KR101940055B1 - Composition for skin moisturizing containing extract of soybean leaf - Google Patents
Composition for skin moisturizing containing extract of soybean leaf Download PDFInfo
- Publication number
- KR101940055B1 KR101940055B1 KR1020180136604A KR20180136604A KR101940055B1 KR 101940055 B1 KR101940055 B1 KR 101940055B1 KR 1020180136604 A KR1020180136604 A KR 1020180136604A KR 20180136604 A KR20180136604 A KR 20180136604A KR 101940055 B1 KR101940055 B1 KR 101940055B1
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- KR
- South Korea
- Prior art keywords
- composition
- soybean
- present
- extract
- skin
- Prior art date
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Classifications
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- A—HUMAN NECESSITIES
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- 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
- A61K8/9783—Angiosperms [Magnoliophyta]
- A61K8/9789—Magnoliopsida [dicotyledons]
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/105—Plant extracts, their artificial duplicates or their derivatives
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/48—Fabaceae or Leguminosae (Pea or Legume family); Caesalpiniaceae; Mimosaceae; Papilionaceae
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
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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
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2200/00—Function of food ingredients
- A23V2200/30—Foods, ingredients or supplements having a functional effect on health
- A23V2200/318—Foods, ingredients or supplements having a functional effect on health having an effect on skin health and hair or coat
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Engineering & Computer Science (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- Natural Medicines & Medicinal Plants (AREA)
- General Health & Medical Sciences (AREA)
- Botany (AREA)
- Mycology (AREA)
- Chemical & Material Sciences (AREA)
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- Biotechnology (AREA)
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- Food Science & Technology (AREA)
- Alternative & Traditional Medicine (AREA)
- Polymers & Plastics (AREA)
- Medical Informatics (AREA)
- Nutrition Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Organic Chemistry (AREA)
- Birds (AREA)
- Cosmetics (AREA)
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Abstract
본 발명은 콩잎 추출물을 포함하는 항산화용, 항노화용, 피부 재생용, 미백용 또는 보습용 조성물에 관한 것이다. 본 발명의 조성물은 신체의 산화를 방지하고, 노화를 방지할 뿐 아니라, 피부 세포를 증식시켜 재생시킬 수 있으며, 피부 미백 기능을 가져서 유용하다. 뿐만 아니라 본 발명의 조성물은 피부 보습 기능을 가진다. 특히 본 발명의 조성물은 항산화능, 항노화능, 피부 재생능, 미백능 또는 보습능에 있어서 콩 추출물보다 더 우수한 효과를 보인다. 본 발명의 조성물은 또한 천연물로부터 얻어지는 자연 유래의 추출물을 포함하여 생물체에 자극이 거의 없다.The present invention relates to a composition for antioxidation, anti-aging, skin regeneration, whitening or moisturizing comprising a soybean leaf extract. The composition of the present invention is useful not only to prevent oxidation of the body, to prevent aging but also to regenerate and regenerate skin cells, and has a skin whitening function. In addition, the composition of the present invention has a skin moisturizing function. In particular, the composition of the present invention shows superior effects to soybean extracts in antioxidant ability, anti-aging ability, skin regeneration ability, whitening ability or moisturizing ability. The composition of the present invention is also free from irritation to organisms including natural extracts obtained from natural products.
Description
본 발명은 콩잎 추출물을 포함하는 항산화용, 항노화용, 피부 재생용, 미백용 또는 보습용 조성물에 관한 것이다.The present invention relates to a composition for antioxidation, anti-aging, skin regeneration, whitening or moisturizing comprising a soybean leaf extract.
인간의 피부는 나이가 들어감에 따라 여러 가지 내적, 외적 요인에 의해 변화를 겪는다. 즉, 내적으로는 신진대사를 조절하는 각종 호르몬의 분비가 감소하고, 면역 세포의 기능과 세포들의 활성이 저하되어 생체에 필요한 면역 단백질 및 생체 구성 단백질들의 생합성이 줄어들게 되며, 외적으로는 오존층 파괴로 인하여 태양 광선 중 지표에 도달하는 자외선의 함량이 증가함에 따라 피부 두께가 감소하고, 주름이 증가되며, 탄력이 감소될 뿐 아니라 기미와 주근깨, 검버섯 또한 증가하는 등 여러 가지 변화를 일으킨다.The human skin undergoes changes due to various internal and external factors as it gets older. In other words, internally, secretion of various hormones that regulate metabolism is reduced, the function of immune cells and the activity of cells are lowered, and the biosynthesis of immune proteins and biocompatible proteins necessary for the living body is reduced, and ozone layer destruction As the amount of ultraviolet rays reaching the surface of sunlight increases, skin thickness decreases, wrinkles increase, elasticity decreases, and various changes such as spots, freckles, and black spots are also caused.
노화가 진행될수록 피부를 구성하는 물질인 콜라겐, 엘라스틴, 히알루론산, 및 당단백질의 함유량 및 배열이 변하거나 감소하는 증상들이 나타나게 되고, 자유 라디칼 및 활성 유해 산소에 의한 산화적 스트레스를 받게 된다. 또한 노화가 진행되거나 자외선에 의해서, 피부를 구성하는 대부분의 세포에서는 염증을 일으킨다고 알려져 있는 전염증성 사이토카인(proinflammatory cytokine)을 생성하는 효소인 사이클로옥시게나제-2(Cox-2, cyclooxygenase)의 생합성이 증가하고, 이들 염증성 인자에 의해 피부조직을 분해하는 효소인 매트릭스 메탈로프로테아제(MMP, Matrix metalloproteinase)의 생합성이 증가하며, iNOS(inducible nitric oxide synthase)에 의한 NO(nitric oxide)생성이 증가하는 것으로 알려져 있다. 즉 자연적으로 진행되는 내인성 노화에 따른 세포 활성의 감소 및 미세염증에 의해 기질물질의 생합성이 감소되고, 여러 가지 유해 환경에 의한 스트레스의 증가 및 태양 광선에 의한 활성 산소 종의 증가와 같은 외적 요인에 의해 분해 및 변성이 가속화되어 피부기질이 파괴되고 얇아지면서 피부 노화의 제반 증상들이 나타나게 된다. 따라서, 이러한 노화의 현상들을 방지하고, 개선시킬 수 있는 활성성분에 대하여 많은 연구가 행해지고 있는 것이 현실이다.As the aging progresses, the content and arrangement of collagen, elastin, hyaluronic acid, and glycoprotein, which constitute the skin, are changed or decreased, and oxidative stress due to free radicals and active noxious oxygen is experienced. Cox-2 (cyclooxygenase), an enzyme that produces proinflammatory cytokines, known to cause inflammation in most of the cells that make up the skin by aging or ultraviolet light The biosynthesis of matrix metalloproteinase (MMP), which is an enzyme that decomposes the skin tissue by these inflammatory factors, is increased, and nitric oxide (NO) production by iNOS (inducible nitric oxide synthase) . In other words, decrease of cellular activity due to natural endogenous aging and decrease of biosynthesis of substrate material by microinflammation, increase of stress due to various harmful environment, increase of active oxygen species by sunlight, As the degradation and denaturation are accelerated, the skin substrate is destroyed and thinned, and various symptoms of skin aging appear. Therefore, it is a reality that many researches have been conducted on active ingredients that can prevent and improve such aging phenomena.
한편, 체내 효소계, 환원대사, 화학약품, 공해물질 및 광화학반응 등의 각종 물리적, 화학적 및 환경적 요인 등에 의하여 생성되는 활성산소는 세포구성 성분들인 지질, 단백질, 당 및 DNA 등에 대하여 비선택적, 비가역적인 파괴작용을 함으로써 세포노화 또는 암을 비롯한 각종 질병을 일으키는 것으로 알려져 있다. 또한 이들 활성산소에 의한 지질과산화의 결과로 생성되는 지질 과산화물을 비롯한 여러 가지 체내 과산화물도 세포에 대한 산화적 파괴를 일으켜 각종 기능장애를 야기함으로써 여러 가지 질병의 원인이 되기도 한다. 따라서, 이와 같은 자유 라디칼을 소거할 수 있는 화합물(free radical scavengers) 또는 과산화물 생성 억제물질과 같은 항산화제들이 이들 산화물들에 기인하는 노화 및 각종 질환의 억제 또는 치료제로서 기대되고 있다. On the other hand, active oxygen generated by various physical, chemical, and environmental factors such as in vivo enzymatic system, reductive metabolism, chemical agents, pollutants and photochemical reaction, etc. is non-selective, irreversible It is known to cause various diseases including cancer or aging cells by destructive action. In addition, lipid peroxides and various peroxides such as lipid peroxides produced as a result of lipid peroxidation by these active oxygen also cause oxidative destruction to cells and cause various dysfunctions and cause various diseases. Therefore, antioxidants such as free radical scavengers or peroxide formation inhibiting substances are expected as agents for inhibiting or treating various diseases caused by these oxides.
