KR20180032765A - A composition for improving, preventing and treating of fatty liver diseases comprising leek extract - Google Patents
A composition for improving, preventing and treating of fatty liver diseases comprising leek extract Download PDFInfo
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- KR20180032765A KR20180032765A KR1020160121810A KR20160121810A KR20180032765A KR 20180032765 A KR20180032765 A KR 20180032765A KR 1020160121810 A KR1020160121810 A KR 1020160121810A KR 20160121810 A KR20160121810 A KR 20160121810A KR 20180032765 A KR20180032765 A KR 20180032765A
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- Prior art keywords
- extract
- fatty liver
- liver disease
- leaf
- preventing
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- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
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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)
- A61K36/88—Liliopsida (monocotyledons)
- A61K36/896—Liliaceae (Lily family), e.g. daylily, plantain lily, Hyacinth or narcissus
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- A—HUMAN NECESSITIES
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- 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
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- A23V2200/00—Function of food ingredients
- A23V2200/30—Foods, ingredients or supplements having a functional effect on health
- A23V2200/3262—Foods, ingredients or supplements having a functional effect on health having an effect on blood cholesterol
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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
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Abstract
Description
본 발명은 대파(leek) 추출물을 유효성분으로 함유함으로써, 비알코올 의존성 지방간 질환을 개선, 예방 또는 치료할 수 있는 조성물에 관한 것이다.The present invention relates to a composition capable of improving, preventing or treating a non-alcohol dependent fatty liver disease by containing a leek extract as an active ingredient.
지방간에서 축적된 지방의 대부분은 중성지방(triglyceride)이며, 지방간은 크게 과음으로 인한 알코올성 지방간 질환(alcoholic fatty liver disease)과 비만, 당뇨병, 고지혈증, 약물 등으로 인한 비알코올성 지방간 질환(nonalcoholic fatty liver disease, NAFLD)으로 나눌 수 있다.Most of the fat accumulated in the liver is triglyceride. The fatty liver is mainly caused by alcoholic fatty liver disease and nonalcoholic fatty liver disease (obesity, diabetes, hyperlipidemia, drug) , NAFLD).
알코올성 지방간 질환은 초기의 단순성 지방간에서 지방간염, 간경변으로 진행된다. 비알코올성 지방간질환은 단순성 지방간에 머물며 병태가 진행되지 않는 것으로 알려져 있었지만, 근래 들어 비알코올성 지방간 질환도 단순성 지방간에서 지방간염이나 간경변으로 병태가 진행되는 경우가 있는 것으로 밝혀졌다.Alcoholic fatty liver disease progresses from simple liver fatty liver to fatty liver and cirrhosis. Nonalcoholic fatty liver disease has been known to remain in simple fatty liver and does not progress. However, recently, it has been found that nonalcoholic fatty liver disease sometimes progresses from simple fatty liver to fatty liver or liver cirrhosis.
최근, 영양상태가 좋아지고 성인병이 늘어감에 따라 지방간 환자가 늘어나는 추세에 있다. 대한간학회에 따르면 최근 10년간 간질환 유병율은 감소하였으나, 지방간 유병율은 20년간 3배나 증가하였으며, 그 중에서도 비알코올성 지방간의 비율이 50%를 초과하여 비알코올성 지방간 비율이 급격히 증가하고 있는 것으로 나타났다.Recently, as the nutritional status improves and the adult diseases increase, the number of patients with fatty liver is increasing. According to the Korean Hepatology Society, the prevalence of liver disease has declined over the past decade, but the prevalence of fatty liver has tripled over 20 years, with the proportion of nonalcoholic fatty liver exceeding 50%, showing a rapid increase in the ratio of nonalcoholic fatty liver.
