KR20210055475A - A composition for improving skin beauty containing glycosaminoglycan - Google Patents
A composition for improving skin beauty containing glycosaminoglycan Download PDFInfo
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- KR20210055475A KR20210055475A KR1020190141970A KR20190141970A KR20210055475A KR 20210055475 A KR20210055475 A KR 20210055475A KR 1020190141970 A KR1020190141970 A KR 1020190141970A KR 20190141970 A KR20190141970 A KR 20190141970A KR 20210055475 A KR20210055475 A KR 20210055475A
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- glycosaminoglycan
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Abstract
Description
본 발명은 글라이코스아미노글리칸을 유효성분으로 함유하는 피부미용증진용 조성물에 관한 것으로서, 더욱 상세하게는 뮤코다당류인 글라이코스아미노글리칸을 함유함으로써, 엘라스틴 분해 및 콜라겐 분해를 억제하며 히알루론산 농도를 크게 증가시켜서 피부의 탄력과 보습의 개선 및 피부주름개선에 도움을 주는 피부미용증진용 조성물에 관한 것이다.The present invention relates to a composition for promoting skin beauty containing glycosaminoglycan as an active ingredient, and more particularly, by containing a mucopolysaccharide glycosaminoglycan, inhibiting elastin degradation and collagen degradation, and hyaluronic acid concentration It relates to a composition for promoting skin beauty that greatly increases the skin elasticity and moisture, and helps to improve skin wrinkles.
피부는 몸의 근육과 기관을 덮어 보호하는 상피조직으로 표피, 진피, 피하지방으로 구성되어 있으며 외부환경으로부터 신체를 보호하고 체온을 조절하며 감각을 느낄 수 있게 하는데 주로 외부의 물리적 손상 및 화학물질로부터 인체를 보호하고, 세균, 곰팡이, 바이러스 등이 피부로 침범하는 것을 방지하며, 수분 손실을 막는 보호 장벽 역할을 한다. 그 중 표피는 3개 층 중 가장 얇은 층으로 피부의 보습 및 보호를 담당하는 중요한 기능을 하며 특히 표피의 각질층에 존재하는 수분에 의하여 피부가 탄력 있고 부드럽게 된다.The skin is an epithelial tissue that covers and protects the muscles and organs of the body. It is composed of the epidermis, the dermis, and the subcutaneous fat. It protects the body from the external environment, regulates the body temperature, and allows the senses to be felt, mainly from external physical damage and chemical substances. It protects the human body, prevents bacteria, fungi, and viruses from invading the skin, and acts as a protective barrier against moisture loss. Among them, the epidermis is the thinnest of the three layers and plays an important role in moisturizing and protecting the skin. In particular, the moisture present in the stratum corneum of the epidermis makes the skin elastic and soft.
또한, 최근에는 소비자의 고기능 화장품에 대한 욕구를 충족시키기 위한 미백, 피부주름 개선 및 억제, 그리고 자외선 차단 기능을 갖는 화장품들이 계속해서 개발되고 있는데, 그 중에서도 특히 주름 개선 및 억제 제품은 피부를 젊게 유지하려는 소비자의 많은 관심을 받는 대상이다.In addition, recently, cosmetics with whitening, skin wrinkle improvement and suppression, and UV protection functions have been continuously developed to satisfy consumers' desire for high-performance cosmetics, and among them, wrinkle improvement and suppression products in particular keep the skin young. It is an object that gets a lot of attention from consumers who want to do it.
나이가 들면서 콜라겐이 부족하거나 질이 저하되어 피부는 탄력을 잃고 주름이 생기게 되는데, 피부의 주름은 피부의 수분 함유량과 콜라겐의 함유량 그리고 면역 기능 등의 여러 요인에 영향을 받으며, 피부의 탄력은 이 콜라겐과 엘라스틴(elastin, 탄력섬유)에 의해서 유지되는 것으로 알려져 있다.With age, collagen deficiency or quality deteriorates, and the skin loses its elasticity and causes wrinkles.The wrinkles of the skin are affected by various factors such as the skin's moisture content, collagen content, and immune function, and the skin's elasticity is affected by this. It is known to be maintained by collagen and elastin (elastin).
생체 내에서 콜라겐과 같은 세포외기질의 합성과 분해는 적절하게 조절되나 노화가 진행되면서 그 합성이 감소하며 콜라겐을 분해하는 효소인 기질 금속 단백질 분해효소(MMP, matrix metalloproteinase)의 발현이 촉진되어 피부의 탄력이 저하되고 주름이 형성된다.Synthesis and decomposition of extracellular matrix such as collagen in vivo is properly regulated, but its synthesis decreases as aging progresses, and the expression of matrix metalloproteinase (MMP), an enzyme that degrades collagen, is promoted in the skin. Elasticity decreases and wrinkles are formed.
한편, 글라이코스아미노글리칸(Glycosaminoglycan, GAG)는 우론산과 글루코사민이 반복되고 부분적으로 설페이트화된 구조를 가지며, 이러한 복잡한 화학구조에 의해 여러 종류의 생물활성을 나타낼 수 있게 된다. 현재까지 알려진 바에 의하면, 이러한 다양한 생물활성은 고유의 특정 서열이 존재함에 기인한 것으로 보인다. GAG류는 생체 내에서 프로테오글리칸(Proteoglycan)의 형태로 존재하며 이러한 프로테오글리칸은 프로테아제와 글리코시다제에 의해 분해되어 당부분이 유리된다. GAG류의 생리적 역할에 대해서 아직까지 많이 밝혀져 있지 않으나, 생체내의 단백질들과 결합하여 생리활성을 나타내는 것으로 보인다.On the other hand, Glycosaminoglycan (GAG) has a structure in which uronic acid and glucosamine are repeated and partially sulfated, and by such a complex chemical structure, various kinds of biological activities can be exhibited. As far as is known, these various biological activities seem to be due to the existence of unique specific sequences. GAGs exist in the form of proteoglycans in vivo, and these proteoglycans are decomposed by proteases and glycosidases to release the sugar part. The physiological role of GAGs has not been elucidated so far, but it appears to exhibit physiological activity by binding to proteins in vivo.
이러한 GAG를 곤충으로부터 제조하는 방법은 한국등록특허 제10-1200402호에 제안되어 있다.A method of manufacturing such GAG from insects is proposed in Korean Patent Registration No. 10-1200402.
종래기술에 의하면, GAG는 서양뒤영벌에서 추출할 수 있는데, 서양뒤영벌(Bombus terrestris)은 벌목 꿀벌과의 곤충으로 유럽이 원산지이며 한국에는 자원곤충(화분매개용)으로 도입되었다. 여왕벌의 몸길이는 20～22㎜이고, 수벌의 몸길이는 14～16㎜이며 일벌의 몸길이는 11～17㎜이다. According to the prior art, GAG can be extracted from the western rear bee, and the western rear bee (Bombus terrestris ) is an insect with the felling bee family, originated in Europe, and was introduced as a resource insect (for pollinators) in Korea. The body length of the queen bee is 20~22㎜, the body length of the male bee is 14~16㎜, and the body length of the worker bee is 11~17㎜.
