KR20190036060A - Skin-lightening Composition Using an Extract of Polygonum amphibium - Google Patents
Skin-lightening Composition Using an Extract of Polygonum amphibium Download PDFInfo
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- KR20190036060A KR20190036060A KR1020170124830A KR20170124830A KR20190036060A KR 20190036060 A KR20190036060 A KR 20190036060A KR 1020170124830 A KR1020170124830 A KR 1020170124830A KR 20170124830 A KR20170124830 A KR 20170124830A KR 20190036060 A KR20190036060 A KR 20190036060A
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- extract
- composition
- skin
- present
- water
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- 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
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- A23V2200/00—Function of food ingredients
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Abstract
Description
본 발명은 물여뀌(Polygonum amphibium) 추출물을 이용한 피부 미백용 조성물에 관한 것이다. The present invention relates to a composition for whitening skin using Polygonum amphibium extract.
현대사회에서 생활수준이 향상됨에 따라 건강뿐만 아니라 미용에 대한 관심도 점차 높아지고 있다. 이 중 자외선, 대기오염 등에 의한 피부손상 및 노화로 인하여 유발되는 기미(melasma), 주근깨(freckle)와 같은 색소 침착형 피부질환 개선과 전반적인 피부톤 개선을 위한 제품 개발이 활발하다. 표피의 기저층에 존재하는 멜라닌(melanin)은 피부색을 결정하는 주요한 색소로서 자외선에 의한 손상을 막고 활성산소를 제거하여 피부를 보호역할을 한다. 그러나 멜라닌 과발현은 피부 흑화, 색소침착, 기미, 주근깨, 피부암 등 피부 건강뿐만 아니라 미적 측면에서도 크게 영향을 받게 된다(Pigment Cell Res 10, 127-138. 1997;Vet Dermatol. 2003 Apr;14(2):57-65.).As living standards have improved in modern society, interest in beauty as well as health has been increasing gradually. Among them, development of products for the improvement of pigmentation-type skin diseases such as melasma and freckle induced by skin damage and aging caused by ultraviolet rays and air pollution and improvement of overall skin tone are actively developed. Melanin, which is present in the basal layer of the epidermis, is a major pigment that determines skin color. It protects skin by preventing ultraviolet damage and removing active oxygen. However, melanin overexpression is greatly affected not only by skin health but also by aesthetic aspects such as skin blackening, pigmentation, stinging, freckles, skin cancer, etc. (Pigment Cell Res 10, 127-138, 1997; Vet Dermatol. 2003 Apr; 14 (2) : 57-65.).
멜라닌은 피부(표피)내 기저층에 존재하는 색세포인 멜라노사이트(melanocyte)에서 합성되며 주변 각질세포(keratinocyte)로 전이되어 사람의 피부색을 나타내는 것으로 알려져 있다. 피부가 햇빛을 받으면 색소 형성 세포인 멜라노사이트에서 화학반응이 일어나 멜라닌이 만들어진다. 멜라닌의 주요한 역할을 피부에서 발생하는 활성산소가 프리라디칼(free radical)을 제거하고, 자외선의 투과를 막아 피부의 내부를 보호하는 것이다(Brain Research Bulletin 10 (6): 847-851, 1983; Mutation Research 571 (1-2): 121-32, 2005; Nature. 22;445(7130):843-50. 2007). It is known that melanin is synthesized in melanocyte, a color cell present in the basal layer in the skin (epidermis), and transformed into peripheral keratinocyte, indicating human skin color. When the skin is exposed to sunlight, a chemical reaction takes place at the melanocyte, a pigmented cell, to produce melanin. The main role of melanin is to remove free radicals from the skin and prevent the penetration of ultraviolet rays to protect the inside of the skin (Brain Research Bulletin 10 (6): 847-851, 1983; Mutation Research 571 (1-2): 121-32, 2005, Nature 22: 445 (7130): 843-50.
멜라닌을 만드는 출발물질은 인체에 정상적으로 존재하는 아미노산의 일종인 티로신(tyrosine)이다. 티로신은 멜라닌세포 내에서 티로시나제(tyrosinase)라는 효소에 의해 산화되어 도파(dihydroxy phenylalanine, DOPA)로 변하고, 도파는 더욱 산화하여 도파퀴논(DOPA-Quinone)을 만들어 낸다. 그리고 도파퀴논이 시스테인(cysteine)을 만나게 되면 시스테이닐도파(cysteinyldopa)가 만들어지고, 그 결과 적갈색의 페오멜라닌(pheomelanin)이 만들어 진다. 이후 생성된 멜라닌은 멜라노소체(melanosome)에 실려 각질 형성 세포로 전달된다(Pigment Cell Res.12(1):4-12.1999.;Cell Mol Biol (Noisy-le-grand).45(7):981-90.1999; An Bras Dermatol.88(1): 76-83. 2013).The starting material for making melanin is tyrosine, a kind of amino acid normally present in the human body. Tyrosine is oxidized by tyrosinase in melanocytes to dihydroxyphenylalanine (DOPA), which is further oxidized to produce dopaquinone (DOPA-Quinone). And when dopaquinone encounters cysteine, a cysteinyldopa is formed, resulting in a reddish brown pheomelanin. The resulting melanin is transferred to keratinocytes by melanosomes (Pigment Cell Res.12 (1): 4-12.1999 .; Cell Mol Biol (Noisy-le-grand) .45 (7): 981 An-Bras Dermatol. 88 (1): 76-83,2013).
색소 침착의 예방 또는 완화를 목적으로 미백 화장품에 배합되는 약제의 기본적인 메커니즘은 티로시나아제 작용 억제, 티로시나아제 생성 억제, 멜라닌 생성 미디에이터의 억제, 멜라닌의 환원 및 강산화 억제, 멜라닌 배설의 촉진, 자외선 차단 등이 있다. 기존의 미백에 관한 연구로는 멜라닌 합성의 과정에 주요한 역할을 하는 티로시나아제의 활성 억제하는 정도를 스크리닝하여 미백소재를 개발하는 경우가 많이 있었지만, 최근에는 멜라닌 생성, 이동, 축척 등 여러 단계에서의 기작을 조절함으로써 다양한 메커니즘을 연구를 통해 보다 효능이 우수한 미백소재를 개발하려는 경우가 증가하고 있다(Lee et al., J Soc Cosmet Scientists Korea 37, 275-822. 2011). 생성된 멜라닌을 환원시키는 미백제로는 비타민C, 글루타티온(glutathione), 아스코르빈산 인산 에스테르 마그네슘 등 항산화제가 알려져 있다(J Cosmet Sci.65(6):365-75. 2014; Int J Cosmet Sci.37(6):567-73. 2015; Yakugaku Zasshi. 128(8):1203-7. 2008; Int J Cosmet Sci.27(3):147-53. 2005; Acta Derm Venereol.63(3):209-14.1983; Skin Pharmacol Appl Skin Physiol.13(3-4):143-9. 2000; J Cosmet Laser Ther.15(2):107-13. 2013.).The basic mechanisms of pharmaceuticals incorporated into whitening cosmetics for the purpose of preventing or alleviating pigmentation are inhibition of tyrosinase action, inhibition of tyrosinase production, inhibition of melanin-producing mediators, inhibition of melanin reduction and strong oxidation, promotion of melanin excretion, And ultraviolet rays. There have been many cases in which a whitening material is developed by screening the degree of inhibition of tyrosinase activity, which plays a major role in the process of melanin synthesis. However, in recent years, (Lee et al., J Soc Cosmet Scientists Korea 37, 275-822, 2011). In this study, we investigated the mechanism of the whitening effect of the whitening agent. Antioxidants such as vitamin C, glutathione, magnesium ascorbyl phosphate and the like are known as whitening agents that reduce the produced melanin (J Cosmet Sci. 65 (6): 365-75. 2014; Int J Cosmet Sci. (6): 567-73, 2015; Yakugaku Zasshi, 128 (8): 1203-7, 2008; Int J Cosmet Sci.27 (3): 147-53 2005; Acta Derm Venereol.63 (3): 209 Skin Pharmacol Appl Skin Physiol. 13 (3-4): 143-9. 2000; J Cosmet Laser Ther. 15 (2): 107-13.