이에 본 발명자들은 콩을 활용하여 소재를 개발하던 중 콩잎 추출물을 피부에 적용하였을 때, 피부의 항산화 기능, 항노화 기능, 피부 재생 기능 및 미백 기능 뿐만 아니라 보습능 역시 증진시킬 수 있는 것을 확인하고 본 발명을 완성하게 되었다.Therefore, the inventors of the present invention found that when the soybean leaf extract was applied to the skin during the development of the material using the soybean, it could enhance not only the skin's antioxidant function, anti-aging function, skin regeneration function and whitening function, but also the moisturizing ability Thereby completing the invention.
본 발명의 목적은 항산화용, 항노화용, 피부 재생용, 미백용 또는 보습용으로 이용할 수 있는 천연 추출물을 포함하는 조성물을 제공하는 데 있다.It is an object of the present invention to provide a composition comprising a natural extract which can be used for antioxidation, anti-aging, skin regeneration, whitening or moisturizing.
상기 목적을 달성하기 위하여, 본 발명은 콩 잎 추출물을 유효성분으로 포함하는 항산화용 조성물을 제공한다.In order to achieve the above object, the present invention provides a composition for antioxidation comprising soybean leaf extract as an active ingredient.
본 발명은 또한 콩 잎 추출물을 유효성분으로 포함하는 항노화용 조성물을 제공하고, 콩 잎 추출물을 유효성분으로 포함하는 피부 재생용 조성물 및 콩 잎 추출물을 유효성분으로 포함하는 미백용 조성물을 제공한다.The present invention also provides an anti-aging composition comprising soybean leaf extract as an active ingredient, and provides a skin regeneration composition containing soybean leaf extract as an active ingredient and a whitening composition comprising soybean leaf extract as an effective ingredient.
본 발명은 또한 콩 잎 추출물을 유효성분으로 포함하는 보습용 조성물을 제공한다.The present invention also provides a moisturizing composition comprising soybean leaf extract as an active ingredient.
본 발명의 조성물은 신체의 산화를 방지하고, 노화를 방지할 뿐 아니라, 피부 세포를 증식시켜 재생시킬 수 있으며, 피부 미백 기능을 가져서 유용하다. 뿐만 아니라 본 발명의 조성물은 피부 보습 기능을 가진다. 특히 본 발명의 조성물은 항산화능, 항노화능, 피부 재생능, 미백능 또는 보습능에 있어서 콩 추출물보다 더 우수한 효과를 보인다. 본 발명의 조성물은 또한 천연물로부터 얻어지는 자연 유래의 추출물을 포함하여 생물체에 자극이 거의 없다.The composition of the present invention is useful not only to prevent oxidation of the body, to prevent aging but also to regenerate and regenerate skin cells, and has a skin whitening function. In addition, the composition of the present invention has a skin moisturizing function. In particular, the composition of the present invention shows superior effects to soybean extracts in antioxidant ability, anti-aging ability, skin regeneration ability, whitening ability or moisturizing ability. The composition of the present invention is also free from irritation to organisms including natural extracts obtained from natural products.
본 명세서에서 '콩'은 그 종류에 제한이 없다. 본 발명의 일 관점인 조성물에 있어서, 구체적으로, 본 명세서의 콩은 서리태, 서목태, 흑태, 청태, 황태, 울타리콩, 강낭콩, 얼룩강낭콩, 적두, 거두, 콩나물콩 및 대두로 구성된 군으로부터 선택되는 어느 하나 이상을 포함할 수 있지만 이에 제한되는 것은 아니다.In this specification, the term 'bean' is not limited. In a composition according to one aspect of the present invention, the soybean of the present specification is selected from the group consisting of Seed Rice, Seomotei, Black Rice, Chungtae, Chrysanthemum, Fence, Soybean, Kidney Bean, But is not limited thereto.
본 명세서에서 '콩잎'는 콩나무의 잎 부분을 의미한다.In the present specification, 'canola' means the leaf part of the bean curd.
본 명세서에서 '콩잎 추출물'은 추출과정에 있어서 예를 들면, 세척하고 건조시킨 콩잎 또는 이를 세말화한 콩잎 가루를 물 또는 유기용매에 넣고 추출하여 침적시킨 후 여과포 여과와 원심분리를 통해 잔사와 여액을 분리하고, 분리된 여액을 감압 농축하여 콩잎 추출물을 얻을 수 있다. 본 발명에 사용 가능한 유기용매는 에탄올, 메탄올, 부탄올, 에테르, 에틸아세테이트, 클로로포름 또는 이들 유기용매와 물의 혼합용매에서 선택될 수 있으며, 원료의 안전성을 고려할 때 바람직하게는 물 또는 30~70% 농도의 에탄올을 사용한다. 상기에서 용매를 이용하여 추출물을 얻은 이후 당업계에 알려진 통상적인 방법으로 상온에서 냉침, 가열 및 여과하여 액상물을 얻을 수 있으며, 또는 추가로 용매를 증발, 분무 건조 또는 동결 건조할 수 있으나, 이에 제한되는 것은 아니며, 당업계에서 일반적으로 사용되는 방법이라면 제한없이 사용하여 얻어질 수 있다.In the present invention, the 'canola leaf extract' is extracted in the extraction process by, for example, washing and drying the soap leaves or the kaolinized kaolin powder in water or an organic solvent, extracting it, and then filtering and separating the residue and the filtrate And the separated filtrate is concentrated under reduced pressure to obtain a soybean leaf extract. The organic solvent usable in the present invention may be selected from ethanol, methanol, butanol, ether, ethyl acetate, chloroform or a mixed solvent of these organic solvents and water. In view of the safety of raw materials, Of ethanol is used. After obtaining the extract by using the solvent, a liquid substance can be obtained by freezing, heating and filtering at room temperature by a conventional method known in the art. Alternatively, the solvent can be evaporated, spray dried or lyophilized But the present invention is not limited thereto, and any method generally used in the art can be used without limitation.
본 발명은 일 관점에서, 콩잎 추출물을 유효성분으로 포함하는 항산화용 조성물에 관한 것이다. In one aspect, the present invention relates to a composition for antioxidation comprising an extract of soybean curd as an active ingredient.
본 명세서에서 '항산화'란 당업계에 알려진 산화 과정을 늦추거나 막거나 또는 예방할 수 있는 효능을 말하는 것으로서 제한이 없다. As used herein, the term " antioxidant " refers to the ability to slow, prevent or prevent the oxidation process known in the art without limitation.
본 발명은 다른 관점에서, 콩잎 추출물을 유효성분으로 포함하는 항노화용 조성물에 관한 것이다.In another aspect, the present invention relates to an anti-aging composition comprising an extract of soybean curd as an active ingredient.
본 명세서에서 '항노화'란 당업계에 알려진 노화 과정을 늦추거나 막거나 또는 예방할 수 있는 효능을 말하는 것으로서, 구체적으로 피부내 콜라게나제의 발현을 효과적으로 억제함으로써 피부 내의 콜라겐 분해를 감소시켜 피부 탄력을 증진시키고 주름을 개선시키는 효능을 의미할 수 있지만 이에 제한되는 것은 아니다.As used herein, the term " anti-aging " refers to an effect of slowing down, preventing or preventing the aging process known in the art. Specifically, it effectively inhibits collagenase expression in the skin, ≪ RTI ID = 0.0 > and / or < / RTI > improving wrinkles.
