WO2017183942A2 - Peptide having efficacy of increasing collagen generation and composition comprising same - Google Patents

Peptide having efficacy of increasing collagen generation and composition comprising same Download PDF

Info

Publication number
WO2017183942A2
WO2017183942A2 PCT/KR2017/004288 KR2017004288W WO2017183942A2 WO 2017183942 A2 WO2017183942 A2 WO 2017183942A2 KR 2017004288 W KR2017004288 W KR 2017004288W WO 2017183942 A2 WO2017183942 A2 WO 2017183942A2
Authority
WO
WIPO (PCT)
Prior art keywords
peptide
composition
aging
less
skin
Prior art date
Application number
PCT/KR2017/004288
Other languages
French (fr)
Korean (ko)
Other versions
WO2017183942A3 (en
Inventor
김상재
Original Assignee
주식회사 젬백스앤카엘
김상재
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by 주식회사 젬백스앤카엘, 김상재 filed Critical 주식회사 젬백스앤카엘
Publication of WO2017183942A2 publication Critical patent/WO2017183942A2/en
Publication of WO2017183942A3 publication Critical patent/WO2017183942A3/en

Links

Images

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/04—Peptides having up to 20 amino acids in a fully defined sequence; Derivatives thereof
    • A61K38/08—Peptides having 5 to 11 amino acids
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/17—Amino acids, peptides or proteins
    • A23L33/18—Peptides; Protein hydrolysates
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
    • A61K8/64—Proteins; Peptides; Derivatives or degradation products thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q19/00—Preparations for care of the skin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q19/00—Preparations for care of the skin
    • A61Q19/08—Anti-ageing preparations
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
    • C07K7/04—Linear peptides containing only normal peptide links
    • C07K7/06—Linear peptides containing only normal peptide links having 5 to 11 amino acids

Definitions

  • the present specification relates to a functional peptide having a collagen production increasing effect and a composition comprising the same, and more particularly, to a functional peptide having a collagen production increasing effect and effective in improving skin wrinkles and preventing skin aging by lowering collagen production. It relates to a peptide composition.
  • Retinol and retinoids are known to be effective in various fields such as psoriasis, aging, cancer, and acne.
  • this retinoid is known to prevent collagen loss by stimulating collagen formation by inhibiting collagenase-1 (MMP-1) when applied to the skin to prevent and restore endogenous and photoaging.
  • MMP-1 collagenase-1
  • TGF- ⁇ Transforming growth factor
  • betulinic acid wild yam extract
  • wild yam extract Korean Patent Publication No. 2009-0055079
  • the above-described skin improvement active ingredients may cause local skin irritation reactions such as erythema, itching, psoriasis, and scaling when applied to the skin, and their stability has not been verified when applied to a living body.
  • Patent Document 1 KR 2009-0055079 A
  • the present inventors have developed a peptide having novel activity and found that it has collagen synthesis efficacy and completed the present invention.
  • One aspect of the present invention is a peptide having at least 80% sequence homology with a peptide consisting of any one of the amino acid sequences represented by SEQ ID NO: 1 to 3, a peptide that is a fragment of the peptide sequence, the peptide sequence or a peptide thereof Provided are peptides or salts thereof that exhibit collagen synthesis activity selected from the group consisting of:
  • Another aspect of the present invention provides an anti-wrinkle and anti-aging composition
  • an anti-wrinkle and anti-aging composition comprising the peptide or a salt thereof as an active ingredient.
  • the present invention has a sequence homology of at least 80% to a peptide consisting of any one of the amino acid sequences represented by SEQ ID NO: 1 to 3, a peptide that is a fragment of the peptide sequence, the peptide sequence or a peptide sequence thereof
  • anti-wrinkle and anti-aging methods comprising applying to a subject in need thereof at least one selected from the group consisting of peptides, and salts thereof.
  • the present invention provides a peptide consisting of any one of amino acid sequences represented by SEQ ID NOs: 1 to 3, peptides which are fragments of the peptide sequences, peptides which are peptides, or fragments thereof, for use in wrinkle improvement and anti-aging. At least one selected from the group consisting of peptides having at least 80% sequence homology with the sequences, and salts thereof.
  • the present invention provides a peptide comprising any one of amino acid sequences represented by SEQ ID NOs: 1 to 3 as an active ingredient for wrinkle improvement and anti-aging, a peptide which is a fragment of the peptide sequence, the peptide sequence or a fragment thereof
  • One or more non-therapeutic cosmetic uses selected from the group consisting of peptides having at least 80% sequence homology with peptide sequences, and salts thereof.
  • the present invention provides a peptide comprising any one of amino acid sequences represented by SEQ ID NOs: 1 to 3 for use in the preparation of a composition for improving wrinkles and preventing aging, a peptide which is a fragment of the peptide sequence, the peptide
  • One or more uses selected from the group consisting of peptides having at least 80% sequence homology with peptide sequences that are sequences or fragments thereof, and salts thereof.
  • the composition may be characterized in that the cosmetic composition for wrinkle improvement and anti-aging.
  • the cosmetic composition may be characterized in that it consists of any one formulation selected from the group consisting of skin, lotion, cream, foundation, essence, gel, pack, foam cleansing, soap and skin ointment. have.
  • the composition may be characterized in that the pharmaceutical composition for wrinkle improvement and anti-aging.
  • the composition may be characterized in that the composition for wrinkle improvement and anti-aging health functional food.
  • Peptides having the sequence of SEQ ID NO according to the present invention have collagen synthesis efficacy, it is expected to provide a wrinkle improvement and anti-aging composition comprising a peptide having collagen synthesis activity.
  • Wrinkle improvement and anti-aging effects according to the present invention refers to the effect of preventing or aging skin by inhibiting or improving the production of skin wrinkles through increased collagen production by the composition according to the present invention.
  • Figure 1 is a human skin fibroblast CCD-986SK cell line, a control (no treatment), a control group treated with TGF- ⁇ 20ng / ml positive control group, peptide according to the invention (CP-54, CP-55 CP-56) each divided into 50 ⁇ M treated group collagen synthesis (collagen synthesis) measured by comparing the results compared to the control (100%) based on the relative graph (%).
  • a peptide having novel activity of collagen synthesis consisting of the sequences of SEQ ID NOs: 1 to 3 is disclosed.
  • the peptides disclosed herein are at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, 99% with each of the peptides selected from SEQ ID NOs: 1-3 It may include a peptide having the above sequence homology.
  • the peptides disclosed herein may include any one peptide selected from 1 to 3 and at least one amino acid, at least two amino acids, at least three amino acids, at least four amino acids, at least five amino acids, at least six amino acids or Seven or more amino acids may comprise a modified peptide.
  • amino acid changes belong to the property that allows the physicochemical properties of the peptide to be altered.
  • amino acid changes can be made, such as improving the thermal stability of the peptide, altering substrate specificity, changing the optimal pH, and the like.
  • Any one peptide selected from SEQ ID NOs: 1 to 3 according to an aspect of the present invention has the advantage of low intracellular toxicity and high in vivo stability.
  • the peptides set forth in SEQ ID NOS: 1 to 3 are shown in Table 1 below.
  • the "name” in Table 1 below is to distinguish peptides.
  • the present invention provides a nucleotide sequence encoding the peptide.
  • composition comprising a peptide comprising any one amino acid sequence selected from SEQ ID NO: 1 to 3 as an active ingredient.
  • composition according to one aspect of the present invention can be applied to all animals including humans, dogs, chickens, pigs, cows, sheep, guinea pigs or monkeys.
  • the external preparation composition for skin is not particularly limited in its formulation, and it is a softening longevity, astringent longevity, nourishing longevity, eye cream, nutrition cream, massage cream, cleansing cream, cleansing foam, cleansing water, powder, Formulations such as essences, packs, and the like.
  • the cosmetic composition according to an aspect of the present invention may be prepared in various forms according to a conventional cosmetic preparation method.
  • the cosmetic composition may be prepared in the form of a cosmetic product containing the peptide, lotion, cream, lotion, etc., which may be diluted with conventional cleansing liquid, astringent liquid, and moisturizing liquid.
  • the cosmetic composition may include conventional adjuvants such as stabilizers, solubilizers, vitamins, pigments, and flavorings commonly used in the field of cosmetic compositions.
  • the cosmetic composition comprising any one of the novel active peptides selected from SEQ ID NOs: 1 to 3 according to an aspect of the present invention may prepare a cosmetic composition by a known cosmetic composition preparation method, and is not limited to general skin cosmetics. Applicable to quasi-drugs and external medicines. These formulations may be prepared in any of the formulations commonly prepared, for example in the form of skins, lotions, creams, foundations, essences, gels, packs, foam cleansing, body oils, lipsticks, mascaras, makeup bases or soaps. It may be prepared as, but is not limited thereto.
  • any one new active peptide selected from SEQ ID NOs: 1 to 3 according to an aspect of the present invention, in addition to any one new active peptide selected from SEQ ID NOs: 1 to 3 other components of the formulation of the cosmetic, According to the purpose of use, those skilled in the art can mix and select the enemy without difficulty.
  • the cosmetic composition of the present invention may be used daily and may also be used for an indefinite period of time.
  • the amount, frequency of use and duration of use may be adjusted according to the age, skin condition or skin type of the user, and the concentration of the peptide.
  • compositions herein can be administered in any manner as desired, for example by oral, rectal, intravenous, arterial, muscular, subcutaneous, intradermal, endometrial or intraventricular injection.
  • Formulations for oral administration of a composition according to one aspect of the invention may be tablets, pills, soft or hard capsules, granules, powders, solutions or emulsions, and formulations for parenteral administration may be injections, drops, lotions, ointments. , Gels, creams, suspensions, emulsions, suppositories, patches or sprays, but is not limited thereto.
  • compositions herein may be used in the form of their pharmaceutically acceptable salts, and may be used alone or in combination with other pharmaceutically active compounds as well as in a suitable collection.
  • the salt is not particularly limited as long as it is pharmaceutically acceptable.
  • hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid, hydrofluoric acid, hydrobromic acid, formic acid acetic acid, tartaric acid, lactic acid, citric acid, fumaric acid, maleic acid, succinic acid, methanesulfonic acid , Benzene sulfonic acid, toluene sulfonic acid, naphthalene sulfonic acid and the like can be used.
  • the concentration of the peptide or salt thereof in the pharmaceutical composition according to one aspect of the present invention may be 1 to 100 ⁇ M.
  • the concentration is at least 1 ⁇ M, at least 3 ⁇ M, at least 5 ⁇ M, at least 7 ⁇ M, at least 10 ⁇ M, at least 15 ⁇ M, at least 20 ⁇ M, at least 25 ⁇ M, at least 30 ⁇ M, at least 35 ⁇ M, at least 40 ⁇ M, at least 50 ⁇ M, at least 60 ⁇ M, at least 70 ⁇ M, 80 ⁇ M or more, 85 ⁇ M or more, 90 ⁇ M or more, or 95 ⁇ M or more.
  • the concentration is 100 ⁇ M or less, 95 ⁇ M or less, 90 ⁇ M or less, 85 ⁇ M or less, 80 ⁇ M or less, 75 ⁇ M or less, 70 ⁇ M or less, 60 ⁇ M or less, 50 ⁇ M or less, 40 ⁇ M or less, 30 ⁇ M or less, 25 ⁇ M or less, 20 ⁇ M or less, 15 ⁇ M or less, 10 ⁇ M Or less, 7 ⁇ M or less, 5 ⁇ M or less, or 2 ⁇ M or less.
  • the concentration may preferably be 20 to 80 ⁇ M, more preferably 50 ⁇ M.
  • the dosage of the peptide or salt thereof according to one aspect of the present invention may be 0.1 ⁇ g / kg to 10 mg / kg.
  • the dosage is at least 0.1 ⁇ g / kg, at least 0.5 ⁇ g / kg, at least 1 ⁇ g / kg, at least 5 ⁇ g / kg, at least 10 ⁇ g / kg, at least 30 ⁇ g / kg, at least 50 ⁇ g / kg, at least 70 ⁇ g / kg , 100 ⁇ g / kg or more, 300 ⁇ g / kg or more, 500 ⁇ g / kg or more, 700 ⁇ g / kg or more, 900 ⁇ g / kg or more, 1 mg / kg or more, 2 mg / kg or more, 3 mg / kg or more, 4 mg / kg or more, 5 mg / kg or more , At least 6 mg / kg, at least 7 mg / kg, at least 8 mg / kg, at least 9 mg / kg, at least 9.5 mg
  • the dosage is 10 mg / kg or less, 9 mg / kg or less, 8 mg / kg or less, 7 mg / kg or less, 6 mg / kg or less, 5 mg / kg or less, 4 mg / kg or less, 3 mg / kg or less, 2 mg / kg or less, 1mg / kg or less, 900 ⁇ g / kg or less, 700 ⁇ g / kg or less, 500 ⁇ g / kg or less, 300 ⁇ g / kg or less, 100 ⁇ g / kg or less, 90 ⁇ g / kg or less, 70 ⁇ g / kg or less, 50 ⁇ g / kg or less, 30 ⁇ g / kg or less, 10 ⁇ g / kg or less, 7 ⁇ g / kg or less, 5 ⁇ g / kg or less, 3 ⁇ g / kg or less, 1 ⁇ g / kg or less, 0.9 ⁇ g / kg or less, 0.7 ⁇ g / kg or less, 0.5 ⁇ g / kg or less, 0.3 ⁇ g / kg or less,
  • the dosage may vary depending on the age, sex, weight, pathology and severity of the subject to be administered, the route of administration or the judgment of the prescriber. Application amount determination based on these factors is within the level of skill in the art.
  • compositions according to one aspect of the invention may include additives such as diluents, excipients, lubricants, binders, disintegrants, buffers, dispersants, surfactants, colorants, flavoring or sweetening agents as needed.
  • additives such as diluents, excipients, lubricants, binders, disintegrants, buffers, dispersants, surfactants, colorants, flavoring or sweetening agents as needed.
  • Compositions according to one aspect of the present invention may be prepared by conventional methods in the art.
  • composition according to an aspect of the present invention may increase collagen production, inhibit collagen production inhibitor secretion, and may inhibit or improve skin wrinkles and prevent aging.
  • Preferred embodiments according to one aspect of the invention include the most optimal mode known to the inventor for carrying out one aspect of the invention. Variations of the preferred embodiments may become apparent to those skilled in the art upon reading the foregoing description. The inventors expect skilled artisans to employ such variations as appropriate, and the inventors expect the various aspects of the invention to be practiced in a manner different from that described herein. Accordingly, this specification includes all modifications and equivalents of the subject matter referred to in the appended claims, as permitted by patent law. Moreover, any combination of the above-mentioned elements within all possible variations is included herein unless expressly stated to the contrary or apparently contradictory in context.
  • Peptides having the novel activity of SEQ ID NO: 1 were prepared according to solid phase peptide synthesis known in the art. Specifically, peptides were synthesized by coupling amino acids one by one from the C-terminus through Fmoc solid phase synthesis (SPPS) using ASP48S (Peptron, Inc., Daejeon, Korea). As follows, the first amino acid at the C-terminus of the peptides was attached to the resin. For example:
  • HBTU [2- (1H-Benzotriazole-1-yl) -1,1,3,3-tetamethylaminium hexafluorophosphate] / HOBt [N-Hydroxxybenzotriazole] / NMM [4-Methylmorpholine] is used as the coupling reagent. It was. Fmoc removal was performed using piperidine in DMF in 20% of DMF.
  • Each peptide was synthesized by repeating a process of reacting the amino acids with each other, washing with a solvent, and then deprotecting the amino acid using the state in which the amino acid protecting group was bound to the solid support.
  • the synthesized peptide was separated from the resin and purified by HPLC. The success of the synthesis was confirmed by LC / MS and lyophilized.
  • Peptides CP-55 and CP-56 were also prepared based on the same method as the preparation of CP-54, in which only the basic skeleton of the peptide was substituted for the corresponding sequence of each peptide.
  • human skin fibroblasts are cultured to improve collagen synthesis effect. After treating TGF- ⁇ (positive control) and each peptide known to be present, relative collagen synthesis of the positive control group and each peptide-treated experimental group was determined based on the control group without any treatment by absorbance measurement method. Calculated.
  • the CCD-986SK (ATCC® CRL-1947 TM ), a human skin fibroblast, was used.
  • the cell lines were placed in DMEM medium containing 10% FBS and penicillin-Streptomycin antibiotics and seeded in a 100 mm culture dish, followed by incubation in a 37 ° C., 5% CO 2 incubator. . While checking the growth rate and state of each cell every day, subcultures were performed at a ratio of 1: 2 to 1: 3 at intervals of 2 to 3 days to maintain a good state of the cells.
  • the cultured CCD-986SK cells were placed in a 24-well plate at 2 ⁇ 10 4 cells per well, and then cultured for 24 hours.
  • the culture medium was removed and washed with PBS, followed by starvation treatment for 2 hours with serum-free medium without serum. Thereafter divided into a control group (no control), a positive control group treated with TGF- ⁇ 20ng / ml and an experimental group treated with the addition of 10 mM peptide so that the final concentration of the peptide according to an aspect of the present invention is 50 ⁇ M each group Was incubated at 5% CO 2 , 37 ° C for 24 hours.
  • the cultures were collected and the amount of collagen was measured using the ELISA (procollagen type IC-peptide EIA kit) method.
  • washing buffer 100 ⁇ l of color reagent (TMBZ) was added thereto, and incubated at room temperature for 15 minutes, followed by 100 ⁇ l of 1N sulfuric acid. It was then measured with an ELISA reader at 450 nm.
  • TMBZ color reagent
  • Collagen synthesis degree derived through the above experimental method was calculated and expressed as a relative value when the control (control) is 100%, as shown in Table 2 below.
  • the peptides CP-54, CP-55, CP-56 having a novel activity according to the present invention showed that the collagen synthesis degree increased by more than 10% compared to the control.
  • the peptide having a novel activity according to an aspect of the present invention and a composition comprising the same show collagen synthesis effects. These effects are to improve the wrinkles and anti-aging of the skin, it can be seen that the peptide and the composition comprising the same according to an aspect of the present invention can have a variety of excellent effects in improving the skin condition.
  • the peptide having a novel activity has collagen synthesis activity, and it is expected that the functional cosmetic composition for wrinkle improvement and anti-aging may be developed using the peptide.

