KR101645238B1 - Cosmetic composition for anti-wrinkle or elasticity of skin - Google Patents

Cosmetic composition for anti-wrinkle or elasticity of skin Download PDF

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KR101645238B1
KR101645238B1 KR1020150158337A KR20150158337A KR101645238B1 KR 101645238 B1 KR101645238 B1 KR 101645238B1 KR 1020150158337 A KR1020150158337 A KR 1020150158337A KR 20150158337 A KR20150158337 A KR 20150158337A KR 101645238 B1 KR101645238 B1 KR 101645238B1
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extract
cosmetic composition
peptide
enzyme
molecular weight
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장애라
김동욱
김희진
길주애
김혜진
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강원대학교산학협력단
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    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00Cosmetics or similar toiletry preparations
    • A61K8/18Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/30Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
    • A61K8/64Proteins; Peptides; Derivatives or degradation products thereof
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00Cosmetics or similar toiletry preparations
    • A61K8/18Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/96Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
    • A61K8/98Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution of animal origin
    • A61K8/981Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution of animal origin of mammals or bird
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61QSPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q19/00Preparations for care of the skin
    • A61Q19/08Anti-ageing preparations

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  • Life Sciences & Earth Sciences (AREA)
  • General Health & Medical Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
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  • Gerontology & Geriatric Medicine (AREA)
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  • Peptides Or Proteins (AREA)
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Abstract

The present invention relates to a cosmetic composition for reducing skin wrinkles, and maintaining or improving skin elasticity, comprising a peptide having an amino acid sequence of SEQ ID NO: 1 separated from the leg bones of horses. The composition exhibits excellent effects in reducing skin wrinkles, and maintaining or improving skin elasticity.

Description

피부주름개선용, 피부탄력유지 또는 개선용 화장료 조성물 {Cosmetic composition for anti-wrinkle or elasticity of skin} [0001] The present invention relates to a cosmetic composition for anti-wrinkle or elasticity of skin,

본 발명은 피부주름개선용, 피부탄력유지 또는 개선용 화장료 조성물에 관한 것으로, 더욱 상세하게는 말 사골로부터 분리한 서열번호 1의 아미노산 서열을 갖는 펩타이드를 함유하는 피부주름개선용, '피부탄력유지 또는 개선용' 화장료 조성물에 관한 것이다. The present invention relates to a cosmetic composition for skin wrinkle improvement and maintenance or improvement of skin elasticity, and more particularly to a cosmetic composition for skin wrinkle improvement which contains a peptide having an amino acid sequence of SEQ ID NO: 1 isolated from horsehair bone, Or improving cosmetic composition.

최근 건강한 삶에 대한 관심이 증가하고, 생활수준의 향상, 여성의 사회진출 증가, 고령화 가속화 등으로 인해, 소비자들의 화장품에 대한 욕구는 아름답게 꾸미기 위한 용도에서 점차 기능적 용도를 요구하는 것으로 변화하고 있다. In recent years, consumers' desire for cosmetics has been changing from requiring use to beauty to increasing functional use due to increased interest in healthy living, improved living standards, increased women's social advancement, and accelerated aging.

이와 같은 시대적 요구에 맞춰, 우수한 기능성을 갖는 원료 소재에 대한 탐색이 광범위하게 일어나고 있는데, 천연 물질에서 우수한 기능성 물질을 찾으려는 노력도 많이 이루어지고 있다.In response to such demands of the times, there has been a wide search for raw materials having excellent functionality, and efforts have been made to find excellent functional materials in natural materials.

펩타이드(peptide)는 아미노산이 2개 이상 결합하여 형성된 물질인데, 화학합성, 효소반응 또는 단백질로부터의 가수분해에 의해 제조된다. 최근 펩타이드 성분을 화장품에 사용하여 피부 주름을 개선하는 연구가 활발하게 진행되고 있다. Peptides are substances formed by combining two or more amino acids, which are produced by chemical synthesis, enzymatic reaction or hydrolysis from proteins. Recently, studies have been actively conducted to improve skin wrinkles by using peptide components in cosmetics.