멜라닌의 생합성에는 여러 종류의 관련 인자들이 관여하기 때문에 단순한 멜라닌 합성 저해 실험 이외에 다양한 실험 방법이 적용되고 있다. 최근 제브라피쉬를 이용한 연구도 활발하게 진행되고 있는데 제브라피쉬는 한번에 100개 이상의 알을 얻을 수 있어 다량의 샘플 확보가 가능하며 투명한 피부를 통해 색소 생성 여부가 바로 관찰 가능하다(Grunwald and Eisen, Nat Rev Genet, 3:717-724,2002).Since various related factors are involved in the biosynthesis of melanin, various experimental methods other than simple inhibition of melanin synthesis have been applied. Recently, studies using zebrafish have been actively carried out. Zebrafish can acquire more than 100 eggs at a time, so that a large amount of samples can be obtained. Transparent skin makes it possible to observe pigmentation (Grunwald and Eisen, Nat Rev Genet. 3: 717-724, 2002).
미백에 대한 관심이 고조됨에 따라 기존의 잘 알려진 물질 이외에 더욱 우수한 효과를 지닌 미백 화장품 개발이 활발하게 이루어지고 있으나 합성미백원료들은 사용 중 분해나 착색, 이취, 안정성 및 안전성 등의 문제가 대두되고 있다. 따라서 세포에 영향을 끼치지 않으며 멜라닌 합성을 감소시키는 천연 미백제에 대한 연구가 활발히 진행되고 있다.As interest in whitening has increased, whitening cosmetics have been actively developed in addition to existing well-known substances. Whilst synthetic whitening materials are in the process of degradation, coloring, odor, stability, and safety, . Therefore, studies on natural whitening agents that do not affect cells and reduce melanin synthesis are actively under way.
본 발명의 목적은 물여뀌 추출물을 이용한 피부 미백용 조성물을 제공하는 데 있다.It is an object of the present invention to provide a skin whitening composition using a water-extract.
본 발명의 다른 목적이나 구체적인 목적은 이하에서 제시될 것이다.Other and further objects of the present invention will be described below.
본 발명은 아래의 실시예 및 실험예에서 확인되는 바와 같이, 물여뀌 추출물이 마우스 유래 멜라노마 세포주인 B16F10에서 멜라닌 합성 저해능을 가지고 제브라피시에서도 멜라닌 생합성 저해능을 가짐을 확인함으로써 완성된 것이다.The present invention was accomplished by confirming that the water extracts have an ability to inhibit melanin synthesis in melanoma cell line B16F10 and inhibit melanin biosynthesis in zebrafish as shown in the following examples and experimental examples.
전술한 바를 고려할 때, 본 발명의 피부 미백용 조성물은 물여뀌 추출물을 유효성분으로 포함함을 특징으로 한다.In view of the above, the skin whitening composition of the present invention is characterized by containing water extracts as an active ingredient.
본 명세서에서, "물여뀌 추출물"이란 추출 대상인 물여뀌 전초, 잎, 줄기, 지상부, 뿌리, 근경, 지하부 또는 이들의 혼합물을 물, 탄소수 1 내지 4의 저급 알콜(메탄올, 에탄올, 부탄올 등), 메틸렌클로라이드, 에틸렌, 아세톤, 헥산, 에테르, 클로로포름, 에틸아세테이트, 부틸아세테이트, N,N-디메틸포름아미드(DMF), 디메틸설폭사이드(DMSO), 1,3-부틸렌글리콜, 프로필렌글리콜 또는 이들의 혼합 용매를 사용하여 침출하여 얻어진 추출물, 이산화탄소, 펜탄 등 초임계 추출 용매를 사용하여 얻어진 추출물 또는 그 추출물을 분획하여 얻어진 분획물을 의미하며, 추출 방법은 활성물질의 극성, 추출 정도, 보존 정도를 고려하여 냉침, 환류, 가온, 초음파 방사, 초임계 추출 등 임의의 방법을 적용할 수 있다. 분획된 추출물의 경우 추출물을 특정 용매에 현탁시킨 후 극성이 다른 용매와 혼합·정치시켜 얻은 분획물, 상기 조추출물을 실리카겔 등이 충진된 칼럼에 흡착시킨 후 소수성 용매, 친수성 용매 또는 이들의 혼합 용매를 이동상으로 하여 얻은 분획물을 포함하는 의미이다. 또한 상기 추출물의 의미에는 동결건조, 진공건조, 열풍건조, 분무건조 등의 방식으로 추출 용매가 제거된 농축된 액상의 추출물 또는 고형상의 추출물이 포함된다. 바람직하게는 추출용매로서 물, 에탄올 또는 이들의 혼합 용매를 사용하여 얻어진 추출물, 특히 추출용매로서 에탄올을 사용하여 얻어진 추출물을 의미한다. 더 바람직하게는 에탄올 추출물의 에탈아세테이트 가용성 추출물, 더욱 바람직하게는 에탄올 추출물을 물에 현탁시킨 후, 노르말 헥산, 디클로로메탄, 에틸아세테이트 및 노르말 부탄올을 이용하여 순차적으로 분획하여 얻어진 각 분획에 따른 추출물, 특히 에틸아세테이트층의 분획물을 의미한다. 여기서 "순차적으로 분획한다"는 것의 의미는 물층의 분획물을 분획 후에도 계속 사용하여 상기 열거된 순서의 용매로 분획한다는 의미이다.As used herein, the term "water-extracted extract" means water, a lower alcohol having 1 to 4 carbon atoms (methanol, ethanol, butanol, etc.), water, (DMF), dimethylsulfoxide (DMSO), 1,3-butylene glycol, propylene glycol, or a mixture thereof. The solvent is preferably selected from the group consisting of methylene chloride, ethylene, acetone, hexane, ether, chloroform, ethyl acetate, butyl acetate, Refers to an extract obtained by leaching using a mixed solvent, an extract obtained by using a supercritical extraction solvent such as carbon dioxide or pentane, or a fraction obtained by fractionating the extract, and the extraction method includes considering the polarity, extraction degree, and preservation degree of the active substance Any method such as cooling, reflux, heating, ultrasonic irradiation, supercritical extraction, and the like can be applied. In the case of the fractionated extract, a fraction obtained by suspending the extract in a specific solvent and mixing and leaving with a solvent having a different polarity, the crude extract is adsorbed on a column packed with silica gel, and then a hydrophobic solvent, a hydrophilic solvent, Quot; means fractions obtained as a mobile phase. Also, the meaning of the extract includes a concentrated liquid extract or a solid extract in which the extraction solvent is removed by a method such as freeze drying, vacuum drying, hot air drying, spray drying and the like. Preferably an extract obtained by using water, ethanol or a mixed solvent thereof as an extraction solvent, especially an extract obtained by using ethanol as an extraction solvent. More preferably, the extract is obtained by suspending ethanol-soluble extracts of ethanol extract, more preferably ethanol extract, in water and sequentially fractionating the mixture using normal hexane, dichloromethane, ethyl acetate and normal butanol, In particular the fraction of the ethyl acetate layer. Here, " sequential fractionation " means that the fraction of the water layer is continuously used after fractionation and is fractionated into the solvent of the above-mentioned order.
