be used as a cosmetic composition for injections through a syringe having a single needle, a syringe having a plurality of needles, a high-speed jet by a patch, a micro needle or a micro jet method, or a shock wave induced by applying a laser beam to an aluminum foil May be injected into the skin through various delivery mechanisms, for example, a device for spraying the drug. According to the method using the laser, since the injection needle is not used, it is possible to prevent the pain at the time of injection and the scar due to the use of the injection needle. According to the injection formulation, the active ingredient contained in the cosmetic composition can be directly delivered to a target site such as skin, thereby maximizing the effect. Further, according to the method using a laser, since the injection needle is not used, it is possible to prevent the pain at the time of injection and the scar due to the use of the injection needle.

The cosmetic composition according to one aspect of the present invention may contain other ingredients, which may be synergistic to the main effect, preferably to the extent that the main effect is not impaired. In addition, the cosmetic composition according to one aspect of the present invention may further contain a moisturizing agent, an emollient agent, a surfactant, an ultraviolet absorber, an antiseptic, a bactericide, an antioxidant, a pH adjuster, an organic or inorganic pigment, a fragrance, . The compounding amount of the above components can be easily selected by those skilled in the art within the range not impairing the object and effect of the present invention, and the amount thereof is 0.01 to 5% by weight, specifically 0.01 to 3% by weight .

The formulation of the food composition according to one aspect of the present invention is not particularly limited, but can be formulated into, for example, tablets, granules, powders, liquids, solid preparations and the like. Each formulation may be blended without difficulty by a person skilled in the art according to the purpose of formulation or use, in addition to the active ingredient, and the synergistic effect may occur when the composition is applied simultaneously with other ingredients.

Determination of the dosage of the active ingredient is within the level of ordinary skill in the art and its daily dose is, for example, 1 μg / kg / day to 10 mg / kg / day, more specifically 10 μg / kg / day Day to 100 mg / kg / day, but it is not limited thereto, and it may be various factors such as the age, health condition, and complication of the subject to be administered ≪ / RTI >

Hereinafter, the constitution and effects of the present invention will be described in more detail with reference to Examples and Experimental Examples. However, the following examples and experimental examples are provided for illustrative purposes only in order to facilitate understanding of the present invention, and the scope and scope of the present invention are not limited thereto.

Example 1 Synthesis of Peptides

Peptides having the amino acid sequences of SEQ ID NO: 1 to SEQ ID NO: 4 were prepared according to a conventional solid phase peptide synthesis method. Specifically, the peptides were synthesized by coupling one amino acid from the C-terminal through Fmoc solid phase peptide synthesis (SPPS) using ASP48S (Peptron, Inc., Daejeon, Korea). The first amino acid at the C-terminus of the peptides attached to the resin was used as follows. For example:

NH ₂ -Lys (Boc) -2-chloro-Trityl Resin

NH ₂ -Ala-2-chloro-Trityl Resin

NH ₂ -Arg (Pbf) -2-chloro-Trityl Resin

Boc, t-Bu (t-butylester), Pbf (2, 2, 4, 6, 6), which are all amino acid sources used for peptide synthesis, are protected by N-term and Fmoc, 7-pentamethyl dihydro-benzofuran-5-sulfonyl). For example:

Fmoc-Pro-OH, Fmoc-Leu-OH, Fmoc-Phe-OH, Fmoc-Phe-OH, Fmoc-Arg- Fmoc-Lys (Boc) -OH, Fmoc-Gln (Trt) -OH, Fmoc-Trp (Boc) -OH, Fmoc-Met-OH, Fmoc- -Asn (Trt) -OH, Fmoc-Tyr (tBu) -OH, Fmoc-Ahx-OH, Trt-Mercaptoacetic acid.

As the coupling reagent, HBTU [2- (1H-Benzotriazole-1-yl) -1,1,3,3-tetramethylaminium hexafluorophosphate] / HOBt [N-Hydroxxybenzotriazole] / NMM [4-Methylmorpholine] Respectively. Fmoc removal was performed using piperidine in DMF in 20% DMF. Cleavage Cocktail [TFA (trifluoroacetic acid) / TIS (triisopropylsilane) / EDT (ethanedithiol) / H ₂ O = 92.5 / 2.5 / 2.5 / 2.5] was used to remove the synthesized peptide from Resin and to remove the protecting group of the residue. Respectively.

