KR20080049332A - Skin-whitening extract from oviductus ranae - Google Patents
Skin-whitening extract from oviductus ranae Download PDFInfo
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- KR20080049332A KR20080049332A KR1020060119772A KR20060119772A KR20080049332A KR 20080049332 A KR20080049332 A KR 20080049332A KR 1020060119772 A KR1020060119772 A KR 1020060119772A KR 20060119772 A KR20060119772 A KR 20060119772A KR 20080049332 A KR20080049332 A KR 20080049332A
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- extract
- skin
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/98—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution of animal origin
- A61K8/987—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution of animal origin of species other than mammals or birds
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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
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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
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
- A61Q19/02—Preparations for care of the skin for chemically bleaching or whitening 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
- 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
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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
- A23V2250/00—Food ingredients
- A23V2250/20—Natural extracts
- A23V2250/204—Animal extracts
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Dermatology (AREA)
- Nutrition Science (AREA)
- Mycology (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Food Science & Technology (AREA)
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Abstract
Description
도 1은 PTU에 대한 멜라닌 세포의 세포생존도 및 멜라닌 생성 억제율을 나타낸 비교도이고,1 is a comparative diagram showing the cell viability and melanin production inhibition rate of melanocytes to PTU,
도 2는 합마유 추출물에 대한 멜라닌 세포의 세포생존도 및 멜라닌 생성 억제율을 나타낸 비교도이다.Figure 2 is a comparative diagram showing the cell viability and melanin production inhibition rate of melanocytes against the extract of the corn oil.
본 발명은 미백용 합마유 추출물 및 이를 포함하는 조성물에 관한 것으로, 보다 상세하게는 개구리 수란관 건조물인 합마유 추출물 및 이를 포함하는 조성물에 관한 것이다.The present invention relates to a whitening seahorse oil extract and a composition comprising the same, and more particularly, to a horseradish oil extract and a composition comprising the same as a dry frog tube.
개구리인 합사마(forest frog, Rana temporaria chensinensis David)는 우리나라의 중요한 야생약용동물로서, 중국임와(Rana temporaria chensinensis David, Hashima-frog), 흑룡강임와(Rana amurensis B.)라는 속칭으로 불리며, 합십마(하쓰마), 홍두전계, 합마(하마), 전계[田鷄, 요녕주요약재(遼寧主要藥材)], 설합[雪蛤, 약재자료휘편(藥材資料彙編)], 합마(하마), 파랍와[빠라와, 길림중초약(吉林中草 藥)]라고도 불린다. 중국임와(Rana temporaria chensinensis David)는 외형은 청개구리와 비슷하고 몸길이가 평균 5 cm이다. 여름철 숲 속에서 활동할 때는 황갈색, 가을, 겨울철은 갈색을 띤다. 주로 길림(吉林), 요녕(遼寧), 흑룡강(黑龍江), 내몽고(內蒙古) 등지에 분포한다. 우리나라에 분포가 비교적 넓으며 주로 장백산지역에서 생산된다. 본초도경[本草圖經, 송(宋)·소송(蘇頌)]의 하마 항에는 "크고 황색이며 흔히 바위틈에 숨어 있으며 기(氣)를 마시고 바람과 이슬을 마시며 잡충을 먹지 않는다; 산합(山蛤, 산하)이라고 부르며 산에서 사는 사람들도 그것을 먹는다; 주로 소아의 피로와 마른 증상(勞瘦) 및 감질(疳疾) 등에 가장 효과가 있다."라고 기술되어 있다. 본초강목[本草綱目, 명(明)·이시진(李時珍)]은 본초도경(本草圖經)의 산합(山蛤)에 대한 서술을 그대로 인용하여 그것을 하마항과 따로 분류하고 그의 형태, 생태, 색깔 면에서는 하마류와 구별이 있음을 나타냈다. Forest frog, Rana temporaria chensinensis David), China imwa (Rana as important wild medicinal animals in Korea temporaria chen sinensis David, Hashima-frog) and Rana amurensis B. are commonly known as Hapma, Hatsuma, Hadama, Hama, and electric field. It is also called Seolgi (雪蛤, medicinal materials 편 料 彙編), Hapma (Hama), and Parapwa (Parawa, Jilin Chinese Herbal Medicine). Chinese Rana temporaria chensinensis David) is similar in appearance to tree frogs and averages 5 cm in length. It is yellowish brown in the summer forest and brown in autumn and winter. It is mainly distributed in Jilin, Liaoning, Heilongjiang and Inner Mongolia. It is relatively wide in Korea and mainly produced in Changbai Mountain. In the Hama Port of the Tosho-cho, there are "large, yellow, often hidden in the cracks of rocks, drinking qi, drinking wind and dew, and not eating insects;山 蛤 (below) and people who live in the mountains eat it; it is most effective in children's fatigue, dry symptoms (및) and sensation (疳 疾). " The main herbaceous tree, Ming and Ishijin, cited the description of the mountainous landscape of the main island as it is, classifies it separately from Hama Port, and its form, ecology, and color. In aspect, it is distinguished from hippopotamus.