본 발명은 또 다른 관점에서, 콩잎 추출물을 유효성분으로 포함하는 피부 재생용 조성물에 관한 것이다. 본 발명의 조성물은 피부 세포의 생장을 촉진시켜 피부를 재생시킬 수 있다. In another aspect, the present invention relates to a composition for regenerating skin comprising an extract of soybean curd as an active ingredient. The composition of the present invention can regenerate the skin by promoting the growth of the skin cells.
본 발명은 또 다른 관점에서, 콩잎 추출물을 유효성분으로 포함하는 미백용 조성물에 관한 것이다. 구체적으로, 본 발명의 조성물은 티로시나제를 저해함으로써 우수한 미백 효과를 제공할 수 있다. In another aspect, the present invention relates to a whitening composition comprising an extract of soybean curd as an active ingredient. Specifically, the composition of the present invention can provide an excellent whitening effect by inhibiting tyrosinase.
본 발명은 또 다른 관점에서 콩잎 추출물을 유효성분으로 포함하는 보습용 조성물에 관한 것이다. 구체적으로, 본 발명의 조성물은 각질 세포의 분화를 촉진하여 우수한 보습 효과를 제공할 수 있다. In another aspect, the present invention relates to a composition for moisturizing comprising an extract of soybean curd as an active ingredient. Specifically, the composition of the present invention promotes the differentiation of keratinocytes and can provide an excellent moisturizing effect.
본 발명의 일 관점인 조성물에 있어서, 상기 조성물은 조성물 총 중량에 대하여 0.001 내지 10 중량%의 콩 잎 추출물을 포함할 수 있다. 조성물 총 중량에 대하여 콩 잎 추출물의 함량이 0.001중량% 미만이면 항산화, 항노화, 미백 및 보습 효과가 미미하고, 10중량%를 초과하면 함유량 증가에 따른 뚜렷한 효과의 증가가 나타나지 않기 때문이다. 상기와 같은 관점에 있어서, 본 발명의 조성물은 조성물 총 중량에 대하여, 0.005~9.5중량%, 0.01~9중량%, 0.03~8.5중량%, 0.05~8중량%, 0.07~7.5중량%, 0.09~7중량%, 0.1~6.5중량%, 0.3~6중량%, 0.5~5.5중량% 또는 0.7~5중량%의 콩잎 추출물을 포함할 수 있다.In one aspect of the present invention, the composition may comprise from 0.001 to 10% by weight of soybean leaf extract, based on the total weight of the composition. If the content of the soybean leaf extract is less than 0.001% by weight based on the total weight of the composition, the antioxidant, anti-aging, whitening and moisturizing effect are insignificant, and if it exceeds 10% by weight, In view of the above, the composition of the present invention is used in an amount of 0.005 to 9.5% by weight, 0.01 to 9% by weight, 0.03 to 8.5% by weight, 0.05 to 8% by weight, 0.07 to 7.5% by weight, 7 to 7 wt%, 0.1 to 6.5 wt%, 0.3 to 6 wt%, 0.5 to 5.5 wt%, or 0.7 to 5 wt% of the soybean leaf extract.
본 발명의 일 관점인 조성물에 있어서, 상기 콩은 서리태 (Seoritae, Glycin max MERR), 서목태 (Seomoktae, Rhynchosia Nolubilis), 흑태 (Black soybean, Glycine max(L.) Merr.), 청태 (blue bean, Glycime max MERR ), 황태 (yellow bean, Glycime max MERR), 울타리콩 (field bean, Vicia faba), 강낭콩 (kidney bean, Phaseolus vulgaris), 얼룩강낭콩 (pinto bean, Phaseolus vulgaris L.), 적두 (small red bean, Vigna angularis), 거두 (small black bean, Phaseolus angularis W.F. WIGHT. ), 콩나물콩 (sprouting bean, Glycine max (L.) Merr.) 및 대두 (soybean, Glycine max)로 구성된 군으로부터 선택되는 어느 하나 이상을 포함할 수 있고, 구체적으로 대두 (soybean, Glycine max)일 수 있지만 이에 제한되는 것은 아니다.In one aspect of the present invention, the soybean is selected from the group consisting of Seoritae, Glycin max MERR, Seomoktae, Rhynchosia Nolubilis , Black soybean, Glycine max (L.) Merr., Blue bean, Glycime max MERR), yellow bean ( Glycime max MERR), field bean ( Vicia faba ), kidney bean ( Phaseolus vulgaris ), pinto bean ( Phaseolus vulgaris L.), small red bean, Vigna angularis , small black bean, Phaseolus angularis WF WIGHT, sprouting bean, Glycine max (L.) Merr. and soybean ( Glycine max ) (Soybean, Glycine max ), but is not limited thereto.
본 발명의 일 관점인 조성물에 있어서, 상기 조성물은 활성 산소종 (Reactive Oxygen Species, ROS)의 생성을 억제시킬 수 있다.In one aspect of the present invention, the composition may inhibit the production of reactive oxygen species (ROS).
본 발명의 일 관점인 조성물에 있어서, 상기 조성물은 콜라게나제 (Collagenase)의 발현 또는 콜라게나제의 활성을 증가시킬 수 있다.In one aspect of the present invention, the composition may increase the expression of collagenase or the activity of collagenase.
본 발명의 일 관점인 조성물에 있어서, 상기 조성물은 피부 세포를 증식시킬 수 있다.In one aspect of the invention, the composition is capable of proliferating skin cells.
본 발명의 일 관점인 조성물에 있어서, 상기 조성물은 티로시나아제 (Tyrosinase)의 발현 또는 티로시나아제의 활성을 저해시킬 수 있다.In one aspect of the present invention, the composition may inhibit the expression of tyrosinase or the activity of tyrosinase.
본 발명의 일 관점인 조성물에 있어서, 상기 조성물은 각질형성세포의 분화를 촉진시킬 수 있다.In one aspect of the present invention, the composition may promote differentiation of keratinocytes.
본 발명의 일 관점인 조성물에 있어서, 상기 조성물은 화장료 조성물을 포함한다.In one aspect of the invention, the composition comprises a cosmetic composition.
본 발명에 따른 화장료 조성물은 국소 적용에 적합한 모든 제형으로 제공될 수 있다. 예를 들면, 용액, 수상에 유상을 분산시켜 얻은 에멀젼, 유상에 수상을 분산시켜 얻은 에멀젼, 현탁액, 고체, 겔, 분말, 페이스트, 포말(foam) 또는 에어로졸 조성물의 제형으로 제공될 수 있다. 이러한 제형의 조성물은 당해 분야의 통상적인 방법에 따라 제조될 수 있다.The cosmetic composition according to the present invention may be provided in all formulations suitable for topical application. For example, it may be provided as a solution, an emulsion obtained by dispersing an oil phase in an aqueous phase, an emulsion obtained by dispersing an oil phase in water, a suspension, a solid, a gel, a powder, a paste, a foam or an aerosol composition. Compositions of such formulations may be prepared according to conventional methods in the art.
본 발명에 따른 화장료 조성물은 상기한 물질 이외에 주 효과를 손상시키지 않는 범위 내에서, 바람직하게는 주 효과에 상승 효과를 줄 수 있는 다른 성분들을 포함할 수 있다. 또한 본 발명에 따른 화장료 조성물은 보습제, 에몰리언트제, 자외선 흡수제, 방부제, 살균제, 산화 방지제, pH 조정제, 유기 및 무기 안료, 향료, 냉감제 또는 제한제를 더 포함할 수 있다. 상기 성분의 배합량은 본 발명의 목적 및 효과를 손상시키지 않는 범위 내에서 당업자가 용이하게 선정 가능하며, 그 배합량은 조성물 전체 중량을 기준으로 0.01 내지 5 중량%, 구체적으로 0.01 내지 3 중량%일 수 있다.The cosmetic composition according to the present invention may contain, in addition to the above-mentioned substances, other ingredients which can give a synergistic effect to the main effect, so long as they do not impair the main effect. The cosmetic composition according to the present invention may further comprise a moisturizing agent, an emollient agent, an ultraviolet absorber, an antiseptic, a bactericide, an antioxidant, a pH adjuster, an organic and inorganic pigment, a fragrance, a cold agent or a limiting agent. The compounding amount of the above components can be easily selected by those skilled in the art within a range not to impair the objects and effects of the present invention. The amount thereof may be 0.01 to 5% by weight, specifically 0.01 to 3% by weight, have.