또 본 명세서에서 "유효성분"이란 단독으로 목적하는 활성을 나타내거나 또는 그 자체는 활성이 없는 담체와 함께 활성을 나타낼 수 있는 성분을 의미한다.In the present specification, the term " active ingredient " alone means an ingredient which exhibits the desired activity or which can exhibit activity together with a carrier which is not itself active.
본 발명의 조성물은 그 유효성분을 용도, 제형, 배합 목적 등에 따라 피부 미백 활성 등을 나타낼 수 있는 한 임의의 양(유효량)으로 포함할 수 있는데, 통상적인 유효량은 조성물 전체 중량을 기준으로 할 때 0.001 중량 % 내지 15 중량 % 범위 내에서 결정될 것이다. 여기서 "유효량"이란 그 적용 대상인 포유동물 바람직하게는 사람에게 의료 전문가, 피부 전문가 등의 제언에 의한 투여 기간 동안 본 발명의 조성물이 투여될 때, 피부 미백 효과 등 의도한 의료적·피부학적 효과를 나타낼 수 있는, 본 발명의 조성물에 포함되는 유효성분의 양을 말한다. 이러한 유효량은 당업자의 통상의 능력 범위 내에서 실험적으로 결정될 수 있다.The composition of the present invention may be contained in any amount (effective amount) as long as it can exhibit skin whitening activity or the like depending on its use, formulation, compounding purpose and the like. A typical effective amount is, Will be determined within the range of 0.001 wt% to 15 wt%. The term " effective amount " as used herein refers to an amount of an effective amount of the composition of the present invention to be administered to a mammal, preferably a human, to a medical professional, a skin specialist, Refers to the amount of the active ingredient contained in the composition of the present invention that can be indicated. Such effective amounts can be determined experimentally within the ordinary skill of those skilled in the art.
본 발명의 피부 미백용 조성물은 유효성분 이외에, 피부 관련 활성을 보강·추가를 위하여 당업계에서 이미 안전성이 검증되고 해당 활성을 갖는 것으로 당업계에 공지된 임의의 화합물이나 천연 추출물을 추가로 포함할 수 있다. The composition for skin whitening of the present invention may further contain any compound or natural extract known in the art as having safety and proven activity in the art to supplement and add skin-related activity in addition to the active ingredient .
구체적으로 피부 미백 성분으로서는 알부틴, 나이아신아마이드, 아스코빌글루코사이드, 알파-비사보롤, 유용성 감초 추출물(Oil Soluble Licorice(Glycyrrhiza) Extract) 등을 들 수 있으며, 또 피부 주름 개선 성분으로서는 레티놀, 레티닐팔미테이트, 아데노신, 폴리에톡실레이티드아마이드 등을 들 수 있고, 자외선 보호 성분으로서는 드로메트리졸, 드로메트리졸트리실록산, 디갈로일트리올리에이트, 디메치코디에칠벤잘말로네이트, 디에칠헥실부타미도트리아존, 소나무 껍질 추출물 등의 복합물, 포스파티딜세린, 핑거루트 추출 분말, 홍삼과 산수유 등의 복합 추출물 등을 들 수 있다. 또 보습 성분으로서는 AP 콜라겐 효소 분해 펩타이드, Collactive 콜라겐 펩타이드, N-아세틸글루코사민, 곤약 감자 추출물, 민들레 등의 복합 추출물, 쌀겨 추출물, 옥수수 배아 추출물, 저분자 콜라겐 펩타이드, 지초 추출 분말, 포스파티딜세린, 히알루론산 등을 들 수 있다. 여타의 피부 미백 성분, 주름 개선 성분, 자외선 보호 성분, 피부 보습 성분들에 대해서는 각국 기능성화장품공전(한국에서는 식약처고시인 "기능성화장품 기준 및 시험방법"임), 각국 건강기능식품공전(한국에서는 식약처고시 "건강기능식품 기준 및 규격임)을 참조할 수 있다. 이러한 성분들은 본 발명의 피부 미백용 조성물에 그 유효성분과 함께 하나 이상 포함될 수 있다.Specific examples of skin whitening ingredients include arbutin, niacinamide, ascorbyl glucoside, alpha-bisabolol and oil soluble licorice (Glycyrrhiza) extract. Examples of the skin wrinkle improving agent include retinol, retinyl palmitate Examples of the ultraviolet protective component include dyes such as drometrizol, drometrizol trisiloxane, dicaloyltriolate, Dimethicyldecibenzalmalonate, diethylhexylbutadamate, Complex extracts such as dithriazone and pine bark extract, phosphatidylserine, finger root extract powder, and complex extracts of red ginseng and mountain juice. Examples of the moisturizing component include AP collagen enzyme digesting peptide, collactive collagen peptide, N-acetyl glucosamine, konjac potato extract, dandelion extract, rice bran extract, corn germ extract, low molecular weight collagen peptide, ground extract powder, phosphatidylserine, hyaluronic acid . For other skin-whitening, wrinkle-reducing, UV-protecting and skin-moisturizing ingredients, the functional cosmetics circulation in each country ("functional cosmetics standards and test methods" Quot; health functional food standards "). These ingredients may be included in the skin whitening composition of the present invention in combination with one or more of their effective ingredients.
본 발명의 피부 미백용 조성물에서 그 유효성분은 그것이 피부 미백 활성을 나타낼 수 있는 한, 용도, 제형, 배합 목적 등에 따라 임의의 양(유효량)으로 포함될 수 있는데, 통상적인 유효량은 조성물 전체 중량을 기준으로 할 때 0.001 중량 % 내지 20.0 중량 % 범위 내에서 결정될 것이다. 여기서 "유효량"이란 피부 미백 효과를 유도할 수 있는 유효성분의 양을 말한다. 이러한 유효량은 당업자의 통상의 능력 범위 내에서 실험적으로 결정될 수 있다. In the skin whitening composition of the present invention, the effective ingredient may be contained in any amount (effective amount) as long as it can exhibit skin whitening activity, depending on the purpose of use, formulation, blending purpose and the like. By weight based on the total weight of the composition. Here, " effective amount " refers to the amount of effective ingredient capable of inducing skin whitening effect. Such effective amounts can be determined experimentally within the ordinary skill of those skilled in the art.
본 발명의 피부 미백용 조성물은 구체적인 양태에 있어서, 화장료 조성물로 파악할 수 있다.The skin whitening composition of the present invention can be identified as a cosmetic composition in a specific embodiment.
본 발명의 조성물이 화장료 조성물로 파악될 경우에도 그 화장료 조성물은 그 용도상, 법률상 임의의 제품 구분을 띨 수 있으며, 구체적으로 피부 미백 등의 용도를 가진 기능성 화장품, 비기능성 일반 화장품 등일 수 있다. 제품 형태에 있어서도 임의의 제품 형태를 띨 수 있는데, 구체적으로 용액, 현탁액, 유탁액, 페이스트, 젤, 크림, 로션, 파우더, 비누, 계면활성제-함유 클렌징, 오일, 분말 파운데이션, 유탁액 파운데이션, 왁스 파운데이션, 스프레이 등으로 제형화될 수 있다. 구체적인 제품 형태에 있어서는 유연 화장수, 영양 화장수, 영양 크림, 마사지 크림, 에센스, 아이 크림, 클렌징 크림, 클렌징 포옴, 클렌징 워터, 팩, 스프레이 또는 파우더의 제형 등일 수 있다.Even when the composition of the present invention is identified as a cosmetic composition, the cosmetic composition may be classified into any product category according to the use of the cosmetic composition. Specifically, the cosmetic composition may be a functional cosmetic having a use such as skin whitening, . Specific examples of the product form include a solution, suspension, emulsion, paste, gel, cream, lotion, powder, soap, surfactant-containing cleansing oil, powder foundation, emulsion foundation, wax Foundation, spray or the like. In a specific product form, it may be a form of flexible lotion, nutritional lotion, nutritional cream, massage cream, essence, eye cream, cleansing cream, cleansing foam, cleansing water, pack, spray or powder.