또한, 곤충으로부터 GAG의 제조방법과 소염, 항당뇨, 항응혈, 항암, 혈관신생억제 용도, 및/또는 혈관확장반응의 기능장애로 인한 질환 치료 및/또는 예방의 의약품 용도가 한국특허공개 제10-2011-0013074호(신규한 곤충글라이코자미노글라이칸의 제조방법)에 개시되어 있으며, 그 이후에 간 보호 식품 조성물(한국특허등록 제10-1869792호) 및 소염 및 관절염 등의 염증개선용 건강기능식품으로의 적용(한국특허등록 제10-1680644호) 등이 개시되었다.In addition, the manufacturing method of GAG from insects, anti-inflammatory, anti-diabetic, anti-coagulant, anti-cancer, anti-angiogenic use, and/or the use of pharmaceuticals for the treatment and/or prevention of diseases caused by dysfunction of the vasodilation reaction are disclosed in Korean Patent Publication No. 10. -2011-0013074 (new method for manufacturing insect glycosaminoglycan), after that, liver protection food composition (Korean Patent Registration No. 10-1869792) and for improving inflammation such as anti-inflammatory and arthritis Application to health functional food (Korean Patent Registration No. 10-1680644) and the like have been disclosed.
그러나 이러한 기존의 GAG에 대한 활용 기술은 아직까지 확실한 치료 효과를 나타내는 제품으로 상업화되지 못하고 있어서 실용화를 위한 연구가 지속되고 있으며, 이에 대한 실제 효능효과를 발휘할 수 있도록 활용성을 충분하게 발휘하도록 제안하지 못하고 있는 문제가 있다.However, these existing technologies for using GAG have not yet been commercialized as products exhibiting a definite therapeutic effect, so research for practical use is continuing, and it is not proposed to demonstrate sufficient utility so that actual efficacy effects can be exhibited. There is a problem that I am not able to do.
특히, 종래기술에서는 GAG를 활용함에 있어서 그 자체로서 유효성분으로서의 기능을 활용하는 기술이 부족하여 이를 다른 담체나 유효성분에 혼합하여 다양한 용도로 사용하고자 하는 시도가 있는 정도에 불과하였다.Particularly, in the prior art, there is a lack of technology that utilizes the function as an active ingredient by itself in utilizing GAG, so there has been only an attempt to use it for various purposes by mixing it with other carriers or active ingredients.
그러므로 종래기술에서는 GAG의 유효성분으로서의 활용성을 제대로 밝혀내지 못한 문제가 있었다.Therefore, in the prior art, there was a problem that the utility of GAG as an active ingredient was not properly revealed.
이와 같은 종래기술의 문제점과 활용성 개선의 문제를 해결하기 위하여, 본 발명은 GAG 자체를 유효성분으로 하는 실효성 있는 새로운 용도의 개발을 해결과제로 한다.In order to solve the problems of the prior art and the problem of improving usability, the present invention makes the development of a new effective use using GAG itself as an active ingredient as a solution.
본 발명자들은 이러한 문제 해결을 위해, GAG의 새로운 용도를 개발하기 위한 연구를 지속적으로 수행하여 GAG가 엘라스틴 분해 및 콜라겐 분해를 억제하며 히알루론산 농도를 증가시키어 피부의 탄력과 보습 및 주름의 개선에 도움을 줌으로써, 피부미용 개선 효과가 있음을 알아내고 본 발명을 완성하였다.In order to solve this problem, the present inventors have continuously conducted research to develop new uses of GAG, so that GAG inhibits elastin degradation and collagen degradation, and increases hyaluronic acid concentration, helping to improve skin elasticity, moisturization, and wrinkles. By giving, it was found that there is an effect of improving skin beauty, and the present invention was completed.
따라서 본 발명의 목적은 GAG의 새로운 용도로서의 피부미용증진용 조성물을 제공하는 것이다.Accordingly, it is an object of the present invention to provide a composition for promoting skin beauty as a new use of GAG.
또한, 본 발명의 다른 목적은 GAG를 함유하는 피부미용증진용 화장료 조성물을 제공하는데 있다.In addition, another object of the present invention is to provide a cosmetic composition for enhancing skin beauty containing GAG.
또한, 본 발명의 또 다른 목적은 GAG를 함유하는 피부미용증진용 건강식품을 제공하는데 있다.In addition, another object of the present invention is to provide a health food for promoting skin beauty containing GAG.
상기와 같은 본 발명의 과제해결을 위하여, 본 발명은 글라이코스아미노글리칸(Glycosaminoglycan, GAG)을 유효성분으로 함유하는 피부미용증진용 조성물을 제공한다.In order to solve the problems of the present invention as described above, the present invention provides a composition for promoting skin beauty containing Glycosaminoglycan (GAG) as an active ingredient.
또한, 본 발명은 글라이코스아미노글리칸(Glycosaminoglycan, GAG)을 유효성분으로 함유하는 피부주름개선용 화장료 조성물을 제공한다.In addition, the present invention provides a cosmetic composition for improving skin wrinkles containing Glycosaminoglycan (GAG) as an active ingredient.
또한, 본 발명은 글라이코스아미노글리칸(Glycosaminoglycan, GAG)을 유효성분으로 함유하는 보습용 화장료 조성물을 제공한다.In addition, the present invention provides a moisturizing cosmetic composition containing Glycosaminoglycan (GAG) as an active ingredient.
또한, 본 발명은 글라이코스아미노글리칸(Glycosaminoglycan, GAG)을 유효성분으로 함유하는 피부미용증진용 건강식품을 제공한다.In addition, the present invention provides a health food for promoting skin beauty containing Glycosaminoglycan (GAG) as an active ingredient.
본 발명의 바람직한 구현예에 따르면, 상기 GAG가 전체 피부미용증진용 조성물 중에 유효성분으로서 0.01-5중량%로 함유하는 것이 바람직하다.According to a preferred embodiment of the present invention, it is preferable that the GAG is contained in an amount of 0.01-5% by weight as an active ingredient in the total skin beauty promotion composition.
본 발명에 의한 GAG을 함유한 피부미용개선용 조성물은 피부에 안전하고, 엘라스틴 분해 및 콜라겐 분해를 억제하며 히알루론산 농도를 증가시켜서 피부의 탄력과 보습에 효과가 있으므로 피부 보습력 유지와 주름개선 등 피부미용개선에 우수한 효과를 발휘한다.The composition for improving skin beauty containing GAG according to the present invention is safe for the skin, inhibits the degradation of elastin and collagen, and increases the concentration of hyaluronic acid to increase the elasticity and moisture of the skin. It exhibits excellent effects in improving beauty.
또한, 이러한 본 발명에 따른 GAG는 기존과는 달리 그 자체로서 피부미용개선에 유효한 활성을 나타내는 것이므로 다른 유효활성 성분을 혼합하여 사용하지 않더라도 그 자체로서 피부미용개선을 위한 조성물로 화장품이나 건강식품으로서 활용이 가능한 효과가 있다.In addition, since the GAG according to the present invention itself exhibits effective activity for improving skin beauty, unlike the existing one, it is itself a composition for improving skin beauty even if other active ingredients are not mixed and used as a cosmetic or health food. There is an effect that can be used.
도 1은 본 발명의 실시예에서 사용한 서양뒤영벌여왕벌에서 유래된 GAG에 대한 Elastase 억제능을 보여주는 그래프이다.
도 2는 본 발명의 실시예에서 사용한 서양뒤영벌여왕벌에서 유래된 GAG의 Collagenase 억제능을 보여주는 그래프이다.
도 3은 본 발명의 실시예에서 사용한 서양뒤영벌여왕벌에서 유래된 GAG의 Hyaluronic acid 생성능을 보여주는 그래프이다.Figure 1 is a graph showing the Elastase inhibitory ability against GAG derived from the western rear young bee queen bee used in the examples of the present invention.