본 발명의 일 관점인 조성물에 있어서, 상기 조성물은 약학 조성물을 포함한다.In one aspect of the invention, the composition comprises a pharmaceutical composition.
본 발명에 따른 조성물을 의약품에 적용할 경우에는, 상기 조성물을 유효성분으로 하여 상용되는 무기 또는 유기의 담체를 가하여 고체, 반고체 또는 액상의 형태로 경구 투여제 혹은 비경구 투여제로 제제화 할 수 있다.When the composition according to the present invention is applied to medicines, it may be formulated into an oral or parenteral dosage form in the form of solid, semi-solid or liquid by adding an inorganic or organic carrier to the composition as an active ingredient.
상기 경구 투여를 위한 제재로서는 정제, 환제, 과립제, 캡슐제, 산제, 세립제, 분제, 유탁제, 시럽제, 펠렛제 등을 들 수 있다. 또한, 상기 비경구 투여를 위한 제재로는 주사제, 점적제, 연고, 로션, 스프레이, 현탁제, 유제, 좌제 등을 들 수 있다. 본 발명의 유효성분을 제제화하기 위해서는 상법에 따라서 실시하면 용이하게 제제화할 수 있으며 계면활성제, 부형제, 착색료, 향신료, 보존료, 안정제, 완충제, 현탁제, 기타 상용하는 보조제를 적당히 사용할 수 있다.Examples of the agent for oral administration include tablets, pills, granules, capsules, powders, fine granules, powders, emulsions, syrups and pellets. Examples of the parenteral administration agent include injections, drops, ointments, lotions, sprays, suspensions, emulsions, suppositories, and the like. In order to formulate the active ingredient of the present invention, it can be easily formulated according to the conventional method. Surfactants, excipients, coloring agents, spices, preservatives, stabilizers, buffering agents, suspending agents and other adjuvants can be suitably used.
본 발명에 따른 상기 약학 조성물은 경구, 비경구, 직장, 국소, 경피, 정맥 내, 근육 내, 복강 내, 피하 등으로 투여될 수 있다.The pharmaceutical composition according to the present invention may be administered orally, parenterally, rectally, topically, transdermally, intravenously, intramuscularly, intraperitoneally, subcutaneously and the like.
본 발명의 약학 조성물의 유효 성분은 투여 받을 대상의 연령, 성별, 체중, 병리 상태 및 그 심각도, 투여 경로 또는 처방자의 판단에 따라 달라질 것이다. 이러한 인자에 기초한 적용량 결정은 당업자의 수준 내에 있으며, 이의 1일 투여 용량은 예를 들어 0.1mg/kg/일 내지 100mg/kg/일, 보다 구체적으로는 5 mg/kg/일 내지 50 mg/kg/일이 될 수 있으나, 이에 제한되는 것은 아니다.The effective ingredients of the pharmaceutical composition of the present invention will vary depending on the age, sex, weight, pathological condition and severity of the subject to be treated, administration route or judgment of the prescriber. Determination of the amount of application based on these factors is within the level of ordinary skill in the art and its daily dose is, for example, from 0.1 mg / kg / day to 100 mg / kg / day, more specifically from 5 mg / kg / day to 50 mg / / Day, but is not limited thereto.
본 발명의 일 관점인 조성물에 있어서, 상기 조성물은 건강식품 조성물을 포함한다. In one aspect of the invention, the composition comprises a health food composition.
*본 발명에 따른 조성물은 포함하는 다양한 형태의 식품 첨가제 또는 기능성 식품을 제공한다. 상기 조성물을 포함하는 발효유, 치즈, 요구르트, 주스, 생균제제, 정제, 과립제, 드링크제, 캐러멜, 다이어트바 등으로 제형화될 수 있고, 통상적인 차 잎 형태나 티백 및 건강보조식품 등으로 가공될 수 있으며, 그 외 다양한 식품 첨가제의 형태로 사용될 수 있다. * The composition according to the present invention provides various types of food additives or functional foods containing them. The composition may be formulated into fermented milk, cheese, yogurt, juice, probiotics, tablets, granules, drinks, caramels, diet bars, and the like, and may be processed into ordinary tea leaves, tea bags, And can be used in the form of various other food additives.
일실시예에서 상기 조성물은, 본 발명이 목적으로 하는 주 효과를 손상시키지 않는 범위 내에서 주 효과에 상승 효과를 줄 수 있는 다른 성분 등을 함유할 수 있다. 예를 들어, 물성 개선을 위하여 향료, 색소, 살균제, 산화방지제, 방부제, 보습제, 점증제, 무기염류, 유화제 및 합성 고분자 물질 등의 첨가제를 더 포함할 수 있다. 그 외에도, 수용성 비타민, 유용성 비타민, 고분자 펩티드, 고분자 다당 및 해초 엑기스 등의 보조 성분을 더 포함할 수도 있다. 상기 성분들은 제형 또는 사용 목적에 따라서 당업자가 어려움 없이 적의 선정하여 배합할 수 있으며, 그 첨가량은 본 발명의 목적 및 효과를 손상시키지 않는 범위 내에서 선택될 수 있다. 예를 들어, 상기 성분들의 첨가량은, 조성물 전체 중량을 기준으로, 0.01~5 중량%, 보다 구체적으로는 0.01~3 중량% 범위일 수 있다.In one embodiment, the composition may contain other ingredients or the like that can give synergistic effects to the main effect within a range that does not impair the intended main effect of the present invention. For example, additives such as perfume, coloring agent, bactericide, antioxidant, preservative, moisturizing agent, thickening agent, inorganic salt, emulsifier and synthetic polymer substance may be further added for improvement of physical properties. In addition, it may further contain auxiliary components such as water-soluble vitamins, oil-soluble vitamins, polymer peptides, polymeric polysaccharides and seaweed extract. The above components may be mixed and selected without difficulty by those skilled in the art depending on the purpose of formulation or use, and the amount thereof may be selected within a range that does not impair the objects and effects of the present invention. For example, the addition amount of the above components may be in the range of 0.01 to 5% by weight, more specifically 0.01 to 3% by weight, based on the total weight of the composition.
본 발명에 따른 조성물의 제형은 용액, 유화물, 점성형 혼합물, 타블렛, 분말 등의 다양한 형태일 수 있으며, 이는 단순 음용, 주사 투여, 스프레이 방식 또는 스퀴즈 방식 등의 다양한 방법으로 투여될 수 있다.The composition according to the present invention may be in various forms such as a solution, an emulsion, a viscous mixture, a tablet, a powder, etc., and may be administered by various methods such as simple drinking, injection administration, spraying or squeezing.
이하, 실시예를 통하여 본 발명을 더욱 상세히 설명하고자 한다. 이들 실시예는 오로지 본 발명을 예시하기 위한 것으로, 본 발명의 범위가 이들 실시예에 의해 제한되는 것으로 해석되지 않는 것은 당업계에서 통상의 지식을 가진 자에게 있어서 자명할 것이다. Hereinafter, the present invention will be described in more detail with reference to Examples. It is to be understood by those skilled in the art that these embodiments are only for illustrating the present invention and that the scope of the present invention is not construed as being limited by these embodiments.