본 발명의 화장료 조성물은 그 유효성분 이외에 화장료 조성물에 통상적으로 이용되는 성분들, 예컨대, 안정화제, 용해화제, 계면활성제, 비타민, 색소 및 항료와 같은 통상적인 보조제, 및 담체를 포함할 수 있다. The cosmetic composition of the present invention may contain, in addition to its active ingredient, conventional additives such as stabilizers, solubilizing agents, surfactants, vitamins, colorants and antioxidants, and carriers commonly used in cosmetic compositions.
본 발명의 제형이 페이스트, 크림 또는 젤인 경우에는 담체 성분으로서 동물성유, 식물성유, 왁스, 파라핀, 전분, 트라칸트, 셀룰로오스 유도체, 폴리에틸렌 글리콜, 실리콘, 벤토나이트, 실리카, 탈크 또는 산화아연 등이 이용될 수 있다.When the formulation of the present invention is a paste, a cream or a gel, an animal oil, vegetable oil, wax, paraffin, starch, tracant, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide may be used as a carrier component .
본 발명의 제형이 파우더 또는 스프레이인 경우에는 담체 성분으로서 락토스, 탈크, 실리카, 알루미늄 히드록시드, 칼슘 실리케이트 또는 폴리아미드 파우더가 이용될 수 있고, 특히 스프레이인 경우에는 추가적으로 클로로플루오로히드로카본, 프로판/부탄 또는 디메틸 에테르와 같은 추진체를 포함할 수 있다.When the formulation of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used as a carrier component. In the case of a spray, in particular, / Propane or dimethyl ether.
본 발명의 제형이 용액 또는 유탁액인 경우에는 담체 성분으로서 용매, 용해화제 또는 유탁화제가 이용되고, 예컨대 물, 에탄올, 이소프로판올, 에틸 카보네이트, 에틸 아세테이트, 벤질 알코올, 벤질 벤조에이트, 프로필렌 글리콜, 1,3-부틸글리콜 오일, 글리세롤 지방족 에스테르, 폴리에틸렌 글리콜, 소르비탄의 지방산 에스테르 등이 이용될 수 있다.When the formulation of the present invention is a solution or an emulsion, a solvent, a dissolving agent or an emulsifying agent is used as a carrier component, and examples thereof include water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, , 3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol, fatty acid esters of sorbitan, and the like.
본 발명의 제형이 현탁액인 경우에는 담체 성분으로서 물, 에탄올 또는 프로필렌 글리콜과 같은 액상의 희석제, 에톡실화 이소스테아릴 알코올, 폴리옥시에틸렌 소르비톨 에스테르, 폴리옥시에틸렌 소르비탄 에스테르와 같은 현탁제, 미소결정성 셀룰로오스, 알루미늄 메타히드록시드, 벤토나이트, 아가 등이 이용될 수 있다.When the formulation of the present invention is a suspension, a carrier, such as water, a liquid diluent such as ethanol or propylene glycol, a suspension such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester, polyoxyethylene sorbitan ester, Cellulose, aluminum metahydroxide, bentonite, agar, etc. may be used.
본 발명의 제형이 계면-활성제 함유 클렌징인 경우에는 담체 성분으로서 지방족 알코올 설페이트, 지방족 알코올 에테르 설페이트, 설포숙신산 모노에스테르, 이세티오네이트, 이미다졸리늄 유도체, 메틸타우레이트, 사르코시네이트, 지방산 아미드 에테르 설페이트, 알킬아미도베타인, 지방족 알코올, 지방산 글리세리드, 지방산 디에탄올아미드, 식물성 유, 라놀린 유도체 또는 에톡실화 글리세롤 지방산 에스테르 등이 이용될 수 있다.When the formulation of the present invention is an interfacial active agent-containing cleansing, the carrier component is selected from aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyltaurate, sarcosinate, fatty acid amide Ether sulfates, alkylamidobetaines, aliphatic alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, lanolin derivatives or ethoxylated glycerol fatty acid esters.
본 발명의 화장료 조성물은 피부 미백 활성을 나타내는 그 유효성분을 포함하는 것을 제외하고는 당업계에 통상적으로 행하여지는 화장료 조성물의 제조방법에 따라 제조할 수 있다.The cosmetic composition of the present invention can be produced according to a method for producing a cosmetic composition which is usually carried out in the art, except that it contains an active ingredient showing skin whitening activity.
본 발명의 조성물은 구체적인 양태에 있어서, 식품 조성물로서 파악할 수 있다. 본 발명의 조성물이 식품 조성물로 파악될 경우 그 용도는 피부 과민 반응 억제 또는 과민 반응 억제로 이해될 수 있다.In a specific embodiment, the composition of the present invention can be identified as a food composition. When the composition of the present invention is identified as a food composition, its use can be understood as inhibiting skin hypersensitivity or hypersensitivity.
본 발명의 식품 조성물은 어떠한 형태로도 제조될 수 있으며, 예컨대 차, 쥬스, 탄산음료, 이온음료 등의 음료류, 우유, 요구르트 등의 가공 유류, 껌류, 떡, 한과, 빵, 과자, 면 등의 식품류, 정제, 캡슐, 환, 과립, 액상, 분말, 편상, 페이스트상, 시럽, 겔, 젤리, 바 등의 건강기능식품 제제류 등으로 제조될 수 있다. The food composition of the present invention can be produced in any form and can be used in various forms such as beverages such as tea, juice, carbonated drink, ionic drink, processed oil such as milk and yogurt, gum, rice cake, Korean confectionery, A food, a health food, a food, a tablet, a capsule, a ring, a granule, a liquid, a powder, a slice, a paste, a syrup, a gel, a jelly and a bar.
또 본 발명의 식품 조성물은 법률상·기능상의 구분에 있어서 제조·유통 시점의 시행 법규에 부합하는 한 임의의 제품 구분을 띨 수 있다. 예컨대 한국 「건강기능식품에관한법률」에 따른 건강기능식품이거나, 한국 「식품위생법」의 식품공전(식약처 고시 「식품의 기준 및 규격」)상 각 식품유형에 따른 과자류, 두류, 다류, 음료류, 특수용도식품 등일 수 있다.In addition, the food composition of the present invention may be classified into any product category as long as it meets the laws and regulations on the time of manufacture and distribution in the legal and functional category. For example, it is a health functional food according to the 「Health Functional Food Act」 in Korea, or a food functional food according to the Korean Food Sanitation Law (Food Standards and Specifications) , Special-purpose food, and the like.