Abstract

The present specification relates to a peptide having novel activity and a composition comprising the same and, more particularly, to a peptide having novel activity exhibiting efficacy in collagen synthesis and a composition for improving wrinkles and preventing aging comprising the same. The peptide having novel activity and the composition comprising the same according to the present invention have a remarkable effect in collagen synthesis, thereby providing a composition for improving wrinkles and preventing aging.

Description

콜라겐 생성 증가 효능을 가지는 펩티드 및 이를 포함하는 조성물Peptides having increased collagen production and compositions comprising the same
본 명세서는 콜라겐 생성 증가 효능을 가지는 기능성 펩티드 및 이를 포함하는 조성물에 관한 것으로, 보다 구체적으로는 콜라겐 생성 증가 효과를 가지는 기능성 펩티드를 포함하며 콜라겐 생성 저하에 의한 피부 주름 개선 및 피부 노화 방지에 효과적인 기능성 펩티드 조성물에 관한 것이다.The present specification relates to a functional peptide having a collagen production increasing effect and a composition comprising the same, and more particularly, to a functional peptide having a collagen production increasing effect and effective in improving skin wrinkles and preventing skin aging by lowering collagen production. It relates to a peptide composition.
피부는 노화 과정으로 인하여 여러 가지 대사 활성이 저하되며, 총체적인 피부 세포의 활성도가 떨어진다. 따라서 이러한 노화로 인하여 콜라겐 생성 능력, 콜라겐의 전사 이후의 일어나는 여러 과정에 변화가 일어나 콜라겐 생성의 감소로 이어지게 된다. 따라서 피부 주름의 발생 원인 중 하나로 피부교원질(콜라겐)의 결핍을 들고 있다. 콜라겐과 엘라스틴은 피부에 탄성과 탄력을 주며 이들이 노화로 약해지면 피부는 쉽게 손상되고 늙는다. 반복적인 자외선 노출로 피부에서의 콜라겐 분해효소들(matrix metalloproteinases, MMPs)이 증가하며, 증가된 콜라겐 분해효소는 피부 콜라겐을 분해하게 된다. 이런 현상의 반복으로 피부 주름이 형성되게 된다.Due to the aging process, the skin is deteriorated in various metabolic activities, and the overall activity of the skin cells is reduced. Therefore, aging causes a change in collagen production ability and various processes occurring after the transcription of collagen, leading to a decrease in collagen production. Therefore, one of the causes of skin wrinkles is holding up the skin collagen (collagen) deficiency. Collagen and elastin give elasticity and elasticity to the skin, and as they weaken with aging, the skin is easily damaged and old. Repeated exposure to ultraviolet light increases matrix metalloproteinases (MMPs) in the skin, and the increased collagen breakdown enzymes break down skin collagen. Repeating this phenomenon leads to skin wrinkles.
조직학적 관점에서 피부노화는 피부의 진피에서 매트릭스를 형성하는 세포 외 조직(extracellular matrix proteins) 조성의 변화로 나타난다. 진피 세포 외 조직 중 콜라겐 단백질들은 피부에 강도와 장력을 부여하며 이로 인해 외부의 자극이나 힘으로부터 피부를 보호하는 역할을 하며 진피층의 90%를 차지하고 있어 콜라겐의 감소는 피부 노화와 주름형성에 밀접한 관련이 있다. 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. Collagen proteins in the dermal extracellular tissue provide strength and tension to the skin, which protects the skin from external irritation or force, and accounts for 90% of the dermal layer.Reducing collagen is closely related to skin aging and wrinkle formation. There is this.
피부 주름 개선 물질로 가장 대표적인 것이 레티놀(retinol)과 레티노이드라고 명명되는 그 유도체들로써 건선, 노화, 암, 여드름 등 다방면에서 좋은 효과를 나타낸다고 알려져 있다. 또한 이 레티노이드는 피부에 도포하였을 때 콜라겐분해효소-1(MMP-1)를 저해 함으로서 콜라겐의 손실을 방지하고 콜라겐 형성을 자극함으로써 내인성 및 광노화를 방지하고 회복시키는 것으로 알려져 있다. 한편, 레티놀 외에는 TGF-β(Transforming growth factor), 베툴린산, 야생 참마 추출물(한국공개특허 제2009-0055079호) 등이 주름개선용 조성물로서 알려져 있다. 그러나, 상기에 기술된 피부개선 유효성분들은 피부에 도포하였을 때 홍반, 가려움, 건선, 스케일링 등의 국소적 피부자극 반응을 일으킬 수 있고, 생체 적용시 그 안정성이 검증되어 있지 않다고 알려져 있다. Retinol and retinoids, the most representative of which is a skin wrinkle improving substance, are known to be effective in various fields such as psoriasis, aging, cancer, and acne. In addition, this retinoid is known to prevent collagen loss by stimulating collagen formation by inhibiting collagenase-1 (MMP-1) when applied to the skin to prevent and restore endogenous and photoaging. On the other hand, other than retinol, TGF-β (Transforming growth factor), betulinic acid, wild yam extract (Korean Patent Publication No. 2009-0055079) and the like is known as a wrinkle improvement composition. However, it is known that the above-described skin improvement active ingredients may cause local skin irritation reactions such as erythema, itching, psoriasis, and scaling when applied to the skin, and their stability has not been verified when applied to a living body.
주름개선 및 노화방지를 위한 콜라겐 합성 효능을 갖는 다양한 펩티드들, 그리고 상기 펩티드의 유도체들을 함유한 조성물들이 보고되고 있지만, 부작용이 적으면서도 콜라겐 합성 효능을 지닌 펩티드 및 이를 포함한 조성물은 아직 보고된 바 없다. 따라서, 이에 대한 연구 필요성이 제기되고 있는 실정이다.Although various peptides and compositions containing derivatives of the peptides have been reported for improving wrinkles and preventing aging, peptides having a collagen synthesis efficacy with little side effects, and compositions containing the same, have not been reported yet. . Therefore, the need for research on this situation is being raised.
[선행기술문헌][Preceding technical literature]
[특허문헌][Patent Documents]
(특허문헌 1) KR 2009-0055079 A(Patent Document 1) KR 2009-0055079 A
이러한 배경하에서 본 발명자들은 신규 활성을 갖는 펩티드를 개발하여 콜라겐 합성 효능이 있음을 발견하고 본 발명을 완성하게 되었다Under these circumstances, the present inventors have developed a peptide having novel activity and found that it has collagen synthesis efficacy and completed the present invention.
본 명세서의 목적은 콜라겐 합성의 신규 활성을 갖는 펩티드 및 이를 포함하는 주름방지 및 노화방지용 조성물을 제공하는데 있다.It is an object of the present specification to provide a peptide having a novel activity of collagen synthesis and an anti-wrinkle and anti-aging composition comprising the same.
본 발명의 일측면은 서열 번호 1 내지 3으로 표시되는 아미노산 서열 중 어느 하나로 구성되는 펩티드, 상기 펩티드 서열의 단편인 펩티드, 상기 펩티드 서열 또는 그의 단편인 펩티드와 80% 이상의 서열 상동성을 갖는 펩티드로 구성된 군으로부터 선택되는 콜라겐 합성 활성을 보이는 펩티드 또는 그의 염을 제공한다.One aspect of the present invention is a peptide having at least 80% sequence homology with a peptide consisting of any one of the amino acid sequences represented by SEQ ID NO: 1 to 3, a peptide that is a fragment of the peptide sequence, the peptide sequence or a peptide thereof Provided are peptides or salts thereof that exhibit collagen synthesis activity selected from the group consisting of:
본 발명의 다른 일측면은, 상기 펩티드 또는 그의 염을 유효성분으로 포함하는 주름개선 및 노화방지용 조성물을 제공한다.Another aspect of the present invention provides an anti-wrinkle and anti-aging composition comprising the peptide or a salt thereof as an active ingredient.
본 발명은 다른 측면에서, 서열 번호 1 내지 3으로 표시되는 아미노산 서열 중 어느 하나로 구성되는 펩티드, 상기 펩티드 서열의 단편인 펩티드, 상기 펩티드 서열 또는 그의 단편인 펩티드 서열과 80% 이상의 서열 상동성을 갖는 펩티드, 및 그들의 염으로 구성된 군으로부터 선택되는 하나 이상을, 이를 필요로 하는 개체에 적용하는 것을 포함하는, 주름개선 및 노화방지 방법을 제공한다.In another aspect, the present invention has a sequence homology of at least 80% to a peptide consisting of any one of the amino acid sequences represented by SEQ ID NO: 1 to 3, a peptide that is a fragment of the peptide sequence, the peptide sequence or a peptide sequence thereof Provided are anti-wrinkle and anti-aging methods comprising applying to a subject in need thereof at least one selected from the group consisting of peptides, and salts thereof.
본 발명은 다른 측면에서, 주름개선 및 노화방지에 사용하기 위한, 서열 번호 1 내지 3으로 표시되는 아미노산 서열 중 어느 하나로 구성되는 펩티드, 상기 펩티드 서열의 단편인 펩티드, 상기 펩티드 서열 또는 그의 단편인 펩티드 서열과 80% 이상의 서열 상동성을 갖는 펩티드, 및 그들의 염으로 구성된 군으로부터 선택되는 하나 이상을 제공한다.In another aspect, the present invention provides a peptide consisting of any one of amino acid sequences represented by SEQ ID NOs: 1 to 3, peptides which are fragments of the peptide sequences, peptides which are peptides, or fragments thereof, for use in wrinkle improvement and anti-aging. At least one selected from the group consisting of peptides having at least 80% sequence homology with the sequences, and salts thereof.
본 발명은 또 다른 측면에서, 주름개선 및 노화방지를 위한 유효성분으로서 서열 번호 1 내지 3으로 표시되는 아미노산 서열 중 어느 하나로 구성되는 펩티드, 상기 펩티드 서열의 단편인 펩티드, 상기 펩티드 서열 또는 그의 단편인 펩티드 서열과 80% 이상의 서열 상동성을 갖는 펩티드, 및 그들의 염으로 구성된 군으로부터 선택되는 하나 이상의 비치료적 화장 용도를 제공한다.In still another aspect, the present invention provides a peptide comprising any one of amino acid sequences represented by SEQ ID NOs: 1 to 3 as an active ingredient for wrinkle improvement and anti-aging, a peptide which is a fragment of the peptide sequence, the peptide sequence or a fragment thereof One or more non-therapeutic cosmetic uses selected from the group consisting of peptides having at least 80% sequence homology with peptide sequences, and salts thereof.