비알코올성 지방간 질환이란 간에 유해할 정도로 인정되는 알코올 섭취 병력이 없음에도 불구하고 간 조직 검사에서 알코올성 간염의 특징적인 소견인 지방성 변화(fatty change, steatosis)와 소엽성간염(lobular hepatitis, steatohepatitis) 등을 나타내는 경우를 말한다. 간의 병리소견은 단순 지방간(non-alcoholic Fatty Liver, NAFL)에서부터 지방간염(non-alcoholic Steatohepatits, NASH), 지방간염과 동반된 섬유화증, 간경변증 등의 다양한 스펙트럼을 나타내는데, 비알코올성 지방간 질환은 이 모두를 포함하는 의미로 사용된다. Nonalcoholic fatty liver disease is a disease characterized by fatty changes (steatosis) and lobular hepatitis (steatohepatitis), which are hallmarks of alcoholic hepatitis in liver biopsy, despite the absence of alcohol- . The pathologic findings of the liver are diverse, ranging from non-alcoholic Fatty Liver (NAFL) to non-alcoholic steatohepatits (NASH), fibrosis associated with hepatitis, and liver cirrhosis. Nonalcoholic fatty liver disease Is used to mean including.
이러한 비알코올성 지방간 질환은 대부분 인슐린 저항성, 비만, 당뇨병, 고지혈증 등의 합병증을 동반한다. 이러한 합병증이 존재하는 경우에는 치료를 실시해야 하는데, 비알코올성 지방간 질환에 대한 치료의 원칙은 식사요법이나 운동요법 등 생활 습관의 개선이지만, 확실하게 실행하기 어려운 것이 현실이다. 특히, 비알코올성 지방간염의 경우 간경변 또는 간세포암으로 진전될 가능성이 높기 때문에 보다 적극적인 약물 치료가 필요하다.Most of these nonalcoholic fatty liver diseases are accompanied by complications such as insulin resistance, obesity, diabetes, and hyperlipemia. In the presence of such complications, treatment should be performed. The principle of treatment for nonalcoholic fatty liver disease is improvement of lifestyle such as diet and exercise therapy, but it is a reality that it is difficult to carry out clearly. In particular, nonalcoholic fatty liver disease is more likely to progress to cirrhosis or hepatocellular carcinoma, so more active medication is needed.
현재까지 지방간의 치료제로는 폴리엔포스파티딜콜린(polyene phosphatidylcholine)이 임상에서 사용되고 있으며, 고지혈증 치료제인 클로피브레이트(clofibrate)로 대표되는 피브레이트(fibrate)계 약제가 지방간에 대한 치료제로 임상에서 사용되고 있다. 이중 피브레이트계 약제는 간장에서 지방간 β산화계 효소를 매개로 지질대사를 개선하는 것으로 생각되고 있다. 그러나 상기 피브레이트계 약제는 간 기능 장애 등의 부작용이 있다는 문제가 있다.To date, polyene phosphatidylcholine has been used clinically as a therapeutic agent for fatty liver. Fibrate-based drugs, such as clofibrate, a therapeutic agent for hyperlipidemia, have been used clinically in the treatment of fatty liver. It is thought that the double fibrate drug improves lipid metabolism through hepatic β - oxidase enzyme in liver. However, the above-mentioned fibrate drugs have a side effect such as liver dysfunction.
따라서, 부작용이 없고 치료 효과가 우수한 새로운 비알코올성 지방간 질환의 예방 및 치료를 위한 치료제의 개발이 필요한 실정이다.Therefore, it is necessary to develop a therapeutic agent for the prevention and treatment of a new nonalcoholic fatty liver disease which has no side effects and excellent therapeutic effect.
본 발명의 목적은 대파(leek) 추출물을 유효성분으로 함유하는 지방간 질환의 개선 또는 예방용 식품 조성물을 제공하는데 있다.It is an object of the present invention to provide a food composition for improving or preventing fatty liver disease containing leek extract as an active ingredient.
또한, 본 발명의 다른 목적은 대파(leek) 추출물을 유효성분으로 함유하는 지방간 질환의 예방 또는 치료용 약학 조성물을 제공하는데 있다.Another object of the present invention is to provide a pharmaceutical composition for preventing or treating fatty liver disease containing leek extract as an active ingredient.
상기한 목적을 달성하기 위한 본 발명의 지방간 질환의 개선 또는 예방용 식품 조성물은 대파(leek) 추출물을 유효성분으로 함유할 수 있다.To achieve the above object, the food composition for improving or preventing fatty liver disease of the present invention may contain leek extract as an active ingredient.
상기 대파 추출물은 대파의 엽초부, 엽신부 또는 이들의 혼합물로 추출된 추출물일 수 있으며, 상기 혼합물은 대파의 엽초부와 엽신부가 1 : 0.3 내지 1의 중량비로 혼합된 혼합물일 수 있다.The lobule extract may be a leaf sheath, a leaf lobe, or a mixture thereof, and the mixture may be a mixture of a leaf sheath and a leaf lobe at a weight ratio of 1: 0.3 to 1.