본 발명의 식품 조성물에는 그 유효성분 이외에 식품첨가물이 포함될 수 있다. 식품첨가물은 일반적으로 식품을 제조, 가공 또는 보존함에 있어 식품에 첨가되어 혼합되거나 침윤되는 물질로서 이해될 수 있는데, 식품과 함께 매일 그리고 장기간 섭취되므로 그 안전성이 보장되어야 한다. 식품의 제조·유통을 규율하는 각국 법률(한국에서는 「식품위생법」임)에 따른 식품첨가물공전에는 안전성이 보장된 식품첨가물이 성분 면에서 또는 기능 면에서 한정적으로 규정되어 있다. 한국 식품첨가물공전(식약처 고시 「식품첨가물 기준 및 규격」)에서는 식품첨가물이 성분 면에서 화학적 합성품, 천연 첨가물 및 혼합 제제류로 구분되어 규정되어 있는데, 이러한 식품첨가물은 기능 면에 있어서는 감미제, 풍미제, 보존제, 유화제, 산미료, 점증제 등으로 구분된다. The food composition of the present invention may contain food additives in addition to the active ingredients thereof. Food additives are generally understood to be substances that are added to foods and mixed or infiltrated into food in the manufacture, processing or preservation of food, and their safety must be ensured since they are ingested daily with food and for long periods of time. In food additives according to the laws of the respective countries ("Food Sanitation Act" in Korea) regulating the manufacture and distribution of food, food additives with safety are specified in terms of ingredient or function. In the Food Additives Code of Korea (Food Additives Standards and Standards), the food additives are classified into chemical compounds, natural additives and mixed preparations in terms of ingredients. Such food additives are classified into sweeteners, flavors Preservatives, emulsifiers, acidulants, and thickeners.
감미제는 식품에 적당한 단맛을 부여하기 위하여 사용되는 것으로, 천연의 것이거나 합성된 것 모두 본 발명의 조성물에 사용할 수 있다. 바람직하게는 천연 감미제를 사용하는 경우인데, 천연 감미제로서는 옥수수 시럽 고형물, 꿀, 수크로오스, 프룩토오스, 락토오스, 말토오스 등의 당 감미제를 들 수 있다. The sweetener is used for imparting a sweet taste suitable for foods, and both natural and synthetic sweeteners can be used in the composition of the present invention. Preferably, natural sweeteners are used. Examples of natural sweeteners include sugar sweeteners such as corn syrup solids, honey, sucrose, fructose, lactose and maltose.
풍미제는 맛이나 향을 좋게 하기 위하여 사용될 수 있는데, 천연의 것과 합성된 것 모두 사용될 수 있다. 바람직하게는 천연의 것을 사용하는 경우이다. 천연의 것을 사용할 경우에 풍미 이외에 영양 강화의 목적도 병행할 수 있다. 천연 풍미제로서는 사과, 레몬, 감귤, 포도, 딸기, 복숭아 등에서 얻어진 것이거나 녹차잎, 둥굴레, 대잎, 계피, 국화 잎, 자스민 등에서 얻어진 것일 수 있다. 또 인삼(홍삼), 죽순, 알로에 베라, 은행 등에서 얻어진 것을 사용할 수 있다. 천연 풍미제는 액상의 농축액이나 고형상의 추출물일 수 있다. 경우에 따라서 합성 풍미제가 사용될 수 있는데, 합성 풍미제는 에스테르, 알콜, 알데하이드, 테르펜 등이 이용될 수 있다. Flavors may be used to enhance taste or flavor, both natural and synthetic. Preferably, a natural one is used. When using natural ones, the purpose of nutritional fortification can be performed in addition to the flavor. Examples of natural flavoring agents include those obtained from apples, lemons, citrus fruits, grapes, strawberries, peaches, and the like, or those obtained from green tea leaves, Asiatica, Daegu, Cinnamon, Chrysanthemum leaves and Jasmine. Also, those obtained from ginseng (red ginseng), bamboo shoots, aloe vera, banks and the like can be used. The natural flavoring agent may be a liquid concentrate or a solid form of extract. Synthetic flavors may be used depending on the case, and synthetic flavors such as esters, alcohols, aldehydes, terpenes and the like may be used.
보존제로서는 소듐 소르브산칼슘, 소르브산나트륨, 소르브산칼륨, 벤조산칼슘, 벤조산나트륨, 벤조산칼륨, EDTA(에틸렌디아민테트라아세트산) 등이 사용될 수 있고, 또 유화제로서는 아카시아검, 카르복시메틸셀룰로스, 잔탄검, 펙틴 등을 들 수 있으며, 산미료로서는 연산, 말산, 푸마르산, 아디프산, 인산, 글루콘산, 타르타르산, 아스코르브산, 아세트산, 인산 등이 사용될 수 있다. 산미료는 맛을 증진시키는 목적 이외에 미생물의 증식을 억제할 목적으로 식품 조성물이 적정 산도로 되도록 첨가될 수 있다.As the preservative, calcium sorbate, sodium sorbate, potassium sorbate, calcium benzoate, sodium benzoate, potassium benzoate and EDTA (ethylenediaminetetraacetic acid) can be used. As the emulsifier, acacia gum, carboxymethyl cellulose, Pectin and the like. As the acidulant, math, malic acid, fumaric acid, adipic acid, phosphoric acid, gluconic acid, tartaric acid, ascorbic acid, acetic acid, phosphoric acid and the like can be used. The acidulant may be added so that the food composition has a proper acidity for the purpose of inhibiting the growth of microorganisms other than the purpose of enhancing the taste.
점증제로서는 현탁화 구현제, 침강제, 겔형성제, 팽화제 등이 사용될 수 있다.Examples of the thickening agent include suspending agents, sedimentation agents, gel-forming agents, bulking agents and the like.
본 발명의 식품 조성물은 전술한 바의 식품첨가물 이외에, 기능성과 영양성을 보충, 보강할 목적으로 당업계에 공지되고 식품첨가물로서 안정성이 보장된 생리활성 물질이나 미네랄류를 포함할 수 있다.The food composition of the present invention may contain physiologically active substances or minerals which are known in the art and which are stable as a food additive in addition to the above-mentioned food additives in order to supplement and supplement functional and nutritional properties.
그러한 생리활성 물질로서는 녹차 등에 포함된 카테킨류, 비타민 B1, 비타민 C, 비타민 E, 비타민 B12 등의 비타민류, 토코페롤, 디벤조일티아민 등을 들 수 있으며, 미네랄류로서는 구연산 칼슘 등의 칼슘 제제, 스테아린산마그네슘 등의 마그네슘 제제, 구연산철 등의 철 제제, 염화 크롬, 요오드칼륨, 셀레늄, 게르마늄, 바나듐, 아연 등을 들 수 있다. Examples of such physiologically active substances include catechins contained in green tea and the like, vitamins such as vitamin B1, vitamin C, vitamin E and vitamin B12, tocopherol, dibenzoyl thiamine, etc. Examples of minerals include calcium preparations such as calcium citrate, magnesium stearate , Iron preparations such as iron citrate, chromium chloride, potassium iodide, selenium, germanium, vanadium, zinc and the like.
본 발명의 식품 조성물에는 전술한 바의 식품첨가물이 제품 유형에 따라 그 첨가 목적을 달성할 수 있는 적량으로 포함될 수 있다.The food composition of the present invention may contain an appropriate amount of the above-mentioned food additives according to the product type so as to achieve the purpose of addition thereof.
본 발명의 식품 조성물에 포함될 수 있는 기타의 식품첨가물과 관련하여서는 각국 식품공전이나 식품첨가물 공전을 참조할 수 있다.With regard to other food additives that may be included in the food composition of the present invention, reference may be made to the Food Code of the respective countries or the Food Additives Code.
본 발명의 피부 미백용 조성물은 구체적인 양태에 있어서, 약제학적 조성물로 파악할 수 있다. The skin whitening composition of the present invention can be identified as a pharmaceutical composition in a specific embodiment.