합사마의 건조시킨 수란관인 합마유는 진귀한 중약재로, <중약지>, <중약대사전>, <전국중초약휘편> 등 여러 저서와 문헌에서는 중국임와(中國林蛙)와 흑룡강임와(黑龍江林蛙)를 합사마유의 동물기원으로 여기고 있다. <중화인민공화국약전>에서는 합마유라는 이름으로 수록하였으며 그 동물기원을 중국임와라 규정하였다. 합마유(Oviductus Ranae: Forest Frog's Oviduct)는 속칭으로 전계유(田鷄油), 합십마유, 합사마유라고도 한다. 합마유는 그 맛이 감(甘), 함(咸)하고 그 성질은 평(平)하다. 그 주성분은 단백질로 알려져 있고 그 외에 수분, 지방, 무질소 유기물, 함질소 유기물, 광물질, 인, 유황 및 여러 가지 종류의 호르몬, 비타민A, D 등으로 밝혀져 있다. 또한 스테로이드 호르몬 성분 연구도 문헌에 밝혀져 있다. 그 기능으로는 보신(補腎) 익정(益精), 양음(陽陰) 윤폐(潤肺)한다. 음허(陰虛) 체약(體弱), 신피(神疲) 핍력(乏力), 심계실명(心悸失眠)에 사용되며 잠잘 때 땀이 그치지 않고 기침하고 각혈할 때 사용한다. 이러한 합마유의 밝혀진 약리방면으로는 항노쇠작용, 진해(鎭咳) 거담(祛痰) 작용, 운동실조에 대한 조절작용, 생장발육 및 성 조숙 촉진작용이 있다.Hap dried horsetail, a dried oak yam, is a rare medicinal herb. Is regarded as the animal origin of hapsa horse oil. The People's Republic of China Pharmacopoeia contained the name of horse oil and defined its origin as China. Horse oil ( Oviductus) Ranae : Forest Frog's Oviduct) is also known as field oil, hapsim horse oil, and hapsa horse oil. The horsetail oil tastes and tastes flat and its properties are flat. Its main constituents are known as proteins and in addition to water, fat, nitrogen-free organics, nitrogen-containing organics, minerals, phosphorus, sulfur and various hormones, vitamins A and D. Steroid hormone component studies are also described in the literature. Its functions are bosin (補腎 精), and yin and yang (yang 윤) lubricity. It is used for yin huo (약), sinpi pip force, and blindness of the heart, and when coughing and bleeding without stopping sweating. The pharmacological aspects of these oils are anti-aging, antitussive expectoration, control of ataxia, growth growth and sexual precoction.
사람의 피부는 노화가 진행됨에 따라서 피부탄력의 상실, 피부 주름의 형성, 피부색의 변화, 피부 베리어 기능상실, 피부면 역기능의 상실 등의 물리 피부화학적인 변화 동반한다. 특히 중금속, 황사, 오존, 자외선, 화장품 기재등과 같은 외부 환경의 자극 물질은 피부 세포의 활성 저하, 피부세포의 괴사, 세포 변성 등을 촉진하여 주름형성촉진, 색소침착, 면역기능의 상실, 염증 증가 등을 촉진하는 주요 인자이다.As the skin ages, the human skin is accompanied by physical and dermochemical changes such as loss of skin elasticity, formation of skin wrinkles, change of skin color, loss of skin barrier function, loss of skin immune function. In particular, irritants in the external environment such as heavy metals, yellow sand, ozone, ultraviolet rays, cosmetics, etc. promote the deactivation of skin cells, necrosis of skin cells, and cell degeneration, which promotes wrinkle formation, pigmentation, loss of immune function, and inflammation. It is the main factor that promotes the increase.
멜라닌은 자연계에 널리 분포하는 페놀류의 생 고분자 물질로 검은 색소와 단백질의 복합체로 환경에 대한 세포의 생존력과 경쟁력을 높여주는 물질이다. 멜라닌 색소의 형성에는 유전적 요인, 호르몬 분비, 스트레스 등과 관련된 생리적 요인 및 자외선 조사 등과 같은 환경적 요인이 영향을 미친다. 멜라닌은 피부에 존재하면서 자외선 등으로부터 신체를 보호하는 중요한 기능이 있지만 멜라닌의 과잉 생산은 인체에 기미, 주근깨 등을 형성하고 피부 노화를 촉진하거나 피부암의 유발에 관여하는 것으로 보고되었다. 또한, 오존층 파괴로 자외선 노출이 심해지면서 발생하는 과잉된 멜라닌 합성은, 인체의 피부 노화에 치명적인 해를 가져오고 있 다. 이에, 강력한 멜라닌 생합성 저해활성 물질의 개발에 많은 노력을 기울이고 있다.Melanin is a biopolymer of phenols widely distributed in nature. It is a complex of black pigment and protein that enhances cell viability and competitiveness for the environment. The formation of melanin pigment is influenced by genetic factors, physiological factors related to hormone secretion, stress, and environmental factors such as ultraviolet irradiation. While melanin is present in the skin and has an important function of protecting the body from ultraviolet rays and the like, the excessive production of melanin has been reported to form blemishes, freckles, etc. in the human body, promote skin aging and induce skin cancer. In addition, excessive melanin synthesis, which is caused by severe ultraviolet exposure due to the ozone layer destruction, has been fatal to human skin aging. Thus, much effort has been made in the development of powerful melanin biosynthesis inhibitors.