본 발명은 또 다른 측면에서, 주름개선 및 노화방지를 위한 조성물의 제조에 사용하기 위한, 서열 번호 1 내지 3으로 표시되는 아미노산 서열 중 어느 하나로 구성되는 펩티드, 상기 펩티드 서열의 단편인 펩티드, 상기 펩티드 서열 또는 그의 단편인 펩티드 서열과 80% 이상의 서열 상동성을 갖는 펩티드, 및 그들의 염으로 구성된 군으로부터 선택되는 하나 이상의 용도를 제공한다.In another aspect, the present invention provides a peptide comprising any one of amino acid sequences represented by SEQ ID NOs: 1 to 3 for use in the preparation of a composition for improving wrinkles and preventing aging, a peptide which is a fragment of the peptide sequence, the peptide One or more uses selected from the group consisting of peptides having at least 80% sequence homology with peptide sequences that are sequences or fragments thereof, and salts thereof.
본 발명의 다른 일측면에 따르면, 상기 조성물은 주름개선 및 노화방지용 화장료 조성물인 것을 특징으로 할 수 있다.According to another aspect of the present invention, the composition may be characterized in that the cosmetic composition for wrinkle improvement and anti-aging.
본 발명의 다른 일측면에 따르면, 상기 화장료 조성물은 스킨, 로션, 크림, 파운데이션, 에센스, 젤, 팩, 폼 클렌징, 비누 및 피부외용 연고로 이루어진 군으로부터 선택된 어느 하나의 제형으로 이루어진 것을 특징으로 할 수 있다.According to another aspect of the present invention, the cosmetic composition may be characterized in that it consists of any one formulation selected from the group consisting of skin, lotion, cream, foundation, essence, gel, pack, foam cleansing, soap and skin ointment. have.
본 발명의 다른 일측면에 따르면, 상기 조성물은 주름개선 및 노화방지용 약학적 조성물인 것을 특징으로 할 수 있다.According to another aspect of the invention, the composition may be characterized in that the pharmaceutical composition for wrinkle improvement and anti-aging.
본 발명의 다른 일측면에 따르면, 상기 조성물은 주름개선 및 노화방지용 건강기능 식품용 조성물인 것을 특징으로 할 수 있다.According to another aspect of the present invention, the composition may be characterized in that the composition for wrinkle improvement and anti-aging health functional food.
본 발명에 따른 서열번호의 서열을 갖는 펩티드들은 콜라겐 합성 효능을 가져, 콜라겐 합성 활성을 지닌 펩티드를 포함하는 주름개선 및 노화방지용 조성물을 제공할 것으로 예상된다. Peptides having the sequence of SEQ ID NO according to the present invention have collagen synthesis efficacy, it is expected to provide a wrinkle improvement and anti-aging composition comprising a peptide having collagen synthesis activity.
본 발명에 따른 주름개선 및 노화방지 효과는 본 발명에 따른 조성물에 의한 콜라겐 생성 증가를 통해, 피부 주름 생성을 억제 또는 개선하여 피부노화를 방지하는 효과를 말한다.Wrinkle improvement and anti-aging effects according to the present invention refers to the effect of preventing or aging skin by inhibiting or improving the production of skin wrinkles through increased collagen production by the composition according to the present invention.
도 1은 인간 피부 섬유아세포(Human skin fibroblast)인 CCD-986SK 세포주에, 아무것도 처리하지 않은 대조군(control), TGF-β를 20ng/ml 처리한 양성대조군, 본 발명에 따른 펩티드(CP-54, CP-55 CP-56)를 각각 50μM 처리한 처리군으로 나누어 콜라겐 합성도 (collagen synthesis)를 측정하여 비교한 결과를 대조군(100%)을 기준으로 상대적(%)으로 나타낸 그래프이다.Figure 1 is a human skin fibroblast CCD-986SK cell line, a control (no treatment), a control group treated with TGF-β 20ng / ml positive control group, peptide according to the invention (CP-54, CP-55 CP-56) each divided into 50μM treated group collagen synthesis (collagen synthesis) measured by comparing the results compared to the control (100%) based on the relative graph (%).
본 명세서는 다양한 변환을 가할 수 있고 여러 가지 실시예를 가질 수 있는 바, 이하, 본 명세서를 보다 구체적으로 설명한다. 그러나, 이는 본 명세서를 특정한 실시 형태에 대해 한정하려는 것이 아니며, 본 명세서의 사상 및 기술 범위에 포함되는 모든 변환, 균등물 내지 대체물을 포함하는 것으로 이해되어야 한다. 본 명세서를 설명함에 있어서 관련된 공지 기술에 대한 구체적인 설명이 본 명세서의 요지를 흐릴 수 있다고 판단되는 경우 그 상세한 설명을 생략한다.The present disclosure may be modified in various ways and may have various embodiments. Hereinafter, the present specification will be described in more detail. However, this is not intended to limit the present specification to specific embodiments, it should be understood to include all transformations, equivalents, and substitutes included in the spirit and scope of the present specification. In describing the present specification, when it is determined that the detailed description of the related known technology may obscure the gist of the present specification, the detailed description thereof will be omitted.
본 발명의 일 측면에서, 서열 번호 1 내지 3의 서열로 구성되는 콜라겐 합성의 신규 활성을 갖는 펩티드를 개시한다.In one aspect of the present invention, a peptide having novel activity of collagen synthesis consisting of the sequences of SEQ ID NOs: 1 to 3 is disclosed.
본 명세서에 개시된 펩티드는 서열 번호 1 내지 3에서 선택된 어느 하나의 펩티드들 각각과 80% 이상, 85% 이상, 90% 이상, 95% 이상, 96% 이상, 97% 이상, 98% 이상, 99% 이상의 서열 상동성을 갖는 펩티드를 포함할 수 있다. The peptides disclosed herein are at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, 99% with each of the peptides selected from SEQ ID NOs: 1-3 It may include a peptide having the above sequence homology.
또한, 본 명세서에 개시된 펩티드는, 1 내지 3에서 선택된 어느 하나의 펩티드들과 1개 이상의 아미노산, 2개 이상의 아미노산, 3개 이상의 아미노산, 4개 이상의 아미노산, 5개 이상의 아미노산, 6개 이상의 아미노산 또는 7개 이상의 아미노산이 변화된 펩티드를 포함할 수 있다. In addition, the peptides disclosed herein may include any one peptide selected from 1 to 3 and at least one amino acid, at least two amino acids, at least three amino acids, at least four amino acids, at least five amino acids, at least six amino acids or Seven or more amino acids may comprise a modified peptide.
본 발명의 일 측면에서, 아미노산 변화는 펩티드의 물리화학적 특성이 변경되도록 하는 성질에 속한다. 예를 들어, 펩티드의 열안정성을 향상시키고, 기질 특이성을 변경시키고, 최적의 pH를 변화시키는 등의 아미노산 변화가 수행될 수 있다.In one aspect of the invention, amino acid changes belong to the property that allows the physicochemical properties of the peptide to be altered. For example, amino acid changes can be made, such as improving the thermal stability of the peptide, altering substrate specificity, changing the optimal pH, and the like.
본 발명의 일 측면에 따른 서열 번호 1 내지 3에서 선택된 어느 하나의 펩티드는 세포 내 독성이 낮고, 생체 내 안정성이 높다는 장점을 가진다. 서열번호 1 내지 3에 기재된 펩티드는 아래 표 1과 같다. 아래 표 1의 "이름"은 펩티드를 구별하기 위해 명명한 것이다. Any one peptide selected from SEQ ID NOs: 1 to 3 according to an aspect of the present invention has the advantage of low intracellular toxicity and high in vivo stability. The peptides set forth in SEQ ID NOS: 1 to 3 are shown in Table 1 below. The "name" in Table 1 below is to distinguish peptides.
서열번호 (SEQ ID NO.)SEQ ID NO. 이름name 서열order 아미노산 길이Amino acid length
1One CP-54CP-54 TSRLRFITSRLRFI 77
22 CP-55CP-55 LRFIPLRFIP 55
33 CP-56CP-56 ALSSRLRGALSSRLRG 88
본 발명은 일 측면에서, 상기 펩티드를 코딩하는 뉴클레오티드 서열을 제공한다.In one aspect, the present invention provides a nucleotide sequence encoding the peptide.
본 발명의 일 측면에서는 서열번호 1 내지 3에서 선택된 어느 하나의 아미노산 서열을 포함하는 펩티드를 유효 성분으로 포함하는 조성물을 제공한다.In one aspect of the invention provides a composition comprising a peptide comprising any one amino acid sequence selected from SEQ ID NO: 1 to 3 as an active ingredient.
본 발명의 일 측면에 따른 조성물은 인간, 개, 닭, 돼지, 소, 양, 기니아피그 또는 원숭이를 포함하는 모든 동물에 적용될 수 있다.The composition according to one aspect of the present invention can be applied to all animals including humans, dogs, chickens, pigs, cows, sheep, guinea pigs or monkeys.
본 발명의 일 측면에 따른 피부 외용제 조성물은 그 제형에 있어서 특별히 한정되는 바가 없으며, 유연화장수, 수렴화장수, 영양화장수, 아이크림, 영양크림, 맛사지크림, 클렌징크림, 클렌징폼, 클렌징워터, 파우더, 엣센스, 팩 등의 제형을 가질 수 있다.The external preparation composition for skin according to an aspect of the present invention is not particularly limited in its formulation, and it is a softening longevity, astringent longevity, nourishing longevity, eye cream, nutrition cream, massage cream, cleansing cream, cleansing foam, cleansing water, powder, Formulations such as essences, packs, and the like.
본 발명의 일 측면에 따른 화장료 조성물은 통상의 화장료 제조방법에 따라, 다양한 형태로 제조될 수 있다. 예를 들어, 상기 화장료 조성물은 상기 펩티드를 함유하는 향장 제품, 화장수, 크림, 로션 등의 형태로 제조될 수 있으며, 이는 통상의 클렌징액, 수렴액 및 보습액으로 희석하여 사용될 수 있다. 또한, 상기 화장료 조성물은 화장료 조성물 분야에서 통상적으로 사용되는 안정화제, 용해화제, 비타민, 안료, 및 향료와 같은 통상적인 보조제를 포함할 수 있다.The cosmetic composition according to an aspect of the present invention may be prepared in various forms according to a conventional cosmetic preparation method. For example, the cosmetic composition may be prepared in the form of a cosmetic product containing the peptide, lotion, cream, lotion, etc., which may be diluted with conventional cleansing liquid, astringent liquid, and moisturizing liquid. In addition, the cosmetic composition may include conventional adjuvants such as stabilizers, solubilizers, vitamins, pigments, and flavorings commonly used in the field of cosmetic compositions.