본 발명의 피부 미백용 조성물이 약제학적 조성물로 파악될 경우 대상 질환은 피부 과색소침착증으로 이해될 수 있으며, 그러한 과색소침작증의 구체적인 예로서는 주근깨, 노인성 반점, 간반, 기미, 갈색 또는 흑점, 일광 색소반, 푸른 흑피증, 칼슘 길항제와 같은 약물 사용 후의 과다색소침착, 조직경화치료법에 따른 과다색소침착 후유증, 임신성 갈색반, 경구 피임약을 복용한 여성에서의 흑피증, 찰상 및 화상을 비롯한 상처 또는 피부염으로 인한 염증 후 과다 색소 침착, 광독성 반응 또는 다른 유사한 소형의 고정 색소성 병변 등이 포함될 수 있으나, 이에 제한되는 것은 아니다.When the composition for skin whitening of the present invention is identified as a pharmaceutical composition, the target disease can be understood as skin hypercholesterolemia. Specific examples of such hypercholesterolaemia include freckles, senile spots, liver, spots, brown or black spots, Hyperpigmentation after drug use such as anti-inflammatory drugs, anti-inflammatory drugs, antioxidants, calcium antagonists, hyperpigmentation sequelae due to tissue hardening therapy, wounds or dermatitis, including pregnancy brown bumps, mastication in women taking oral contraceptives, scratches and burns , Post-inflammatory hyperpigmentation due to inflammation, phototoxic response or other similar small fixed pigmented lesions, and the like.
본 발명의 약제학적 조성물은 유효성분 이외에 약제학적으로 허용되는 담체를 포함하여 당업계에 공지된 통상의 방법으로 투여 경로에 따라 경구용 제형 또는 비경구용 제형으로 제조될 수 있다. 여기서 투여 경로는 국소 경로, 경구 경로, 정맥 내 경로, 근육 내 경로, 및 점막 조직을 통한 직접 흡수를 포함하는 임의의 적절한 경로일 수 있으며, 두 가지 이상의 경로를 조합하여 사용할 수도 있다. 두 가지 이상 경로의 조합의 예는 투여 경로에 따른 두 가지 이상의 제형의 약물이 조합된 경우로서 예컨대 1차로 어느 한 약물은 정맥 내 경로로 투여하고 2차로 다른 약물은 국소 경로로 투여하는 경우이다. The pharmaceutical composition of the present invention may be prepared into oral formulations or parenteral formulations according to the route of administration by conventional methods known in the art, including pharmaceutically acceptable carriers in addition to the active ingredient. Where the route of administration may be any suitable route including local routes, oral routes, intravenous routes, intramuscular routes, and direct absorption through mucosal tissues, and combinations of two or more routes may be used. An example of a combination of two or more routes is a combination of two or more formulations of the drug according to the route of administration, for example, one drug is administered intravenously and another drug is administered via a local route.
약학적으로 허용되는 담체는 투여 경로나 제형에 따라 당업계에 주지되어 있으며, 구체적으로는 "대한민국약전"을 포함한 각국의 약전을 참조할 수 있다. Pharmaceutically acceptable carriers are well known in the art depending on the route of administration and formulation, and specific reference may be made to the pharmacopoeia of each country, including the " Korean Pharmacopoeia ".
본 발명의 약제학적 조성물이 경구용 제형으로 제조될 경우, 적합한 담체와 함께 당업계에 공지된 방법에 따라 분말, 과립, 정제, 환제, 당의정제, 캡슐제, 액제, 겔제, 시럽제, 현탁액, 웨이퍼 등의 제형으로 제조될 수 있다. 이때 적합한 담체의 예로서는 락토스, 글루코스, 슈크로스, 덱스트로스, 솔비톨, 만니톨, 자일리톨 등의 당류, 옥수수 전분, 감자 전분, 밀 전분 등의 전분류, 셀룰로오스, 메틸셀룰로오스, 에틸셀룰로오스, 나트륨 카르복시메틸셀룰로오스, 하이드록시프로필메틸셀룰로오스 등의 셀룰로오스류, 폴리비닐 피롤리돈, 물, 메틸히드록시벤조에이트, 프로필히드록시벤조에이트, 마그네슘 스테아레이트, 광물유, 맥아, 젤라틴, 탈크, 폴리올, 식물성유, 에탄올, 그리세롤 등을 들 수 있다. 제제화활 경우 필요에 따라적절한 결합제, 윤활제, 붕해제, 착색제, 희석제 등을 포함시킬 수 있다. 적절한 결합제로서는 전분, 마그네슘 알루미늄 실리케이트, 전분페리스트, 젤라틴, 메틸셀룰로스, 소듐 카복시메틸셀룰로스, 폴리비닐피롤리돈, 글루코스, 옥수수 감미제, 소듐 알지네이트, 폴리에틸렌 글리콜, 왁스 등을 들 수 있고, 윤활제로서는 올레산나트륨, 스테아르산나트륨, 스테아르산마그네슘, 벤조산나트륨, 초산나트륨, 염화나트륨, 실리카, 탈쿰, 스테아르산, 그것의 마그네슘염과 칼슘염, 폴리데틸렌글리콜 등을 들 수 있으며, 붕해제로서는 전분, 메틸 셀룰로스, 아가(agar), 벤토나이트, 잔탄 검, 전분, 알긴산 또는 그것의 소듐 염 등을 들 수 있다. 또 희석제로서는 락토즈, 덱스트로즈, 수크로즈, 만니톨, 소비톨, 셀룰로스, 글라이신 등을 들 수 있다. When the pharmaceutical composition of the present invention is prepared into an oral formulation, it may be formulated into powder, granules, tablets, pills, sugar tablets, capsules, solutions, gels, syrups, suspensions, wafers And the like. Examples of suitable carriers include starches such as lactose, glucose, sucrose, dextrose, sorbitol, mannitol and xylitol, corn starch, potato starch and wheat starch, cellulose, methylcellulose, ethylcellulose, sodium carboxymethylcellulose, Hydroxypropylmethylcellulose and the like; polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, magnesium stearate, mineral oil, malt, gelatin, talc, polyol, vegetable oil, ethanol Serol, and the like. In case of formulation, suitable binders, lubricants, disintegrants, coloring agents, diluents and the like may be included as needed. Examples of suitable binders include starch, magnesium aluminum silicate, starch pellets, gelatin, methyl cellulose, sodium carboxymethyl cellulose, polyvinyl pyrrolidone, glucose, corn sweetener, sodium alginate, polyethylene glycol, wax and the like. Examples of the disintegrating agent include starch, methylcellulose, magnesium stearate, magnesium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride, silica, talcum, stearic acid, Agar, bentonite, xanthan gum, starch, alginic acid or its sodium salt, and the like. Examples of the diluent include lactose, dextrose, sucrose, mannitol, sorbitol, cellulose, glycine and the like.
본 발명의 약제학적 조성물이 비경구용 제형으로 제조될 경우, 적합한 담체와 함께 당업계에 공지된 방법에 따라 주사제, 경피 투여제, 비강 흡입제 및 좌제의 형태로 제제화될 수 있다. 주사제로 제제화할 경우 적합한 담체로서는 수성 등장 용액 또는 현탁액을 사용할 수 있으며, 구체적으로는 트리에탄올 아민이 함유된 PBS(phosphate buffered saline)나 주사용 멸균수, 5% 덱스트로스 같은 등장 용액 등을 사용할 수 있다. 경피 투여제로 제제화할 경우 연고제, 크림제, 로션제, 겔제, 외용액제, 파스타제, 리니멘트제, 에어롤제 등의 형태로 제제화할 수 있다. 비강 흡입제의 경우 디클로로플루오로메탄, 트리클로로플루오로메탄, 디클로로테트라플루오로에탄, 이산화탄소 등의 적합한 추진제를 사용하여 에어로졸 스프레이 형태로 제제화할 수 있으며, 좌제로 제제화할 경우 그 담체로는 위텝솔(witepsol), 트윈(tween) 61, 폴리에틸렌글리콜류, 카카오지, 라우린지, 폴리옥시에틸렌 소르비탄 지방산 에스테르류, 폴리옥시에틸렌 스테아레이트류, 소르비탄 지방산 에스테르류 등을 사용할 수 있다.When the pharmaceutical composition of the present invention is prepared into a parenteral dosage form, it may be formulated in the form of an injection, transdermal drug delivery, nasal aspirate and suppository together with a suitable carrier according to methods known in the art. As the carrier suitable for injection preparation, aqueous isotonic solutions or suspensions may be used. Specifically, PBS (phosphate buffered saline) containing triethanolamine, sterile water for injection, and isotonic solution such as 5% dextrose may be used . When formulated with a transdermal preparation, it can be formulated in the form of ointments, creams, lotions, gels, external liquids, pastes, liniments, and air-lozenges. Nasal inhalers may be formulated in the form of aerosol sprays using suitable propellants such as dichlorofluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide, etc., and when formulated as a suppository, witepsol, tween 61, polyethylene glycols, cacao butter, laurin, polyoxyethylene sorbitan fatty acid esters, polyoxyethylene stearates, and sorbitan fatty acid esters.