상기 대파 추출물은 물, 탄소수 1 내지 4의 저급알코올 또는 이들의 혼합용매로 추출된 것일 수 있다.The Mallow extract may be extracted with water, a lower alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof.
상기 혼합용매는 20 내지 80 부피%의 메탄올, 에탄올, 부탄올 또는 프로판올 수용액일 수 있으며, 바람직하게는 20 내지 80 부피%의 에탄올 수용액일 수 있다.The mixed solvent may be 20 to 80% by volume of methanol, ethanol, butanol or propanol aqueous solution, preferably 20 to 80% by volume of ethanol aqueous solution.
상기 지방간 질환은 비알코올 의존성 지방간 질환일 수 있다.The fatty liver disease may be a non-alcohol dependent fatty liver disease.
또한, 상기한 다른 목적을 달성하기 위한 본 발명의 지방간 질환의 예방 또는 치료용 약학 조성물은 대파(leek) 추출물을 유효성분으로 함유할 수 있다.In order to achieve the above and other objects, the present invention provides a pharmaceutical composition for preventing or treating fatty liver disease, which comprises leek extract as an active ingredient.
상기 대파 추출물은 20 내지 80 부피%의 에탄올 수용액으로 추출된 것일 수 있다.The Mallow extract may be extracted with 20 to 80% by volume aqueous ethanol solution.
본 발명의 지방간 질환의 개선, 예방 또는 치료용 조성물은 지방축적, 지질축적으로 의한 간조직의 변형 등 비알콜성 지방간으로 인한 병리학적 소견을 완화시킬 수 있으므로 비알코올성 지방간 질환의 예방 또는 치료에 현저한 효과가 있다.The composition for improving, preventing or treating fatty liver disease of the present invention can alleviate the pathological findings due to non-alcoholic fatty liver such as fat accumulation and modification of liver tissue due to lipid accumulation, and thus the composition for preventing or treating non-alcoholic fatty liver disease It is effective.
또한, 본 발명은 천연물질인 대파 추출물을 유효성분으로 포함하여 부작용이 없으며 저렴하므로 비알코올성 지방간 질환에 장기적으로 사용할 수 있다.In addition, the present invention can be used for a non-alcoholic fatty liver disease for a long period of time because it has no side effects and is inexpensive because it contains a natural ingredient, Daechi Extract, as an effective ingredient.
도 1은 오일 레드 O 용액으로 염색된 대조군, OA군, OA+실시예 1 내지 OA+실시예 6 및 OA+비교예 1의 HepG2 세포를 용해한 후 측정한 흡광도 그래프이다.
도 2는 대조군, OA군 및 상기 OA군에 실시예 1에 따라 제조된 추출물로 처리한 군(OA+실시예 1)의 HepG2 세포에 축적된 지방을 적색가시화상현미경으로 촬영한 사진이다.
도 3은 실시예 3의 추출물을 투여한 시료 투여군, 정상군 및 대조군의 마우스를 희생시켜 간을 촬영한 사진이다.
도 4는 12주 동안 실시예 3의 추출물을 투여한 시료 투여군, 정상군 및 대조군 마우스의 체중을 측정한 그래프이다.
도 5a는 실시예 3의 추출물을 투여한 시료 투여군, 정상군 및 대조군 마우스를 희생시킨 후 추출된 간 조직의 무게를 나타낸 그래프이며, 도 5b는 상기 각 군에서 추출된 간 조직을 촬영한 사진이다.
도 6a는 실시예 3의 추출물을 투여한 시료 투여군, 정상군 및 대조군 마우스를 희생시킨 후 추출된 부고환지방의 무게를 나타낸 그래프이며, 도 5b는 상기 각 군에서 추출된 부고환지방을 촬영한 사진이다.
도 7a 내지 7f는 실시예 3의 추출물을 투여한 시료 투여군, 정상군(N) 및 대조군(C) 마우스에서 채혈한 혈액의 Alkaline phosphatase (ALP), Alanine aminotransaminase (ALT), Aaspartate aminotransferase (AST), Total cholesterol (TC), Low density lipoprotein-cholesterol (LDL-C), High density lipoprotein-cholesterol (HDL-C) 및 Total protein 수치를 나타낸 그래프이다.
