KR20170128726A - Cosmetics composition for anti-aging comprising agar-derived neoagarotetraose - Google Patents
Cosmetics composition for anti-aging comprising agar-derived neoagarotetraose Download PDFInfo
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- KR20170128726A KR20170128726A KR1020160058916A KR20160058916A KR20170128726A KR 20170128726 A KR20170128726 A KR 20170128726A KR 1020160058916 A KR1020160058916 A KR 1020160058916A KR 20160058916 A KR20160058916 A KR 20160058916A KR 20170128726 A KR20170128726 A KR 20170128726A
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- cosmetic composition
- neoagarotetraose
- skin
- agar
- aging
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Images
Classifications
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/02—Cosmetics or similar toiletry preparations characterised by special physical form
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- 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
- A61K8/72—Cosmetics or similar toiletry preparations characterised by the composition containing organic macromolecular compounds
- A61K8/73—Polysaccharides
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- 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
- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
- A61K8/9706—Algae
- A61K8/9717—Rhodophycota or Rhodophyta [red algae], e.g. Porphyra
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- 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
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Animal Behavior & Ethology (AREA)
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- Veterinary Medicine (AREA)
- Engineering & Computer Science (AREA)
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- Biotechnology (AREA)
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- Dermatology (AREA)
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Abstract
Description
본 발명은 네오아가로테트라오스를 함유하는 피부노화 개선용 화장료 조성물에 관한 것으로, 더욱 구체적으로 한천을 DagA 효소 반응시켜 제조한 네오아가로테트라오스를 함유하는 피부노화 개선용 화장료 조성물에 관한 것이다.The present invention relates to a cosmetic composition for improving skin aging which contains neoagarotetraose, and more specifically to a cosmetic composition for improving skin aging containing neoagarotetraose produced by DagA enzyme reaction of agar.
피부는 신체의 가장 바깥에 존재하는 기관으로, 외부 환경으로부터 우리 몸을 보호하는 역할을 수행한다. 그러나 피부는 다른 기관들에 비해 외부 자극에 많이 노출되어 있어 쉽게 공격받을 수 있고, 특히 그 중에서도 얼굴 피부의 경우에는 자외선 및 공해 물질 등에 직접적으로 노출되어 있어 조기에 주름이 발생하기 쉬운 특징을 갖는다.Skin is the outermost organ of the body and plays a role in protecting our body from external environment. However, skin is exposed to external stimuli more easily than other organs, and can be easily attacked. Especially, face skin is directly exposed to ultraviolet rays and pollutants, and therefore, wrinkles are easily generated in the early stage.
일반적으로 피부 노화는 크게 두 가지, 외인성 노화와 내인성 노화로 구분되는데, 내인성 노화는 연대기적 노화라고도 일컬어지며 주로 시간의 흐름에 의해 발생한다. 이에 반해 외인성 노화는 여러 가지 외부 요인들에 의해 발생하는 현상으로 특히 자외선에 의한 광노화가 대표적으로 알려져 있다.In general, skin aging can be classified into two types, extrinsic aging and endogenous aging. Endogenous aging is also called chronological aging and is mainly caused by the passage of time. On the other hand, extrinsic aging is a phenomenon caused by various external factors, and photo-aging due to ultraviolet rays is typically known.
여러 피부노화 유발 요인들에 의해 나타나는 주름, 탄력 감소, 피부 쳐짐 및 건조 현상 등은 대부분 세포외기질(extracellular matrix) 단백질의 변화로 인해 발생하는 현상으로써, 주로 진피층에 존재하는 결합조직인 콜라겐, 엘라스틴, 히아루론산 등의 파괴가 원인으로 지목된다. 특히 이 중에서도 matrix metalloproteinases(MMPs)가 기질단백질을 분해하는 대표적인 효소로 알려져 있다. MMP는 단 한 번의 자외선 조사에 의해서도 그 활성이 현저히 증가되어 콜라겐을 비롯한 여러 기질단백질을 분해함으로써 주름 형성을 가속화한다.The wrinkles, elasticity reduction, skin burning and drying phenomena caused by various skin aging factors are mostly caused by changes of extracellular matrix proteins, and are mainly caused by collagen, elastin, It is pointed out as a cause of destruction of hyaluronic acid and the like. Especially, matrix metalloproteinases (MMPs) are known as the most important enzymes to degrade matrix proteins. The activity of MMP is greatly increased by a single ultraviolet irradiation, and it accelerates the formation of wrinkles by decomposing a plurality of substrate proteins including collagen.