본 발명의 일 측면에 따른 서열번호 1 내지 3에서 선택된 어느 하나의 신규 활성 펩티드를 포함하는 화장료 조성물은 이미 알려진 화장료 조성물 제조방법에 의해 화장료 조성물을 제조할 수 있고, 일반적인 피부 화장료에 한정되지 않고 의약부외품, 외용 의약품에도 적용할 수 있다. 이들의 제형은 통상적으로 제조되는 어떠한 제형으로도 제조될 수 있으며, 예를 들어, 스킨, 로션, 크림, 파운데이션, 에센스, 젤, 팩, 폼클린징, 바디오일, 립스틱, 마스카라, 메이크업베이스 또는 비누형태로 제조할 수 있으나, 이에 한정되는 것은 아니다. The cosmetic composition comprising any one of the novel active peptides selected from SEQ ID NOs: 1 to 3 according to an aspect of the present invention may prepare a cosmetic composition by a known cosmetic composition preparation method, and is not limited to general skin cosmetics. Applicable to quasi-drugs and external medicines. These formulations may be prepared in any of the formulations commonly prepared, for example in the form of skins, lotions, creams, foundations, essences, gels, packs, foam cleansing, body oils, lipsticks, mascaras, makeup bases or soaps. It may be prepared as, but is not limited thereto.
본 발명의 일 측면에 따른 서열번호 1 내지 3에서 선택된 어느 하나의 신규 활성 펩티드를 포함하는 화장료 조성물에 있어서, 서열번호 1 내지 3에서 선택된 어느 하나의 신규 활성 펩티드 이외에 다른 성분들은 기타 화장료의 제형, 사용목적 등에 따라 당업자가 어려움 없이 적의 선정하여 배합할 수 있다.In the cosmetic composition comprising any one new active peptide selected from SEQ ID NOs: 1 to 3 according to an aspect of the present invention, in addition to any one new active peptide selected from SEQ ID NOs: 1 to 3 other components of the formulation of the cosmetic, According to the purpose of use, those skilled in the art can mix and select the enemy without difficulty.
본 발명의 화장료 조성물은 매일 사용할 수 있으며 또한 정해지지 않은 기간 동안에도 사용할 수 있다. 바람직하게는 사용자의 연령, 피부상태 또는 피부타입, 펩티드의 농도에 따라 사용량, 사용횟수 및 기간을 조절할 수 있다.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, frequency of use and duration of use may be adjusted according to the age, skin condition or skin type of the user, and the concentration of the peptide.
본 명세서의 조성물은 목적하는 바에 따라 모든 방식으로 투여할 수 있는데, 예를 들면, 경구, 직장, 정맥, 동맥, 근육, 피하, 피내, 자궁내경막 또는 뇌실내 주사에 의해 투여될 수 있다. 본 발명의 일 측면에 따른 조성물의 경구 투여를 위한 제형은 정제, 환제, 연질 또는 경질 캅셀제, 과립제, 산제, 액제 또는 유탁제일 수 있고, 비경구 투여를 위한 제형은 주사제, 점적제, 로션, 연고, 겔, 크림, 현탁제, 유제, 좌제, 패취 또는 분무제일 수 있으나, 이에 제한되는 것은 아니다. The compositions herein can be administered in any manner as desired, for example by oral, rectal, intravenous, arterial, muscular, subcutaneous, intradermal, endometrial or intraventricular injection. Formulations for oral administration of a composition according to one aspect of the invention may be tablets, pills, soft or hard capsules, granules, powders, solutions or emulsions, and formulations for parenteral administration may be injections, drops, lotions, ointments. , Gels, creams, suspensions, emulsions, suppositories, patches or sprays, but is not limited thereto.
본 명세서의 조성물의 투여 형태는 이들의 약학적으로 허용 가능한 염의 형태로도 사용될 수 있고, 또한 단독으로 또는 타 약학적 활성 화합물과 결합뿐만 아니라 적당한 집합으로 사용될 수 있다. 상기 염으로는 약학적으로 허용되는 것이면 특별히 한정되지 않으며, 예를 들어 염산, 황산, 질산, 인산, 불화수소산, 브롬화수소산, 포름산 아세트산, 타르타르산, 젖산, 시트르산, 푸마르산, 말레산, 숙신산, 메탄술폰산, 벤젠술폰산, 톨루엔술폰산, 나프탈렌술폰산 등을 사용할 수 있다. Dosage forms of the compositions herein may be used in the form of their pharmaceutically acceptable salts, and may be used alone or in combination with other pharmaceutically active compounds as well as in a suitable collection. The salt is not particularly limited as long as it is pharmaceutically acceptable. For example, hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid, hydrofluoric acid, hydrobromic acid, formic acid acetic acid, tartaric acid, lactic acid, citric acid, fumaric acid, maleic acid, succinic acid, methanesulfonic acid , Benzene sulfonic acid, toluene sulfonic acid, naphthalene sulfonic acid and the like can be used.
본 발명의 일 측면에 따른 약학 조성물 내의 상기 펩티드 또는 그의 염의 농도는 1 내지 100 μM일 수 있다. 일 구현 예로서, 상기 농도는 1μM 이상, 3μM 이상, 5μM 이상, 7μM 이상, 10μM 이상, 15μM 이상, 20μM 이상, 25μM 이상, 30μM 이상, 35μM 이상, 40μM 이상, 50μM 이상, 60μM 이상, 70μM 이상, 80μM 이상, 85μM 이상, 90μM 이상, 또는 95μM 이상일 수 있다. 다른 예로서, 상기 농도는 100μM 이하, 95μM 이하, 90μM 이하, 85μM 이하, 80μM 이하, 75μM 이하, 70μM 이하, 60μM 이하, 50μM 이하, 40μM 이하, 30μM 이하, 25μM 이하, 20μM 이하, 15μM 이하, 10μM 이하, 7μM 이하, 5μM 이하, 또는 2μM 이하일 수 있다. 상기 농도는 바람직하게는 20 내지 80 μM, 보다 바람직하게는 50μM 일 수 있다.The concentration of the peptide or salt thereof in the pharmaceutical composition according to one aspect of the present invention may be 1 to 100 μM. In one embodiment, the concentration is at least 1 μM, at least 3 μM, at least 5 μM, at least 7 μM, at least 10 μM, at least 15 μM, at least 20 μM, at least 25 μM, at least 30 μM, at least 35 μM, at least 40 μM, at least 50 μM, at least 60 μM, at least 70 μM, 80 μM or more, 85 μM or more, 90 μM or more, or 95 μM or more. As another example, the concentration is 100 μM or less, 95 μM or less, 90 μM or less, 85 μM or less, 80 μM or less, 75 μM or less, 70 μM or less, 60 μM or less, 50 μM or less, 40 μM or less, 30 μM or less, 25 μM or less, 20 μM or less, 15 μM or less, 10 μM Or less, 7 μM or less, 5 μM or less, or 2 μM or less. The concentration may preferably be 20 to 80 μM, more preferably 50 μM.
본 발명의 일측면에 따른 상기 펩티드 또는 그의 염의 투여량은 0.1 μg/kg 내지 10 mg/kg일 수 있다. 일 구현 예로서, 상기 투여량은 0.1μg/kg 이상, 0.5μg/kg 이상, 1μg/kg 이상, 5μg/kg 이상, 10μg/kg 이상, 30μg/kg 이상, 50μg/kg 이상, 70μg/kg 이상, 100μg/kg 이상, 300μg/kg 이상, 500μg/kg 이상, 700μg/kg 이상, 900μg/kg 이상, 1mg/kg 이상, 2mg/kg 이상, 3mg/kg 이상, 4mg/kg 이상, 5mg/kg 이상, 6mg/kg 이상, 7mg/kg 이상, 8mg/kg 이상, 9mg/kg 이상, 9.5mg/kg 이상, 또는 9.8mg/kg 이상일 수 있다. 다른 구현 예로서, 상기 투여량은 10mg/kg 이하, 9mg/kg 이하, 8mg/kg 이하, 7mg/kg 이하, 6mg/kg 이하, 5mg/kg 이하, 4mg/kg 이하, 3mg/kg 이하, 2mg/kg 이하, 1mg/kg 이하, 900μg/kg 이하, 700μg/kg 이하, 500μg/kg 이하, 300μg/kg 이하, 100μg/kg 이하, 90μg/kg 이하, 70μg/kg 이하, 50μg/kg 이하, 30μg/kg 이하, 10μg/kg 이하, 7μg/kg 이하, 5μg/kg 이하, 3μg/kg 이하, 1μg/kg 이하, 0.9μg/kg 이하, 0.7μg/kg 이하, 0.5μg/kg 이하, 0.3μg/kg 이하, 0.2μg/kg 이하, 0.15μg/kg 이하, 또는 0.12μg/kg 이하일 수 있다. 상기 투여량은 바람직하게는 0.001 내지 1 mg/kg일 수 있다.The dosage of the peptide or salt thereof according to one aspect of the present invention may be 0.1 μg / kg to 10 mg / kg. In one embodiment, the dosage is at least 0.1 μg / kg, at least 0.5 μg / kg, at least 1 μg / kg, at least 5 μg / kg, at least 10 μg / kg, at least 30 μg / kg, at least 50 μg / kg, at least 70 μg / kg , 100 μg / kg or more, 300 μg / kg or more, 500 μg / kg or more, 700 μg / kg or more, 900 μg / kg or more, 1 mg / kg or more, 2 mg / kg or more, 3 mg / kg or more, 4 mg / kg or more, 5 mg / kg or more , At least 6 mg / kg, at least 7 mg / kg, at least 8 mg / kg, at least 9 mg / kg, at least 9.5 mg / kg, or at least 9.8 mg / kg. In another embodiment, the dosage is 10 mg / kg or less, 9 mg / kg or less, 8 mg / kg or less, 7 mg / kg or less, 6 mg / kg or less, 5 mg / kg or less, 4 mg / kg or less, 3 mg / kg or less, 2 mg / kg or less, 1mg / kg or less, 900μg / kg or less, 700μg / kg or less, 500μg / kg or less, 300μg / kg or less, 100μg / kg or less, 90μg / kg or less, 70μg / kg or less, 50μg / kg or less, 30μg / kg or less, 10μg / kg or less, 7μg / kg or less, 5μg / kg or less, 3μg / kg or less, 1μg / kg or less, 0.9μg / kg or less, 0.7μg / kg or less, 0.5μg / kg or less, 0.3μg / kg or less, 0.2 μg / kg or less, 0.15 μg / kg or less, or 0.12 μg / kg or less. The dosage may preferably be from 0.001 to 1 mg / kg.
상기 투여량은 투여 받을 대상의 연령, 성별, 체중, 병리 상태 및 그 심각도, 투여 경로 또는 처방자의 판단에 따라 달라질 수 있다. 이러한 인자에 기초한 적용량 결정은 당업자의 수준 내에 있다.The dosage may vary depending on the age, sex, weight, pathology and severity of the subject to be administered, the route of administration or the judgment of the prescriber. Application amount determination based on these factors is within the level of skill in the art.
본 발명의 일 측면에 따른 조성물은 필요에 따라 희석제, 부형제, 활택제, 결합제, 붕해제, 완충제, 분산제, 계면 활성제, 착색제, 향료 또는 감미제 등의 첨가제를 포함할 수 있다. 본 발명의 일 측면에 따른 조성물은 당업계의 통상적인 방법에 의해 제조될 수 있다.Compositions according to one aspect of the invention may include additives such as diluents, excipients, lubricants, binders, disintegrants, buffers, dispersants, surfactants, colorants, flavoring or sweetening agents as needed. Compositions according to one aspect of the present invention may be prepared by conventional methods in the art.