그런데, 화학합성된 펩타이드의 경우, 화학합성에 대한 소비자들의 거부감으로 인해, 활발히 사용되지 못하고 있는 실정이고, 효소반응의 경우, 고가의 효소를 사용하기 때문에 경제적이지 못한 문제점이 있다. 따라서, 단백질로부터 가수분해하여 펩타이드를 저렴하게 회수할 수 있는 방법이 대안으로 떠오르고 있다.In the case of chemically synthesized peptides, however, they are not actively used due to the rejection of chemical synthesis. In the case of enzyme reaction, expensive enzymes are used, which is not economical. Therefore, a method of hydrolyzing the protein and recovering the peptide at low cost has emerged as an alternative.

한편, 말의 뼈는 국내에서 흔히 사용되지는 않지만, 인터넷 시장 혹은 제주지역에서 소비되고 있는 실정이다. 한의학에서는 말의 뼈가 의약소재로 사용되기도 하였다. 중화 본초에서는 말의 뼈가 성질이 달고 차며, 동의보감에서도 신경통, 관절염, 이명 등에 효과가 있고, 말의 머리뼈는 머리와 귀에 생긴 염증을 제어한다고 보고되어 있다. On the other hand, horse bones are not commonly used in Korea, but they are consumed in the internet market or Jeju area. In Oriental medicine, horse bones were also used as medicinal materials. In Chinese herb bases, horse bones are characterized by their properties. In Dongbokgi, neuralgia, arthritis and tinnitus are also effective. Horses' bones are reported to control inflammation in the head and ear.

그런데, 말의 뼈와 관련된 연구는 주로 골다공증, 뼈 성장 등에 관해 이루어졌을 뿐, 화장품 소재로서 연구된 바는 거의 없다. 특히, 말의 사골을 화장품 소재로 개발한 연구는 전혀 보고된 바 없다.However, studies related to horse bones have mainly focused on osteoporosis, bone growth, and have not been studied as cosmetic materials. In particular, no research has been reported on the development of horse bones as cosmetic materials.

대한민국 특허등록번호 제10-0553174호 (등록일자 2006. 02. 10)에는, 사람 SNAP-25의 C-말단에서 유래한 펩타이드에 PTD 펩티드(Protein Transduction Domain)가 결합된 융합 펩타이드로서, 높은 생체내 투과성과 안정성을 가지며, 주름살 형성에 관련된 신경전달물질인 카테콜아민의 분비를 억제하여 우수한 피부 주름 개선 효과를 보이는 펩타이드가 기재되어 있다.Korean Patent Registration No. 10-0553174 (filed on Mar. 02, 2006) discloses a fusion peptide in which a PTD peptide (Protein Transduction Domain) is bound to a peptide derived from the C-terminal of human SNAP-25, A peptide having permeability and stability and suppressing secretion of catecholamine, which is a neurotransmitter related to wrinkle formation, and showing excellent skin wrinkle-reducing effect. 대한민국 특허등록번호 제10-0844386호 (등록일자 2008. 07. 01)에는, 오징어 콜라겐으로부터 피부주름 억제활성을 갖는 펩타이드를 제조하는 방법에 관한 것으로서, 종래 산 및 염기에 의한 가수분해시 단점인 장치의 부식문제, 폐수 발생 문제, 효소에 의한 가수분해시 단점인 염의 다량 발생 문제 등을 해결하고, 또한 간단한 효소 처리 후 초음파를 병행하여 빠른 시간 내에 고수율로 주름억제용 향장소제 펩타이드를 제조할 수 있는 방법이 기재되어 있다.Korean Patent Registration No. 10-0844386 (registered on Jan. 07, 2008) discloses a process for producing a peptide having skin wrinkle inhibitory activity from squid collagen, which is a disadvantage in the hydrolysis with acids and bases The problem of the generation of waste water, the problem of generation of a large amount of salt, which is a disadvantage in the hydrolysis by enzymes, and the like, and after ultrasonication after simple enzyme treatment, the peptides A method is described.