[실시예 1] 콩잎 추출물의 제조[Example 1] Production of soybean leaf extract
콩잎을 정제수로 세척하고 건조시킨 다음 세말화하였다. 상기 콩잎 가루 100g을 70% 에탄올 수용액 1리터에 넣고 냉각 콘덴서가 달린 추출기에서 12시간 끓여서 추출한 후, 300 메쉬 여과포로 여과하였다. 그 후, 상기 여과액을 4-15 ℃도에서 7일간 방치하여 숙성시킨 후 와트만 2번 여과지로 여과시켰다. 상기 추출액을 3 리터 분액 깔때기에 넣고 에칠아세테이트 1리터를 가한 후, 흔들어서 섞어준 다음 두 층으로 완전히 분리되면 상층(에틸아세테이트 층)을 취한다. 하층(물층)을 다시 분액 깔대기로 두 번 더 추출한다. 각각의 분리된 상층을 모두 합한 뒤 냉각 콘덴서가 달린 증류 장치를 이용하여 50℃로 감압 농축하고 건조하였다.The soybean leaves were washed with purified water, dried and then hemihalized. 100 g of the bean leaf powder was placed in 1 liter of a 70% aqueous ethanol solution, boiled for 12 hours in an extractor equipped with a condenser, and then filtered through a 300-mesh filter cloth. Thereafter, the filtrate was treated with 4-15 And allowed to stand for 7 days at < RTI ID = 0.0 > 0 C < / RTI > for aging and then filtered through Watson # 2 filter paper. Add the above extract to a 3-liter separatory funnel, add 1 liter of ethylacetate, shake and mix, then take the upper layer (ethyl acetate layer) when complete separation into two layers. The lower layer (water layer) is extracted twice more with a separatory funnel. The separated upper layers were combined and concentrated under reduced pressure at 50 ° C using a distillation apparatus equipped with a cooling condenser and dried.
[비교실시예 1] 콩 추출물의 제조[Comparative Example 1] Preparation of soybean extract
콩을 정제수로 세척하고 건조시킨 다음 세말화하였다. 상기 콩 가루 100g을 70% 에탄올 수용액 1리터에 넣고 냉각 콘덴서가 달린 추출기에서 12시간 끓여서 추출한 후, 300 메쉬 여과포로 여과하였다. 그 후, 상기 여과액을 4-15 ℃도에서 7일간 방치하여 숙성시킨 후 와트만 2번 여과지로 여과시켰다. 상기 추출액을 3 리터 분액 깔때기에 넣고 에칠아세테이트 1리터를 가한 후, 흔들어서 섞어준 다음 두 층으로 완전히 분리되면 상층(에틸아세테이트 층)을 취한다. 하층(물층)을 다시 분액 깔대기로 두 번 더 추출한다. 각각의 분리된 상층을 모두 합한 뒤 냉각 콘덴서가 달린 증류 장치를 이용하여 50℃로 감압 농축하고 건조하였다.The beans were washed with purified water, dried and then chased. 100 g of soybean flour was placed in 1 liter of a 70% aqueous ethanol solution, which was boiled in an extractor equipped with a condenser for 12 hours and then filtered through a 300-mesh filter cloth. Thereafter, the filtrate was treated with 4-15 And allowed to stand for 7 days at < RTI ID = 0.0 > 0 C < / RTI > for aging and then filtered through Watson # 2 filter paper. Add the above extract to a 3-liter separatory funnel, add 1 liter of ethylacetate, shake and mix, then take the upper layer (ethyl acetate layer) when complete separation into two layers. The lower layer (water layer) is extracted twice more with a separatory funnel. The separated upper layers were combined and concentrated under reduced pressure at 50 ° C using a distillation apparatus equipped with a cooling condenser and dried.
[시험예 1] 활성 산소종(ROS: reactive oxygen species) 생성 억제 효과[Test Example 1] Inhibitory effect on reactive oxygen species (ROS) production
사람의 표피 조직에서 분리한 각질형성세포(keratinocyte)를 24웰 플레이트의 각 웰에 5×104개를 넣고 24시간 동안 부착시켰다. 16시간 후, 콩잎 추출물을 1%로 처리하였다. 이 때 비교를 위하여 콩 추출물을 1% 처리하였으며, 대조군은 콩잎 추출물을 처리하지 않았다. 2시간 뒤에 배양액을 제거한 후, 각 웰에 100㎕의 인산완충염액(PBS)을 넣었다. 이 각질형성세포에 자외선 B(UVB) 램프(Model: K5T8, UV B 15 W, Sankyo Dennki社, Japan)를 이용하여 자외선 30mJ/㎠를 조사한 다음, PBS를 덜어내고 각 웰에 각질형성세포 배양액 200㎕를 첨가하였다. 여기에 콩잎 추출물 및 콩 추출물을 다시 처리하고 일정 시간대별로 자외선 자극에 의해 증가한 활성 산소종의 양을 정량하였다. ROS의 양은 ROS에 의해 산화되는 DCF-DA(dichlorofluorescin diacetate)의 형광을 측정하는 Tan의 방법을 참고하여 정량하였으며(Tan et al., 1998, J. Cell Biol. Vol. 141, pp1423-1432), 비히클만 처리한 대조군의 ROS에 대한 비율을 산출한 결과를 하기 표 1에 나타내었다.Keratinocytes isolated from human epidermal tissue were added to each well of a 24-well plate at 5 × 10 4 for 24 hours. After 16 hours, the bean leaf extract was treated with 1%. For comparison, the soybean extract was treated with 1% and the control group was not treated with the soybean leaf extract. After 2 hours, the culture solution was removed, and 100 μl of phosphate buffered saline (PBS) was added to each well. This keratinocyte was irradiated with ultraviolet rays of 30 mJ / cm 2 using an ultraviolet B (UVB) lamp (Model: K5T8, UVB 15 W, Sankyo Dennki, Japan), and then PBS was removed. The keratinocyte culture medium 200 ≪ / RTI > The soybean extract and soybean extract were treated again and the amount of active oxygen species increased by ultraviolet stimulation was determined at predetermined time intervals. The amount of ROS was determined by reference to Tan's method of measuring the fluorescence of DCF-DA (dichlorofluorescin diacetate) oxidized by ROS (Tan et al., 1998, J. Cell Biol. Vol. 141, pp1423-1432) The ratios of the vehicle-treated control to the ROS were calculated. The results are shown in Table 1 below.
상기 표 1과 같이, 본 발명의 콩잎 추출물은 자외선에 의해 피부 세포 손상을 일으키는 것으로 알려진 ROS의 생성을 콩 추출물보다 더 효과적으로 억제하는 것으로 확인되었다. 아울러, 본 발명의 콩잎 추출물은 자외선으로 자극을 가한 후의 ROS 양을 자외선을 가하지 않은 경우와 비슷하게 억제하는 것으로 보아, 항산화 효능이 매우 뛰어남을 알 수 있었다. 그러므로, 본 발명에 의한 콩 잎 추출물이 산화를 억제하고 노화를 막음으로써 모공이 넓어지는 것을 예방할 수 있으며, 자극에 대하여 피부를 방어함으로써 피부 트러블을 개선시킬 수 있음을 확인하였다.As shown in Table 1, it was confirmed that the soybean leaf extract of the present invention inhibits the production of ROS, which is known to cause skin cell damage by ultraviolet light, more effectively than soybean extract. In addition, the extract of canola leaves of the present invention inhibits the amount of ROS after UV stimulation, which is similar to the case of not irradiating ultraviolet rays, indicating that the antioxidative effect is excellent. Therefore, it has been confirmed that the soybean leaf extract according to the present invention can prevent the wrinkling of pores by inhibiting oxidation, preventing aging, and improving the skin troubles by protecting the skin against irritation.
[시험예 2] 콜라게나제 생성 억제 효과[Test Example 2] Collagenase inhibitory effect
상기 실시예 1 및 비교실시예 1에서 제조한 콩잎 추출물 및 콩 추출물의 콜라게나제 (collagenase) 생성 억제능을 토코페롤(비교예 1) 및 EGCG(비교예 2)와 비교하여 측정하였다. 토코페롤 및 EGCG는 피부의 표피 세포를 재생시켜 피부의 노화를 방지하는 기능이 있는 것으로 알려진 물질이다. The ability of the soybean leaf extract and soybean extract prepared in Example 1 and Comparative Example 1 to inhibit collagenase production was measured in comparison with tocopherol (Comparative Example 1) and EGCG (Comparative Example 2). Tocopherol and EGCG are substances known to regenerate epidermal cells of the skin and prevent aging of the skin.