세포의 노화는 기질의 노화를 일으키는 시작점으로써 노화된 세포가 주변에 미치는 부정적인 영향이 최종적으로 기질의 노화를 야기한다는 점이 널리 알려져 있다. 세포의 노화를 확인하는 가장 대표적인 바이오마커로는 노화된 세포에서만 발현되는 senescence-associated (SA)-β galactosidase를 염색하는 방법이 있다.Aging of cells is a starting point for aging of the substrate, and it is widely known that the negative effects of aging cells on the environment ultimately cause aging of the substrate. One of the most representative biomarkers for confirming senescence of cells is dyeing senescence-associated (SA) -β galactosidase, which is expressed only in aged cells.
그동안 주름개선을 위한 소재로는 주로 비타민C, α-토코페롤, 레티놀 및 그의 유도체 등이 화장품 및 의약품에 배합되어 이용되어 왔으나 이들은 제형에 배합되었을 경우 변취 현상이 발생하거나 화학적 안정성이 좋지 못하다는 단점이 있다.In the meantime, vitamin C, α-tocopherol, retinol and derivatives thereof have been used in cosmetics and medicines for improving wrinkles. However, when they are formulated in the formulations, they have a disadvantage that they are not removed or chemical stability is poor have.
이러한 단점을 극복하기 위해 최근 천연물 유래 물질들을 이용한 주름개선용 화장료 조성물의 개발이 각광받고 있으며 이의 발굴을 필요로 한다.In order to overcome these disadvantages, the development of a cosmetic composition for wrinkle improvement using natural materials derived from natural materials has been attracting attention and needs to be discovered.
한천(Agar)은 오래전부터 식품첨가물, 의약품, 가축사료 및 공업원료 등으로 널리 이용되고 있는 대표적인 해조류 유래 다당류로, 국내의 경우 해마다 그 생산량이 약 3,600톤에 이르고 있는 비교적 풍부한 수산자원의 하나이다. 그러나 실제 이용 면에서는 전체 생산량의 일부만이 단순가공 처리되어 값싼 원료로 사용될 뿐이며, 그 나머지는 대부분 방치되고 있어 부존자원량에 비해 부가가치가 매우 낮은 실정이다. 따라서 풍부한 국내 한천의 새로운 용도개발과 부가가치 향상에 관한 연구가 크게 요구되고 있는 실정이다.Agar is a representative seaweed-derived polysaccharide that has been widely used in food additives, pharmaceuticals, livestock feed and industrial materials for a long time. In Korea, agar is one of the relatively abundant fisheries resources that produce about 3,600 tons per year. However, in actual use, only a part of the total production is simply processed and used as cheap raw materials, and the rest of the production is largely neglected. Therefore, there is a great demand for research on the development of new applications of agar and the improvement of added value.
한천은 꼬시래기(Gracilariales)목과 우뭇가사리(Gelidiales)목의 홍조류의 세포벽에서 발견되는 다당류로서 아가로스(agarose)와 아가로펙틴(agaropectin)으로 구성된다. 아가로스는 3-O-linked β-D-galactopyranose와 4-O-linked 3,6-anhydro-α-L-galactopyranose 잔기가 교차되어 구성된 자연의 선형 다당류로 겔(gel)화력이 강한 반면, 아가로펙틴은 황산에스테르(sulfate esters), 피루브산염 아세탈(pyruvate acetal), 메틸 에테르(methyl ethers)로 치환된 유사한 골격으로 구성되어 겔화력이 약하다. 특히 아가로스는 β-1,4 결합에 작용하는 베타-아가레이스(β-agarase)에 의해서 네오아가로테트라오스(neoagarotetraose)를 거쳐 네오아가로비오스(neoagarobiose)로 분해되고 계속해서 α-1,3 결합에 작용하는 알파-아가레이즈(α-agarase)에 의해서 갈락토스(D-galactose)와 3,6-anhydro-L-galactose로 최종 분해된다.Agar consists of agarose and agaropectin, polysaccharides found in the cell walls of red algae in the necks of Gracilariales and Gelidiales. Agarose is a natural linear polysaccharide composed of 3-O-linked β-D-galactopyranose and 4-O-linked 3,6-anhydro-α-L-galactopyranose residues crossed with each other, Lopectin is composed of similar skeletons substituted with sulfate esters, pyruvate acetal, and methyl ethers and has weak gelation power. Particularly, agarose is decomposed into neoagarobiose via neoagarotetraose by β-agarase acting on β-1,4 bond, and then α-1, 3-anhydro-L-galactose by galactose (D-galactose) and 3,6-anhydro-L-galactose by the alpha-agarase acting on the enzyme.