본 발명의 일 측면에 따른 조성물은 콜라겐 생성을 증가시키고, 콜라겐 생성 억제 인자 분비를 저해시켜 피부 주름 생김을 억제 또는 개선시킬 수 있으며 노화를 방지할 수 있다.The composition according to an aspect of the present invention may increase collagen production, inhibit collagen production inhibitor secretion, and may inhibit or improve skin wrinkles and prevent aging.
본 명세서에서 사용된 용어들은 특정 구체예들을 설명하기 위한 목적으로만 의도된 것이지 본 발명을 한정하고자 하는 의도가 아니다. 명사 앞에 개수가 생략된 용어는 수량을 제한하고자 하는 것이 아니라 언급된 명사 물품이 하나 이상 존재하는 것을 나타내는 것이다. 용어 "포함하는", "갖는", 및 "함유하는"은 열린 용어로 해석된다(즉, "포함하지만 이에 한정되지는 않는"의 의미). The terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. The term without the number before the noun is not intended to limit the quantity but rather to the presence of one or more of the mentioned noun articles. The terms "comprising", "having", and "comprising" are interpreted as open terms (ie, meaning "including but not limited to").
수치의 범위를 언급하는 것은 단지 그 범위 내에 속하는 각각의 별개의 수치들을 개별적으로 언급하는 것을 대신하는 쉬운 방법이기 때문이며, 그것이 아님이 명시되어 있지 않는, 각 별개의 수치는 마치 개별적으로 명세서에 언급되어 있는 것처럼 본 명세서에 통합된다. 모든 범위의 끝 값들은 그 범위 내에 포함되며 독립적으로 조합 가능하다. Reference to a range of numbers is simply an easy way to substitute for referring to each individual number within the range individually, and unless stated otherwise, each individual number is referred to individually as if It is incorporated herein as is. The end values of all ranges fall within that range and can be combined independently.
본 명세서에 언급된 모든 방법들은 달리 명시되어 있거나 문맥에 의해 명백히 모순되지 않는 한 적절한 순서로 수행될 수 있다. 어느 한 실시예 및 모든 실시예 또는 예시적 언어 (예컨대, "~과 같은")를 사용하는 것은, 청구범위에 포함되어 있지 않는 한, 단지 본 발명의 여러 측면을 더 잘 기술하기 위함이지 본 발명의 범위를 제한하고자 함이 아니다. 명세서의 어떤 언어도 어떤 비청구된 구성요소를 본 발명의 일 측면에 따른 실시에 필수적인 것으로 해석되어서는 아니된다. 다른 정의가 없는 한, 본 명세서에 사용되는 기술적 및 과학적 용어들은 본 발명이 속하는 기술 분야에서 통상의 지식을 갖는 사람에 의해 통상 이해되는 것과 같은 의미를 갖는다. All methods mentioned herein may be performed in the proper order unless otherwise specified or clearly contradicted by context. The use of one embodiment and all embodiments or example language (eg, “such as”) is merely intended to better describe various aspects of the invention, unless the claims include it. It is not intended to limit the scope of. No language in the specification should be construed as essential to the practice of any aspect of the invention. Unless defined otherwise, technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.
본 발명의 일 측면에 따른 바람직한 구체예들은 본 발명의 일 측면을 수행하기 위해 발명자에게 알려진 가장 최적의 모드를 포함한다. 바람직한 구체예들의 변이들이 앞선 기재를 읽으면 당업자에게 명백하게 될 수 있다. 본 발명자들은 당업자들이 그러한 변이를 적절히 이용하길 기대하고, 발명자들은 본 명세서에 기재된 것과 다른 방식으로 본 발명의 여러 측면이 실시되기를 기대한다. 따라서, 본 명세서는, 특허법에 의해 허용되는 것과 같이, 첨부된 특허청구범위에서 언급된 발명의 요지의 균등물 및 모든 변형들을 포함한다. 더욱이, 모든 가능한 변이들 내에서 상기 언급된 구성요소들의 어떤 조합이라도 여기서 반대로 명시하거나 문맥상 명백히 모순되지 않는 한 본 명세서에 포함된다. 본 명세서는 예시적인 구체예들을 참조하여 구체적으로 나타내어지고 기술되었지만, 당업자들은 하기 청구범위에 의해 정의되는 발명의 정신 및 범위를 벗어나지 않고서도 형태 및 디테일에서 다양한 변화가 행해질 수 있음을 잘 이해할 것이다 . Preferred embodiments according to one aspect of the invention include the most optimal mode known to the inventor for carrying out one aspect of the invention. Variations of the preferred embodiments may become apparent to those skilled in the art upon reading the foregoing description. The inventors expect skilled artisans to employ such variations as appropriate, and the inventors expect the various aspects of the invention to be practiced in a manner different from that described herein. Accordingly, this specification includes all modifications and equivalents of the subject matter referred to in the appended claims, as permitted by patent law. Moreover, any combination of the above-mentioned elements within all possible variations is included herein unless expressly stated to the contrary or apparently contradictory in context. Although the specification has been particularly shown and described with reference to exemplary embodiments, those skilled in the art will understand that various changes in form and detail may be made without departing from the spirit and scope of the invention as defined by the following claims .
이하, 실시예 및 실험예를 들어 본 발명의 일 측면 상 구성 및 효과를 보다 구체적으로 설명한다. 그러나 아래 실시예 및 실험예는 본 발명의 일 측면에 대한 이해를 돕기 위해 예시의 목적으로만 제공된 것일 뿐 본 발명의 범주 및 범위가 그에 의해 제한되는 것은 아니다.Hereinafter, the configuration and effects on one aspect 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 only for the purpose of illustration in order to facilitate understanding of one aspect of the present invention, and the scope and scope of the present invention is not limited thereto.
[실시예 1]Example 1
신규 활성을 갖는 펩티드의 합성 Synthesis of Peptides with Novel Activity
서열번호 1의 신규 활성을 지닌 펩티드(이하 CP-54이라 명명함)를 종래에 알려진 고상 펩티드 합성법에 따라 제조하였다. 구체적으로, 펩티드들은 ASP48S(Peptron, Inc., 대한민국 대전)를 이용하여 Fmoc 고상 합성법(solid phase peptide synthesis, SPPS)을 통해 C-말단부터 아미노산 하나씩 커플링함으로써 합성하였다. 다음과 같이, 펩티드들의 C-말단의 첫번째 아미노산이 수지에 부착된 것을 사용하였다. 예컨대 다음과 같다:Peptides having the novel activity of SEQ ID NO: 1 (hereinafter referred to as CP-54) were prepared according to solid phase peptide synthesis known in the art. Specifically, peptides were synthesized by coupling amino acids one by one from the C-terminus through Fmoc solid phase synthesis (SPPS) using ASP48S (Peptron, Inc., Daejeon, Korea). As follows, the first amino acid at the C-terminus of the peptides was attached to the resin. For example:
NH2-S(tBu)-2-chloro-Trityl ResinNH 2 -S (tBu) -2-chloro-Trityl Resin
NH2-Ala-2-chloro-Trityl ResinNH 2 -Ala-2-chloro-Trityl Resin
NH2-Arg(Pbf)-2-chloro-Trityl ResinNH 2 -Arg (Pbf) -2-chloro-Trityl Resin
펩티드 합성에 사용한 모든 아미노산 원료는 N-term이 Fmoc으로 보호(protection)되고, 잔기는 모두 산에서 제거되는 Trt, Boc, tBu (t-butylester), Pbf (2,2,4,6,7-pentamethyl dihydro-benzofuran-5-sulfonyl) 등으로 보호된 것을 사용하였다. 예컨대 다음과 같다: All amino acid feedstocks used for peptide synthesis were protected with N-term Fmoc and all residues removed from acid Trt, Boc, tBu (t-butylester), Pbf (2,2,4,6,7- pentamethyl dihydro-benzofuran-5-sulfonyl) and the like were used. For example:
Fmoc-Ala-OH, Fmoc-Arg(Pbf)-OH, Fmoc-Glu(OtBu)-OH, Fmoc-Pro-OH, Fmoc-Leu-OHFmoc-Ala-OH, Fmoc-Arg (Pbf) -OH, Fmoc-Glu (OtBu) -OH, Fmoc-Pro-OH, Fmoc-Leu-OH
커플링 시약(coupling reagent)으로는 HBTU[2-(1H-Benzotriazole-1-yl)-1,1,3,3-tetamethylaminium hexafluorophosphate] / HOBt [N-Hydroxxybenzotriazole] /NMM [4-Methylmorpholine] 를 사용하였다. Fmoc 제거는 20%의 DMF 중 피페리딘(piperidine in DMF)을 이용하였다. 합성된 펩티드를 Resin에서 분리 및 잔기의 보호기 제거에는 절단 칵테일(cleavage Cocktail) [TFA (trifluoroacetic acid) /TIS (triisopropylsilane) / EDT (ethanedithiol) / H2O=92.5/2.5/2.5/2.5] 를 사용하였다.HBTU [2- (1H-Benzotriazole-1-yl) -1,1,3,3-tetamethylaminium hexafluorophosphate] / HOBt [N-Hydroxxybenzotriazole] / NMM [4-Methylmorpholine] is used as the coupling reagent. It was. Fmoc removal was performed using piperidine in DMF in 20% of DMF. Cleavage Cocktail (TFA (trifluoroacetic acid) / TIS (triisopropylsilane) / EDT (ethanedithiol) / H 2 O = 92.5 / 2.5 / 2.5 / 2.5] was used to isolate the synthesized peptide from Resin and remove the protecting group of residues. It was.
아미노산 보호기가 결합된 출발 아미노산이 고상 지지체에 결합되어 있는 상태를 이용하여 여기에 해당 아미노산들을 각각 반응시키고 용매로 세척한 후 탈보호하는 과정을 반복함으로써 각 펩티드를 합성하였다. 합성된 펩티드를 수지로부터 끊어낸 후 HPLC로 정제하고, 합성 성공여부를 LC/MS로 확인하고 동결 건조하였다. Each peptide was synthesized by repeating a process of reacting the amino acids with each other, washing with a solvent, and then deprotecting the amino acid using the state in which the amino acid protecting group was bound to the solid support. The synthesized peptide was separated from the resin and purified by HPLC. The success of the synthesis was confirmed by LC / MS and lyophilized.
본 실시예에 사용된 펩티드에 대해 고성능 액체 크로마토그래피 결과, 모든 펩티드의 순도는 95% 이상이었다.High performance liquid chromatography on the peptides used in this example showed that all peptides had a purity of at least 95%.
예컨대, 펩티드 CP-54 제조에 관한 구체적인 과정을 설명하면 다음과 같다. For example, the specific procedure for preparing peptide CP-54 is as follows.
1) 커플링 1) Coupling
NH2-I-2-chloro-Trityl Resin에 보호된 아미노산(8당량)와 커플링 시약 HBTU(8당량)/HOBt(8당량)/NMM(16당량) 을 DMF에 녹여서 첨가한 후, 상온에서 2시간 동안 반응하고 DMF, MeOH, DMF 순으로 세척하였다.Amino acid (8 equivalents) protected by NH 2 -I-2-chloro-Trityl Resin and coupling reagent HBTU (8 equivalents) / HOBt (8 equivalents) / NMM (16 equivalents) were dissolved in DMF and added at room temperature. The reaction was carried out for 2 hours and washed in the order of DMF, MeOH, and DMF.