본 발명은 피부주름개선 또는 피부탄력증진 효과가 있는 펩타이드를 천연물로부터 발굴하여 화장료 조성물로 개발하여 제공하고자 한다. The present invention is to develop a peptide composition having a skin wrinkle improving effect or skin elasticity improving effect as a cosmetic composition by extracting it from a natural product.

본 발명은 서열번호 1에 기재된 아미노산 서열을 갖는 펩타이드를 유효성분으로 함유하는 것을 특징으로 하는 피부주름개선용 화장료 조성물을 제공한다. 또한, 본 발명은 서열번호 1에 기재된 아미노산 서열을 갖는 펩타이드를 유효성분으로 함유하는 것을 특징으로 하는 피부탄력유지 또는 개선용 화장료 조성물을 제공한다.The present invention provides a cosmetic composition for improving skin wrinkles, which comprises a peptide having the amino acid sequence of SEQ ID NO: 1 as an active ingredient. The present invention also provides a cosmetic composition for skin elasticity maintenance or improvement, which comprises a peptide having the amino acid sequence of SEQ ID NO: 1 as an active ingredient.

하기 본 발명의 실험에 의할 경우, 서열번호 1에 기재된 아미노산 서열을 갖는 펩타이드는 56.07%의 콜라게나아제 저해 활성을 나타내어, 우수한 주름개선 효과와 '피부탄력유지 또는 개선 효과'를 나타내는 소재임을 알 수 있었다. According to the experiment of the present invention as described below, the peptide having the amino acid sequence shown in SEQ ID NO: 1 showed collagenase inhibitory activity of 56.07%, indicating excellent wrinkle improving effect and 'skin elasticity maintenance or improvement effect' I could.

콜라게나아제는 콜라겐을 분해하는 효소인데, 콜라겐은 대부분 진피층에 존재하며, 피부 전체 건조 중량의 약 70~80%를 차지하고 있다. 콜라겐은 섬유아세포에서 생합성되는데, 자연노화 및 광노화에 의해서 그 양이 감소하며, 20세에서 80세에 이르는 동안 65%의 감소가 일어난다고 알려져 있다(Shuster S., 1975, British journal of determatology, 93(6): 639-643). 또한, 콜라겐의 감소는 피부의 주름 형성과 밀접한 연관이 있는 것으로 알려져 있다.Collagenase is an enzyme that degrades collagen. Most collagen is present in the dermis and accounts for about 70-80% of the dry weight of the skin. Collagen is biosynthesized in fibroblasts, reduced in amount by natural aging and photoaging, and is known to decrease by 65% during the period from 20 to 80 years of age (Shuster S., 1975, British journal of determatology, 93 (6): 639-643). In addition, collagen reduction is known to be closely related to the wrinkling of the skin.

한편, 본 발명의 피부주름개선용 또는 '피부탄력유지 또는 개선용' 화장료 조성물에 있어서, 상기 주름개선 또는 '피부탄력유지 또는 개선'은 바람직하게 콜라게나아제 효소 활성의 억제로 말미암은 것일 수 있다. On the other hand, in the cosmetic composition for skin wrinkle improvement or skin elasticity maintenance or improvement according to the present invention, the wrinkle improvement or 'skin elasticity maintenance or improvement' may preferably be due to inhibition of collagenase enzyme activity.

한편, 본 발명의 피부주름개선용 또는 '피부탄력유지 또는 개선용' 화장료 조성물에 있어서, 상기 서열번호 1의 아미노산 서열을 갖는 펩타이드는, 일 예로 말 사골로부터 유래한 것일 수 있다. Meanwhile, in the cosmetic composition for improving skin wrinkles or 'skin elasticity maintenance or improvement' of the present invention, the peptide having the amino acid sequence of SEQ ID NO: 1 may be one derived from horse bone bone.