멜라닌의 생합성 대사는 피부암과 관련하여 최근 집중적으로 연구되고 있고 이로부터 다양한 멜라닌 생성 저해제 등이 개발되어 의약품 산업에서 피부 질환 치료제, 화장품 산업에서 기미, 주근깨 등의 예방 및 치료용 피부 미백제 또는 식품 산업에서 갈변 방지제 등으로 적용되고 있다. 또한 최근에는 환경문제와 관련하여 상기의 예방 및 치료제의 수요가 급격히 증가하고 있다.The biosynthetic metabolism of melanin has recently been intensively researched in connection with skin cancer, and various melanin-producing inhibitors have been developed from this, and thus, in the pharmaceutical industry, the skin lightening agent in the pharmaceutical industry, the cosmetic industry in the skin lightening agent for the prevention and treatment of blemishes and freckles, or in the food industry It is applied as a browning inhibitor. In recent years, the demand for the above-mentioned prophylactic and therapeutic agents is rapidly increasing in relation to environmental problems.
멜라닌 생합성 속도조절단계의 효소인 타이로시나제(tyrosinase)를 직접 억제한다. 타이로시나제는 구리결합 효소로서, 동물, 식물, 미생물 및 사람 등에 널리 분포하고 모노하이드록시 또는 디하이드록시페닐알라닌(dihydroxy-phenylalanine, DOPA) 등의 페놀 화합물에서 호기적 산화를 촉진하고, 자외선에 심하게 노출된 피부에 멜라닌 토스를 침착시켜 피부의 노화와 손상을 유발시키는 작용을 한다. It directly inhibits tyrosinase, an enzyme of melanin biosynthesis rate regulation. Tyrosinase is a copper-binding enzyme, widely distributed in animals, plants, microorganisms and humans, and promotes aerobic oxidation in phenolic compounds such as monohydroxy or dihydroxyphenylalanine (DOPA). Melanin toss is deposited on heavily exposed skin, causing aging and damage to the skin.
지금까지 멜라닌 생성 저해제의 연구는 주로 타이로시나제 저해제를 개발하는 방향으로 집중되어 왔고, 대표적인 타이로시나제 저해제로는 타이로시나제 활성부위의 구리 이온에 대한 킬레이트 형성물질, 퀴논류를 페놀류로 환원시키는 아스코빅산 등의 환원제 그리고 타이로시나제 자체를 변성시키는 비설파이트 제제 등을 들 수 있다.Until now, research on melanogenesis inhibitors has been mainly focused on developing tyrosinase inhibitors. Representative tyrosinase inhibitors include chelates, quinones, and chelates for copper ions in the active region of tyrosinase. And reducing agents such as ascorbic acid to be reduced, and bisulfite preparations that denature tyrosinase itself.
상기와 같이, 타이로시나제 저해제가 다양한 미백제 등으로 개발되어 현재 사용되고 있지만, 여러 가지 문제점도 동시에 제기되고 있다. 실제로, 기미, 주근 깨, 반점 및 임신기 과색소 침착과 같은 과잉 색소증 치료에 국부적으로 사용되고 있는 4-히드록시아니솔 및 하이드로퀴논 등은 강력한 멜라닌 생성 저해활성은 있으나 동시에 색소세포의 변성 또는 치사를 유발하고 세포 본래의 기능을 손상시키는 등의 부작용을 나타낸다. 특히 하이드로퀴논 계열의 화합물은 멜라닌 생합성을 저해하는 미백용 크림으로 개발되어 사용되었으나, 세포 독성으로 인한 피부 자극 또는 피부병을 유발하는 것으로 알려져 현재 일부 국가에서만 사용이 허가되고 있는 실정이다.As described above, although tyrosinase inhibitors have been developed and used in various whitening agents and the like, various problems have been raised at the same time. Indeed, 4-hydroxyanisole and hydroquinone, which are used locally to treat hyperpigmentation such as blemishes, freckles, spots and gestational hyperpigmentation, have potent melanogenesis-inhibiting activities but at the same time prevent pigmentation or lethality. Side effects such as causing and impairing the intrinsic function of the cell. In particular, hydroquinone-based compounds have been developed and used as a whitening cream that inhibits melanin biosynthesis, but are known to cause skin irritation or dermatosis due to cytotoxicity, and are currently used only in some countries.
더불어, 멜라닌 생성 저해제로서 파라메톡시페놀(p-methoxy phenol), 코직산(kojic acid) 또는 알부틴(arbutin) 등이 사용되어 왔으나, 이들은 불충분한 미백효과, 피부에 대한 안전성 문제, 화장료에 배합시 제형화 및 제형내 안정성 문제 등으로 인해 그 사용이 제한되고 있는 실정이다.In addition, wateuna as melanin production inhibitors include para-methoxyphenol (p -methoxy phenol), kojic acid (kojic acid) or arbutin (arbutin) is used, all of which are sufficient whitening effect, safety problems for the skin, when formulated in cosmetic formulations The use is limited due to problems in the formulation and stability in the formulation.