2) Fmoc 탈보호 2) Fmoc deprotection
20%의 DMF 중의 피페리딘(piperidine in DMF)을 가하고 상온에서 5분 간 2회 반응하고 DMF, MeOH, DMF 순으로 세척하였다.Piperidine in DMF at 20% was added thereto, reacted twice at room temperature for 5 minutes, and washed in the order of DMF, MeOH, and DMF.
3) 1과 2의 반응을 반복적으로 하여 펩티드 기본 골격 NH2-T-S(tBu)-R(Pbf)-L-R(tBu)-F-I-2-chloro-Trityl Resin)을 만들었다. 3) The reaction of 1 and 2 was repeated to make peptide basic skeleton NH2-T-S (tBu) -R (Pbf) -L-R (tBu) -F-I-2-chloro-Trityl Resin).
4) 절단(cleavage): 합성이 완료된 펩티드 수지(resin)에 절단 칵테일(cleavage cocktail) 을 가하여 펩티드를 수지에서 분리하였다.4) Cleavage: The peptide cocktail was separated from the resin by adding a cleavage cocktail to the synthesized peptide resin.
5) 얻어진 혼합물에 냉각 디에틸 에테르를 가한 후, 원심 분리하여 얻어진 펩티드를 침전시킨다.5) After cooling diethyl ether is added to the obtained mixture, the obtained peptide is precipitated by centrifugation.
6) 위의 과정에서 얻어진 조(crude) 펩티드를 Prep HPLC를 이용하여 분리 정제하였다. 컬럼은 Vydac Everest C18 column (250 mm × 22 mm, 10 μm)를 이용하였다. Eluent는 0.1% (v/v) 트리플루오로아세트산(trifluoroacetic acid)가 함유된 10~75% (v/v) 워터-아세토니트릴(water-acetonitrile) 선형 그레디언트(linear gradient)를 이용하였다. 6) The crude peptide obtained in the above procedure was separated and purified using Prep HPLC. The column used a Vydac Everest C18 column (250 mm × 22 mm, 10 μm). Eluent used 10-75% (v / v) water-acetonitrile linear gradients containing 0.1% (v / v) trifluoroacetic acid.
7) 분리된 펩티드가 원하는 서열대로 합성이 되었는지를 LC/MS를 (Agilent HP1100 series) 이용하여 확인하였다. 7) LC / MS (Agilent HP1100 series) was confirmed whether the isolated peptide was synthesized according to the desired sequence.
8) 분자량이 확인된 펩티드를 다시 분석용 HPLC를 이용하여 95% 순도이상으로 분리 정제된 것을 확인하고 동결건조 과정을 거쳐 흰색 파우더로 제조하였다.8) The molecular weight was confirmed that the peptide was separated and purified to 95% purity or more using analytical HPLC, and then prepared as a white powder through a lyophilization process.
펩티드 CP-55와 CP-56도 상기 CP-54 제조와 같은 방법을 기본으로, 펩티드의 기본 골격만 각 펩티드의 해당 서열에 맞게 치환되어 제조하였다.Peptides CP-55 and CP-56 were also prepared based on the same method as the preparation of CP-54, in which only the basic skeleton of the peptide was substituted for the corresponding sequence of each peptide.
[실시예 2]Example 2
신규 활성을 갖는 펩티드들의 콜라겐 생성 증대 효과 측정Determination of the Collagen Production Enhancement Effect of Novel Peptides
신규 활성을 갖는 펩티드 CP-54, CP-55 및 CP-56 과 이를 포함한 조성물이 주름개선을 위한 콜라겐 생성 효과가 있는지 검증하기 위하여, 인간 피부 섬유아세포(human skin fibroblast)를 배양하여 콜라겐 합성 효과가 있는 것으로 알려진 TGF-β(양성대조군) 및 각각의 펩티드를 처리한 뒤, 흡광도 측정 방법을 통하여, 아무것도 처리하지 않은 대조군을 기준으로 양성대조군과 각 펩티드 처리 실험군의 상대 콜라겐 합성도(collagen synthesis)를 계산하였다.In order to verify whether the peptides CP-54, CP-55 and CP-56 having a novel activity and the composition containing the same have collagen production effect for wrinkle improvement, human skin fibroblasts are cultured to improve collagen synthesis effect. After treating TGF-β (positive control) and each peptide known to be present, relative collagen synthesis of the positive control group and each peptide-treated experimental group was determined based on the control group without any treatment by absorbance measurement method. Calculated.
실험 세포주 배양Experiment cell line culture
세포내의 콜라겐 생성 양을 비교하기 위하여, 인간 피부 섬유아세포주 (human skin fibroblast)인 CCD-986SK (ATCC® CRL-1947™)를 사용하였다. 상기 세포주를 10% FBS와 페니실린-스트렙토마이신(Penicillin-Streptomycin) 항생제가 첨가된 DMEM 배지에 넣어 100mm 배양 접시로 접종(seeding) 한 후, 37°C, 5% CO2 인큐베이터 (incubator)에서 배양하였다. 각 세포의 성장 속도 및 상태를 매일 확인하면서 2~3일 간격으로 1:2~1:3의 비율로 계대 배양 (subculture)하여 세포의 상태를 양호하게 유지하였다.To compare the amount of collagen production in cells, the CCD-986SK (ATCC® CRL-1947 ™ ), a human skin fibroblast, was used. The cell lines were placed in DMEM medium containing 10% FBS and penicillin-Streptomycin antibiotics and seeded in a 100 mm culture dish, followed by incubation in a 37 ° C., 5% CO 2 incubator. . While checking the growth rate and state of each cell every day, subcultures were performed at a ratio of 1: 2 to 1: 3 at intervals of 2 to 3 days to maintain a good state of the cells.
실험을 위하여 상기 배양된 CCD-986SK 세포를 24-웰 플레이트에 웰 당 2 x 104 개씩 넣은 후, 24 시간 동안 배양하였다.For the experiment, the cultured CCD-986SK cells were placed in a 24-well plate at 2 × 10 4 cells per well, and then cultured for 24 hours.
실험 방법Experiment method
배양 배지를 제거하고 PBS로 세척 한 후에 혈청을 첨가하지 않은 무혈청 배지로 2시간 동안 무영양(starvation) 처리하였다. 그 후 아무것도 처리하지 않는 대조군(control), TGF-β 20ng/ml을 처리한 양성대조군 및 본 발명의 일 측면에 따른 펩티드의 최종 농도가 50μM이 되도록 10mM의 펩티드를 가하여 처리한 실험군으로 나누어 각 군을 5% CO2, 37℃에서 24시간 배양하였다.The culture medium was removed and washed with PBS, followed by starvation treatment for 2 hours with serum-free medium without serum. Thereafter divided into a control group (no control), a positive control group treated with TGF-β 20ng / ml and an experimental group treated with the addition of 10 mM peptide so that the final concentration of the peptide according to an aspect of the present invention is 50μM each group Was incubated at 5% CO 2 , 37 ° C for 24 hours.
배양액을 수거하여 콜라겐 양을 ELISA (procollagen type IC-peptide EIA kit) 방법을 이용하여 측정하였다.The cultures were collected and the amount of collagen was measured using the ELISA (procollagen type IC-peptide EIA kit) method.
ELISA 방법은 ELISA plate에 배양액을 희석하여 20 ㎕를 넣고 항체-POD 컨쥬게이트 용액 100 ㎕를 함께 넣은 다음 37℃에서 3시간 배양하였다.In the ELISA method, 20 μl of the culture solution was diluted in an ELISA plate, 100 μl of the antibody-POD conjugate solution was added together, and then incubated at 37 ° C. for 3 hours.
워싱 버퍼(washing buffer)로 4회 세척 한 후에 발색시약 (TMBZ) 100 ㎕를 넣고 상온에서 15분간 배양한 다음 1N 황산 100 ㎕를 넣었다. 그 후에 450 nm에서 ELISA 리더(reader)로 측정하였다.After washing four times with a washing buffer (washing buffer), 100 μl of color reagent (TMBZ) was added thereto, and incubated at room temperature for 15 minutes, followed by 100 μl of 1N sulfuric acid. It was then measured with an ELISA reader at 450 nm.
실험 결과 및 분석Experiment Results and Analysis
상기 실험 방법을 통하여 도출된 콜라겐 합성도는 대조군(control)을 100%라 했을 때 그 상대적인 값으로 계산하여 나타내었으며 다음 표 2 와 같다.Collagen synthesis degree derived through the above experimental method was calculated and expressed as a relative value when the control (control) is 100%, as shown in Table 2 below.
처리군Treatment group 콜라겐 합성도 (%)Collagen Synthesis (%)
ControlControl 100100
TGF-β 20 ng/mlTGF-β 20 ng / ml 112.5112.5
CP-54 50 μMCP-54 50 μM 112.2112.2
CP-55 50 μMCP-55 50 μM 124.8124.8
CP-56 50 μMCP-56 50 μM 118.9118.9
측정 및 계산 결과, 본발명에 따른 신규 활성을 갖는 펩티드 CP-54, CP-55, CP-56 처리시, 대조군에 비하여 10% 이상 콜라겐 합성도가 증가하는 것이 나타났다.As a result of the measurement and calculation, the peptides CP-54, CP-55, CP-56 having a novel activity according to the present invention showed that the collagen synthesis degree increased by more than 10% compared to the control.
이를 통하여 신규 활성을 갖는 펩티드 CP-54, CP-55, CP-56이 인간 피부 세포에서 콜라겐 생성을 증대시키는 효과가 있는 것을 알 수 있었다.It was found that peptides CP-54, CP-55, and CP-56 having novel activities have an effect of increasing collagen production in human skin cells.
상기 실험 결과를 통하여, 본 발명의 일 측면에 따른 신규 활성을 갖는 펩티드 및 이를 포함하는 조성물은 콜라겐 합성 효과를 보이는 것을 알 수 있다. 이 효과들은 피부의 주름개선 및 노화방지 효과를 줄 수 있는 것으로, 본 발명의 일 측면에 따른 펩티드 및 이를 포함한 조성물이 피부 상태 개선에서 다양하고 우수한 효과를 가질 수 있음을 알 수 있다.Through the above experimental results, it can be seen that the peptide having a novel activity according to an aspect of the present invention and a composition comprising the same show collagen synthesis effects. These effects are to improve the wrinkles and anti-aging of the skin, it can be seen that the peptide and the composition comprising the same according to an aspect of the present invention can have a variety of excellent effects in improving the skin condition.
본 발명의 일 측면에 따른 신규 활성을 갖는 펩티드가 콜라겐 합성 활성이 있음을 알 수 있으며, 이를 이용하여 주름개선 및 노화방지용 기능성 화장료 조성물 개발이 가능할 것으로 기대된다.It can be seen that the peptide having a novel activity according to an aspect of the present invention has collagen synthesis activity, and it is expected that the functional cosmetic composition for wrinkle improvement and anti-aging may be developed using the peptide.