Figure 2 is a graph showing the collagenase inhibitory ability of GAG derived from the western rear young bee queen bee used in the examples of the present invention.
3 is a graph showing the hyaluronic acid-producing ability of GAG derived from the western rear young bee queen bee used in the examples of the present invention.
이하, 본 발명을 하나의 구현예로서 더욱 상세하게 설명하면 다음과 같다.Hereinafter, the present invention will be described in more detail as one embodiment as follows.
본 발명은 기존에 항암 등의 효과가 있는 것으로 알려져 있는 글라이코스아미노글리칸; GAG)의 새로운 용도에 관한 것이다.The present invention is a glycosaminoglycan known to have an effect such as anticancer in the past; GAG) relates to a new use.
특히, 본 발명은 GAG를 단순하게 다른 유효성분과 혼합하여 사용하는 것이 아니라 본질적으로 GAG 단독으로도 유효성분으로서 기능하도록 하는 것으로서, GAG에 대한 유효성분으로서 용도를 새롭게 제시하고자 한다.In particular, the present invention does not simply use GAG by mixing it with other active ingredients, but essentially allows GAG to function as an active ingredient alone, and to propose a new use as an active ingredient for GAG.
일반적으로 GAG는 의약 용도로서 소염, 항당뇨, 항응혈, 항암, 혈관신생억제 용도, 및/또는 혈관확장반응의 기능장애로 인한 질환 치료 및/또는 예방의 의약품이나 지방 축적 억제용 식품 등으로 알려져 있으나, 피부미용증진을 위한 유효성분으로서의 실질적인 용도에 관해서는 전혀 제안된 바 없다.In general, GAG is known for pharmaceutical use, anti-inflammatory, anti-diabetic, anti-coagulation, anti-cancer, anti-angiogenesis, and/or medicine for the treatment and/or prevention of diseases caused by dysfunction of the vasodilation reaction, or food for suppressing fat accumulation. However, no proposal has been made regarding its practical use as an active ingredient for promoting skin beauty.
많은 의약품이나 건강식품이 개발되고 있지만 이러한 의약이나 식품용 성분들이 피부미용증진이나 피부미용개선에 효과가 있는 것을 발명하는 것은 어려운 작업 중의 하나이다.Although many medicines and health foods are being developed, it is one of the difficult tasks to invent that these medicines or food ingredients are effective in promoting skin beauty or improving skin beauty.
본 발명에서는 기존에 피부미용증진용 효과가 밝혀지지 않은 GAG 성분을 피부미용증진에 대한 새로운 효과를 밝혀낸 것으로서, 특히 피부의 보습과 주름개선에 효과가 뚜렷함을 새롭게 밝혀냈다.In the present invention, the GAG ingredient, which has not been previously found to have an effect for promoting skin beauty, was found to have a new effect on promoting skin beauty, and in particular, it was newly revealed that it has a distinct effect on moisturizing the skin and improving wrinkles.
피부는 매우 민감한 부분이며, 특히 안면 피부의 경우 다른 부위에 비해 매우 민감하여 이러한 피부에 미용개선효과를 발현하는 물질을 찾아내는 것은 어려운 작업이 수반된다. 피부의 주름은 피부의 수분 함유량과 콜라겐의 함유량 그리고 면역 기능 등의 여러 요인에 영향을 받으므로, 엘라스틴 분해 및 콜라겐 분해를 억제하며 히알루론산 농도를 현저하게 증가시켜서 피부의 탄력과 보습의 개선에 도움을 줌으로써, 피부 주름이 개선될 수 있다. The skin is a very sensitive part, and in particular, the facial skin is very sensitive compared to other parts, so finding a substance that expresses the cosmetic improvement effect on such skin involves a difficult task. Skin wrinkles are affected by various factors such as the skin's moisture content, collagen content, and immune function. Therefore, it helps to improve skin elasticity and moisturization by suppressing elastin and collagen breakdown and increasing hyaluronic acid concentration remarkably. By giving the skin wrinkles can be improved.
본 발명에 따르면, 본 발명에서 제조된 GAG가 엘라스틴 분해 및 콜라겐 분해를 억제하며 히알루론산 농도를 크게 증가시킨다는 사실을 최초로 밝혀냈다. According to the present invention, it was discovered for the first time that the GAG prepared in the present invention inhibits the degradation of elastin and collagen and greatly increases the hyaluronic acid concentration.
따라서 본 발명은 GAG를 엘라스틴 분해 억제, 콜라겐 분해 억제 및 고농도의 히알루론산을 유효성분으로 함유하는 피부미용증진용 조성물을 제공한다. 특히, GAG를 엘라스틴 분해 및 콜라겐 분해 억제와 고농도의 히알루론산을 유효성분으로 하여 피부 주름개선용 화장료 조성물로 적용하는 경우에 탁월한 효능을 보인다. 그 이외에도 피부 보습용으로도 우수한 효과가 있고, 이너뷰티로 먹는 화장품이나 피부미용증진을 위한 건강식품으로도 적용이 가능하다.Accordingly, the present invention provides a composition for promoting skin beauty containing GAG as an active ingredient in inhibiting elastin decomposition, inhibiting collagen decomposition, and high concentration of hyaluronic acid. In particular, it shows excellent efficacy when GAG is applied as a cosmetic composition for improving skin wrinkles with suppression of elastin and collagen degradation and high concentration of hyaluronic acid as an active ingredient. In addition, it has an excellent effect for moisturizing the skin, and it can be applied as a cosmetic to be eaten as an inner beauty or as a health food for promoting skin beauty.
GAG는 다양한 천연자원에서 추출하여 얻을 수 있는데, 예컨대 바람직하게는 곤충에서 추출하여 얻을 수 있다.GAG can be obtained by extracting from various natural resources, for example, preferably obtained by extracting from insects.
본 발명의 바람직한 구현예에 따르면, 바람직하게는 본 발명의 목적에 가장 부합되는 GAG 추출물은 서양뒤영벌여왕벌에서 얻은 GAG가 특히, 엘라스틴 분해, 콜라겐 분해 및 히알루론산 농도 증가 효과가 우수함을 확인하였다.According to a preferred embodiment of the present invention, it was confirmed that the GAG extract, which is the most suitable for the purpose of the present invention, is that the GAG obtained from the western rear bee queen bee is particularly effective in decomposing elastin, decomposing collagen, and increasing hyaluronic acid concentration.
서양뒤영벌은 농촌에서 화분매개용으로 많이 사용되고 있으므로 이를 공급하는 양봉업체도 발달되어 있어 수집하기가 용이한 장점이 있으며, 특히 여왕벌은 수벌이나 일벌에 비해 몸집이 매우 커서 사용하기가 용이한 장점이 있다.Western rear bees are widely used as pollen media in rural areas, so beekeeping companies that supply them are also developed, so they are easy to collect. In particular, queen bees have the advantage of being very large and easy to use compared to male bees and worker bees. .
또한, 폐기되는 벌을 사용할 수 있어 구입비용이 저렴하고 폐기되는 곤충 부산물을 재활용하는 것이므로 환경적으로도 매우 바람직하다.In addition, since it is possible to use the discarded bees, the purchase cost is low, and the discarded insect by-products are recycled, which is very desirable environmentally.