한편, 본 발명에 따른 화장료 조성물은, 상기 펩타이드 외에 본 발명이 목적으로 하는 주 효과를 손상시키지 않는 범위 내에서 주 효과에 상승효과를 줄 수 있는 다른 성분 등을 함유하는 것도 무방하다.On the other hand, the cosmetic composition according to the present invention may contain, in addition to the above-mentioned peptides, other components or the like which can give synergy to the main effect within a range not impairing the intended main effect of the present invention.

한편, 본 발명에 따른 조성물은, 화장 분야에서 통상적으로 사용되는 보조제, 예컨대 친수성 또는 친유성 겔화제, 친수성 또는 친유성 활성제, 보존제, 항산화제, 용매, 방향제, 충전제, 차단제, 안료, 흡취제 및 염료를 함유할 수 있다. 이들 다양한 보조제의 양은 당해 분야에서 통상적으로 사용되는 양이며, 예컨대 조성물 총중량에 대해 0.001 내지 30중량%가 사용될 수 있다. 다만, 어떠한 경우라도 보조제 및 그 비율은 본 발명에 따른 화장료 조성물의 바람직한 성질에 악영향을 미치지 않도록 선택되어야 한다.On the other hand, the composition according to the present invention can be used as an auxiliary agent commonly used in cosmetics such as a hydrophilic or lipophilic gelling agent, a hydrophilic or lipophilic active agent, a preservative, an antioxidant, a solvent, a fragrance, a filler, May contain a dye. The amount of these various adjuvants is an amount ordinarily used in the art, for example, 0.001 to 30% by weight based on the total weight of the composition may be used. However, in any case, the adjuvants and their proportions should be chosen so as not to adversely affect the desired properties of the cosmetic composition according to the invention.

한편, 본 발명의 화장료 조성물 제형은 특정한 것에 한정되는 것은 아니고, 일 예로 화장수, 젤, 수용성 리퀴드, 크림, 에센스, 수중유(O/W)형 및 유중수(W/O)형의 에멀젼, 연고로 이루어진 기초화장료 제형, 수중유형 및 유중수형 메이크업베이스, 파운데이션 제형 중에서 선택되는 어느 하나인 것이 좋다. 또한, 이는 선택적으로 에어로졸 형태로 피부에 적용될 수 있고, 젤 또는 크림의 형태일 수도 있으며, 피부용 케어 제품 및/또는 메이크업 제품으로 사용될 수 있다.On the other hand, the cosmetic composition composition of the present invention is not limited to a specific one, and examples thereof include emulsions such as lotion, gel, water-soluble liquid, cream, essence, O / W type and w / A foundation cosmetic formulation, an underwater type and a water-in-oil type make-up base, and a foundation formulation. It can also be applied to the skin, optionally in the form of an aerosol, in the form of a gel or cream, and as a skin care and / or makeup product.

본 발명의 서열번호 1에 기재된 아미노산 서열을 갖는 펩타이드는 높은 콜라게나아제 저해 활성을 나타내는 바, 우수한 주름개선, '피부탄력유지 또는 개선'의 효과를 발휘한다. The peptide having the amino acid sequence of SEQ ID NO: 1 of the present invention exhibits a high collagenase inhibitory activity and exhibits excellent wrinkle improvement and 'skin elasticity maintenance or improvement' effects.

도 1은 말 사골 추출물의 콜라게나아제 저해 활성을 측정한 그래프이다.
도 2는 겔 여과 크로마토그래피를 이용하여 분획한 펩타이드 분획물의 분포도이다.
도 3은 '도 2'에서 분획한 분획물의 콜라게나아제 억제 효과를 측정한 결과이다.
FIG. 1 is a graph showing collagenase inhibitory activity of the horse bovine bone extract. FIG.
Figure 2 is a distribution diagram of peptide fractions fractionated using gel filtration chromatography.
FIG. 3 shows the results of measuring the inhibitory effect of collagenase on the fractions fractionated in FIG. 2. FIG.