Each of the peptides was synthesized by repeating the steps of reacting corresponding amino acids with a starting amino acid having an amino acid protecting group bonded thereto on a solid support, washing with a solvent, followed by deprotection. The synthesized peptide was cleaved from the resin and purified by HPLC. The synthesis was confirmed by MS and lyophilized.

A specific procedure is as follows, taking as an example a peptide having 16 amino acid sequences of EARPALLTSRLRFIPK (SEQ ID NO: 6) as a telomerase-derived peptide.

1) Coupling

The protected amino acid (8 eq) and the coupling reagent HBTU (8 eq) / HOBt (8 eq) / NMM (16 eq) were dissolved in DMF and added to NH ₂ -Lys (Boc) -2-chloro-Trityl Resin , And reacted at room temperature for 2 hours and washed with DMF, MeOH and DMF in that order.

2) Fmoc deprotection

Piperidine in DMF in 20% DMF was added, and the reaction was carried out at room temperature for 5 minutes twice, followed by washing with DMF, MeOH and DMF.

3) Repeating the reaction of 1 and 2 to obtain a peptide basic skeleton NH2-E (OtBu) -AR (Pbf) -PALLT (tBu) -S (tBu) -R (Pbf) LR (Pbf) 2-chloro-Trityl Resin).

4) Cleavage: Cleavage cocktail was added to the synthesized peptide Resin to separate the peptide from Resin.

5) Add Cooling diethyl ether to the obtained mixture and centrifuge to precipitate the obtained peptide.

6) After purification by Prep-HPLC, molecular weight was confirmed by LC / MS and frozen to prepare powder.

Example 2: Formulation of formulation

The lyophilized powder-form peptide obtained according to the method described in Example 1 was dissolved in phosphate buffered saline (PBS) and used. (Purity: 97.3%, content: 85.3%) against the purity of the peptide was prepared. Phosphate buffer solution (PBS) was used as an excipient immediately before administration to prepare a test solution at a concentration of 40 mM.

Example 3: Effect of enhancing collagen biosynthesis of peptides

1) Cell culture

Human dermal fibroblast (Gibco Cat. Number: C-013-5C) was prepared to confirm the effect of increasing the collagen biosynthesis of the peptides. The prepared human dermal fibroblasts were cultured in Medium 106 medium at 37 ° C and 5% CO ₂ by adding LSGS (Low Serum Growth Supplement), and subcultured by trypsinization, and 4 to 10 generation cells were used for the experiment .

2) Confirming the effect of increasing collagen synthesis

The human dermal fibroblast cultured as described above was added to a 24-well plate at 5 × 10 ⁴ / well and cultured for 14 to 16 hours. To remove the culture media and processed for each 5 μM, 50 μM, 250 μM peptide in Medium 106 medium without addition of LSGS (Low Serum Growth Supplement) After washing with phosphate buffered solution _{(PBS) 5% CO 2,} 37 Lt; 0 > C for 24 hours. The culture broth was collected and the amount of collagen was measured by ELISA (Procollagen type IC-peptide EIA kit). The ELISA method is as follows. Antibody-POD conjugated solution (100 μl) was added to 96-wells and incubated at 37 ° C for 3 hours. After removing the culture medium from the 96-wells and washing with washing buffer, 100 μl of color development reagent (TMBZ) was added and incubated at room temperature for 15 minutes. Then 100 μl of 1 N sulfuric acid was added. And then measured with an ELISA reader at 450 nm. The experimental results are shown in Fig.

As shown in FIG. 1, the peptides pep-RIA-109, pep-RIA-129, pep-RIA-134 and pep-RIA-136 increased collagen synthesis to 119%, 114%, 118% and 123% at 50 μM You can tell. That is, when the peptide according to one aspect of the present invention is not added (control) at 100%, the synthesis of collagen is increased by about 114 to 123% in all cases where the peptide is added at a concentration of 50 μM Respectively. This means that peptides according to one aspect of the present invention can increase collagen biosynthesis, thereby improving skin wrinkles and elasticity.

Example 4: Cytotoxicity test of peptides

To confirm the cytotoxicity of peptides, human dermal fibroblast (Gibco Cat. No. C-013-5C) was added to a 96-well plate at 1 × 10 ⁴ cells per well and cultured for 12 hours.