또한, 멜라닌 생성을 저해하기 위한 목적으로 비타민 C 및 그 유도체 등이 사용되어 왔으나, 이들 역시 타이로시나제 저해 활성이 낮다는 문제가 있다. 따라서, 세포독성이 낮고, 소량에 의해서도 타이로시나제 활성 및 멜라닌 생성을 저해할 수 있는 새로운 저해제의 개발이 시급한 실정이다(Chen J & Agric J, Food, 39, 1991).In addition, vitamin C and derivatives thereof have been used for the purpose of inhibiting melanin production, but they also have a problem of low tyrosinase inhibitory activity. Therefore, there is an urgent need to develop new inhibitors that have low cytotoxicity and that can inhibit tyrosinase activity and melanin production even with a small amount (Chen J & Agric J, Food , 39, 1991).
한편, 기존 미백 효능 물질들 이외에 천연물 중에서 미백 활성성분을 찾기 위한 연구가 계속 이루어지고 있는데, 그 중 상백피(일본공개특허 소55-44375, 소64-26507, 소64-83009, 평1-25687, 평5-139950, 한국공개특허 제1992-002109호, 제1997-021273호), 감초(일본공개특허 소60-214721, 소63-23809, 소64-63506, 평1- 149706, 한국공개특허 제1992-002109호, 제1997-025601호)등 다수의 식물추출물이 타이로시나제에 작용하여 멜라닌 생성을 억제한다는 사실이 밝혀졌으나, 이들 역시 안전성, 안정성, 변색 가능성 등의 측면에서 화장품이나 의약품에 유효 농도 이상으로 사용하는 데는 많은 문제점이 있으며 만족할 만한 효과를 내지 못하는 실정이다. 그리고, 현재까지 개구리의 수란관(이하, 합마유)의 추출물을 포함하는 멜라닌 생성에 대하여 저해효과를 나타내는 조성물에 관한 특허는 없는 실정이다.On the other hand, research to find the whitening active ingredient in natural products in addition to the existing whitening efficacy substances are continuously being made, among them, Sangbaekpi (Japanese Patent Publication No. 55-44375, So64-26507, So64-83009, Pyeong1-25687, Hei 5-139950, Korean Laid-Open Patent Nos. 1992-002109, 1997-021273), Licorice (Japanese Laid-Open Patent Nos. 60-214721, Small 63-23809, Small 64-63506, Flat 1-149706, Korean Laid-Open Patent No. Many plant extracts, such as 1992-002109 and 1997-025601), have been shown to act on tyrosinase to inhibit melanin production, but they are also used in cosmetics and pharmaceuticals in terms of safety, stability, and discoloration potential. There are many problems in using above the effective concentration and the situation does not produce a satisfactory effect. In addition, there is no patent on a composition showing an inhibitory effect on the production of melanin including the extract of the oviparous duct of the frog (hereinafter referred to as yam oil).
이에, 본 발명자들은 종래의 멜라닌 생성 저해제보다 우수한 물질을 찾기 위해 노력하던 중, 합마유 추출물이 멜라닌 생성과 관련하여 우수하고 독성이 없는 저해제로서의 작용할 수 있음을 알아내고, 미백용 조성물로 사용할 수 있음을 확인함으로써 본 발명을 완성하였다.Thus, the inventors of the present invention, while trying to find a material superior to the conventional melanin inhibitors, finds that the yam oil extract can act as a good and non-toxic inhibitor with respect to melanin production, can be used as a whitening composition By confirming that the present invention was completed.
본 발명의 목적은 합사마의 수란관을 이용하여, 독성이 적고, 기존의 피부 미백물질 보다 우수한 멜라닌 생성 억제 효과를 갖는 합마유 추출물을 제공하는 것이다.It is an object of the present invention to provide a yam oil extract using the oocyte's oviduct, which has less toxicity and has an effect of inhibiting melanin production than conventional skin whitening substances.
상기 목적을 달성하기 위하여, 본 발명은 멜라닌 생성 억제용 합마유 추출물을 제공한다.In order to achieve the above object, the present invention provides melanin extract for inhibiting melanin production.
또한, 본 발명은 합마유 추출물을 유효성분으로 함유하는 멜라닌 생성 억제 및 피부 미백용 조성물을 제공한다.In another aspect, the present invention provides a composition for inhibiting melanin production and skin whitening containing the extract of the yam oil as an active ingredient.
이하, 본 발명을 상세히 설명한다.Hereinafter, the present invention will be described in detail.
본 발명은 멜라닌 생성 억제용 합마유 추출물을 제공한다.The present invention provides melanin extract for inhibiting melanin production.