이하, 본 발명의 내용을 하기 실시예 및 실험예를 통해 더욱 상세히 설명하고자 한다. 다만, 본 발명의 권리범위가 하기 실시예 및 실험예에만 한정되는 것은 아니고, 그와 등가의 기술적 사상의 변형까지를 포함한다. Hereinafter, the present invention will be described in more detail with reference to the following Examples and Experimental Examples. However, the scope of the present invention is not limited to the following embodiments and experimental examples, and includes modifications of equivalent technical ideas.

[[ 제조예Manufacturing example 1: 말 사골 추출물 제조] 1: Preparation of horse bone meal extract]

깨끗이 세척된 말 사골 1 kg을 찬물에 16시간 동안 침수시켜 핏물을 제거하였다. 그 후, 3 L의 물을 넣고 끓는 물에서 30분간 끓인 후, 물을 제거하고 말 사골을 찬물로 깨끗이 세척하였다. 그 후, 이물질이 제거된 말 사골에 6 L의 물을 넣고 1차(8시간), 2차(8시간), 3차(8시간)로 나누어 가열하여 젤라틴 함유 추출물을 제조하였다. 상기 젤라틴 함유 추출물을 거즈로 각각 여과하여 동결 건조한 다음 1차 추출물, 2차 추출물, 3차 추출물을 동일한 양 즉, 각각의 추출물을 100 g씩 혼합하여 말 사골 가수분해물 300 g을 제조하였다. 그 후, 상기 말 사골 가수분해물을 냉동하여 지방을 제거한 후, 동결건조하였다. 상기 동결건조된 말 사골 가수분해물 100 g에 증류수 800 ㎖를 넣고 80℃에서 1시간 동안 교반한 후, 1M NaOH를 첨가하여 pH를 8.0으로 조정하였다. 그 후, 판크립신(Pancripsin, 판크레아틴의 제품명, 비젼바이오켐) 0.2 g을 넣고, 50℃에서 4시간 동안 효소분해를 하여 말 사골 추출물을 수득하였다. 그 후, 10분 동안 90℃를 유지하여 효소를 불활성화하였다. One kilogram of cleanly washed horseshoe was immersed in cold water for 16 hours to remove blood. After that, 3 L of water was added, boiled in boiling water for 30 minutes, then water was removed, and the horse bones were cleanly washed with cold water. Then, 6 L of water was added to the horseshoe bone with the foreign substances removed, and the mixture was heated for the first (8 hours), the second (8 hours) and the third (8 hours) to prepare a gelatin-containing extract. The gelatin-containing extracts were each filtered with gauze, lyophilized, and then 100 g of each of the first extract, the second extract and the third extract were mixed with each other to prepare 300 g of horsehide hydrolyzate. Thereafter, the horse bone hydrolyzate was frozen to remove fat, followed by lyophilization. 800 ml of distilled water was added to 100 g of the lyophilized horse bone hydrolyzate, and the mixture was stirred at 80 ° C for 1 hour. Then, the pH was adjusted to 8.0 by adding 1 M NaOH. Thereafter, 0.2 g of Pancripsin (product name of pancreatin, Vision BioChem) was added, and enzymatic degradation was performed at 50 캜 for 4 hours to obtain a horse bark extract. Thereafter, the enzyme was inactivated by keeping it at 90 DEG C for 10 minutes.

한편, 상기 효소분해된 말 사골 추출물을 멤브레인 여과 장치를 이용하여 3 KDa 이상의 고분자 분해물, 3 KDa 이하의 저분자 분해물로 분리하였다. 그 후, 상기 말 사골 가수분해물을 'HM4(hydrolysates)', 3 KDa 이상의 말 사골 고분자 효소 분해물을 'HO4(hydrolysates over 3 KDa)', 3 KDa 이하의 말 사골 저분자 효소 분해물을 'HL4(hydrolysates less than 3 KDa)'로 지칭한 후, 하기 실시예에서 사용하였다.On the other hand, the enzyme-degraded horse bone bark extract was separated into polymer decomposition products of 3 KDa or more and low molecular decomposition products of 3 KDa or less by using a membrane filtration apparatus. Thereafter, the hydrolyzate of the horse bone hydrolyzate is referred to as' HM4 (hydrolysates) ', the hydrolyzate over 3 KDa hydrogel oxide hydrolyzate of 3 KDa or more and the hydrogel oxide hydrolyzate hydrolysates less than 3 KDa as' HL4 < / RTI > than 3 KDa), which was used in the following examples.