After the culture medium was removed, the cells were washed with PBS. Then, each of the 500 μM peptides was treated with Medium 106 medium without LSGS (Low Serum Growth Supplement) and cultured at 37 ° C. for 24 hours at 5% CO ₂ . After the 24 hours, MTT solution (3- (4,5-Dimethylthiazol-2 -yl) -2,5-diphenyltetrazoliumbromide; 5 mg / ml) was added to 10 ㎕ for 2 hours in 5% CO _2, 37 ℃ Lt; / RTI > The medium was removed and 100 μl of DMSO (dimethylsulfoxide) was added. After shaking at room temperature for 10 minutes, the absorbance was measured at 570 nm using a microplate reader. The cell viability (%) was confirmed using the measured values.

The survival rate of the cells was calculated by the following equation (1).

OD ₅₇₀ treated: 570 nm absorbance of the peptide treated wells

OD ₅₇₀ untreated: absorbance at 570 nm in the untreated well

OD ₅₇₀ blank: 570 nm absorbance of cell-cultured wells

Experimental results As shown in Fig. 2, the peptides were incubated at 5 [mu] M, 50 [mu] M. It can be confirmed that no cytotoxicity is induced in all cases where 250 μM is added.

Cosmetic Formulation Example: Softening Longevity and Cream Production

As a formulation, a lotion and cream containing a peptide of the present invention as an active ingredient were prepared according to a conventional method with the following composition.