따라서 본 발명의 바람직한 예로서, GAG를 서양뒤영벌여왕벌에서 추출한 것을 피부미용증진용 조성물의 유효성분으로 사용할 수 있다.Therefore, as a preferred example of the present invention, the extract of GAG from the western rear young bee queen bee can be used as an active ingredient in a composition for promoting skin beauty.
또한, 본 발명은 GAG 추출을 위해, 바람직하게는 예컨대 서양뒤영벌여왕벌을 유기용매에 침지하여 지방류를 제거하고 잔유물을 건조하는 단계, 건조된 잔유물을 150~500㎛로 분쇄하는 단계, 위 단계의 분쇄물을 단백분해제로 단백질을 분해하고 이를 제거하는 단계, 단백질이 제거된 잔유물을 에탄올에 침전시키고 세척하는 단계를 포함하는 방법으로 서양뒤영벌여왕벌로부터 GAG을 제조할 수 있다. 그러므로 본 발명은 이러한 GAG 추출물의 제조방법을 포함한다.In addition, the present invention is for GAG extraction, preferably, for example, immersing the queen bee in an organic solvent to remove fats and drying the residue, the step of pulverizing the dried residue to 150 ~ 500㎛, the pulverization of the above step It is possible to prepare GAG from the western rear bee queen bee by a method including the step of decomposing the protein with water with a proteolytic agent and removing the protein, and precipitating and washing the protein-removed residue in ethanol. Therefore, the present invention includes a method for preparing such a GAG extract.
본 발명에 따른 피부미용증진용 조성물에 함유되는 서양뒤영벌여왕벌로부터 제조될 수 있는 글라이코스아미노글리칸(GAG)은 예컨대 한국등록특허 제10-1200402호(신규한 곤충 글라이코자미노글라이칸의 제조방법)에 의하여 제조할 수도 있으나, 구체적으로 다음 방법으로 제조하여 사용하는 것이 바람직하다.Glycosaminoglycan (GAG), which can be prepared from the Western posterior young bee queen bee contained in the composition for enhancing skin beauty according to the present invention, is, for example, Korean Patent No. 10-1200402 (Preparation of novel insect glycosaminoglycan Method), but specifically, it is preferable to manufacture and use by the following method.
(1) 서양뒤영벌여왕벌을 세척하고 건조한 후 1차 분쇄하는 단계;(1) washing and drying the western rear young bee queen bee, followed by primary pulverization;
(2) 위 분쇄물을 에탄올로 추출하여 지방류를 제거하고 잔사물은 건조하여 평균직경 150~500㎛ 로 분쇄하는 단계;(2) extracting the pulverized product with ethanol to remove fat, and drying the residue and pulverizing the average diameter of 150 to 500 µm;
(3) 위 잔사물에 5-15배 부피의 탄산나트륨, 좋기로는 pH 8-10, 더욱 좋기로는 pH 8.5-9.5인 탄산나트륨 을 넣어 교반한 후 단백질 분해효소로 단백질을 분해하는 단계; (3) adding 5-15 times the volume of sodium carbonate, preferably pH 8-10, more preferably pH 8.5-9.5 sodium carbonate, to the above residue, stirring, and decomposing the protein with a proteolytic enzyme;
(4) 위 분해액에 Trichloroacetic acid 를 첨가하고 침전을 유도한 후 분해단백질을 제거하는 단계;(4) Trichloroacetic in gastric digestive solution adding acid and inducing precipitation, and then removing the decomposed protein;
(5) 위 상등액에 에탄올과 초산나트륨을 첨가하여 침전시키고 원심분리 하여 펠릿을 회수하는 단계;(5) adding ethanol and sodium acetate to the supernatant for precipitation and centrifuging to recover the pellet;
(6) 위 펠릿을 에탄올로 세척하고 건조하여 서양뒤영벌여왕벌 GAG을 얻는 단계.(6) The above pellet was washed with ethanol and dried to obtain GAG.
위 1단계는 단순하게 이물질을 제거하는 것만이 아니라, 1차 분쇄하여 지방류 제거가 용이하도록 하는 것이다. 일반적으로 곤충에는 지방류 함량이 높고 이에 따라 필요로 하는 성분의 추출이 어려운 점이 있어 산업적으로는 지방류 제거가 매우 중요한 문제가 된다. 기존에는 이에 대한 특별한 해결책이 제시되지 아니하고 바로 다음 단계인 분쇄를 시행하였다. 그러나 본 발명은 바람직하게도 이러한 종래 기술의 문제를 해결하기 위해 곤충의 수집단계에서 많이 혼입되어 있는 이물질들을 제거하고 또한 1차 간헐분쇄를 통하여 크기를 작게 하여 용매와 닿는 면적을 넓히고 내부로의 흡수를 용이하게 하여 지방류가 매우 효율적으로 제거되도록 한 것이다.The first step above is not just to remove foreign substances, but to make it easier to remove fats by first pulverizing. In general, insects have a high content of fats and thus it is difficult to extract necessary components, and therefore, removal of fats is a very important problem in the industrial sense. Previously, no specific solution was suggested, and the next step, crushing, was implemented. However, the present invention preferably removes foreign substances mixed in the insect collection step in order to solve the problems of the prior art, and also reduces the size through the first intermittent pulverization, thereby increasing the area in contact with the solvent and reducing absorption into the interior. This is to facilitate the removal of fats very efficiently.
위 2단계에서 잔사물의 평균직경은 150㎛ (통상 100mesh)보다 작으면 분쇄하기가 매우 어렵고 손실율도 높아지며, 500㎛ (통상 35mesh)보다 크면 효소분해가 늦어질 수 있는데, 더욱 좋기로는 350~500㎛ (통상 35~45mesh)이 더 바람직하다. In the second step above, if the average diameter of the residue is less than 150㎛ (usually 100mesh), it is very difficult to crush and the loss rate is high, and if it is larger than 500㎛ (typically 35mesh), the enzyme decomposition may be delayed. 500 µm (usually 35 to 45 mesh) is more preferable.
본 발명에 따르면, 이러한 분쇄과정에서 본 발명에 비해 더 작은 크기로 분쇄하는 경우, 예컨대 100mesh 크기보다 더 작은 입경으로 분쇄하는 경우 그 제조과정에서 곤충 특성상 건조가 어려운데다가 분쇄공정이 매우 어려운 문제가 있을 뿐만 아니라 분쇄에 필요한 공정시간도 오래 소요되며, 입자가 너무 작아서 오히려 효소분해가 늦어질 염려가 있다. 또한, 원심분리시 상등액에 유입될 염려가 크기 때문에 분리도 어려워서 전제적으로 오히려 공정상 불리하고 비경제적이며, 수율도 저하되는 요인으로 작용한다. 그러므로 본 발명에서는 이러한 문제 해결을 위해 잔사물의 평균입경, 바람직하게는 입경을 100mesh보다 크게 하는 것이 바람직하다.According to the present invention, in the case of grinding to a size smaller than that of the present invention in this grinding process, for example, when grinding to a particle size smaller than 100 mesh, drying is difficult due to the nature of insects during the manufacturing process, and the grinding process is very difficult. In addition, the processing time required for pulverization takes a long time, and there is a fear that the enzymatic decomposition may be delayed because the particles are too small. In addition, since there is a high risk of flowing into the supernatant during centrifugation, separation is also difficult, which is rather disadvantageous and uneconomical in the process, and it acts as a factor that lowers the yield. Therefore, in the present invention, in order to solve this problem, it is preferable to increase the average particle diameter of the residue, preferably larger than 100 mesh.