Claims (9)

  1. 서열 번호 1 내지 3으로 표시되는 아미노산 서열 중 어느 하나로 구성되는 펩티드, 상기 펩티드 서열의 단편인 펩티드, 상기 펩티드 서열 또는 그의 단편인 펩티드와 80% 이상의 서열 상동성을 갖는 펩티드로 구성된 군으로부터 선택되는 콜라겐 합성 활성을 보이는 펩티드 또는 그의 염.Collagen selected from the group consisting of a peptide consisting of any one of the amino acid sequences represented by SEQ ID NOs: 1 to 3, a peptide that is a fragment of the peptide sequence, a peptide having 80% or more sequence homology with the peptide sequence or a peptide that is a fragment thereof Peptides or salts thereof that exhibit synthetic activity.
  2. 제 1항에 따른 상기 펩티드를 코딩하는 뉴클레오티드 서열.A nucleotide sequence encoding the peptide according to claim 1.
  3. 제 1항에 따른 펩티드 또는 그의 염을 유효성분으로 포함하는 주름개선 및 노화방지 기능성 조성물.Wrinkle improvement and anti-aging functional composition comprising the peptide according to claim 1 or a salt thereof as an active ingredient.
  4. 제 3항에 있어서, 상기 기능성 조성물 내의 상기 펩티드 또는 그의 염의 농도는 20 내지 80 μM인, 기능성 조성물.The functional composition of claim 3, wherein the concentration of the peptide or salt thereof in the functional composition is 20-80 μM.
  5. 제 3항에 있어서, 상기 펩티드 또는 그의 염의 투여량은 0.001 내지 10 mg/kg인, 기능성 조성물.The functional composition of claim 3, wherein the dosage of the peptide or salt thereof is from 0.001 to 10 mg / kg.
  6. 제 3항에 있어서, 상기 조성물은 주름개선 및 노화방지용 화장료 조성물인 것을 특징으로 하는 주름개선 및 노화방지 기능성 조성물.According to claim 3, wherein the composition is anti-wrinkle and anti-aging functional composition, characterized in that the cosmetic composition for wrinkle improvement and anti-aging.
  7. 제 6항에 있어서, 상기 화장료 조성물은 스킨, 로션, 크림, 파운데이션, 에센스, 젤, 팩, 폼 클렌징, 비누 및 피부외용 연고로 이루어진 군으로부터 선택된 어느 하나의 제형으로 이루어진 것을 특징으로 하는 특징으로 하는 주름개선 및 노화방지 기능성 조성물.According to claim 6, wherein the cosmetic composition wrinkles, characterized in that consisting of any one selected from the group consisting of skin, lotion, cream, foundation, essence, gel, pack, foam cleansing, soap and skin ointment. Improved and anti-aging functional composition.
  8. 제 3항에 있어서, 상기 조성물은 주름개선 및 노화방지용 약학적 조성물인 것을 특징으로 하는 주름개선 및 노화방지 기능성 조성물.According to claim 3, wherein the composition is wrinkle improvement and anti-aging functional composition characterized in that the pharmaceutical composition for anti-wrinkle and anti-aging.
  9. 제 3항에 있어서, 상기 조성물은 주름개선 및 노화방지용 건강기능 식품용 조성물인 것을 특징으로 하는 주름개선 및 노화방지 기능성 조성물.According to claim 3, The composition is wrinkle improvement and anti-aging functional composition, characterized in that the composition for wrinkle improvement and anti-aging health functional food.
PCT/KR2017/004288 2016-04-21 2017-04-21 Peptide having efficacy of increasing collagen generation and composition comprising same WO2017183942A2 (en)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
KR20160048749 2016-04-21
KR10-2016-0048749 2016-04-21