도 8은 실시예 3의 추출물을 투여한 시료 투여군, 정상군 및 대조군 마우스에서 측정한 동맥경화 지수를 나타낸 그래프이다.FIG. 1 is a graph of absorbance of a control, OA group, OA + Example 1 to OA + Example 6, and OA + Comparative Example 1 HepG2 cells stained with Oil Red O solution.
FIG. 2 is a photograph of the fat accumulated in HepG2 cells of the group treated with the extract prepared according to Example 1 (OA + Example 1) in the control group, the OA group and the OA group by a red visible-light microscope.
FIG. 3 is a photograph of a liver taken at the sacrifice of a mouse administered with a sample administered with the extract of Example 3, a normal group, and a control group.
FIG. 4 is a graph showing body weight of a sample-administered group, a normal group, and a control mouse in which the extract of Example 3 was administered for 12 weeks.
FIG. 5A is a graph showing the weights of liver tissues extracted after sacrifice of a sample administration group, a normal group and a control mouse to which the extract of Example 3 was administered, and FIG. 5B is a photograph of liver tissues extracted from the above groups .
FIG. 6A is a graph showing the weight of epididymal fat extracted after sacrifice of a sample administration group, a normal group and a control mouse to which the extract of Example 3 was administered, and FIG. 5B is a photograph of epididymal fat extracted from each of the above groups .
FIGS. 7A to 7F are graphs showing changes in the concentrations of Alkaline phosphatase (ALP), alanine aminotransaminase (ALT), and Aaspartate aminotransferase (AST) in the blood collected from the sample administration group, the normal group (N) (TC), low density lipoprotein-cholesterol (LDL-C), high density lipoprotein-cholesterol (HDL-C) and total protein levels.
8 is a graph showing the arteriosclerosis index measured in the sample administration group, the normal group and the control mouse to which the extract of Example 3 was administered.
본 발명은 대파(leek) 추출물을 유효성분으로 함유함으로써, 비알코올 의존성 지방간 질환을 개선, 예방 또는 치료할 수 있는 조성물에 관한 것이다.
The present invention relates to a composition capable of improving, preventing or treating a non-alcohol dependent fatty liver disease by containing a leek extract as an active ingredient.
이하, 본 발명을 상세하게 설명한다. Hereinafter, the present invention will be described in detail.
본 발명의 지방간 질환을 개선, 예방 또는 치료할 수 있는 조성물은 대파(leek) 추출물을 유효성분으로 함유한다.The composition capable of improving, preventing or treating the fatty liver disease of the present invention contains leek extract as an active ingredient.
상기 대파는 백합과에 속하는 다년생 초본으로서 뿌리와 잎으로 구분되며, 상기 잎은 짧고 연백(軟白)되어서 굵고 긴 백경부(白莖部)를 형성하는 엽초부(葉초部)와, 잎의 끝이 뾰족한 속이 빈 원통모양으로 녹색(綠色)이며 표면에 납질을 띠고 있는 엽신부(葉身部)로 나뉜다. The green leaf is a perennial plant belonging to the genus Liliaceae and is divided into roots and leaves. The leaves are short and soft white, and have a leaf sheath portion forming a thick and long white neck portion, This pointed hollow cylinder is greenish (綠色) and has a leaf on the surface.
본 발명은 대파의 뿌리를 제외한 대파의 엽초부, 엽신부 또는 이들의 혼합물을 이용하며, 바람직하게는 엽초부와 엽신부가 1 : 0.3 내지 1의 중량비, 더욱 바람직하게는 1 : 0.5 내지 0.8의 중량비로 혼합된 혼합물을 이용한다. The present invention utilizes a leaf sheath, a leaf blade or a mixture thereof of the root, excluding roots of the root, and preferably the weight ratio of the leaf sheath portion and the leaf blade portion is 1: 0.3 to 1, more preferably 1: 0.5 to 0.8 Is used.
상기 엽초부와 엽신부를 단독으로 사용하더라도 원하는 효과를 얻을 수 있지만, 엽초부와 엽신부를 상기의 함량으로 혼합하여 사용하면 단독으로 사용하는 경우보가 더욱 우수한 효과를 보인다.Although the desired effect can be obtained even when the leaf sheath part and the leaf take up part are used alone, when the leaf sheath part and the leaf take up part are mixed with each other in the above amount, the effect is more excellent when used alone.