2.5%의 우태아 혈청이 함유된 DMEM(Dulbecco's Modified Eagle's Media) 배지가 들어 있는 96공 평판배양기(96-well microtiter plate)에 인간의 섬유아세포를 5,000 세포/웰(well)이 되도록 넣고, 90% 정도 자랄 때까지 배양하였다. 그 후 무혈청 DMEM 배지에서 24시간 배양한 다음, 무혈청 DMEM 배지에 녹여진 상기 실시예 1의 콩잎 추출물 100ug/ml농도, 비교예 1의 콩 추출물 100ug/ml농도, 토코페롤 및 EGCG 각각을 10-4 몰농도로 24시간 동안 처리한 후, 세포배양액을 채취하였다. Human fibroblasts were plated at 5,000 cells / well in a 96-well microtiter plate containing DMEM (Dulbecco's Modified Eagle's Media) medium containing 2.5% fetal bovine serum, Lt; / RTI > until growth. Then the serum-free one in the DMEM culture medium cultured for 24 hours, and then serum-free the exemplary binary dissolved in DMEM culture medium in Example 1 of kongip extract bean extract of 100ug / ml concentration, comparative example 1 100ug / ml concentration, tocopherol and EGCG, respectively 10 - 4- molar concentration for 24 hours, and then the cell culture solution was collected.
콜라게나제 측정기구 (아머샴파마샤 사, 미국)를 이용하여 채취한 세포배양액의 콜라게나제 생성 정도를 측정하였다. 먼저 1차 콜라게나제 항체가 균일하게 도포된 96-공 평판(96-well plate)에 채취된 세포 배양액을 넣고 3시간 동안 항원-항체 반응을 항온조에서 실시하였다. The degree of collagenase production in the cell culture medium was measured using a collagenase measuring device (Amersham Pharmacia, USA). First, the cell culture solution collected in a 96-well plate uniformly coated with the primary collagenase antibody was added and the antigen-antibody reaction was performed in a thermostatic chamber for 3 hours.
3시간 후 발색단이 결합된 2차 콜라겐 항체를 96-웰 플레이트 (96-well plate)에 넣고 다시 15분간 반응시켰다. 15분 후, 발색 유발물질을 넣어 실온에서 15분간 발색시키고, 다시 1M 황산을 넣어 반응 (발색)을 중지시키면 반응액의 색깔은 노란색을 띠게 되는데, 반응의 진행 정도에 따라 노란색의 정도가 다르게 나타남을 확인하였다.After 3 hours, the second collagen antibody bound to the chromophore was put into a 96-well plate and reacted for another 15 minutes. After 15 minutes, the color development inducing substance was added, coloring was performed at room temperature for 15 minutes, and 1 M sulfuric acid was added to stop the reaction (color development). The color of the reaction solution became yellow, and the degree of yellow was different depending on the progress of the reaction Respectively.
노란색을 띠는 96-웰 플레이트 (96-well plate)의 흡광도를 흡광계를 이용하여 405nm에서 측정하였고, 하기 수학식 1에 의해 콜라게나제의 합성 정도를 계산하였다. 이때 조성물을 처리하지 않은 군의 채취된 세포배양액의 반응 흡광도를 대조군으로 하였다. 즉, 비처리군에서의 콜라게나제의 발현 정도를 100으로 하고, 이에 대비하여 시험물질을 처리한 군에서의 콜라게나제의 발현 정도를 구하였으며, 그 결과는 하기 표 2에 나타내었다. The absorbance of a yellow 96-well plate was measured at 405 nm using a spectrophotometer, and the degree of synthesis of collagenase was calculated by the following equation (1). At this time, the reaction absorbance of the cell culture obtained from the untreated group was used as a control. That is, the degree of expression of collagenase in the untreated group was set at 100, and the degree of expression of collagenase in the group treated with the test substance was determined, and the results are shown in Table 2 below.
[수학식 1][Equation 1]
콜라게나제 발현 정도(%)=물질 처리 세포군의 흡광도/대조군의 흡광도 X 100(%) = Absorbance of the substance-treated cell group / absorbance of the control group X 100
상기 표 2에서 알 수 있는 바와 같이, 콩잎 추출물은 콩 추출물보다 콜라게나제를 더 효과적으로 억제하였으며, 항산화 물질로 알려져 있는 토코페롤보다 콜라게나제의 발현 억제능이 우수함을 확인할 수 있었다. As can be seen from Table 2, the soybean leaf extract inhibited the collagenase more effectively than the soybean extract, and it was confirmed that the tocopherol, which is known as an antioxidant, is superior in inhibiting the expression of collagenase.
[시험예 3] 세포 증식 증가 효과[Test Example 3] Effect of increasing cell proliferation
피부 재생 효과로써 피부세포(Skin cell) 증식효과를 보기 위해 인체 정상 섬유아세포(fibroblast)를 96-웰 마이크로 플레이트(96-wellmicroplate)의 각 웰(well)에 1×104 세포가 되도록 접종하여 디엠이엠(DMEM : Dulbecco's Modified Eagle's Medium) 배지에서 24시간 배양하였다. 배양 후 콩잎 추출물 과 콩 추출물을 최종농도 100 μg/ml이 되도록 조정한, 혈청이 없는 디엠이엠 배지로 교체한 후 24시간 더 배양하였다. 3-(4,5-디메틸씨아졸-2-일)-2,5-디페닐테트라졸리움(MTT: (3-(4,5-dimdimeth-thiazol-2-yl)2,5-diphenyl tetrazolium : 5mg/ml) 10㎕씩 첨가하고 4시간 방치 후, 배지 부분을 버렸다. 각 웰당 100㎕의 디메틸 설폭사이드 용액을 가하여 마이크로플레이트 리더로 570nm에서 흡광도를 측정하였다.Human skin fibroblasts were inoculated into each well of a 96-well microplate so as to have a skin cell reproduction effect as a skin regeneration effect so as to be 1 × 10 4 cells. And cultured in DMEM (Dulbecco's Modified Eagle's Medium) for 24 hours. After the cultivation, the soybean leaf extract and the soybean extract were replaced with serum-free DMEM medium adjusted to a final concentration of 100 μg / ml, and cultured for another 24 hours. 3- (4,5-Dimethylthiazol-2-yl) 2,5-diphenyl tetrazolium (MTT: 5 mg / ml) was added to each well and allowed to stand for 4 hours, after which the medium was discarded. 100 μl of dimethyl sulfoxide solution was added to each well and the absorbance was measured at 570 nm with a microplate reader.
상기 과정의 시험을 실시하여 하기 수학식 2로부터 세포증식 효과를 구해 그 결과를 하기 표 3에 나타내었다.Tests of the above procedure were conducted to determine the cell proliferation effect from the following equation (2), and the results are shown in Table 3 below.
[수학식 2]&Quot; (2) "
세포증식 효과(%)=[(추출물처리시 흡광도-대조군 흡광도)/대조군 흡광도]X100Cell proliferation effect (%) = [(absorbance at the time of extract treatment - control absorbance) / control absorbance] X100
농도(μg/ml)Test substance
Concentration (μg / ml)
상기 표 3에서 보는 바와 같이 콩잎 추출물은 콩 추출물보다 더 우수하게 정상 섬유아세포를 증식하는 것으로 확인되었다. As shown in Table 3, it was confirmed that the soybean leaf extract proliferated better than the soybean extract.
[시험예 4] 티로시나제 저해 효과Test Example 4: Effect of tyrosinase inhibition
티로시나제 효소는 버섯류(Mushroom)로부터 추출된 것으로 시그마(SIGMA)사의 것을 사용하였다. 먼저 기질인 티로신을 증류수에 용해시켜 0.3mg/ml의 용액으로 만들고 그 용액을 1.0ml씩 시험관에 넣은 다음, 여기에 포타슘-포스페이트 완충용액(0.1mol 농도, pH 6.8) 1.0ml 및 증류수 0.7ml를 첨가하였다.The tyrosinase enzyme was extracted from Mushroom and was obtained from Sigma (SIGMA). First, the substrate, tyrosine, is dissolved in distilled water to make a 0.3 mg / ml solution. The solution is put into a test tube in an amount of 1.0 ml. Then, 1.0 ml of potassium phosphate buffer solution (0.1 mol concentration, pH 6.8) and 0.7 ml of distilled water .