한편, 방선균인 스트렙토마이세스 시리칼라(Streptomyces coelicolor) A3는 한천을 분해하는 세포외 아가레이스를 생산한다고 알려져 있으며, 이 아가레이스는 dagA 유전자에 의해서 코드되어 있다. 방선균에서는 dagA 유전자가 그 기능이 유일하게 알려진 베타-아가레이스 유전자로서 방선균에서의 아가레이스 생산 연구에 있어 중요한 위치를 갖는다. 특히 스트렙토마이세스 시리칼라는 방선균의 분자생물학적 연구에 가장 널리 사용되는 균주로서 2002년 영국의 Sanger centre에서 염색체 DNA 서열이 분석되어 공개되어 있다(Bantley et al., 2002, Nature).Streptomyces coelicolor A3, actinomycetes, is known to produce agarase-degrading extracellular agarase, which is encoded by the dagA gene. In actinomycetes, the dagA gene has an important role in the study of agarase production in actinomycetes as a beta-agarase gene whose function is the only known. Streptomyces sericola is the most widely used strain for the molecular biology of actinomycetes. In 2002, the chromosomal DNA sequence was analyzed at the Sanger center in England (Bantley et al., 2002, Nature).
공개특허 10-2013-0030955에서 발명자는 효소적인 방법을 이용하여 내-형(endo-type) 베타-아가레이스인 스트렙토마이세스 시리칼라 dagA 유전자의 발현시스템을 구축하고, dagA가 암호화하는 단백질인 DagA 단백질이 아가 또는 아가로스와 반응하여 네오아가로테트라오스 및 네오아가로헥사오스를 생산함을 확인하였다.In the patent document 10-2013-0030955, the inventors constructed an expression system of the dagA gene of the endo-type beta-agarase, Streptomyces sericola, using an enzymatic method, and found that the expression of DagA It was confirmed that the protein reacts with agar or agarose to produce neoagarotetraose and neoagarohexaose.
이에, 본 발명자들은 상기 종래기술을 통해 발명한 네오아가로테트라오스의 신규 효능 발굴을 위해 예의 연구노력한 결과, 한천을 DagA 효소 반응시켜 조제한 한천 유래 네오아가로테트라오스를 유효성분으로 함유하는 화장료 조성물의 경우, 세포증식 촉진 및 세포노화 억제를 통해 내·외부 요인에 의한 피부노화를 효과적으로 개선할 수 있음을 확인하고, 본 발명을 완성하게 되었다.The inventors of the present invention have made intensive researches for discovering the novel efficacy of neoagarotetraose invented through the above-mentioned prior art. As a result, they have found that cosmetic composition containing agar-derived neoagarotetraose prepared by agar agar- The present inventors have confirmed that skin aging caused by internal and external factors can be effectively improved through promotion of cell proliferation and inhibition of cell senescence, thus completing the present invention.
따라서, 본 발명의 주된 목적은 피부의 세포증식 또는 세포 재생을 촉진시킴으로써 피부주름을 개선시켜 피부 노화를 개선하는 효과를 갖는 네오아가로테트라오스를 함유하는 피부노화 개선용 화장료 조성물을 제공하는 데 있다.Accordingly, it is a main object of the present invention to provide a cosmetic composition for improving skin aging containing neoagarotetraose, which has an effect of improving skin wrinkles and promoting skin aging by promoting cell proliferation or cell regeneration of skin .