[[ 실시예Example 1: 말 사골 추출물의  1: Horseshoe extract 콜라게나아제Collagenase 저해활성 측정] Measurement of inhibitory activity]

본 실험예에서는 말 사골 추출물의 피부 주름 개선 효과를 확인하고자 콜라게나아제(collagenase) 저해활성을 측정하였다.In this experiment, collagenase inhibitory activity was measured in order to confirm the effect of improving the wrinkles of the skin of the horseshoe crab extract.

시료로서 1, 10, 25, 50, 100, 200 ㎎/㎖의 농도로 고정한 3 KDa 이하의 말 사골 저분자 효소 분해물을 이용하였다.A low molecular weight enzyme hydrolyzate of 3 KDa or less fixed at a concentration of 1, 10, 25, 50, 100 and 200 mg / ml was used as a sample.

0.3 mg/㎖ 콜라게나아제 발색단(Colaagenase Chromophore) 기질 0.25 ㎖에 농도별로 시료 0.1 ㎖를 넣고, 0.2 mg/㎖ 콜라게나아제(Collagenase) 효소를 0.15 ㎖ 첨가하여 실온에서 20분간 방치한 후, 6% 시트릭산(citric acid) 0.5 ㎖를 넣어 효소반응을 정지시켰다. 그 후, 에틸아세테이트(ethylacetate) 1.5 ㎖를 첨가하여 상등액을 320 nm에서 흡광도를 측정하였다. 콜라게나아제(Collagenase) 저해활성은 하기 수학식 2를 이용하여 계산하였다.0.1 mg of each sample was added to 0.25 ml of a 0.3 mg / ml colaagenase chromophore substrate, 0.15 ml of a 0.2 mg / ml collagenase enzyme was added, and the mixture was allowed to stand at room temperature for 20 minutes, The enzyme reaction was stopped by adding 0.5 ml of citric acid. Then, 1.5 ml of ethylacetate was added and the supernatant was measured for absorbance at 320 nm. Collagenase inhibitory activity was calculated using the following equation (2).

Figure 112015110005568-pat00001
Figure 112015110005568-pat00001

실험결과, 농도 의존적으로 콜라게나아제(Collagenase) 저해활성이 높게 나타났다. 특히, 농도 200 mg/㎖일 때, 98.35%로 가장 높은 저해 활성을 나타내었다(도 1).As a result, collagenase inhibitory activity was high in a concentration-dependent manner. In particular, when the concentration was 200 mg / ml, the highest inhibitory activity was shown at 98.35% (FIG. 1).

상기와 같은 결과로부터, 말 사골 추출물의 우수한 피부 주름 개선 효과 및 피부 노화 억제 효과를 확인할 수 있었다.From the results described above, it was confirmed that the extract of horsehair bark extract had excellent skin wrinkle improving effect and skin aging inhibiting effect.

[[ 실시예Example 2: 32: 3 KDaKDa 이하의 말 사골  Below 저분자Low molecule 효소  enzyme 분해물로부터From the degradation product 콜라게나아제Collagenase 억제 기능성 펩타이드의 분리] Isolation of inhibitory functional peptides]

3 KDa 이하의 말 사골 저분자 효소 분해물을 0.15 N NaCl이 포함되어 있는 50 mM 포스페이트 버퍼 (pH 7.0)로 평형화시킨 'Superdex™ Peptide 1//300 GL'에 주입하여 분획별로 분리하고, 280 nm에서 흡광도를 측정하였다. 분획별로 얻어진 펩타이드는 투석막을 사용하여 투석시킨 후, 동결건조하여 사용하였다. 3 KDa or less was injected into Superdex Peptide 1 // 300 GL, which was equilibrated with 50 mM phosphate buffer (pH 7.0) containing 0.15 N NaCl, and the fractions were separated into fractions. The absorbance at 280 nm Were measured. The peptides obtained for each fraction were dialyzed using a dialysis membrane and then lyophilized to use.