본 발명의 콩잎 추출물과 비교실시예 1의 콩 추출물을 에탄올 용액에 최종 농도 1 mg/ml 농도로 혼합하여 준비한 시료액 0.2ml를 반응액에 넣은 다음 37℃ 항온조에서 10분 동안 반응시켰다. 이때 대조군은 각 시료액 대신 용매만을 0.2ml 넣은 것으로 준비하였고, 양성대조군으로는 아스코르브산을 사용하였다. 이 반응액에 2500유닛/ml의 티로시나제 용액 0.1ml씩을 넣고 다시 37℃ 항온조에서 10분 동안 반응시켰다. 이 반응액이 든 시험관을 얼음물속에 넣어서 급냉시켜 반응을 중지시키고 광전분광분석계로 파장 475nm에서의 흡광도를 측정하였으며, 그 결과를 하기 표 2에 나타내었다. 각각의 티로시나제 저해 효과는 하기 수학식 3로 산출하였다.0.2 ml of the sample liquid prepared by mixing the soybean leaf extract of the present invention and the soybean extract of Comparative Example 1 in an ethanol solution at a final concentration of 1 mg / ml was added to the reaction solution and reacted for 10 minutes in a 37 ° C thermostatic bath. At this time, the control group was prepared by adding 0.2 ml of a solvent instead of each sample solution, and ascorbic acid was used as a positive control group. 0.1 ml of tyrosinase solution of 2500 units / ml was added to the reaction solution, followed by reaction at 37 ° C for 10 minutes. The reaction tube was immersed in ice water to quench the reaction, and absorbance at a wavelength of 475 nm was measured with a photoelectron spectrometer. The results are shown in Table 2 below. Each tyrosinase inhibitory effect was calculated by the following equation (3).
[수학식 3]&Quot; (3) "
표 4와 같이, 본 발명에 의한 콩잎 추출물은 티로시나제 억제율이 콩 추출물 보다 우수 할뿐만 아니라 공지된 티로시나제 저해제인 아스코르브산보다 훨씬 높아 미백 효과가 매우 우수함을 알 수 있다.As shown in Table 4, it can be seen that the soybean leaf extract according to the present invention is superior in the inhibition rate of tyrosinase to soybean extract, and is much higher than ascorbic acid, which is a known tyrosinase inhibitor, so that the whitening effect is excellent.
[시험예 5] 피부 보습 시험[Test Example 5] Skin moisturizing test
일차 배양한 인간의 각질형성세포를 배양용 플라스크에 넣어 바닥에 부착시킨 후, 콩잎 추출물과 콩 추출물을 1 mg/ml 농도로 배양액에 첨가하여, 세포가 바닥 면적의 80~90% 정도 자랄 때까지 5일간 배양하였다. 이 세포를 수확하여 PBS (phosphate buffered saline)로 세척한 뒤, 2% SDS (Sodium Dodecyl Sulfate)와 20mM 농도의 DTT (Dithiothreitol)를 함유한 10mM 농도의 트리스-염산 완충액 (Tris-HCl, pH 7.4) 1㎖를 가하여 3분간 소니케이션 (sonication)하고 난 후, 10분간 끓였다. 그 후, 이를 1200rpm으로 30분간 원심 분리하고 침전물을 분리하여 PBS 1㎖에 현탁시킨 후, 340nm에서 흡광도를 측정하였다.Primary cultured human keratinocytes were placed in a culture flask and adhered to the bottom. Then, the soybean-leaf extract and the soybean extract were added to the culture solution at a concentration of 1 mg / ml, until the cells reached 80 to 90% And cultured for 5 days. The cells were harvested and washed with PBS (phosphate buffered saline). Then, 10 mM Tris-HCl buffer (Tris-HCl, pH 7.4) containing 2% SDS (sodium dodecyl sulfate) and 20 mM DTT (Dithiothreitol) 1 ml was added thereto, sonicated for 3 minutes, and boiled for 10 minutes. Thereafter, it was centrifuged at 1200 rpm for 30 minutes, the precipitate was separated, suspended in 1 ml of PBS, and the absorbance at 340 nm was measured.
이와 별도로 상기 소니케이션 후의 용액 일부를 취하여 단백질 함량을 측정하여, 세포 분화 정도의 평가시 기준으로 삼았다. 저칼슘(0.03mM) 처리군과 고칼슘(1.2mM) 처리군을 각각 음성/양성 대조군으로 하고, 저칼슘 농도에 시험물질을 첨가하여 실시한 시험 결과를 하기 표 5에 나타내었다.Separately, a portion of the solution after the sonication was taken to measure the protein content and used as a criterion when evaluating the degree of cell differentiation. The test results are shown in Table 5 below, in which low calcium (0.03 mM) and high calcium (1.2 mM) treatment groups were used as negative / positive control groups and low calcium concentration test substances were added.
상기 표 5에 나타난 바와 같이, 각질형성세포 분화시 생성되는 CE (Cornified Envelop)의 양을 측정하여 세포분화 촉진효과를 비교한 결과, 본 발명에 의한 콩잎 추출물은 콩 추출물에 비해 각질형성세포에서 분화를 촉진하는 것을 알 수 있었다. As shown in Table 5, when the amount of CE (cornified envelope) produced during keratinocyte differentiation was measured to compare cell differentiation promoting effects, It can be seen that the soybean leaf extract according to the present invention promotes differentiation from keratinocytes in comparison with soybean extract.
[실시예 7] 자극감 시험[Example 7]
공지된 미백 물질인 코직산과 본 발명에서 유효성분으로 사용되는 콩잎 추출물의 사용성을 비교하기 위하여, 따가움, 화끈거림 등의 자극감에 민감한 패널 15명을 대상으로 따가움, 화끈거림 등 자극감의 정도를 실험하였다.In order to compare the usability of kojic acid, which is a known whitening substance, with the use of an extract of canola leaves used as an active ingredient in the present invention, the degree of irritation such as stinginess and burning was examined in 15 panel members susceptible to stimuli such as stinging and burning .
피험자에게 코직산(kojic acid, YM chemical 통해 구입)과 상기 실시예 1에서 얻은 콩잎 추출물을 각각 0.5㎖씩 좌우를 무작위로 바꾸어 적용하여 문지르고 0.1점 단위로 하여 0~3.0 사이의 점수를 매기도록 하였다. 그 결과는 하기 표 6에 나타내었다.The subjects were randomly assigned 0.5ml each of kojic acid (purchased from YM chemical) and the luciferase extract obtained in Example 1 at random, and rubbed and rubbed, and the score was scored between 0 and 3.0 in units of 0.1 point. The results are shown in Table 6 below.
<평가기준><Evaluation Criteria>
0 ~ 0.4: 자극 없음0 to 0.4: No stimulation
0.5 ~ 1.0: 약간 자극이 있음0.5 to 1.0: slight irritation
1.1 ~ 2.0: 보통 정도의 자극이 있음1.1 to 2.0: normal irritation
2.1 ~ 3.0: 자극이 심함2.1 ~ 3.0: Severe irritation
상기 표 6에서 알 수 있는 바와 같이, 코직산의 경우는 따가움, 화끈거림이 어느 정도 있어 보통 약간 느낄 수 있는 정도의 자극감이 있는 것으로 나타났다. 반면, 본 발명에서 사용되는 콩잎 추출물은 화끈거림과 따가움 모두 거의 느낄 수 없을 정도여서 대체로 자극이 없는 것으로 나타냈다.이하, 본 발명에 따른 조성물의 제형 예를 설명하나, 약학 조성물 및 화장료 조성물은 여러 가지 제형으로 응용 가능하며, 이는 본 발명을 한정하고자 함이 아닌 단지 구체적으로 설명하고자 함이다.As can be seen in Table 6, in the case of kojic acid, it was found that there was some degree of stinging and burning, and there was a feeling of irritation to the extent that it could feel a slight feeling. The composition of the composition according to the present invention will be described below, but the pharmaceutical composition and the cosmetic composition may be variously applied to various kinds of cosmetic compositions. It is to be understood that the present invention is not limited thereto but is only specifically described.
[제제예 1] 연질캅셀제[Formulation Example 1] Soft capsule
콩잎 추출물 150 mg, 팜유 2 mg, 팜경화유 8 mg, 황납 4 mg 및 레시틴 6 mg을 혼합하고, 통상의 방법에 따라 1 캡슐당 400 mg씩 충진하여 연질캅셀을 제조하였다.Soft bean capsule was prepared by mixing 150 mg of soybean leaf extract, 2 mg of palm oil, 8 mg of palm kernel oil, 4 mg of yellow radish and 4 mg of lecithin and 400 mg per capsule according to a conventional method.