합마유 추출을 위한 용매는 물, 알코올 또는 이의 혼합물이고 바람직하게는 알콜은 C1~C3의 저급알콜, 보다 바람직하게는 물로서, 본 발명자들은 합마유에 물을 혼합하여 추출한 후, 여과지로 여과하여 합마유 추출물을 수득하였다.The solvent for extracting the corn oil is water, alcohol or a mixture thereof. Preferably, the alcohol is C 1 ~ C 3 lower alcohol, more preferably water. Filtration yielded the horse oil extract.
본 발명자들은 합마유 추출물이 멜라닌 생성 억제에 나타내는 효과를 알아보기 위하여, 멜라닌 세포주를 대상으로 동물세포 실험을 하였다.The present inventors conducted animal cell experiments on melanin cell lines to determine the effect of the extract of the corn oil on melanin production.
그 결과, 합마유 추출물과 대조군[페닐티오우레아(phenylthiourea; 이하, PTU)]을 비교할 때, 10 ㎍/㎖ 추출물에서 멜라닌 생성량이 74%, PTU 대조군에서 81%로 나타나, 합마유 추출물을 사용하였을 때 멜라닌 생성 억제효과가 더 좋았다(표 1 참고).As a result, when comparing the yam oil extract with the control (phenylthiourea (PTU)], melanin production was 74% in the 10 ㎍ / ㎖ extract, 81% in the PTU control, it would be used When the melanin production inhibitory effect was better (see Table 1).
본 발명의 상기의 멜라닌 세포주를 대상으로 합마유 추출물이 세포 독성에 미치는 영향을 알아본 결과, PTU와 달리 100 ㎍/㎖의 높은 농도에서도 세포 독성이 없는 것으로 나타났다(도 1, 도 2 참고).As a result of examining the effect of the yam oil extract on the cytotoxicity of the melanin cell line of the present invention, it was found that there is no cytotoxicity at a high concentration of 100 ㎍ / ㎖ unlike PTU (see Fig. 1, 2).
따라서, 본 발명의 합마유 추출물은 기존의 멜라닌 생성 억제제에 비해 억제효과도 높고 세포 독성도 없는 효과적인 미백제임을 확인하였다.Therefore, it was confirmed that the yam oil extract of the present invention is an effective whitening agent having a high inhibitory effect and no cytotoxicity, compared to the conventional melanin inhibitor.
또한, 본 발명은 합마유 추출물을 유효성분으로 함유하는 멜라닌 생성 억제및 피부 미백용 조성물을 제공한다.In another aspect, the present invention provides a composition for inhibiting melanin production and skin whitening containing the extract of the yam oil as an active ingredient.
본 발명의 합마유 추출물을 유효성분으로 함유하는 화장료 조성물로 제조되는 화장품은 일반적인 유화 제형 및 가용화 제형의 형태로 제조할 수 있다. 유화 제형의 화장품으로는 영양화장수, 크림, 에센스 등이 있으며, 가용화 제형의 화장품으로는 유연화장수가 있다. 또한, 본 발명의 합마유 추출물을 함유하는 화장품 이외에도 피부과학적으로 허용가능한 매질 또는 기제를 함유함으로써 피부과학 분야에서 통상적으로 사용되는 국소적용 또는 전신적용할 수 있는 보조제 형태로 제조될 수 있다.Cosmetics prepared with a cosmetic composition containing the yam oil extract of the present invention as an active ingredient may be prepared in the form of a general emulsion formulation and solubilized formulation. Cosmetics of the emulsified formulations include nutrient cosmetics, creams, essences, etc., and cosmetics of the solubilized formulations are flexible cosmetics. In addition to the cosmetic containing the extract of the seahorse oil of the present invention by containing a dermatologically acceptable medium or base can be prepared in the form of a topical or systemic application commonly used in the field of dermatology.
적합한 화장품의 제형으로는 예를 들면 용액, 겔, 고체 또는 반죽 무수 생성물, 수상에 유상을 분산시켜 얻은 에멀젼, 현탁액, 마이크로에멀젼, 마이크로캡슐, 미세과립구 또는 이온형(리포좀), 비이온형의 소낭 분산제의 형태, 크림, 스킨, 로션, 파우더, 연고, 스프레이 또는 콘실 스틱(conceal stick)의 형태로 제공될 수 있다. 또한, 포말(foam)의 형태 또는 압축된 추진제를 더 함유한 에어로졸 조성물의 형태로도 제조될 수 있다.Suitable cosmetic formulations include, for example, emulsions, suspensions, microemulsions, microcapsules, microgranules or ionic (liposomes), nonionic vesicles obtained by dispersing an oil phase in a solution, gel, solid or pasty anhydrous product, aqueous phase, for example. It may be provided in the form of a dispersant, cream, skin, lotion, powder, ointment, spray or conceal stick. It may also be prepared in the form of a foam or in the form of an aerosol composition further containing a compressed propellant.