본 발명의 위 3단계에서 단백질분해효소는 예컨대 Alcalase (Alkaline Protease)를 사용하며 실온이상의 온도에서 1~7일 반응시킬 수 있다. In the above step 3 of the present invention, the protease is, for example, Alcalase (Alkaline Protease), and can be reacted for 1 to 7 days at a temperature above room temperature.
본 발명의 피부미용증진용 조성물은, GAG을 유효성분으로 함유하기 위해서 전체 조성물 총중량 대비 0.01~5.0 중량%로 사용할 수 있다. 만일, 본 발명의 피부미용증진용 조성물을 화장품으로 구성하는 경우 경제적인 면을 고려하면 바람직하게는 0.01-1.0중량%, 더욱 바람직하게는 0.03~0.3 중량% 포함할 수 있다. 또한, 화장품 중에서 경구 투여되는 이너뷰티용 화장품이나 건강식품 등으로 적용하여 유효성분으로 함유하도록 하기 위해서는 상기보다 과량, 예컨대 0.3-5.0중량%로 함유하도록 할 수 있다. 바람직한 예로서 GAG가 5-1000㎍/ml, 좋기로는 100-700㎍/ml, 더욱 좋기로는 200-700㎍/ml로 함유하는 것이 바람직하다.The composition for promoting skin beauty of the present invention may be used in an amount of 0.01 to 5.0% by weight based on the total weight of the total composition in order to contain GAG as an active ingredient. If the composition for promoting skin beauty of the present invention is composed of a cosmetic, it may include preferably 0.01 to 1.0% by weight, more preferably 0.03 to 0.3% by weight, considering the economic aspect. In addition, in order to be included as an active ingredient by applying to cosmetics for inner beauty or health food that are orally administered among cosmetics, it may be contained in an excess of, for example, 0.3-5.0% by weight. As a preferred example, it is preferable to contain GAG at 5-1000 μg/ml, preferably 100-700 μg/ml, more preferably 200-700 μg/ml.
본 발명에 따르면, GAG가 피부미용증진용 조성물의 유효성분으로 사용되기 위해서 총 중량대비 0.01 중량% 미만 포함되는 경우 엘라스틴 분해 및 콜라겐 분해 억제와 히알루론산 함량 증가 효과가 나타나지 않는 문제가 있으며, 전체 용량 중에 대해 5.0 중량% 초과인 경우 건조 피부를 가진 사람에게 피부 가려움 등 부작용이 발생될 수 있다.According to the present invention, when GAG is contained in less than 0.01% by weight of the total weight in order to be used as an active ingredient in a composition for promoting skin beauty, there is a problem that the effect of inhibiting elastin decomposition and collagen decomposition and increasing hyaluronic acid content does not appear. If it exceeds 5.0% by weight, side effects such as skin itching may occur in people with dry skin.
본 발명의 화장료 조성물은 당 업계에서 통상적으로 제조되는 모든 제형, 예를 들면, 용액, 현탁액, 페이스트, 겔, 크림, 로션, 팩, 폼 등으로 제조될 수 있으나, 이에 한정하는 것은 아니다. 이들 각 제형으로 이루어진 화장품은 통상의 화장료 조성물에 사용되는 점증제, 산화방지제, 보존제, 피부 보습제, 피부 유연화제, 계면활성제, L-멘톨, 모발 컨디셔닝제, 기포방지제, 향료 등을 더 포함할 수 있다.The cosmetic composition of the present invention may be prepared in all formulations conventionally prepared in the art, for example, solutions, suspensions, pastes, gels, creams, lotions, packs, foams, etc., but is not limited thereto. Cosmetics made of each of these formulations may further include thickeners, antioxidants, preservatives, skin moisturizers, skin softeners, surfactants, L-menthol, hair conditioning agents, anti-bubble agents, fragrances, etc. used in conventional cosmetic compositions. have.
또한, 피부미용증진용 건강식품으로서는 액제, 엑기스, 고형제, 정제, 분말제, 그래뉼, 캡슐 등으로 사용될 수 있다.In addition, as a health food for promoting skin beauty, it can be used as a liquid, extract, solid, tablet, powder, granule, capsule, or the like.
본 발명에 따른 피부미용증진용 조성물을 화장품으로 사용하는 경우 그 화장품 제형이 페이스트, 크림 또는 겔인 경우에는 담체 성분으로서 동물성유, 식물성유, 왁스, 파라핀, 전분, 셀룰로오스 유도체, 폴리에틸렌 글리콜, 실리콘, 벤토나이트, 실리카, 탈크 또는 산화아연 등이 이용될 수 있다.When the composition for promoting skin beauty according to the present invention is used as a cosmetic, when the cosmetic formulation is a paste, cream or gel, animal oil, vegetable oil, wax, paraffin, starch, cellulose derivative, polyethylene glycol, silicone, bentonite as carrier components are used as carrier components. , Silica, talc, or zinc oxide may be used.
본 발명에 따른 피부미용증진용 조성물을 이용한 화장품의 제형이 용액 또는 유탁액의 경우에는 담체 성분으로서 용매, 용매화제 또는 유탁화제가 사용될 수 있고, 예컨대 물, 에탄올, 이소프로판올, 에틸카보네이트, 에틸아세테이트, 벤질알코올, 벤질벤조에이트, 프로필렌글리콜, 1,3-부틸글리콜 오일, 글리세롤 지방족에스테르, 폴리에틸렌글리콜 또는 소르비탄의 지방산에스테르가 있다.In the case of a solution or emulsion in the formulation of a cosmetic product using the composition for promoting skin beauty according to the present invention, a solvent, a solvating agent or an emulsifying agent may be used as a carrier component, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, Benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol or fatty acid ester of sorbitan.
본 발명에 따른 피부미용증진용 조성물을 이용한 화장품의 제형이 현탁액인 경우에는 담체 성분으로서 물, 에탄올 또는 프로필렌글리콜과 같은 액상 희석제, 에톡실화이소스테아릴알코올, 폴리옥시에틸렌소르비톨에스테르 및 폴리옥시에틸렌소르비탄에스테르와 같은 현탁제, 미소결정성 셀룰로오스, 알루미늄 메타히드록시드, 벤토나이트, 아가 또는 트라칸트 등이 이용될 수 있다.When the cosmetic formulation using the composition for promoting skin beauty according to the present invention is a suspension, liquid diluents such as water, ethanol or propylene glycol, ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorb as carrier components Suspension agents such as non-ester, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar or tracanth, and the like may be used.
또한, 본 발명에 따른 피부미용증진용 건강식품은 다양한 제형으로 제조될 수 있으며, 추가적으로 다른 성분들을 복합하여 사용할 수도 있다. In addition, the health food for promoting skin beauty according to the present invention may be prepared in various formulations, and may additionally be used in combination with other ingredients.
이하, 본 발명을 실시 예를 통하여 보다 구체적으로 설명하지만, 이는 본 발명을 보다 상세하게 설명하기 위한 것으로, 본 발명의 권리범위가 이에 의해 한정되는 것은 아니다. Hereinafter, the present invention will be described in more detail through examples, but this is for describing the present invention in more detail, and the scope of the present invention is not limited thereto.