본 발명의 한 양태에 따르면, 본 발명은 한천 유래의 네오아가로테트라오스(neoagarotetraose)를 함유하는 피부노화 개선용 화장료 조성물을 제공한다.According to one aspect of the present invention, there is provided a cosmetic composition for improving skin aging comprising agar-derived neoagarotetraose.
본 발명에 있어서, 상기 네오아가로테트라오스는 한천을 DagA 효소 반응시켜 제조한 것을 특징으로 한다.In the present invention, the neoagarotetraose is characterized in that agar is produced by DagA enzyme reaction.
상기 용어‘네오아가로테트라오스’는 아가로스(agarose)가 베타-아가라아제(β-agarase)의해 분해되어 생성되는 물질로서, 본원발명에서는 한국등록특허 제1302655호의 제조방법으로 제조되었다. 네오아가로테트라오스는 종래에 항비만, 항당뇨, 면역기능 증강 및 항암 작용 등과 같은 효과가 알려져 있으나, 본원발명과 같이 네오아가로테트라오스의 피부노화 개선 효과에 대해서는 알려진바 없다. The term " neoagarotetraose " is a substance produced by degradation of agarose by beta-agarase. In the present invention, the neoagarotetraose was prepared by the method of Korean Patent No. 1302655. Neoagarotetraose has been conventionally known to have effects such as anti-obesity, anti-diabetic, immune function enhancement, and anti-cancer effect. However, the effect of neoagarotetraose on improving skin aging is not known.
본 발명에 있어서, 상기 네오아가로테트라오스는 화장료 조성물 총 중량 대비 0.01 내지 50 중량% 함유하는 것을 특징으로 한다. 상기 범위를 벗어날 경우 제형화가 용이하지 않기 때문에 바람직하지 않다.In the present invention, the neoagarotetraose is contained in an amount of 0.01 to 50% by weight based on the total weight of the cosmetic composition. Outside of the above range, it is not preferable since the formulation is not easy.
본 발명에 있어서, 상기 화장료 조성물은 피부 주름을 개선시키는 것을 특징으로 한다. In the present invention, the cosmetic composition is characterized by improving skin wrinkles.
또한, 상기 화장료 조성물은 피부의 세포증식 또는 세포재생을 촉진시킴으로써 피부주름개선 효과를 나타내는 것을 특징으로 한다. Further, the cosmetic composition is characterized by exhibiting skin wrinkle-improving effect by promoting cell proliferation or cell regeneration of the skin.
구체적으로, 상기 피부노화 또는 주름 생성은 외인성 요인 및 내인성 요인에 의해 유발되는 것으로, 좀 더 구체적으로는 상기 피부노화는 내ㆍ외부 요소의 변화에 의한 자극이 피부조직을 구성하고 있는 성분을 변화시켜 주름을 유발하는 것을 말한다. 여기에는 자외선, 공해, 담배연기, 화학물질 등에 의한 외부 자극에 의해 유도되는 조기노화 증상뿐만 아니라, 나이가 들어감에 의해 피부세포의 증식이 감소함에 따라 발생하는 자연노화 현상을 포함한다.Specifically, the skin aging or wrinkle formation is caused by an extrinsic factor and an intrinsic factor. More specifically, the aging of the skin changes a component constituting the skin tissue by stimulation caused by a change in inner and outer factors It refers to causing wrinkles. These include not only premature aging symptoms induced by external stimulation by ultraviolet rays, pollution, tobacco smoke, chemicals, etc., but also natural aging phenomenon caused by decreasing proliferation of skin cells due to aging.