또한, 본 발명의 화장료 조성물은 합마유 추출물에 추가로 지방 물질, 유기 용매, 용해제, 농축제 및 겔화제, 연화제, 항산화제, 현탁화제, 안정화제, 발포제(foaming agent), 방향제, 계면활성제, 물, 이온형 또는 비이온형 유화제, 충전제, 금속이온 봉쇄제 및 킬레이트화제, 보존제, 비타민, 차단제, 습윤화제, 필수 오일, 염료, 안료, 친수성 또는 친유성 활성제, 지질 소낭 또는 화장품에 통상적으로 사용되는 임의의 다른 성분과 같은 화장품학 또는 피부과학 분야에서 통상적으로 사용되는 보조제를 함유할 수 있다. 그리고, 상기의 성분들은 피부과학 분야에서 일반적으로 사용되는 양으로 도입될 수 있다.In addition, the cosmetic composition of the present invention, in addition to the oil extract of the oil, fatty substances, organic solvents, solubilizers, thickening and gelling agents, emollients, antioxidants, suspending agents, stabilizers, foaming agents, fragrances, surfactants, Commonly used in water, ionic or nonionic emulsifiers, fillers, metal ion sequestrants and chelating agents, preservatives, vitamins, blockers, wetting agents, essential oils, dyes, pigments, hydrophilic or lipophilic active agents, lipid vesicles or cosmetics And any other ingredients that may be used, such as auxiliaries commonly used in the cosmetic or dermatological arts. In addition, the above components may be introduced in an amount generally used in the dermatology field.
본 발명의 화장료 조성물을 첨가할 수 있는 제품으로는, 예를 들어, 수렴화장수, 유연화장수, 영양화장수, 각종 크림, 에센스, 팩, 파운데이션 등과 같은 화장품류와 클렌징, 세안제, 비누, 트리트먼트, 미용액 등이 있다.Examples of products to which the cosmetic composition of the present invention may be added include, for example, cosmetics such as astringent cosmetics, soft cosmetics, nourishing cosmetics, various creams, essences, packs, foundations, and the like, cleansing agents, soaps, treatments, and essences. Etc.
본 발명의 화장료 조성물의 구체적인 제형으로서는 스킨로션, 스킨 소프너, 스킨토너, 아스트린젠트, 로션, 밀크로션, 모이스처 로션, 영양로션, 맛사지크림, 영양크림, 모이스처 크림, 핸드크림, 에센스, 영양에센스, 팩, 비누, 샴푸, 클렌징폼, 클렌징로션, 클렌징크림, 바디로션, 바디클렌저, 유액, 프레스파우더, 루스파우더, 아이섀도 등의 제형을 포함한다.Specific formulations of the cosmetic composition of the present invention include skin lotion, skin softener, skin toner, astringent, lotion, milk lotion, moisturizing lotion, nutrition lotion, massage cream, nutrition cream, moisture cream, hand cream, essence, nutrition essence, pack, Formulations such as soaps, shampoos, cleansing foams, cleansing lotions, cleansing creams, body lotions, body cleansers, emulsions, press powders, loose powders, eye shadows and the like.
이하, 본 발명을 실시예에 의해 상세히 설명하다. 단 하기 실시예는 본 발명을 예시하는 것일 뿐, 본 발명의 내용을 한정하지는 않는다.Hereinafter, the present invention will be described in detail by way of examples. However, the following examples are merely to illustrate the present invention, but not to limit the content of the present invention.
<< 실시예Example 1> 1> 합마유Yam oil 추출물의 제조 Preparation of Extract
합마유 추출을 위한 용매는 물로서, 본 발명자들은 합마유 1 g(중국 길림성 화전시)을 물 500 ㎖과 혼합하여 소니케이션(sonication) 방법으로 1 시간씩 2회 추출한 후, 여과지로 여과하였다. 이 여액을 회전 진공 농축기로 감압 농축하고, 결과물을 급속 냉동냉장고에 하루 동안 방치하여 동결 건조함으로써 합마유-물 추출물 70.8 mg을 얻었다.The solvent for the extraction of the corn oil is water, and the present inventors mixed 1 g of horse oil (Jilin, China) with 500 ml of water, extracted twice by sonication method, and filtered through filter paper. The filtrate was concentrated under reduced pressure with a rotary vacuum concentrator, and the resultant was left in a quick freezer for one day and lyophilized to obtain 70.8 mg of horse oil-water extract.
<< 실시예Example 2> 초대 2> invite 멜라닌세포의Melanocyte 배양 culture
다자란 정상 C57/6 쥐의 피부에서 초대 멜라닌 세포를 분리하였다(Laurence et al., Mol. Cell. Biol., 24, 3396-3403, 2004). 상기 분리된 초대 멜라닌 세포를, 10% FBS(fetal bovine serum, Gibco, Invitrogen, USA)와 200 nM 포르볼 12-마이리스테이트-13-아세테이트(phorbol 12-myristate-13-acetate; PMA, Sigma)를 넣은 RPMI1640 배지에서 배양하였다. 100π 조직배양접시에 하나의 웰 당 5×105개의 세포를 넣고 37oC, 5% CO2의 조건을 갖춘 CO2 세포배양기에서 3일 내지 4일 배양하였다.Primary melanocytes were isolated from the skin of mature C57 / 6 mice (Laurence et al., Mol. Cell. Biol., 24, 3396-3403, 2004). The isolated primary melanocytes were treated with 10% FBS (fetal bovine serum, Gibco, Invitrogen, USA) and 200 nM phorbol 12-myristate-13-acetate (PMA, Sigma). Cultured in RPMI1640 medium. 5 × 10 5 cells per well were placed in a 100π tissue culture dish and cultured for 3-4 days in a CO 2 cell incubator equipped with 37 ° C. and 5% CO 2 .