제조예 : 글라이코스아미노글리칸(GAG)의 제조Preparation Example: Preparation of glycosaminoglycan (GAG)
서양뒤영벌여왕벌 1㎏을 체를 이용하여 세척 후 Dry-Oven 에서 건조시킨 후에 간헐분쇄 하고, 위 분쇄물을 3배 부피의 에탄올로 추출하였으며, 추출 잔사를 Dry-Oven에서 건조시키고 분쇄기로 35mesh (500㎛)가 될 때까지 분쇄하였다. After washing with a sieve, 1 kg of queen bee of Western back young bee was washed with a sieve, dried in Dry-Oven, intermittently pulverized, and the pulverized product was extracted with 3 times the volume of ethanol, and the extracted residue was dried in Dry-Oven and 35 mesh (500 meshes) Μm).
분쇄된 잔사 100g 에 10배 부피의 탄산나트륨(pH 9.0)을 넣어 교반한 후, 잔사 중량의 1/10 부피의 알칼레이스(Alcalase) 분말을 첨가하고 3일간 정치 배양하였다. After 10 times the volume of sodium carbonate (pH 9.0) was added to 100 g of the pulverized residue and stirred, 1/10 volume of Alcalase powder was added to the residue, followed by incubation for 3 days.
배양물을 원심분리를 통해 침전물과 상층으로 분리하고, 용액에 존재하는 단백질 및 효소의 불활성화를 위해서 트리클로로아세트산(Trichloroacetic acid)을 최종농도 5%가 되도록 처리하고 침전을 유도한다.The culture was separated into a precipitate and an upper layer through centrifugation, and trichloroacetic acid was treated to a final concentration of 5% for inactivation of proteins and enzymes present in the solution, and precipitation was induced.
상등액에 2배 부피의 에탄올을 첨가하고 최종농도 1% 되도록 초산나트륨(Sodium acetate)을 첨가하여 침전시켰으며, 10,000 rpm에서 5분간 원심분리하여 Pellet만 회수하였다.Two times the volume of ethanol was added to the supernatant, sodium acetate was added to make the final concentration 1%, and precipitated, and only pellets were recovered by centrifugation at 10,000 rpm for 5 minutes.
회수된 Pellet을 95% 에탄올로 2회 세척한 후 66℃에서 3일간 건조시키어 서양뒤영벌여왕벌 GAG을 얻었으며 최종 수득율은 원물대비 2.6%이었다. 이를 50㎎/㎖로 희석하여 효과평가의 시료로 사용하였다. The recovered pellets were washed twice with 95% ethanol and then dried at 66° C. for 3 days to obtain GAG, and the final yield was 2.6% of the original product. This was diluted to 50 mg/ml and used as a sample for effect evaluation.
비교제조예 : 글라이코스아미노글리칸(GAG)의 제조Comparative Preparation Example: Preparation of glycosaminoglycan (GAG)
제조예의 방법 중 서양뒤영벌여왕벌의 세척 및 간헐분쇄를 하지 않았고, 건조 후 분쇄기로 200mesh가 될 때까지 분쇄한 것을 제외하고는 동일한 방법으로 제조하였으며 최종수득율은 원물대비 1.6% 이었다.Among the methods of Preparation Example, the Western rear young bee queen bee was not washed and intermittently pulverized, and was prepared by the same method, except that it was pulverized with a pulverizer until 200 mesh after drying, and the final yield was 1.6% compared to the original product.
실험예 : 글라이코스아미노글리칸(GAG)의 피부미용증진 효과 평가Experimental Example: Evaluation of the skin beauty enhancing effect of glycosaminoglycan (GAG)
(1) Elastase 활성 억제능 평가(1) Elastase activity inhibition ability evaluation
상기 제조예에서 제조된 GAG의 피부미용증진(주름개선) 효과를 평가하기 위해 아데노신을 비교물질로 하여 Elastase 활성 억제능을 측정하였다. In order to evaluate the effect of enhancing skin beauty (wrinkling improvement) of the GAG prepared in Preparation Example, the ability to inhibit Elastase activity was measured using adenosine as a comparative material.
0.1M Tris-HCl buffer (pH 8.0)에 농도별(125㎍/㎖, 250㎍/㎖, 500㎍/㎖)로 희석한 시료 220 ㎕와 2 ㎎/㎖ N-succinyl-Ala-Ala-Ala-ρ-nitroanilide 20 ㎕를 첨가하여 25℃ 조건에서 10분간 반응시켰다. 1 ㎍/㎖의 porcine pancreatic elastase (PPE) 20 ㎕를 첨가하여 25℃ 조건에서 20분간 반응시킨 다음 얼음위에서 반응을 종결시킨 후 microplate reader를 이용하여 405 ㎚에서 흡광도를 측정하였으며, Elastase 억제능은 시료 처리구와 무처리구의 흡광도 감소율로 나타내었다. 220 µl of sample diluted by concentration (125µg/ml, 250µg/ml, 500µg/ml) in 0.1M Tris-HCl buffer (pH 8.0) and 2 µg/ml N-succinyl-Ala-Ala-Ala- 20 µl of ρ-nitroanilide was added and reacted for 10 minutes at 25°C. After adding 20 µl of 1 µg/ml porcine pancreatic elastase (PPE) and reacting for 20 minutes at 25°C, the reaction was terminated on ice, and the absorbance was measured at 405 ㎚ using a microplate reader. It was expressed as the absorbance reduction rate of the and untreated groups.
억제능(%) = 1 - (시료첨가구의 흡광도/무첨가구의 흡광도)×100 Inhibitory ability (%) = 1-(absorbance of sample added/absorbance of no additive)×100
Elastase 억제능 평가 결과, 아래 표 1 및 도 1에서와 같이 GAG는 비교물질인 아데노신에 비하여 500㎍/㎖에서 약 2배 가까운 Elastase 억제 활성을 나타내었다. Elastase inhibitory ability evaluation results, as shown in Table 1 and Figure 1 below, GAG exhibited an Elastase inhibitory activity nearly twice as high at 500 µg/ml compared to adenosine, which is a comparative substance.
(아데노신)Control
(Adenosine)
(2) Collagenase 활성 억제능 평가(2) Evaluation of collagenase activity inhibitory ability
상기 제조예에서 제조된 GAG의 피부미용증진 효과를 평가하기 위해 아데노신을 비교물질로 하여 Collagenase 활성 억제능을 측정하였다. In order to evaluate the skin beauty enhancing effect of the GAG prepared in Preparation Example, the ability to inhibit collagenase activity was measured using adenosine as a comparative material.
시료를 농도별(125㎍/㎖, 250㎍/㎖, 500㎍/㎖)로 희석한 후 Collagenase Activity Assay kit(Abcam)을 사용하여 측정하였다. Assay buffer 88 ㎕에 농도별로 희석한 시료 2 ㎕와 Collagenase 10 ㎕를 추가한 후 100 ㎕의 Collagenase Reaction Mix를 첨가하여 반응시킨 후 microplate reader를 이용하여 345 ㎚에서 흡광도를 측정하였으며, Collagenase 억제능은 시료 처리구와 무처리구의 흡광도 감소율로 나타내었다.Samples were diluted by concentration (125 µg/ml, 250 µg/ml, 500 µg/ml) and measured using a Collagenase Activity Assay kit (Abcam). After adding 2 µl of sample diluted by concentration and 10 µl of Collagenase to 88 µl of assay buffer, 100 µl of Collagenase Reaction Mix was added to react, and absorbance was measured at 345 ㎚ using a microplate reader. It was expressed as the absorbance reduction rate of the and untreated groups.