본 발명의 실험예에 따르면, 피부의 진피를 구성하는 섬유아세포에 네오아가로테트라오스를 처리한 경우에 세포 증식이 촉진되는 것을 확인하였으며, 임의로 긁어 상처를 낸 섬유아세포에 네오아가로테트라오스를 처리한 경우에서는 세포재생이 가속화됨을 확인할 수 있었다. 또한, 과도한 세포분열로 인해 상적인 세포분열 과정이 망가진 노화 섬유아세포에 네오아가로테트로오스를 처리한 경우에서는 SA(senescence-associated)-β galactosidase를 발현하는 세포의 수가 현저히 감소함을 확인하였다. 이러한 결과로 볼 때, 본원발명의 네오아가로테트라오스를 함유하는 화장료 조성물의 경우, 세포증식 또는 세포재생을 촉진시킴으로써 피부 주름 또는 피부노화를 개선할 수 있음을 알 수 있다(실험예 1-3 참조).According to the experimental example of the present invention, it was confirmed that cell proliferation was promoted when neuroagarotetraose was treated with fibroblasts constituting the dermis of the skin, and neoagarotetraose was randomly scratched and wounded It was confirmed that cell regeneration was accelerated in the case of treatment. In addition, it was confirmed that the number of cells expressing senescence-associated -β-galactosidase was significantly reduced when neoagarotetrose was treated with aging fibroblasts, which had undergone a cell division process due to excessive cell division . From these results, it can be seen that the cosmetic composition containing neoagarotetraose of the present invention can improve skin wrinkles or skin aging by promoting cell proliferation or cell regeneration (Experimental Examples 1-3 Reference).
본 발명에 있어서, 상기 조성물은 당업계에서 통장적으로 제조되는 어떠한 제형으로도 제조될 수 있으며, 바람직하게는 스킨로션, 스킨소프너, 스킨토너, 아스트린젠트, 로션, 밀크로션, 모이스쳐 로션, 영양로션, 맛사지크림, 영양크림, 모이스처크림, 핸드크림, 파운데이션, 에센스, 영양에센스, 팩, 비누, 클렌징폼, 클렌징로션, 클렌징크림, 바디로션 또는 바디클린저로 구성된 제형에서 선택된 하나 이상의 제형인 것을 특징으로 한다.According to the present invention, the composition may be prepared in any form commonly used in the art and is preferably a skin lotion, a skin softener, a skin toner, an astringent, a lotion, a milk lotion, a moisturizing lotion, Wherein the at least one formulation is at least one selected from the group consisting of a massager cream, a nutritional cream, a moisturizing cream, a hand cream, a foundation, an essence, a nutritional essence, a pack, a soap, a cleansing foam, a cleansing lotion, a cleansing cream, a body lotion or a body cleanser .
이상 설명한 바와 같이, 본 발명의 네오아가로테트라오스를 유효성분으로 함유하는 화장료 조성물은 콜라겐을 생성하는 섬유아세포의 증식은 촉진시키고, 노화된 세포의 성장을 회복시킴으로써 피부노화 또는 주름을 개선하는 데 우수한 효과를 나타낸다.As described above, the cosmetic composition containing neoagarotetraose as an active ingredient of the present invention promotes the proliferation of collagen-producing fibroblasts and restores the growth of aged cells, thereby improving skin aging or wrinkles And exhibits excellent effects.
도 1은 네오아가로테트라오스의 세포증식 촉진능을 확인한 실험예 1에 대한 결과이다.
도 2는 손상을 유도한 섬유아세포에서 네오아가로테트라오스의 세포재생 촉진능을 확인한 실험예 2에 대한 결과이다.
도 3은 노화된 섬유아세포에서 네오아가로테트라오스의 SA-β galactosidase 발현 억제 효능을 확인한 실험예 3에 대한 결과이다.FIG. 1 shows the results of Experimental Example 1 in which neoagarotetraose promotes cell proliferation.
FIG. 2 shows the results of Experimental Example 2 in which the cell regeneration promoting ability of neoagarotetraose was confirmed in the fibroblasts in which damage was induced.
FIG. 3 shows the results of Experimental Example 3, which confirmed the inhibitory effect of neoagarotetraose on the expression of SA-β galactosidase in senescent fibroblasts.
이하, 실시예를 통하여 본 발명을 더욱 상세히 설명하기로 한다. 이들 실시예는 단지 본 발명을 예시하기 위한 것이므로, 본 발명의 범위가 이들 실시예에 의해 제한되는 것으로 해석되지는 않는다.Hereinafter, the present invention will be described in more detail with reference to Examples. These embodiments are only for illustrating the present invention, and thus the scope of the present invention is not construed as being limited by these embodiments.