<< 실험예Experimental Example 1> 1> 합마유Yam oil 추출물이 멜라닌 생성에 미치는 영향 Effect of Extracts on Melanin Production
실시예 2에 의해 배양한 멜라닌 세포를 24-웰 조직배양접시에 하나의 웰 당 105개씩 넣고 하루 동안 CO2 세포배양기에 넣어두었다. 각 웰에 매일 RPMI1640 배지 990 ㎕와 합마유 추출물 10 ㎕(PBS에 1 ㎍, 10 ㎍, 100 ㎍의 합마유 추출물 첨가)를 3일 동안 첨가하였다.The melanocytes cultured in Example 2 were placed in a 24-well tissue culture dish with 10 5 cells per well and placed in a CO 2 cell culture medium for one day. To each well were added 990 μl of RPMI1640 medium and 10 μl of horse oil extract (addition of 1 μg, 10 μg, 100 μg of horse oil extract to PBS) for 3 days.
CO2 세포배양기에 하루 동안 방치한 상기의 세포주 내의 멜라닌 양을 호소 이(Hosoi J et al. Cancer Res, 45, 1474-1478, 1985)의 방법을 이용하여 측정하였다. 구체적으로, 각 웰에서 배지를 제거한 후 PBS로 세척하고, 멜라닌을 용해하기 위하여 1N NaOH 1 ㎖을 첨가한 후, 이를 ELISA를 이용하여 405 ㎚에서 흡광도를 측정하였다. 멜라닌 양은 추출물을 첨가하지 않은 음성 대조군의 멜라닌 양에 대한 백분율로 나타내었다. 양성 표준 대조군(Positive standard control)으로는 타이로시나제 활성을 억제함으로써 멜라닌화를 막는 억제제로 알려진 PTU(phenylthiourea; 동경화성공업주식회사, 일본)를 사용하였다.The amount of melanin in the cell line left for one day in the CO 2 cell culture was measured using the method of Hosoi J et al. Cancer Res , 45, 1474-1478, 1985. Specifically, the medium was removed from each well, washed with PBS, 1 mL of 1N NaOH was added to dissolve melanin, and the absorbance was measured at 405 nm using an ELISA. The melanin amount was expressed as a percentage of the melanin amount of the negative control without the extract. As a positive standard control, PTU (phenylthiourea; Tokyo Chemical Industries, Ltd.), which is known as an inhibitor of melanogenesis by inhibiting tyrosinase activity, was used.
실험결과는 평균표준편차로 표시하였으며, 각 실험군의 수치결과는 대조군에 대한 백분율로 나타내었다. 각 군 간의 통계적 유의성에 대한 검증은 스튜던트의 t-검정(student's t-test)을 이용하였다.The experimental results are expressed as average standard deviation, and the numerical results of each experimental group are expressed as a percentage of the control group. Student's t- test was used to verify statistical significance between groups.
그 결과, 표 1에 나타난 바와 같이, 합마유 추출물 10 ㎍/㎖ 에서 멜라닌 생성량이 74%, 대조군에서 81%, 1 ㎍/㎖에서 멜라닌 생성량이 90%, 대조군에서 99%로, 합마유 추출물을 사용하였을 때 멜라닌 생성 억제효과가 더 좋았다(표 1, 도 1 및 도 2).As a result, as shown in Table 1, melanin production was 74% at 10 μg / ml of the horse oil extract, 81% at the control group, melanin production was 90% at 1 μg / ml, and 99% at the control group. The melanin production inhibitory effect was better when used (Table 1, Figure 1 and Figure 2).
<< 실험예Experimental Example 2> 2> 합마유Yam oil 추출물이 세포증식에 미치는 영향 Effect of Extracts on Cell Proliferation
합마유 추출물에 대한 세포생존도 측정을 통한 세포 독성검사를 위하여, 상기의 실험예 1에서 서술한 대로 준비한 세포주에 합마유 추출물 및 PTU로 농도별 처리를 수행하였다.For cytotoxicity test by measuring the cell viability of the extract of the corn oil, the cell lines prepared as described in Experimental Example 1 above were treated with the concentration of the corn oil extract and PTU.