상기 엽초부와 엽신부를 혼합하여 사용하는 경우에, 상기 엽초부를 기준으로 엽신부의 함량이 상기 하한치 미만인 경우에는 상기 범위내로 사용하는 경우 및 엽초부와 엽신부를 단독으로 사용하는 경우보다도 저하된 효과를 보이며, 상기 상한치 초과인 경우에는 독성을 유발할 수 있다. When the leaf sheath part and the leaf lance part are used in combination, when the content of the leaf lance part is less than the lower limit value based on the leaf sheath part, the use is made within the above-mentioned range, and the case where the leaf sheath part and the leaf lance part are used alone And if it exceeds the upper limit value, toxicity may be caused.
상기 대파의 엽초부, 엽신부 또는 이들의 혼합물은 추출용매와 1 : 5 내지 25, 바람직하게는 1 : 8 내지 15의 중량비로 혼합하여 20 내지 40 ℃에서 추출함으로써 추출물을 제조한다. 상기 대파의 엽초부, 엽신부 또는 이들의 혼합물과 추출용매의 중량비가 상기 범위를 벗어나는 경우에는 추출물에 대파의 엽초부, 엽신부 또는 이들의 혼합물의 유효성분이 적은 양으로 추출될 수 있다. 또한, 추출온도가 상기 하한치 미만인 경우에는 대파의 엽초부, 엽신부 또는 이들의 혼합물의 유효성분이 추출되지 않을 수 있으며, 상기 상한치 초과인 경우에는 대파의 엽초부, 엽신부 또는 이들의 혼합물의 유효성분이 적은 양으로 추출되고 특히, 80 ℃이상이면 추출물이 변질될 수 있다. The extract is prepared by mixing the leaf herb, leaf bud or mixture thereof with an extraction solvent at a weight ratio of 1: 5 to 25, preferably 1: 8 to 15, and extracting at 20 to 40 ° C. If the weight ratio of the leafhopper, leaf blade or mixture thereof to the extraction solvent is out of the above range, the effective amount of the leaf herb, leaf blade or mixture thereof can be extracted in the extract in a small amount. When the extraction temperature is lower than the lower limit value, the active ingredient of the leaf sheath part, leaf tip part or mixture thereof may not be extracted. If the extraction temperature is higher than the upper limit value, the effective part of the leaf sheath part, Extracts may be deteriorated if they are extracted in small quantities, especially above 80 ° C.
상기 대파의 엽초부, 엽신부 또는 이들 혼합물의 추출물을 추출하는 추출용매로는 지방간 질환의 개선, 예방 또는 치료에 바람직하게 작용할 수 있도록 물, 탄소수 1 내지 4의 저급알코올, 또는 이들의 혼합용매를 사용할 수 있다. 상기 혼합용매로는 특별히 한정하는 것은 아니지만, 바람직하게는 20 내지 80 부피%의 메탄올, 에탄올, 부탄올 또는 프로판올 수용액, 더욱 바람직하게는 60 내지 80 부피%의 에탄올 수용액으로 추출된 추출물을 들 수 있다.The extraction solvent for extracting the leaf herb, leaf tip or mixture of the above-mentioned meals may be water, a lower alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof so as to be preferably used for the improvement, prevention or treatment of fatty liver disease Can be used. The mixed solvent is not particularly limited, but an extract obtained by extracting with 20 to 80% by volume of an aqueous solution of methanol, ethanol, butanol or propanol, more preferably 60 to 80% by volume of ethanol is preferable.
본 명세서에서 대파의 엽초부, 엽신부 또는 이들의 혼합물을 언급하면서 사용되는 용어 '추출물'은 추출용매를 처리하여 얻은 조추출물뿐만 아니라 대파의 엽초부, 엽신부 또는 이들의 혼합물 추출물의 가공물도 포함한다. 예를 들어, 대파의 엽초부, 엽신부 또는 이들 혼합물의 추출물은 감압 증류 및 동결 건조 또는 분무 건조 등과 같은 추가적인 과정에 의해 분말 상태로 제조될 수 있다.The term " extract " used in reference to the leaf sheath part, leaf lance part or mixture thereof in the present specification includes not only the crude extract obtained by treating the extracting solvent but also the processed products of the leaf sheath part, leaf lobe part or mixture thereof do. For example, extracts of leafhopper, leafhopper, or mixture thereof may be prepared in powder form by an additional process such as vacuum distillation and freeze drying or spray drying.