억제능(%) = 1 - (시료첨가구의 흡광도/무첨가구의 흡광도) ×100Inhibitory ability (%) = 1-(absorbance of sample added/absorbance of no additive) ×100
Collagenase 활성 억제능 평가 결과, 아래 표 2 및 도 2에서와 같이 GAG의 Collagenase 억제 활성도 아데노신에 비하여 500㎍/㎖에서 약2배 가까운 우수한 활성을 나타내었다.As a result of evaluating the ability to inhibit collagenase activity, as shown in Table 2 and FIG. 2 below, the collagenase inhibitory activity of GAG also showed an excellent activity of about twice as much at 500 μg/ml compared to adenosine.
(3) Hyaluronic acid 생성능 평가(3) Hyaluronic acid production ability evaluation
상기 제조예에서 제조된 GAG의 피부미용증진 효과를 평가하기 위해 레티노산을 비교물질로 하여 Hyaluronic acid 생성능을 측정하였다. In order to evaluate the skin beauty enhancing effect of the GAG prepared in Preparation Example, the ability to produce hyaluronic acid was measured using retinoic acid as a comparative material.
시료를 농도별(25㎍/㎖, 50㎍/㎖, 100㎍/㎖, 200㎍/㎖, 300㎍/㎖, 400㎍/㎖, 500㎍/㎖)로 처리하여 히알루론산 생성 정도를 ELISA (Enzyme-linked immunosorbent assay) Kit를 통해 확인하였다.Samples were treated by concentration (25 µg/ml, 50 µg/ml, 100 µg/ml, 200 µg/ml, 300 µg/ml, 400 µg/ml, 500 µg/ml) to determine the level of hyaluronic acid production by ELISA ( Enzyme-linked immunosorbent assay) Kit.
HaCaT 세포를 6-웰 플레이트에 웰 당 1.0 × 106 세포로 접종하여 24시간 경과 후, 시료를 농도별로 처리한 후 24시간 동안 5% CO2가 공급되는 37℃의 항온 세포배양기에서 배양하였다. 세포배양 후, 히알루론산-ELISA Kit (Corgenix Inc., USA)를 이용하여 제조사의 프로토콜에 따라 HaCaT 세포가 분비하는 배양액내의 히알루론산의 양을 측정하였다.HaCaT cells were inoculated into a 6-well plate at 1.0 × 10 6 cells per well, and after 24 hours, the samples were treated by concentration and cultured in an incubator at 37° C. supplied with 5% CO 2 for 24 hours. After cell culture, the amount of hyaluronic acid in the culture medium secreted by HaCaT cells was measured using a hyaluronic acid-ELISA Kit (Corgenix Inc., USA) according to the manufacturer's protocol.
그 히알루론산 생성능 측정 결과, 아래 표 3 및 도 3에 나타난 바와 같이 GAG을 100㎍/㎖이상 처리한 군에서 대조군(Retinoic Acid, 1μmole)에 비해 증가된 히알루론산 생성을 확인하였으며, 이에 따라 보습 및 주름개선 효과가 있음을 확인하였다.As a result of measuring the hyaluronic acid production ability, as shown in Table 3 and Fig. 3 below, it was confirmed that hyaluronic acid production increased compared to the control (Retinoic Acid, 1 μmole) in the group treated with GAG of 100 µg/ml or more, and accordingly moisturizing and It was confirmed that there is an effect of improving wrinkles.
(㎍/㎖)Sample treatment concentration
(㎍/mL)
위와 같이 GAG는 Elastase 활성 억제능, Collagenase 활성 억제능 및 Hyaluronic acid 생성능을 가짐으로써 주름개선 등 피부미용에 탁월한 효과가 있는 것으로 확인되었다.As described above, GAG was found to have excellent effects on skin care such as wrinkle improvement by having the ability to inhibit Elastase activity, inhibit collagenase activity, and generate hyaluronic acid.
실시예 1 : GAG를 포함하는 피부미용증진용 화장품(겔제) 제조Example 1: Preparation of cosmetics (gels) for promoting skin beauty containing GAG
다음과 같은 조성의 겔제를 일반적인 겔제의 제조방법과 동일한 방법으로 제조하였다.A gel formulation having the following composition was prepared in the same manner as a general gel formulation method.
글리세린
유동파라핀
디에톡시에틸 석시네이트
클로르페네신
트리에탄올아민
페닐 트리메치콘
코폴리머
Disodium EDTA
부틸렌 글라이콜
정제수
감초뿌리추출물
GAG Alcohol
glycerin
Floating paraffin
Diethoxyethyl succinate
Chlorphenesin
Triethanolamine
Phenyl trimethicone
Copolymer
Disodium EDTA
Butylene glycol
Purified water
Licorice Root Extract
GAG
2.0
8.0
0.5
0.3
0.5
0.5
0.2
0.1
3.0
83.5
0.1
0.31.0
2.0
8.0
0.5
0.3
0.5
0.5
0.2
0.1
3.0
83.5
0.1
0.3
실시예 2 : GAG를 포함하는 피부미용증진용 화장품(크림제) 제조Example 2: Preparation of cosmetics (cream agent) for skin beauty promotion containing GAG
다음과 같은 조성의 크림제를 일반적인 크림제의 제조방법과 동일한 방법으로 제조하였다.A cream formulation having the following composition was prepared in the same manner as a general cream formulation.
호호바 골덴
아르간 오일
로즈힙 오일
마카데미어 넛 오일
올리브 유화 왁스
몬타 왁스 68
씨벅톤 오일
세라마이드
어성초추출물
감초뿌리추출물
나프리
GAGRose Premium Water
Jojoba Golden
Argan oil
Rosehip oil
Macadameer Nut Oil
Olive oil painting wax
Monta wax 68
Seabuckton Oil
Ceramide
Eoseongcho Extract
Licorice Root Extract
Napri
GAG
10.0
6.0
6.0
6.0
5.0
2.0
2.0
2.0
2.0
1.0
1.0
0.556.5
10.0
6.0
6.0
6.0
5.0
2.0
2.0
2.0
2.0
1.0
1.0
0.5
실험예 2 : GAG의 안전성 평가Experimental Example 2: Safety evaluation of GAG
안전성 시험에는 상기 제조예에서 제조된 GAG의 농도가 600㎍/㎖인 시료를 사용하였다. In the safety test, a sample having a concentration of 600 µg/ml of GAG prepared in Preparation Example was used.
(1) 인체피부 일차 자극 시험(1) Human skin primary stimulation test
미국화장품협회 가이드라인에 따라 단회 첩포 시험 방법(48hours Single Patch Test)에 따라 시험하였으며 Frosch & Kligman 법을 반영한 기준에 따라 평가하였다. 시험 결과 피부 반응이 관찰되지 않았으며 따라서 GAG는 인체피부의 일차 자극 측면에서 저자극 범주의 물질로 판단되었다.It was tested according to the 48hours Single Patch Test according to the guidelines of the American Cosmetics Association and evaluated according to the criteria reflecting the Frosch & Kligman method. As a result of the test, no skin reaction was observed, and therefore GAG was judged to be a substance in the hypoallergenic category in terms of primary irritation of human skin.