상기의 준비된 세포를 세포생존도 측정용 염색제인 크리스탈 바이올렛(crystal violet; 이하, CV)으로 염색한 후, 실험군과 합마유 추출물 및 PTU를 넣지 않은 음성대조군의 비교를 통해 살아있는 세포의 비율을 나타내었다. 구체적으로, 합마유 추출물을 처리한 조직배양접시의 배지를 제거한 후 PBS로 세척하고, 각 웰에 크리스탈 바이올렛 용액(CV 0.1%, 10% EtOH, 89.9% PBS) 200 ㎕을 처리하고, 5분 정도 실온에서 방치한 후 이를 물로 2번 씻어 낸다. 그런 다음 각 웰 당 1 ㎖ EtOH를 넣고 실온에서 10분 정도 휘저음 한다. 이를 ELISA로 CV의 흡수파장인 570 ㎚에서 측정하였다. 또한, 실험결과는 실험예 1과 동일한 방법으로 통계적으로 처리하였다.The prepared cells were stained with crystal violet (CV), which is a dye for measuring cell viability, and the ratio of living cells was shown by comparison between the experimental group and the negative control group without the horse oil extract and PTU. . Specifically, after removing the medium of the tissue culture plate treated with the yam oil extract, washed with PBS, 200 μl of a crystal violet solution (CV 0.1%, 10% EtOH, 89.9% PBS) to each well, and about 5 minutes After standing at room temperature, wash it twice with water. Then add 1 ml EtOH for each well and stir for 10 minutes at room temperature. This was measured at 570 nm, the absorption wavelength of CV by ELISA. In addition, the experimental results were statistically treated in the same manner as in
그 결과, 표 1, 도 1 및 도 2에 나타난 바와 같이, 합마유 추출물 100 ㎍/㎖ 으로 처리하면, 원래의 수준의 세포생존도를 유지(98%)하고 있는 반면, PTU처리 대조군은 82% 까지 세포생존도가 떨어짐으로 고농도 PTU처리에 의한 독성을 나타내고 있다.As a result, as shown in Table 1, Figures 1 and 2, when treated with 100 μg / ml of the horsetail oil extract, the original level of cell viability was maintained (98%), while the PTU-treated control group was 82% Toxicity by the high concentration of PTU treatment has been shown to decrease cell viability.
따라서, 본 발명의 합마유 추출물은 고농도(100 ㎍/㎖)에서도 높은 수준의 세포생존도를 유지하고 있고, 10 ㎍/㎖의 저농도에서도 PTU에 비해 향상된 멜라닌 생성 억제 효과를 나타냄을 확인하였다. 즉, 멜라닌 세포에 합마유 추출물을 처리하였을 때 그 처리 농도가 높아져도 세포에 영향을 주지 않으므로 이는 합마유 추출물이 세포 독성을 가지지 않는 것을 말한다.Therefore, it was confirmed that the yam oil extract of the present invention maintains a high level of cell viability even at high concentrations (100 μg / ml), and shows an improved melanin production inhibitory effect compared to PTU even at low concentrations of 10 μg / ml. In other words, when treated with melanin extract to the melanocytes does not affect the cells even if the treatment concentration is high, this means that the extract is not cytotoxic.
본 발명의 합마유 추출물 및 이를 포함하는 조성물은 멜라닌 생성 억제 효과가 월등하고 세포독성을 나타내지 않으므로, 세포독성이 있는 기존의 미백용 조성물을 대체하여 활용가능하다.Synthetic oil extract of the present invention and the composition comprising the same is excellent in melanin production inhibitory effect and does not exhibit cytotoxicity, it is possible to replace the existing whitening composition having cytotoxicity can be utilized.
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101698683B (en) * | 2009-09-21 | 2011-09-28 | 宁波大学 | Method for extracting polysaccharides from ovidutus ranae |
| CN102885296A (en) * | 2011-07-21 | 2013-01-23 | 成都永康制药有限公司 | Formula of dietary supplement for female health care |
| CN103330212A (en) * | 2013-07-17 | 2013-10-02 | 辽宁宏宇生物科技有限公司 | Rana japonica oil composite electuary and preparation method thereof |
| CN104664396A (en) * | 2013-11-28 | 2015-06-03 | 宽甸满族自治县光太药材有限公司 | Instant rana japonica oil and radix polygonati officinalis tin and preparation method thereof |
| CN104824665A (en) * | 2015-05-04 | 2015-08-12 | 王伟国 | Preparation method of health Chinese forest frog oil and vitamin E lozenge |
| CN106389172A (en) * | 2016-04-30 | 2017-02-15 | 孟令刚 | Cosmetic containing herba cistanche and oviductus ranae |
-
2006
- 2006-11-30 KR KR1020060119772A patent/KR20080049332A/en not_active Application Discontinuation
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101698683B (en) * | 2009-09-21 | 2011-09-28 | 宁波大学 | Method for extracting polysaccharides from ovidutus ranae |
| CN102885296A (en) * | 2011-07-21 | 2013-01-23 | 成都永康制药有限公司 | Formula of dietary supplement for female health care |
| CN103330212A (en) * | 2013-07-17 | 2013-10-02 | 辽宁宏宇生物科技有限公司 | Rana japonica oil composite electuary and preparation method thereof |
| CN104664396A (en) * | 2013-11-28 | 2015-06-03 | 宽甸满族自治县光太药材有限公司 | Instant rana japonica oil and radix polygonati officinalis tin and preparation method thereof |
| CN104824665A (en) * | 2015-05-04 | 2015-08-12 | 王伟国 | Preparation method of health Chinese forest frog oil and vitamin E lozenge |
| CN106389172A (en) * | 2016-04-30 | 2017-02-15 | 孟令刚 | Cosmetic containing herba cistanche and oviductus ranae |
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