또한, 본 발명의 대파의 엽초부, 엽신부 또는 이들 혼합물의 추출물은 광의로는 대파의 엽초부, 엽신부 또는 이들이 혼합물을 동물에게 투여할 수 있도록 제형화된 대파의 엽초부, 엽신부 또는 이들 혼합물의 가공물, 예컨대, 대파의 엽초부, 엽신부 또는 이들의 혼합물 분말도 포함하는 의미를 갖는다. 비록 본 발명에서 대파의 엽초부, 엽신부 또는 이들 혼합물의 추출물로 실험을 진행하긴 하였으나, 대파의 엽초부, 엽신부 또는 이들 혼합물의 가공물과 같은 형태로도 목적하는 효과를 달성할 수 있음은 당업자라면 예상 가능할 것이다.In addition, the extract of the leafhopper, leafhopper, or mixture thereof of the present invention may be in the form of a leafhopper, a leafhopper, or a leafhopper of a green leaf formulated to be administered to an animal such as a leafhopper, a leafhopper, But also to include workpieces of the mixture, for example, the leaf sheath parts of green tea, leaf green tea, or mixtures thereof. Although the present invention has been carried out with the extracts of leaf, leaf or mixture thereof in the present invention, the desired effect can be achieved in the form of a leaf sheath, a leaf or a mixture thereof, It would be possible to predict.
한편, 본 명세서에서 용어 '유효성분으로 함유하는'이란 대파의 엽초부, 엽신부 또는 이들의 혼합물 추출물의 효능 또는 활성을 달성하는 데 충분한 양을 포함하는 것을 의미한다. 일예로, 상기 대파의 엽초부, 엽신부 또는 이들 혼합물의 추출물은 10 내지 1500 ㎍/㎖, 바람직하게는 100 내지 1000 ㎍/㎖의 농도로 사용된다. 대파의 엽초부, 엽신부 또는 이들 혼합물의 추출물은 천연물로서 과량 투여하여도 인체에 부작용이 없으므로 본 발명의 조성물 내에 포함되는 대파의 엽초부, 엽신부 또는 이들의 혼합물 추출물의 양적 상한은 당업자가 적절한 범위 내에서 선택하여 실시할 수 있다.
As used herein, the term " comprising as an active ingredient " is meant to include an amount sufficient to achieve efficacy or activity of a leaf sheath, leaf bud or mixture thereof. For example, the extracts of leafhopper, leafhopper, or mixture thereof are used at a concentration of 10 to 1500 μg / ml, preferably 100 to 1000 μg / ml. The quantitative upper limit of the leaf extract, leaf extract or mixture of the extracts of the green meats contained in the composition of the present invention is not particularly limited as long as those skilled in the art And can be selected within the range.
본 발명의 약제학적 조성물은 상기 유효 성분 이외에 약제학적으로 적합하고 생리학적으로 허용되는 보조제를 사용하여 제조될 수 있으며, 상기 보조제로는 부형제, 붕해제, 감미제, 결합제, 피복제, 팽창제, 윤활제, 활택제 또는 향미제 등을 사용할 수 있다.The pharmaceutical composition of the present invention can be prepared by using pharmaceutically acceptable and physiologically acceptable adjuvants in addition to the above-mentioned active ingredients. Examples of the adjuvants include excipients, disintegrants, sweeteners, binders, coating agents, swelling agents, lubricants, A lubricant or a flavoring agent can be used.
상기 약제학적 조성물은 투여를 위해서 상기 기재한 유효 성분 이외에 추가로 약제학적으로 허용 가능한 담체를 1종 이상 포함하여 약제학적 조성물로 바람직하게 제제화할 수 있다.The pharmaceutical composition may be formulated into a pharmaceutical composition containing at least one pharmaceutically acceptable carrier in addition to the above-described active ingredients for administration.