Publications (2)

Publication Number Publication Date
WO2017183942A2 true WO2017183942A2 (en) 2017-10-26
WO2017183942A3 WO2017183942A3 (en) 2018-08-02

Family

ID=60116231

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/KR2017/004288 WO2017183942A2 (en) 2016-04-21 2017-04-21 Peptide having efficacy of increasing collagen generation and composition comprising same

Country Status (1)

Country Link
WO (1) WO2017183942A2 (en)

Family Cites Families (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP4439510B2 (en) * 2005-03-22 2010-03-24 ロート製薬株式会社 New peptide
KR100721144B1 (en) * 2006-03-22 2007-05-25 (주)아르떼르화장품 Cosmetic composition for caring wrinkle containing peptide combination
US9527888B2 (en) * 2012-05-11 2016-12-27 Gemvax & Kael Co., Ltd. Anti-inflammatory peptides and composition comprising the same
WO2016006733A1 (en) * 2014-07-09 2016-01-14 강원대학교산학협력단 Composition containing fibroblast growth factor-derived peptide fragments for wound healing or inhibiting skin wrinkle formation
JP7000161B2 (en) * 2015-05-26 2022-01-19 ジェムバックス アンド カエル カンパニー，リミティド New peptide and composition containing it

Also Published As

Publication number Publication date
WO2017183942A3 (en) 2018-08-02

Similar Documents

Publication Publication Date Title
WO2021033829A1 (en) Peptide having activity of improving skin condition and use thereof
WO2017155232A2 (en) Peptide exhibiting hair growth promoting activity and/or melanin production promoting activity and use thereof
WO2016190660A1 (en) Novel peptide and composition containing the same
WO2016088968A1 (en) Peptide having activity to improve skin condition and use thereof
BR122020010541B1 (en) PEPTIDE HAVING AN ACTIVITY OF A GROWTH FACTOR (GF) AND DERIVATIVE OF GF AND PHARMACEUTICAL COMPOSITION FOR THE IMPROVEMENT OF A SKIN CONDITION AND FOR THE TREATMENT OF AN INJURY
WO2021033831A1 (en) Peptide having skin whitening activity and use thereof
WO2017176087A1 (en) Peptide having effects of increasing telomerase activity and extending telomere, and composition containing same
WO2019045248A2 (en) Peptide exhibiting wrinkle-improving activity and uses thereof
EP3815674A1 (en) Composition for muscle relaxation
KR20190011207A (en) Tyrosinase antisense oligonucleotides
WO2017116138A1 (en) Melanin-inhibiting peptide and composition containing same
WO2017034301A1 (en) Peptide having the activity of epithelial growth factor, and use therefor
WO2021153946A1 (en) Peptide inhibiting formation of snare complex and use thereof
WO2017119756A1 (en) Wrinkle-improving, anti-aging, whitening, anti-inflammatory functional novel peptide, and composition containing same
WO2018034453A1 (en) Conjugate of minoxidil and peptide
WO2017183942A2 (en) Peptide having efficacy of increasing collagen generation and composition comprising same
WO2022124776A1 (en) Composition for whitening or wrinkle reduction, comprising alloferon-pantothenic acid complex as active ingredient
WO2021033833A1 (en) Peptide having skin whitening activity and use thereof
US11771636B2 (en) Composition for promoting hair growth comprising adiponectin-derived peptide
WO2022050634A1 (en) Peptide, and cosmetic composition and pharmaceutical composition comprising same
WO2023055007A1 (en) Peptide having anti-aging activity, and use thereof
KR102346006B1 (en) Pentapeptide derived from spider web and composition for anti-aging containing the same
WO2021251790A1 (en) Deoxycholic acid-peptide conjugate having anti-obesity activity and use thereof
WO2023054817A1 (en) Peptide having anti-aging activity, and use thereof
KR101054519B1 (en) Human Growth Hormone-Derived Peptides and Uses thereof

Legal Events

Date Code Title Description
NENP Non-entry into the national phase

Ref country code: DE

121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 17786210

Country of ref document: EP

Kind code of ref document: A2

122 Ep: pct application non-entry in european phase

Ref document number: 17786210

Country of ref document: EP

Kind code of ref document: A2