[Formulation Example 1] Flexible longevity

 [Formulation Example 2] Cream

<110> KAEL-GEMVAX CO., LTD.          KIM, Sangjae <120> Composition for preventing and improving skin wrinkles <130> 13P393 / IND <160> 6 <170> PatentIn version 3.2 <210> 1 <211> 4 <212> PRT <213> Homo sapiens <400> 1 Leu Thr Ser Arg   One <210> 2 <211> 3 <212> PRT <213> Homo sapiens <400> 2 Arg Pro Ala   One <210> 3 <211> 7 <212> PRT <213> Homo sapiens <400> 3 Pro Ala Leu Leu Thr Ser Arg   1 5 <210> 4 <211> 5 <212> PRT <213> Homo sapiens <400> 4 Pro Ala Leu Leu Thr   1 5 <210> 5 <211> 1132 <212> PRT <213> Homo sapiens <400> 5 Met Pro Arg Ala Pro Arg Cys Arg Ala Val Arg Ser Leu Leu Arg Ser   1 5 10 15 His Tyr Arg Glu Val Leu Pro Leu Ala Thr Phe Val Arg Arg Leu Gly              20 25 30 Pro Gln Gly Trp Arg Leu Val Gln Arg Gly Asp Pro Ala Ala Phe Arg          35 40 45 Ala Leu Val Ala Gln Cys Leu Val Cys Val Pro Trp Asp Ala Arg Pro      50 55 60 Pro Pro Ala Ala Pro Ser Phe Arg Gln Val Ser Cys Leu Lys Glu Leu  65 70 75 80 Val Ala Arg Val Leu Gln Arg Leu Cys Glu Arg Gly Ala Lys Asn Val                  85 90 95 Leu Ala Phe Gly Phe Ala Leu Leu Asp Gly Ala Arg Gly Gly Pro Pro             100 105 110 Glu Ala Phe Thr Thr Ser Val Arg Ser Tyr Leu Pro Asn Thr Val Thr         115 120 125 Asp Ala Leu Arg Gly Ser Gly Ala Trp Gly Leu Leu Leu Arg Arg Val     130 135 140 Gly Asp Asp Val Leu Val His Leu Leu Ala Arg Cys Ala Leu Phe Val 145 150 155 160 Leu Val Ala Pro Ser Cys Ala Tyr Gln Val Cys Gly Pro Pro Leu Tyr                 165 170 175 Gln Leu Gly Ala Ala Thr Gln Ala Arg Pro Pro His Ala Ser Gly             180 185 190 Pro Arg Arg Arg Leu Gly Cys Glu Arg Ala Trp Asn His Ser Val Arg         195 200 205 Glu Ala Gly Val Pro Leu Gly Leu Pro Ala Pro Gly Ala Arg Arg Arg     210 215 220 Gly Gly Ser Ala Ser Arg Ser Leu Pro Leu Pro Lys Arg Pro Arg Arg 225 230 235 240 Gly Ala Ala Pro Glu Pro Glu Arg Thr Pro Val Gly Gln Gly Ser Trp                 245 250 255 Ala His Pro Gly Arg Thr Arg Gly Pro Ser Asp Arg Gly Phe Cys Val             260 265 270 Val Ser Pro Ala Arg Pro Ala Glu Glu Ala Thr Ser Leu Glu Gly Ala         275 280 285 Leu Ser Gly Thr Arg His Ser Ser Ser Val Gly Arg Gln His His     290 295 300 Ala Gly Pro Pro Ser Thr Ser Arg Pro Pro Arg Pro Trp Asp Thr Pro 305 310 315 320 Cys Pro Pro Val Tyr Ala Glu Thr Lys His Phe Leu Tyr Ser Ser Gly                 325 330 335 Asp Lys Glu Gln Leu Arg Pro Ser Phe Leu Leu Ser Ser Leu Arg Pro             340 345 350 Ser Leu Thr Gly Ala Arg Arg Leu Val Glu Thr Ile Phe Leu Gly Ser         355 360 365 Arg Pro Trp Met Pro Gly Thr Pro Arg Arg Leu Pro Arg Leu Pro Gln     370 375 380 Arg Tyr Trp Gln Met Arg Pro Leu Phe Leu Glu Leu Leu Gly Asn His 385 390 395 400 Ala Gln Cys Pro Tyr Gly Val Leu Leu Lys Thr His Cys Pro Leu Arg                 405 410 415 Ala Ala Val Thr Pro Ala Ala Gly Val Cys Ala Arg Glu Lys Pro Gln             420 425 430 Gly Ser Val Ala Ala Pro Glu Glu Glu Asp Thr Asp Pro Arg Arg Leu         435 440 445 Val Gln Leu Leu Arg Gln His Ser Ser Pro Trp Gln Val Tyr Gly Phe     450 455 460 Val Arg Ala Cys Leu Arg Arg Leu Val Pro Pro Gly Leu Trp Gly Ser 465 470 475 480 Arg His Asn Glu Arg Arg Phe Leu Arg Asn Thr Lys Lys Phe Ile Ser                 485 490 495 Leu Gly Lys His Ala Lys Leu Ser Leu Gln Glu Leu Thr Trp Lys Met             500 505 510 Ser Val Arg Asp Cys Ala Trp Leu Arg Arg Ser Pro Gly Val Gly Cys         515 520 525 Val Pro Ala Ala Glu His Arg Leu Arg Glu Glu Ile Leu Ala Lys Phe     530 535 540 Leu His Trp Leu Met Ser Val Tyr Val Val Glu Leu Leu Arg Ser Phe 545 550 555 560 Phe Tyr Val Thr Glu Thr Thr Phe Gln Lys Asn Arg Leu Phe Phe Tyr                 565 570 575 Arg Lys Ser Val Trp Ser Lys Leu Gln Ser Ile Gly Ile Arg Gln His             580 585 590 Leu Lys Arg Val Gln Leu Arg Glu Leu Ser Glu Ala Glu Val Arg Gln         595 600 605 His Arg Glu Ala Arg Pro Ala Leu Leu Thr Ser Arg Leu Arg Phe Ile     610 615 620 Pro Lys Pro Asp Gly Leu Arg Pro Ile Val Asn Met Asp Tyr