약제학적 조성물의 구체적인 제제화와 관련하여서는 당업계에 공지되어 있으며, 예컨대 문헌[Remington's Pharmaceutical Sciences(19th ed., 1995)] 등을 참조할 수 있다. 상기 문헌은 본 명세서의 일부로서 간주 된다.The formulation of pharmaceutical compositions is well known in the art and can be found, for example, in Remington ' s Pharmaceutical Sciences (19th ed., 1995). This document is considered part of this specification.
본 발명의 약제학적 조성물의 바람직한 투여량은 환자의 상태, 체중, 성별, 연령, 환자의 중증도, 투여 경로에 따라 1일 0.001mg/kg ~ 10g/kg 범위, 바람직하게는 0.001mg/kg ~ 1g/kg 범위일 수 있다. 투여는 1일 1회 또는 수회로 나누어 이루어질 수 있다. 이러한 투여량은 어떠한 측면으로든 본 발명의 범위를 제한하는 것으로 해석되어서는 아니 된다. The preferred dosage of the pharmaceutical composition of the present invention is 0.001 mg / kg to 10 g / kg per day, preferably 0.001 mg / kg to 1 g / day, depending on the patient's condition, body weight, sex, age, / kg < / RTI > The administration can be carried out once or several times a day. Such dosages should in no way be construed as limiting the scope of the invention.
전술한 바와 같이, 본 발명에 따르면 물여뀌 추출물을 이용한 피부 미백용 조성물을 제공할 수 있다.As described above, according to the present invention, it is possible to provide a skin whitening composition using a water-extracting extract.
본 발명의 피부 미백용 조성물은 식품, 화장품, 약품 등으로 제품화될 수 있다.The skin whitening composition of the present invention can be commercialized into foods, cosmetics, medicines and the like.
도 1은 물여뀌 추출물과 그 분획물의 제조 과정 모식도이다.
도 2 내지 도 4는 물여뀌 추출물과 그 분획물의 마우스 유래 멜라노마 세포주인 B16F10에서의 멜라닌 합성 저해능을 나타낸 결과이다.
도 5는 에틸아세테이트 분획물의 마우스 유래 멜라노마 세포주인 B16F10에서의 세포독성을 나타낸 결과이다.
도 6은 에틸아세테이트 분획물의 제브라피시에서의 멜라닌 합성 저해능을 나타낸 결과이다.FIG. 1 is a schematic view showing the production process of water-extracted extract and its fractions.
FIGS. 2 to 4 show the results of inhibition of melanin synthesis in B16F10, a melanoma cell line derived from mouse, of water extract and its fractions.
Fig. 5 shows the cytotoxicity of the ethyl acetate fraction in mouse-derived melanoma cell line B16F10.
Figure 6 shows the results of the inhibition of melanin synthesis in zebrafish of the ethyl acetate fraction.
이하 본 발명을 실시예 및 실험예를 참조하여 설명한다. 그러나 본 발명의 범위가 이러한 실시예 및 실험예에 한정되는 것은 아니다.Hereinafter, the present invention will be described with reference to Examples and Experimental Examples. However, the scope of the present invention is not limited to these examples and experimental examples.
<< 실시예Example > 물여뀌 추출물> Water extract 의of 제조 Produce
물여뀌(전초) 분말에 20배 중량의 100% 에탄올(100% ethanol, HPLC grade)를 첨가하여 24시간 침출한 뒤 와트만(whatman paper) 여과지 No.2로 여과하고 그 여과액을 감압농축하여 물여뀌 추출물을 제조하였다. 다음 그 에탄올 추출물을 물에 현탁시키고 여기에 동량의 노르말 헥산을 첨가하여 분획한 후 그 증류수 증에 동량의 클로로포름, 동량의 에틸아세테이트, 동량의 노르말 부탄올을 가하여 순차적으로 분획하여 각 분획물을 얻었다. 상기 추출물과 각 분획물을 얻는 과정의 모식도를 그 수율과 함께 도 1에 나타내었다. 20% by weight of 100% ethanol (100% ethanol, HPLC grade) was added to the powdered water (outpost) powder, and the mixture was leached for 24 hours. Then, the solution was filtered through Whatman paper filter paper No. 2 and the filtrate was concentrated under reduced pressure Water extracts were prepared. Next, the ethanol extract was suspended in water, and the same amount of normal hexane was added to the fraction. The same amount of chloroform, the same amount of ethyl acetate and the same amount of normal butanol were added to the distilled water, and the fractions were sequentially fractionated. A schematic diagram of the process for obtaining the extract and each fraction is shown in FIG. 1 together with the yield.
<< 실험예Experimental Example > 미백 활성 실험> Whitening activity experiment
<< 실험예Experimental Example 1> 멜라닌 합성 저해능 평가 1> Melanin synthesis inhibition evaluation
C57BL/6 마우스에서 유래한 멜라노마 세포주인 B16F10을 이용하여 물여뀌 추출물의 멜라난 합성 저해능을 분석하였다. The melanin synthesis inhibitory effect of water extract was investigated using B16F10 melanoma cell line derived from C57BL / 6 mouse.
B16F10 세포의 배양은 10% fetal bovine serum, streptomycin-penicillin (100 mg/mL)과 100 nM α-melanocyte stimulating hormone이 포함된 DMEM 배지를 사용하여 37℃, 5% CO2의 환경에서 세포주를 배양하여 사용하였다. 멜라닌 생합성 저해 활성을 보기 위한 구체적 실험방법은 B16F10 세포 1×105개를 24-well plate의 각 well에 분주하여 하룻밤 배양하여 실험에 사용하였다. 각 샘플을 농도별로 배지에 처리하여 3일 동안 배양했다. 이후 phosphate-buffered saline으로 세척하고 1N NaOH를 200㎕씩 각 well에 처리하여 용해시켰다. 용해된 세포를 96 well plate로 옮겨 microplate reader를 사용하여 optical density를 405 nm에서 측정하고 melanin 생성 억제율은 시료 무처리군인 음성 대조군과 대비해 백분율로 계산하고 결과를 도 2 내지 도 4에 나타내었다.B16F10 cells were cultured in DMEM medium containing 10% fetal bovine serum, streptomycin-penicillin (100 mg / mL) and 100 nM α-melanocyte stimulating hormone at 37 ° C and 5% CO 2 Respectively. Specific experimental methods for inhibiting melanin biosynthesis were 1 × 10 5 B16F10 cells in each well of a 24-well plate and cultured overnight. Each sample was treated with a concentration-dependent medium and cultured for 3 days. After washing with phosphate-buffered saline, 200 μl of 1N NaOH was dissolved in each well. The melted cells were transferred to a 96-well plate, and the optical density was measured at 405 nm using a microplate reader. The inhibition rate of melanin production was calculated as a percentage of the negative control group as the untreated group. The results are shown in FIG. 2 to FIG.