상기 약제학적 조성물의 제제 형태는 과립제, 산제, 정제, 피복정, 캡슐제, 좌제, 액제, 시럽, 즙, 현탁제, 유제, 점적제 또는 주사 가능한 액제 등이 될 수 있다. 예를 들어, 정제 또는 캡슐제의 형태로의 제제화를 위해, 유효 성분은 에탄올, 글리세롤, 물 등과 같은 경구, 무독성의 약제학적으로 허용 가능한 불활성 담체와 결합될 수 있다. 또한, 원하거나 필요한 경우, 적합한 결합제, 윤활제, 붕해제 및 발색제 또한 혼합물로 포함될 수 있다. 적합한 결합제는 이에 제한되는 것은 아니나, 녹말, 젤라틴, 글루코스 또는 베타-락토오스와 같은 천연 당, 옥수수 감미제, 아카시아, 트래커캔스 또는 소듐올레이트와 같은 천연 및 합성 검, 소듐 스테아레이트, 마그네슘 스테아레이트, 소듐 벤조에이트, 소듐 아세테이트, 소듐 클로라이드 등을 포함한다. 붕해제는 이에 제한되는 것은 아니나, 녹말, 메틸 셀룰로스, 아가, 벤토니트, 잔탄 검 등을 포함한다.The pharmaceutical composition may be in the form of granules, powders, tablets, coated tablets, capsules, suppositories, liquids, syrups, juices, suspensions, emulsions, drops or injectable solutions. For example, for formulation into tablets or capsules, the active ingredient may be combined with an oral, non-toxic pharmaceutically acceptable inert carrier such as ethanol, glycerol, water, and the like. Also, if desired or necessary, suitable binders, lubricants, disintegrants and coloring agents may also be included as a mixture. Suitable binders include, but are not limited to, natural sugars such as starch, gelatin, glucose or beta-lactose, natural and synthetic gums such as corn sweeteners, acacia, tracker candles or sodium oleate, sodium stearate, magnesium stearate, sodium Benzoate, sodium acetate, sodium chloride, and the like. Disintegrants include, but are not limited to, starch, methyl cellulose, agar, bentonite, xanthan gum and the like.
액상 용액으로 제제화되는 조성물에 있어서 허용 가능한 약제학적 담체로는, 멸균 및 생체에 적합한 것으로서, 식염수, 멸균수, 링거액, 완충 식염수, 알부민 주사용액, 덱스트로즈 용액, 말토 덱스트린 용액, 글리세롤, 에탄올 및 이들 성분 중 1 성분 이상을 혼합하여 사용할 수 있으며, 필요에 따라 항산화제, 완충액, 정균제 등 다른 통상의 첨가제를 첨가할 수 있다. 또한 희석제, 분산제, 계면활성제, 결합제 및 윤활제를 부가적으로 첨가하여 수용액, 현탁액, 유탁액 등과 같은 주사용 제형, 환약, 캡슐, 과립 또는 정제로 제제화할 수 있다.Acceptable pharmaceutical carriers for compositions that are formulated into a liquid solution include sterile solutions suitable for the living body such as saline, sterile water, Ringer's solution, buffered saline, albumin injection solution, dextrose solution, maltodextrin solution, glycerol, One or more of these components may be mixed and used. If necessary, other conventional additives such as an antioxidant, a buffer, and a bacteriostatic agent may be added. In addition, diluents, dispersants, surfactants, binders, and lubricants may be additionally added to formulate into injectable solutions, pills, capsules, granules or tablets such as aqueous solutions, suspensions, emulsions and the like.
더 나아가 해당분야의 적절한 방법으로 Remington's Pharmaceutical Science, Mack Publishing Company, Easton PA에 개시되어 있는 방법을 이용하여 각 질환에 따라 또는 성분에 따라 바람직하게 제제화할 수 있다.Further, it can be suitably formulated according to each disease or ingredient, using the method disclosed in Remington's Pharmaceutical Science, Mack Publishing Company, Easton PA as an appropriate method in the field.
본 발명의 약제학적 조성물은 경구 또는 비경구로 투여할 수 있고, 비경구 투여인 경우에는 정맥내 주입, 피하 주입, 근육 주입, 복강 주입, 경피 투여 등으로 투여할 수 있으며, 바람직하게는 경구 투여이다.The pharmaceutical composition of the present invention can be administered orally or parenterally. In the case of parenteral administration, the composition can be administered by intravenous injection, subcutaneous injection, muscle injection, intraperitoneal injection, transdermal administration, etc., .