Val Val 625 630 635 640 Gly Ala Arg Thr Phe Arg Arg Glu Lys Arg Ala Glu Arg Leu Thr Ser                 645 650 655 Arg Val Lys Ala Leu Phe Ser Val Leu Asn Tyr Glu Arg Ala Arg Arg             660 665 670 Pro Gly Leu Leu Gly Ala Ser Val Leu Gly Leu Asp Asp Ile His Arg         675 680 685 Ala Trp Arg Thr Phe Val Leu Arg Val Arg Ala Gln Asp Pro Pro Pro     690 695 700 Glu Leu Tyr Phe Val Lys Val Asp Val Thr Gly Ala Tyr Asp Thr Ile 705 710 715 720 Pro Gln Asp Arg Leu Thr Glu Val Ile Ala Ser Ile Ile Lys Pro Gln                 725 730 735 Asn Thr Tyr Cys Val Arg Arg Tyr Ala Val Val Gln Lys Ala Ala His             740 745 750 Gly His Val Arg Lys Ala Phe Lys Ser His Val Ser Thr Leu Thr Asp         755 760 765 Leu Gln Pro Tyr Met Arg Gln Phe Val Ala His Leu Gln Glu Thr Ser     770 775 780 Pro Leu Arg Asp Ala Val Valle Glu Gln Ser Ser Ser Leu Asn Glu 785 790 795 800 Ala Ser Ser Gly Leu Phe Asp Val Phe Leu Arg Phe Met Cys His His                 805 810 815 Ala Val Arg Ile Arg Gly Lys Ser Tyr Val Gln Cys Gln Gly Ile Pro             820 825 830 Gln Gly Ser Ile Leu Ser Thr Leu Leu Cys Ser Leu Cys Tyr Gly Asp         835 840 845 Met Glu Asn Lys Leu Phe Ala Gly Ile Arg Arg Asp Gly Leu Leu Leu     850 855 860 Arg Leu Val Asp Asp Phe Leu Leu Val Thr Pro His Leu Thr His Ala 865 870 875 880 Lys Thr Phe Leu Arg Thr Leu Val Arg Gly Val Pro Glu Tyr Gly Cys                 885 890 895 Val Val Asn Leu Arg Lys Thr Val Val Asn Phe Pro Val Glu Asp Glu             900 905 910 Ala Leu Gly Gly Thr Ala Phe Val Gln Met Pro Ala His Gly Leu Phe         915 920 925 Pro Trp Cys Gly Leu Leu Leu Asp Thr Arg Thr Leu Glu Val Gln Ser     930 935 940 Asp Tyr Ser Ser Tyr Ala Arg Thr Ser Ile Arg Ala Ser Leu Thr Phe 945 950 955 960 Asn Arg Gly Phe Lys Ala Gly Arg Asn Met Arg Arg Lys Leu Phe Gly                 965 970 975 Val Leu Arg Leu Lys Cys His Ser Leu Phe Leu Asp Leu Gln Val Asn             980 985 990 Ser Leu Gln Thr Val Cys Thr Asn Ile Tyr Lys Ile Leu Leu Leu Gln         995 1000 1005 Ala Tyr Arg Phe His Ala Cys Val Leu Gln Leu Pro Phe His Gln Gln    1010 1015 1020 Val Trp Lys Asn Pro Thr Phe Phe Leu Arg Val Ile Ser Asp Thr Ala 1025 1030 1035 1040 Ser Leu Cys Tyr Ser Ile Leu Lys Ala Lys Asn Ala Gly Met Ser Leu                1045 1050 1055 Gly Ala Lys Gly Ala Gly Pro Leu Pro Ser Glu Ala Val Gln Trp            1060 1065 1070 Leu Cys His Gln Ala Phe Leu Leu Lys Leu Thr Arg His Arg Val Thr        1075 1080 1085 Tyr Val Pro Leu Leu Gly Ser Leu Arg Thr Ala Gln Thr Gln Leu Ser    1090 1095 1100 Arg Lys Leu Pro Gly Thr Thr Leu Thr Ala Leu Glu Ala Ala Ala Asn 1105 1110 1115 1120 Pro Ala Leu Pro Ser Asp Phe Lys Thr Ile Leu Asp                1125 1130 <210> 6 <211> 16 <212> PRT <213> Homo sapiens <400> 6 Glu Ala Arg Pro Ala Leu Leu Thr Ser Arg Leu Arg Phe Ile Pro Lys   1 5 10 15

Claims (8)

As the active ingredient,
A peptide comprising an amino acid sequence of any one of SEQ ID NO: 1 to SEQ ID NO: 4, a peptide having 80% or more of sequence homology with the peptide sequence, or a fragment thereof,
Wherein the peptide or fragment thereof has collagen biosynthesis capability. The composition for preventing or improving skin wrinkles according to claim 1, wherein the fragment is a peptide consisting of three or more amino acids. The composition for preventing or improving skin wrinkles according to claim 1, wherein the peptide is a peptide consisting of an amino acid sequence of any one of SEQ ID NOS: 1 to 4. The composition according to claim 1, wherein the peptide is derived from human telomerase. The composition according to claim 1, wherein the composition is an external preparation for skin. The composition according to claim 1, wherein the composition is a cosmetic composition. The composition according to claim 1, wherein the composition is a pharmaceutical composition. The composition according to claim 1, wherein the composition is a food composition.

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