실험결과, 10개의 분획으로 분리한 분획물을 수득하였으며 (도 2), 농도 1 mg/ml일 때, 3, 4, 5번 분획에서, 각각 56.07, 53.69, 49.39%로 다른 분획물에 비해 유의적으로 높은 콜라게나아제 저해 활성을 확인할 수 있었다 (도 3). As a result of the experiment, fractions separated into 10 fractions were obtained (FIG. 2). The fractions were 56.07, 53.69 and 49.39% in the fractions 3, 4 and 5 at the concentration of 1 mg / High collagenase inhibitory activity was confirmed (FIG. 3).

이중 가장 높은 활성을 보인 3번 분획물에 대해 시퀀스 분석을 수행한 결과, 'GPAGPTGPVGKDGR'의 아미노산 서열을 갖는 것으로 확인되었고, 분자량이 1,265.66 Da으로 확인되었다. As a result of sequence analysis of the fraction 3 having the highest activity, it was confirmed to have the amino acid sequence of 'GPAGPTGPVGKDGR' and its molecular weight was found to be 1,265.66 Da.

<110> KNU-Industry Cooperation Foundation <120> Cosmetic composition for anti-wrinkle or elasticity of skin <130> AP-2015-0223 <160> 1 <170> KopatentIn 2.0 <210> 1 <211> 14 <212> PRT <213> Equus caballus <400> 1 Gly Pro Ala Gly Pro Thr Gly Pro Val Gly Lys Asp Gly Arg 1 5 10 <110> KNU-Industry Cooperation Foundation <120> Cosmetic composition for anti-wrinkle or elasticity of skin <130> AP-2015-0223 <160> 1 <170> Kopatentin 2.0 <210> 1 <211> 14 <212> PRT <213> Equus caballus <400> 1 Gly Pro Ala Gly Pro Thr Gly Pro Val Gly Lys Asp Gly Arg   1 5 10

Claims (6)