실시예 1: 네오아가로테트로오스의 제조Example 1: Preparation of neoagarotetroose
본 발명에서 사용된 네오아가로테트로오스는 종래의 공개특허 10-2013-0030955에 기술된 방법을 통해 제조된 것을 사용하였다.The neoagarotetrons used in the present invention were those prepared through the method described in the prior art Patent No. 10-2013-0030955.
구체적으로, 스트렙토마이세스 시리칼라 A3(2)(Streptomyces coelicolor A3(2)) 유래 DagA 단백질을 아가로오스와 반응시켜 얻은 네오아가로테트로오스를 PBS(phosphate buffer saline)에 1 mg/ml 및 10 mg/ml의 농도로 용해시켜 이하의 실험예에 사용하였다.Specifically, neoagarotetrose obtained by reacting DagA protein derived from Streptomyces coelicolor A3 (2) with agarose was dissolved in PBS (phosphate buffer saline) at 1 mg / ml and 10 mg / ml and used in the following experimental examples.
실험예 1: 네오아가로테트로오스의 세포증식 촉진능 평가Experimental Example 1: Evaluation of cell proliferation promoting ability of neoagarotetroose
실시예 1의 네오아가로테트로오스의 세포증식 촉진 효과를 평가하기 위해 사람의 섬유아세포 세포주(Human dermal fibroblast, Hs68)에서의 세포수 변화를 측정하였다.Cellular changes in human dermal fibroblast (Hs68) were measured to evaluate cell proliferation promoting effect of neoagarotetroose of Example 1.
먼저 Hs68 섬유아세포를 96 well plate에 2x104의 수만큼 분주한 후, 37℃, 5% CO2 조건의 배양기에서 24시간 동안 배양하였다. 이후 네오아가로테트로오스를 포함하지 않은 배지와 1㎍/㎖ 또는 10㎍/㎖ 포함하는 배지로 교체해 준 다음 24시간 동안 추가로 배양하였다. 24시간 후 각 well의 배지를 제거하고 Dulbecco's phosphate buffered saline(DPBS)을 이용한 washing 과정을 1회 거친 다음 각 well당 10㎕의 CCK-8(cell counting kit-8, Dojindo, 일본) 시약을 포함하는 배지로 교체하여 37℃에서 4시간 동안 반응시켰다. 이후 UV-Vis spectrophotometer를 이용하여 450nm에서의 흡광도를 측정하였다. 세포의 생존율은 네오아가로테트로오스를 포함하지 않은 배지를 처리한 경우를 대조군으로 하여 이의 평균 흡광도 값에 대한 백분율로 나타내었다.First, Hs68 fibroblasts were divided into 2 × 10 4 cells in a 96-well plate, and cultured in an incubator at 37 ° C. and 5% CO 2 for 24 hours. Thereafter, the medium was replaced with a medium containing no neoagarotetroose and a medium containing 1 μg / ml or 10 μg / ml, followed by further incubation for 24 hours. After 24 hours, the culture medium of each well was removed, and the plate was washed once with Dulbecco's phosphate buffered saline (DPBS). Then, 10 μl of CCK-8 (cell counting kit-8, Dojindo, Japan) And the cells were reacted at 37 캜 for 4 hours. The absorbance at 450 nm was then measured using a UV-Vis spectrophotometer. The survival rate of the cells was expressed as a percentage of the average absorbance value as a control when the medium containing no neoagarotetrose was treated.
그 결과, 도면 1에 나타낸 바와 같이 네오아가로테트로오스(NAT)를 처리한 경우에서 세포증식이 촉진됨을 확인하였다.As a result, it was confirmed that cell proliferation was promoted when neoagarotetrone (NAT) was treated as shown in Fig.
실험예 2: 네오아가로테트로오스의 세포재생 촉진능 평가Experimental Example 2: Evaluation of cell regeneration promoting ability of neoagarotetroose
실시예 1의 네오아가로테트로오스의 세포재생 촉진 효과를 평가하기 위해 실험예 1에서와 동일 세포를 사용하여 실험을 진행하였다.Experiments were carried out using the same cells as in Experimental Example 1 to evaluate the cell regeneration promoting effect of neoagarotetroose of Example 1.