[제제예 2] 정제[Formulation Example 2] Tablets
콩잎 추출물 150 mg, 포도당 100 mg, 홍삼추출물 50 mg, 전분 96 mg 및 마그네슘 스테아레이트 4 mg을 혼합하고 30% 에탄올을 40 mg 첨가하여 과립을 형성한 후, 60℃에서 건조하고 타정기를 이용하여 정제로 타정하였다.The granules were prepared by mixing 150 mg of soybean leaf extract, 100 mg of glucose, 50 mg of red ginseng, 96 mg of starch and 4 mg of magnesium stearate and 40 mg of ethanol was added thereto, followed by drying at 60 ° C., Respectively.
[제제예 3] 과립제[Formulation Example 3] Granules
콩잎 추출물 150 mg, 포도당 100 mg, 홍삼추출물 50 mg 및 전분 600 mg을 혼합하고 30% 에탄올을 100 mg 첨가하여 과립을 형성한 후, 60℃에서 건조하여 과립을 형성한 다음 포에 충진하였다. 내용물의 최종 중량은 1 g으로 하였다.The granules were formed by mixing 150 mg of soybean leaf extract, 100 mg of glucose, 50 mg of red ginseng extract and 600 mg of starch, and 100 mg of 30% ethanol was added thereto, followed by drying at 60 ° C to form granules. The final weight of the contents was 1 g.
[제제예 4] 드링크제[Formulation Example 4] Drinking agent
콩잎 추출물 150 mg, 포도당 10 g, 홍삼추출물 50 mg, 구연산 2 g 및 정제수 187.8 g을 혼합하고 병에 충진하였다. 내용물의 최종 용량은 200 ml로 하였다.150 mg of soybean leaf extract, 10 g of glucose, 50 mg of red ginseng extract, 2 g of citric acid and 187.8 g of purified water were mixed and filled in a bottle. The final volume of the contents was adjusted to 200 ml.
[제제예 5] 건강 식품의 제조[Formulation Example 5] Preparation of health food
콩잎 추출물 .............................. 1000 ㎎ Soybean leaf extract .............................. 1000 mg
비타민 혼합물 Vitamin mixture
비타민 A 아세테이트.......................70 ㎍ Vitamin A Acetate ....................... 70 ㎍
비타민 E ............................................ 1.0 ㎎ Vitamin E ............................................ 1.0 mg
비타민 B1........................................... 0.13 ㎎ Vitamin B1 ........................................... 0.13 mg
비타민 B2 .......................................... 0.15 ㎎ Vitamin B2 .......................................... 0.15 mg
비타민 B6........................................... 0.5 ㎎ Vitamin B6 ........................................... 0.5 mg
비타민 B12......................................... 0.2 ㎍ Vitamin B12 .............................. 0.2 g
비타민 C............................................. 10 ㎎ Vitamin C ............................................. 10 mg
비오틴.................................................. 10 ㎍ Biotin ................................................. 10 [mu] g
니코틴산아미드.................................. 1.7 ㎎ Nicotinic acid amide .................................. 1.7 mg
엽산...................................................... 50 ㎍ Folic acid ................................................. ..... 50 μg
판토텐산 칼슘.................................... 0.5 ㎎ Calcium pantothenate .................................... 0.5 mg
무기질 혼합물 Mineral mixture
황산제1철.......................................... 1.75 ㎎ Ferrous sulfate .......................................... 1.75 mg
산화아연.............................................. 0.82 ㎎ Zinc oxide ............................................ 0.82 mg
탄산마그네슘...................................... 25.3 ㎎ Magnesium carbonate ...................................... 25.3 mg
제1인산칼륨.......................................... 15 ㎎ Potassium Phosphate ......................................... 15 mg
제2인산칼슘.......................................... 55 ㎎ Secondary calcium phosphate ...................................... 55 mg
구연산칼륨............................................ 90 ㎎ Potassium citrate ............................................ 90 mg
탄산칼슘.............................................. 100 ㎎ Calcium carbonate .............................................. 100 mg
염화마그네슘..................................... 24.8 ㎎ Magnesium chloride ..................................... 24.8 mg
상기의 비타민 및 미네랄 혼합물의 조성비는 비교적 건강식품에 적합한 성분을 바람직한 실시예로 혼합 조성하였지만, 그 배합비를 임의로 변형 실시하여도 무방하며, 통상의 건강식품 제조방법에 따라 상기의 성분을 혼합한 다음, 과립을 제조하고, 통상의 방법에 따라 건강식품 조성물 제조에 사용할 수 있다.Although the composition ratio of the above-mentioned vitamin and mineral mixture is comparatively mixed with a composition suitable for health food as a preferred embodiment, the compounding ratio may be arbitrarily modified, and the above ingredients are mixed according to a conventional method for producing healthy foods , Granules can be prepared and used in the manufacture of health food compositions according to conventional methods.
[제제예 6] 건강 음료의 제조 [Formulation Example 6] Preparation of health drink
콩잎 추출물............................ 1000 ㎎ Canned Bean Extract ............................ 1000 mg
구연산..................................................... 1000 ㎎ Citric acid ................................................. ... 1000 mg
올리고당..................................................... 100 g oligosaccharide................................................. .... 100 g
매실농축액..................................................... 2 g Plum concentrate ................................................ ..... 2 g
타우린............................................................ 1 g Taurine ................................................. ........... 1 g
정제수를 가하여 전체......................... 900 ㎖ Purified water was added to the mixture to complete 900 ml
통상의 건강음료 제조방법에 따라 상기의 성분을 혼합한 다음, 약 1시간 동안 85℃에서 교반 가열한 후, 만들어진 용액을 여과하여 멸균된 2ℓ용기에 취득하여 밀봉 멸균한 뒤 냉장 보관한 다음 본 발명의 건강음료 조성물 제조에 사용한다.The above components were mixed according to a conventional health drink manufacturing method, and the mixture was heated for about 1 hour at 85 DEG C with stirring, and the solution thus prepared was filtered to obtain a sterilized 2-liter container, which was sealed and sterilized, ≪ / RTI >
상기 조성비는 비교적 기호음료에 적합한 성분을 바람직한 실시예로 혼합 조성하였지만 수요계층이나, 수요국가, 사용 용도 등 지역적, 민족적 기호도에 따라서 그 배합비율을 임의로 변형 실시하여도 무방하다. 본 발명이 속한 기술 분야에서 통상의 지식을 가진 자라면 상기 내용을 바탕으로 본 발명의 범주 내에서 다양한 응용 및 변형을 행하는 것이 가능할 것이다. Although the composition ratio is relatively mixed with a component suitable for a favorite drink, it is also possible to arbitrarily modify the blending ratio according to the regional or national preference such as the demand level, the demanding country, the use purpose, and the like. It will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention as defined by the appended claims.
[제형예 1] 로션[Formulation Example 1] Lotion
[제형예 2] 크림[Formulation Example 2] Cream
[제형예 3] 팩[Formulation Example 3] Pack
[제형예 4] 미용액[Formulation Example 4]
[제형예 5] 연고[Formulation Example 5] Ointment
[제형예 6] 연고[Formulation Example 6] ointment
이상으로 본 발명 내용의 특정한 부분을 상세히 기술하였는 바, 당업계의 통상의 지식을 가진 자에게 있어서 이러한 구체적 기술은 단지 바람직한 실시태양일 뿐이며, 이에 의해 본 발명의 범위가 제한되는 것이 아닌 점은 명백할 것이다. 따라서, 본 발명의 실질적인 범위는 첨부된 청구항들과 그것들의 등가물에 의하여 정의된다고 할 것이다.While the present invention has been particularly shown and described with reference to specific embodiments thereof, those skilled in the art will readily appreciate that many modifications are possible in the exemplary embodiments without materially departing from the novel teachings and advantages of this invention. something to do. Accordingly, the actual scope of the present invention will be defined by the appended claims and their equivalents.
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