(2) 인체피부 광독성 시험(2) Human skin phototoxicity test
미국화장품협회 가이드라인에 따라 단회 첩포 시험 방법(24hours Single Patch Test)에 따라 시험하였으며, 광(UVA) 조사 후 30분, 24시간, 48시간 경과 시에 Frosch & Kligman 법을 반영한 기준에 따라 피부반응을 비교하여 평가하였다. 시험 결과 40%에서 미세한 홍반이 있었으나 24시간 이내에 모두 소실되었다. 따라서 GAG는 인체피부의 광독성 측면에서 안전한 물질로 판단되었다.It was tested according to the 24hours Single Patch Test according to the guidelines of the American Cosmetics Association, and skin reaction according to the criteria reflecting the Frosch & Kligman method at 30 minutes, 24 hours, and 48 hours after irradiation with light (UVA). Were compared and evaluated. As a result of the test, there was a minute erythema in 40%, but all disappeared within 24 hours. Therefore, GAG was judged to be a safe substance in terms of phototoxicity of human skin.
(3) 안자극 평가 시험(3) Eye irritation evaluation test
유럽동물대체시험법검증센터의 유럽유해물질관리지침에 따라 시험 및 판정을 하였으며, 시험 결과 판정기준에 따른 평균 반응도는 0.5~3.6, 더욱 바람직하게는 1.8로 나타났다. 따라서 GAG는 안점막 자극의 수준이 저자극 범주의 물질로 판단되었다.Tests and judgments were made according to the European Hazardous Substances Management Guidelines of the European Animal Substitute Test Method Verification Center, and as a result of the test, the average reactivity according to the criteria was 0.5 to 3.6, more preferably 1.8. Therefore, GAG was judged to be a substance in the hypoallergenic category with the level of ocular mucosal irritation.
위와 같이 본 발명에 따르면, GAG은 인체 피부 및 안점막에 대한 안전성이 확인되었으므로 화장품의 유효성분 원료로 사용될 수 있는 것이 확인되었다.As described above, according to the present invention, it was confirmed that GAG can be used as an active ingredient in cosmetics since it has been confirmed that it is safe for human skin and ocular mucosa.
본 발명에 따른 GAG를 유효성분으로 함유하는 피부미용증진용 조성물은 Elastase 활성 억제능, Collagenase 활성 억제능, Hyaluronic acid 생성능이 모두 현저하게 증가하는 것으로 확인되었으며, 인체피부에 안전한 물질임이 확인되어 피부미용증진용 화장품이나 피부미용증진용 건강식품, 의약품 등으로 사용이 가능하다.The composition for promoting skin beauty containing GAG according to the present invention was confirmed to significantly increase the ability to inhibit Elastase activity, inhibit collagenase activity, and generate hyaluronic acid, and it was confirmed that it is a safe substance for human skin. It can be used as cosmetics, health foods for promoting skin beauty, and medicines.
Claims (10)
A composition for promoting skin beauty containing Glycosaminoglycan (GAG) as an active ingredient.
The composition for enhancing skin beauty according to claim 1, wherein the glycosaminoglycan (GAG) is extracted from an insect.
The composition for enhancing skin beauty according to claim 1, wherein the glycosaminoglycan (GAG) is extracted from the western rear young bee queen bee.
The composition for enhancing skin beauty according to claim 1, wherein the glycosaminoglycan (GAG) is contained in an amount of 0.01-5.0% by weight.
서양뒤영벌여왕벌을 에탄올로 추출하여 지방류를 제거하고 잔사물은 건조하여 평균직경 150～500㎛ 로 분쇄하는 단계;
위 분쇄한 잔사물에 10배 부피의 탄산나트륨을 넣어 교반한 후 단백질분해효소로 단백질을 분해하는 단계;
위 단백질 분해된 분해액에 Trichloroacetic acid 를 첨가하고 침전을 유도한 후 분해단백질을 제거하는 단계;
위 분해단백질이 제거된 상등액에 에탄올과 초산나트륨을 첨가하여 침전시키고 원심분리 하여 펠릿을 회수하는 단계;
위 펠릿을 에탄올로 세척하고 건조하여 서양뒤영벌여왕벌 글라이코스아미노글리칸을 얻는 단계;
로 제조한 것임을 특징으로 하는 피부미용증진용 조성물.
The method according to claim 1, Glycosaminoglycan (GAG) is
Extracting the western rear young bee queen bee with ethanol to remove fats and drying the residue and pulverizing it to an average diameter of 150∼500㎛;
Adding 10 times the volume of sodium carbonate to the pulverized residue, stirring, and decomposing the protein with protease;
Trichloroacetic in gastric protein digestion solution adding acid and inducing precipitation, and then removing the decomposed protein;
Sedimentation by adding ethanol and sodium acetate to the supernatant from which the above digested protein has been removed and centrifuging to recover the pellet;
Washing the above pellet with ethanol and drying to obtain glycosaminoglycans of the queen bee and queen bee;
A composition for promoting skin beauty, characterized in that it is made of.
The method according to claim 1, characterized in that Glycosaminoglycan (GAG) is prepared through the step of pulverizing the residue obtained by primary pulverization after washing, drying and pulverizing the queen bee to an average diameter of 150 to 500 µm. A composition for promoting skin beauty.
A cosmetic composition for improving skin wrinkles containing Glycosaminoglycan (GAG) as an active ingredient.
A moisturizing cosmetic composition containing Glycosaminoglycan (GAG) as an active ingredient.
The cosmetic composition according to claim 7 or 8, wherein the composition is in the form of a solution, suspension, paste, gel, cream, lotion, pack or foam.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR102446952B1 (en) * | 2022-05-17 | 2022-09-23 | 주식회사 베니슈 | Mask pack using snake venom peptide |
| KR102496019B1 (en) * | 2022-10-06 | 2023-02-08 | 박미영 | Cosmetic composition for skin regeneration and anti-wrinkle with improved skin permeability and component stability and manufacturing methods thereof |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20110013074A (en) | 2009-07-31 | 2011-02-09 | 대한민국(농촌진흥청장) | Novel use of glycosaminoglycans derived from insect and a preparation method of the glycosaminoglycans |
| KR101200402B1 (en) | 2011-08-26 | 2012-11-14 | 대한민국 | A novel method of producing glycosaminoglycans derived from insect |
| KR101680644B1 (en) | 2014-12-18 | 2016-11-30 | 대한민국 | N-Glycan Purifed from Queen Bees and Use Thereof |
| KR101869792B1 (en) | 2017-06-27 | 2018-06-21 | 대한민국 | The glycosaminoglycan derived from queen of Bumbus ignitus and its inhibition activity of body fat accumulation |
-
2019
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Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20110013074A (en) | 2009-07-31 | 2011-02-09 | 대한민국(농촌진흥청장) | Novel use of glycosaminoglycans derived from insect and a preparation method of the glycosaminoglycans |
| KR101200402B1 (en) | 2011-08-26 | 2012-11-14 | 대한민국 | A novel method of producing glycosaminoglycans derived from insect |
| KR101680644B1 (en) | 2014-12-18 | 2016-11-30 | 대한민국 | N-Glycan Purifed from Queen Bees and Use Thereof |
| KR101869792B1 (en) | 2017-06-27 | 2018-06-21 | 대한민국 | The glycosaminoglycan derived from queen of Bumbus ignitus and its inhibition activity of body fat accumulation |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR102446952B1 (en) * | 2022-05-17 | 2022-09-23 | 주식회사 베니슈 | Mask pack using snake venom peptide |
| KR102496019B1 (en) * | 2022-10-06 | 2023-02-08 | 박미영 | Cosmetic composition for skin regeneration and anti-wrinkle with improved skin permeability and component stability and manufacturing methods thereof |
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