서열번호 1에 기재된 아미노산 서열로 구성되는 펩타이드를 유효성분으로 함유하며,
상기 펩타이드는 말 사골로부터 유래한 것으로 분자량이 3 KDa 이하인 펩타이드인 것을 특징으로 하는 피부주름개선용 화장료 조성물.
A peptide comprising the amino acid sequence of SEQ ID NO: 1 as an active ingredient,
Wherein the peptide is derived from horseshoe bone and has a molecular weight of 3 KDa or less.
제1항에 있어서,
상기 주름개선은,
콜라게나아제 효소 활성의 억제로 말미암은 것을 특징으로 하는 피부주름개선용 화장료 조성물.
The method according to claim 1,
The wrinkle-
A cosmetic composition for improving skin wrinkles characterized by inhibition of collagenase enzyme activity.
말 사골을 열수 추출하여 젤라틴 함유 추출물을 제조한 후,
상기 젤라틴 함유 추출물에서 지방을 제거하고 pH를 7.0~9.0으로 조정한 후, 단백질 분해효소를 첨가하여 효소분해하여 말 사골 추출물을 수득한 후,
상기 효소분해된 말 사골 추출물을 멤브레인 여과장치를 거쳐 분자량이 3 KDa 이하인 말 사골 저분자 효소 분해물을 수득한 후 분획하여 '서열번호 1에 기재된 아미노산 서열로 구성되는 펩타이드'를 수득하는 과정을 포함하는 것을 특징으로 하는 피부주름개선용 화장료 조성물의 제조방법.
After extracting the gelatin-containing extract by hydrothermal extraction of horseshoe crab,
After removing the fat from the gelatin-containing extract and adjusting the pH to 7.0 to 9.0, proteolytic enzyme was added to the extract to enzymatically decompose the extract,
The process comprises the step of obtaining a horse-bovine low molecular weight enzyme hydrolyzate having a molecular weight of 3 KDa or less through a membrane filtration apparatus and fractionating the enzyme-hydrolyzed horse bone bark extract to obtain a peptide consisting of the amino acid sequence of SEQ ID NO: 1 Wherein the cosmetic composition is a cosmetic composition for improving skin wrinkles.
서열번호 1에 기재된 아미노산 서열로 구성되는 펩타이드를 유효성분으로 함유하며,
상기 펩타이드는 말 사골로부터 유래한 것으로 분자량이 3 KDa 이하인 펩타이드인 것을 특징으로 하는 피부탄력유지 또는 개선용 화장료 조성물.
A peptide comprising the amino acid sequence of SEQ ID NO: 1 as an active ingredient,
Wherein the peptide is derived from a horseshoe bone and has a molecular weight of 3 KDa or less.
제4항에 있어서,
상기 피부탄력유지 또는 개선은,
콜라게나아제 효소 활성의 억제로 말미암은 것을 특징으로 하는 피부탄력유지 또는 개선용 화장료 조성물.
5. The method of claim 4,
The skin elasticity maintenance or improvement may be performed,
A cosmetic composition for maintaining or improving skin elasticity characterized by inhibition of collagenase enzyme activity.
말 사골을 열수 추출하여 젤라틴 함유 추출물을 제조한 후,
상기 젤라틴 함유 추출물에서 지방을 제거하고 pH를 7.0~9.0으로 조정한 후, 단백질 분해효소를 첨가하여 효소분해하여 말 사골 추출물을 수득한 후,
상기 효소분해된 말 사골 추출물을 멤브레인 여과장치를 거쳐 분자량이 3 KDa 이하인 말 사골 저분자 효소 분해물을 수득한 후 분획하여 '서열번호 1에 기재된 아미노산 서열로 구성되는 펩타이드'를 수득하는 과정을 포함하는 것을 특징으로 하는 피부탄력유지 또는 개선용 화장료 조성물의 제조방법.
After extracting the gelatin-containing extract by hydrothermal extraction of horseshoe crab,
After removing the fat from the gelatin-containing extract and adjusting the pH to 7.0 to 9.0, proteolytic enzyme was added to the extract to enzymatically decompose the extract,
The process comprises the step of obtaining a horse-bovine low molecular weight enzyme hydrolyzate having a molecular weight of 3 KDa or less through a membrane filtration apparatus and fractionating the enzyme-hydrolyzed horse bone bark extract to obtain a peptide consisting of the amino acid sequence of SEQ ID NO: 1 Wherein the cosmetic composition is a cosmetic composition.
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KR101814744B1 (en) 2016-10-25 2018-01-03 천유화장품(주) Manufacturing method of composition for inhibiting skin aging using donkey skin
KR20180080736A (en) * 2017-01-04 2018-07-13 강원대학교산학협력단 Cosmetic composition with small molecule gelatin peptide from donkey bone for anti-wrinkle and anti-aging of skin
KR102657790B1 (en) 2023-11-23 2024-04-17 주식회사 뜰꽃 Cosmetic composition for improving skin elasticity containing natural extracts

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KR101814744B1 (en) 2016-10-25 2018-01-03 천유화장품(주) Manufacturing method of composition for inhibiting skin aging using donkey skin
KR20180080736A (en) * 2017-01-04 2018-07-13 강원대학교산학협력단 Cosmetic composition with small molecule gelatin peptide from donkey bone for anti-wrinkle and anti-aging of skin
KR102657790B1 (en) 2023-11-23 2024-04-17 주식회사 뜰꽃 Cosmetic composition for improving skin elasticity containing natural extracts

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