Hs68 섬유아세포를 5x105 cells/well의 농도로 6 well plate에 분주한 후, 37℃, 5% CO2 조건의 배양기에서 24시간 동안 배양하여 100% confluent한 상태를 만들어 주었다. 이후 pipette tip을 이용하여 well의 가운데를 가로지르도록 임의로 긁어 상처를 내고, 네오아가로테트로오스를 포함하지 않은 배지와 1㎍/㎖ 또는 10㎍/㎖ 포함하는 배지로 교체해 준 다음 24시간 동안 추가로 배양하였다. 24시간 후 네오아가로테트로오스를 포함하지 않은 경우와 포함한 경우에서 세포에 임의로 낸 상처가 어느 정도 회복되었는지를 현미경으로 촬영하였다.Hs68 fibroblasts were plated at a density of 5 × 10 5 cells / well in a 6-well plate and cultured in an incubator at 37 ° C and 5% CO 2 for 24 hours to make 100% confluent. Thereafter, the wound was scratched randomly to cross the center of the well using a pipette tip, and the medium was replaced with a medium containing no neoagarotetroose and a medium containing 1 μg / ml or 10 μg / ml. After 24 hours And further cultured. After 24 hours, the extent of injury to the cells was examined microscopically for the case of neoagarotetrose not containing or the case of containing.
그 결과, 도면 2에 나타낸 바와 같이 네오아가로테트로오스를 처리하지 않은 경우에 비해 처리한 경우에서 세포재생이 더욱 촉진됨을 확인하였다.As a result, as shown in Fig. 2, it was confirmed that cell regeneration was further promoted in the case of treatment as compared with the case in which neoagarotetrose was not treated.
실험예 3: 네오아가로테트로오스의 세포노화 억제능 평가Experimental Example 3: Evaluation of inhibition of cell senescence of neoagarotetroose
실시예 1의 네오아가로테트로오스의 세포노화 억제 효과를 평가하기 위해 실험예 1에서와 동일 세포를 사용하여 실험을 진행하였다. 단지 시간의 흐름에 따른 노화세포 모델을 구축하기 위해 계대수가 낮은(PD 10 이하) 세포와 계대수가 높은(PD 25 이상) 세포를 각각 어린(young) 모델과 노화된(old) 모델로 정의하고 실험을 진행하였다.Experiments were carried out using the same cells as in Experimental Example 1 to evaluate the cell aging inhibitory effect of neoagarotetrose of Example 1. In order to construct a model of aging cells over time, we define cells with low (
먼저 낮은 계대수의 어린 세포모델과 높은 계대수의 노화된 세포 모델을 각각 2x105 cells/well의 농도로 6 well plate에 분주한 후 37℃, 5% CO2 조건의 배양기에서 24시간 동안 배양하였다. 이후 네오아가로테트로오스를 포함하지 않은 배지와 1㎍/㎖ 또는 10㎍/㎖ 포함하는 배지로 교체해 준 다음 72시간 동안 추가로 배양하였다. 72시간 이후 배양 배지를 제거하고 DPBS washing 과정을 3회 거친 다음 Cellular senescence assay kit(CBA-230, Cell biolabs, 미국)를 이용하여 노화에 의해 성장이 멈춘 세포에서 특이적으로 발현되는 SA-β galactosidase 효소에 대한 염색을 진행하였다.First, low-passaged young cell models and high-passaged aged cell models were plated in 6-well plates at a concentration of 2 × 10 5 cells / well and cultured in an incubator at 37 ° C. and 5% CO 2 for 24 hours . Thereafter, the medium was replaced with a medium containing no neoagarotetroose and a medium containing 1 μg / ml or 10 μg / ml, followed by further incubation for 72 hours. After 72 hours, the culture medium was removed and DPBS washing procedure was performed three times. Cell-senescence assay kit (CBA-230, Cell biolabs, USA) was used to elucidate the specific expression of SA-β galactosidase And staining for the enzyme proceeded.