도 2 및 도 3에 에탄올 추출물과 그 각 분획물을 50 ㎍/ml 농도로 처리하였을 때의 멜라닌 생합성 저해 활성을 나타내었고, 도 4에 가장 활성이 우수한 에틸아세테이트 분획물의 처리 농도별 멜라닌 생합성 저해 활성을 나타내었다. 에틸아세테이트 분획물은 농도 의존적으로 멜라닌 생합성 저해 활성을 보였고 최고 처리 농도인 50 ㎍/ml에서 32.6%의 저해 활성을 보였다. 양성대조군으로는 알부틴(Arbutin)을 사용하였다.FIG. 2 and FIG. 3 show melanin biosynthesis inhibitory activity when the ethanol extract and its fractions were treated at a concentration of 50 μg / ml. FIG. 4 shows the activity of inhibiting the melanin biosynthesis by the treatment concentration of the ethyl acetate fraction having the highest activity Respectively. Ethyl acetate fraction showed melanin biosynthesis inhibitory activity in a concentration - dependent manner and showed 32.6% inhibition activity at the highest treatment concentration of 50 ㎍ / ml. Arbutin was used as a positive control.
도 5에는 MTT assay에 의한 세포독성 결과를 나타내었는데, 최고 처리 농도인 50 ㎍/ml에서도 특별한 세포독성을 나타내지 않았다.FIG. 5 shows cytotoxicity results by the MTT assay. No cytotoxicity was observed even at the highest treatment concentration of 50 μg / ml.
<< 실험예Experimental Example 2> 2> 제브라피시에서의 멜라닌 합성 저해능 평가Evaluation of melanin synthesis inhibition in zebrafish
가장 높은 미백활성을 나타낸 물여뀌 유래 에틸아세테이트 분획물을 이용하여 제브라피시(zebrafish) 멜라닌 합성 저해 실험을 하였다. 성숙 제브라피시 암수를 알 채취 전날 알 채취용 수조에 넣고 다음날 광주기 시기 1-2시간 이후에 알을 채취하였다. 채취된 알은 제브라피시 배아용 배지(zebrafish embryo medium)에 넣고 24시간 발생 시킨 후, 각 샘플을 농도별로 처리하였다. 샘플 처리 시 코리온(chorion)의 샘플 투과 정도를 알 수 없기에 각 알의 코리온에 구멍을 내어 처리 하였고, 샘플 처리 후 24시간 이후 샘플의 독성이 나타난 알은 제거 하고, 나머지 알의 코리온을 완전 제거 후 2번째 샘플을 처리 하였다. 샘플 처리 48 시간 이후에 대조구 대비 샘플 처리구의 색소 발생 정도를 실체현미경으로 관찰하였다.The inhibition of melanin synthesis in zebrafish was performed using ethyl acetate fraction derived from water, which showed the highest whitening activity. Mature zebrafish ginsengs were picked in the sampling tank for sampling the day before harvest and eggs were collected 1-2 hours after the gwangju period. The harvested eggs were placed in a zebrafish embryo medium for 24 hours, and each sample was treated by concentration. Since the degree of permeation of the chorion sample was not known at the time of the sample treatment, it was treated by perforating the corions of each egg. After 24 hours after the sample treatment, the eggs showing toxicity of the sample were removed, The second sample was processed after complete removal. After 48 hours of the sample treatment, the degree of pigmentation of the sample treated with the control was observed with a stereomicroscope.
그 결과, 도 6에서와 같이 시료를 처리하지 않은 대조구(Control)에 비해 물여뀌 유래 에틸아세테이트 분획물을 50 ㎍/ml을 처리한 제브라피시의 요크(yolk) 부분의 색소 침착이 저해 될 뿐만 아니라 전체적으로 멜라닌의 합성이 저해됨을 확인하였다. As a result, as shown in FIG. 6, the pigmentation of the yolk portion of the zebrafish treated with 50 μg / ml of ethyl acetate fraction derived from water was inhibited as compared with the control without the sample (control) It was confirmed that the synthesis of melanin was inhibited.
Claims (6)
A composition for skin whitening comprising an extract of water as an active ingredient.
상기 물여뀌 추출물은 물여뀌 전초를 물, 에탄올 또는 그 혼합 용매로 추출하여 얻은 것을 특징으로 하는 피부 미백용 조성물.
The method according to claim 1,
Wherein the water-extracted extract is obtained by extracting water-containing herbs with water, ethanol or a mixed solvent thereof.
상기 상기 물여뀌 추출물은 에틸아세테이트 가용성 추출물인 것인 특징으로 하는 피부 미백용 조성물.
The method according to claim 1,
Wherein the water-extracting extract is an ethyl acetate-soluble extract.
상기 조성물은 화장료 조성물인 것을 특징으로 하는 피부 미백용 조성물.
4. The method according to any one of claims 1 to 3,
Wherein the composition is a cosmetic composition.
상기 조성물은 식품 조성물인 것을 특징으로 하는 피부 미백용 조성물.
4. The method according to any one of claims 1 to 3,
Wherein the composition is a food composition.
상기 조성물은 약제학적 조성물인 것을 특징으로 하는 피부 미백용 조성물.
4. The method according to any one of claims 1 to 3,
Wherein the composition is a pharmaceutical composition.
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Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
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KR102268130B1 (en) * | 2020-02-24 | 2021-06-22 | 이화여자대학교 산학협력단 | Cosmetic Composition comprising Polygonum vellardii All Extract |
KR20220164274A (en) * | 2021-06-04 | 2022-12-13 | 재단법인 제주테크노파크 | Skin-lightening Composition Using an Extract of Schizophragma hydrangeoides |
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KR20080105426A (en) * | 2007-05-31 | 2008-12-04 | 주식회사 코리아나화장품 | Cosmetic composition comprising the extract of persicaria blumei as active ingredient |
KR20110130132A (en) * | 2010-05-27 | 2011-12-05 | 이수제약(주) | Antioxidant composition containing p. nepalensis and p. japonica extracts |
KR20140083866A (en) * | 2012-12-26 | 2014-07-04 | 한국식품연구원 | Pharmaceutical or food composition comprising Persicaria posumbu |
KR20140101570A (en) * | 2013-02-12 | 2014-08-20 | 서울과학기술대학교 산학협력단 | Cosmetic composition for reinforcing skin barrier containing Persicaria hydropiper L. extract, and the liposome for enhancing transdermal delivery of Persicaria hydropiper L. extract |
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KR20080105426A (en) * | 2007-05-31 | 2008-12-04 | 주식회사 코리아나화장품 | Cosmetic composition comprising the extract of persicaria blumei as active ingredient |
KR20110130132A (en) * | 2010-05-27 | 2011-12-05 | 이수제약(주) | Antioxidant composition containing p. nepalensis and p. japonica extracts |
KR20140083866A (en) * | 2012-12-26 | 2014-07-04 | 한국식품연구원 | Pharmaceutical or food composition comprising Persicaria posumbu |
KR20140101570A (en) * | 2013-02-12 | 2014-08-20 | 서울과학기술대학교 산학협력단 | Cosmetic composition for reinforcing skin barrier containing Persicaria hydropiper L. extract, and the liposome for enhancing transdermal delivery of Persicaria hydropiper L. extract |
Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
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KR102268130B1 (en) * | 2020-02-24 | 2021-06-22 | 이화여자대학교 산학협력단 | Cosmetic Composition comprising Polygonum vellardii All Extract |
KR20220164274A (en) * | 2021-06-04 | 2022-12-13 | 재단법인 제주테크노파크 | Skin-lightening Composition Using an Extract of Schizophragma hydrangeoides |
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