그 결과, 도면 3에 나타낸 바와 같이 노화된 세포(old)에서 노화되지 않은 세포(young)에 비해 SA-β galactosidase를 발현하는 세포의 수가 월등히 많은 것과 비교하여, 노화된 세포에 네오아가로테트로오스를 처리한 경우에서는 SA-β galactosidase 발현 세포의 수가 현저히 감소함을 확인하였다.As a result, as shown in FIG. 3, compared with the case where the number of cells expressing SA-β galactosidase is much higher than that of undigested cells (aged), the aged cells have neoagarotetrons The number of SA-β galactosidase-expressing cells was significantly reduced in the case of treatment with osse.
제형예 1: 토너제의 제조Formulation Example 1: Preparation of Toner
상기 실시예 1에서 얻은 성분을 함유하는 토너제를 하기의 표 1에 나타낸 조성성분 및 조성비에 따라 통상적인 방법으로 제조하였다.Toners containing the components obtained in Example 1 were prepared in a conventional manner according to the compositional components and composition ratios shown in Table 1 below.
제형예 2: 로션제의 제조Formulation Example 2: Preparation of lotion agent
상기 실시예 1에서 얻은 성분을 함유하는 로션제를 하기의 표 2에 나타낸 조성성분 및 조성비에 따라 통상적인 방법으로 제조하였다.A lotion agent containing the components obtained in Example 1 was prepared by a conventional method according to the compositional components and composition ratios shown in Table 2 below.
제형예Formulation Example 3: 크림제의 제조 3: Manufacture of cream agent
상기 실시예 1에서 얻은 성분을 함유하는 크림제를 하기의 표 3에 나타난 조성성분 및 조성비에 따라 통상적인 방법으로 제조하였다.A creaming agent containing the ingredients obtained in Example 1 was prepared in a conventional manner according to the composition components and composition ratios shown in Table 3 below.
Claims (6)
A cosmetic composition for improving skin aging comprising agar-derived neoagarotetraose.
[Claim 3] The cosmetic composition for improving skin aging according to claim 1, wherein the neoagarotetraose is produced by DagA enzyme reaction of agar.
The cosmetic composition for improving skin aging according to claim 1, wherein the neoagarotetraose is contained in an amount of 0.01 to 50% by weight based on the total weight of the cosmetic composition.
The cosmetic composition for improving skin aging according to claim 1, wherein the cosmetic composition improves skin wrinkles.
The cosmetic composition for improving skin aging according to claim 1, wherein the cosmetic composition promotes skin cell proliferation or cell regeneration, thereby exhibiting skin wrinkle-improving effect.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN109731001A (en) * | 2019-02-13 | 2019-05-10 | 蓝脑科技(厦门)有限公司 | The newly new application of fine jade oligosaccharides and the composition containing new fine jade oligosaccharides |
| CN110960439A (en) * | 2019-12-31 | 2020-04-07 | 蓝脑科技(厦门)有限公司 | Agar oligosaccharide-oligopeptide composition, preparation method and composition for repairing skin inflammation |
| CN115386547A (en) * | 2022-10-09 | 2022-11-25 | 谷文忠 | Application of oligosaccharide in promoting proliferation of hUC-MSCs and preparing corresponding culture medium |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109731001A (en) * | 2019-02-13 | 2019-05-10 | 蓝脑科技(厦门)有限公司 | The newly new application of fine jade oligosaccharides and the composition containing new fine jade oligosaccharides |
| CN110960439A (en) * | 2019-12-31 | 2020-04-07 | 蓝脑科技(厦门)有限公司 | Agar oligosaccharide-oligopeptide composition, preparation method and composition for repairing skin inflammation |
| CN110960439B (en) * | 2019-12-31 | 2022-05-10 | 蓝脑科技(厦门)有限公司 | Agar oligosaccharide-oligopeptide composition, preparation method and composition for repairing skin inflammation |
| CN115386547A (en) * | 2022-10-09 | 2022-11-25 | 谷文忠 | Application of oligosaccharide in promoting proliferation of hUC-MSCs and preparing corresponding culture medium |
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