본 발명의 약제학적 조성물의 적합한 투여량은 제제화 방법, 투여 방식, 환자의 연령, 체중, 성, 병적 상태, 음식, 투여 시간, 투여 경로, 배설 속도 및 반응 감응성과 같은 요인들에 의해 다양하며, 보통으로 숙련된 의사는 소망하는 치료 또는 예방에 효과적인 투여량을 용이하게 결정 및 처방할 수 있다. 본 발명의 바람직한 구현예에 따르면, 본 발명의 약제학적 조성물의 1일 투여량은 0.001-10 g/kg이다.The appropriate dosage of the pharmaceutical composition of the present invention varies depending on factors such as the formulation method, administration method, age, body weight, sex, pathological condition, food, administration time, administration route, excretion rate and responsiveness of the patient, Usually, a skilled physician can readily determine and prescribe dosages effective for the desired treatment or prophylaxis. According to a preferred embodiment of the present invention, the daily dosage of the pharmaceutical composition of the present invention is 0.001-10 g / kg.
본 발명의 약제학적 조성물은 약제학적으로 허용되는 담체 및/또는 부형제를 이용하여 제제화함으로써 단위 용량 형태로 제조되거나 또는 다용량 용기 내에 내입시켜 제조될 수 있다. 이때 제형은 오일 또는 수성 매질중의 용액, 현탁액 또는 유화액 형태이거나 엑스제, 분말제, 과립제, 정제 또는 캅셀제 형태일 수도 있으며, 분산제 또는 안정화제를 추가적으로 포함할 수 있다.The pharmaceutical composition of the present invention can be prepared in unit dose form by formulating it with a pharmaceutically acceptable carrier and / or excipient or can be manufactured by inserting it into a multi-dose container. The formulations may be in the form of solutions, suspensions or emulsions in oils or aqueous media, or in the form of excipients, powders, granules, tablets or capsules, and may additionally contain dispersing or stabilizing agents.
또한, 본 발명은 대파의 엽초부, 엽신부 또는 이들 혼합물의 추출물을 유효성분으로 함유하는 지방간 질환의 개선, 예방 또는 치료용 식품 조성물을 제공한다.In addition, the present invention provides a food composition for improving, preventing or treating fatty liver disease comprising as an active ingredient an extract of a leaf sheath, a leaf lobe or a mixture thereof.
본 발명에 따른 식품 조성물은 상기 약제학적 조성물과 동일한 방식으로 제제화되어 기능성 식품으로 이용하거나, 각종 식품에 첨가할 수 있다. 본 발명의 조성물을 첨가할 수 있는 식품으로는 예를 들어, 음료류, 알코올 음료류, 과자류, 다이어트바, 유제품, 육류, 초코렛, 피자, 라면, 기타 면류, 껌류, 아이스크림류, 비타민 복합제, 건강보조식품류 등이 있다.The food composition according to the present invention can be formulated in the same manner as the above pharmaceutical composition and used as a functional food or added to various foods. Foods to which the composition of the present invention can be added include, for example, beverages, alcoholic beverages, confectioneries, diet bars, dairy products, meat, chocolates, pizza, ram noodles, other noodles, gums, ice cream, .
본 발명의 식품 조성물은 유효성분으로서 대파의 엽초부, 엽신부 또는 이들 혼합물의 추출물뿐만 아니라, 식품 제조 시에 통상적으로 첨가되는 성분을 포함할 수 있으며, 예를 들어, 단백질, 탄수화물, 지방, 영양소, 조미제 및 향미제를 포함한다. 상술한 탄수화물의 예는 모노사카라이드, 예를 들어, 포도당, 과당 등; 디사카라이드, 예를 들어 말토스, 슈크로스, 올리고당 등; 및 폴리사카라이드, 예를 들어 덱스트린, 사이클로덱스트린 등과 같은 통상적인 당 및 자일리톨, 소르비톨, 에리트리톨 등의 당알콜이다. 향미제로서 천연 향미제 [타우마틴, 스테비아 추출물 (예를 들어 레바우디오시드 A, 글리시르히진 등]) 및 합성 향미제(사카린, 아스파르탐 등)를 사용할 수 있다. 예컨대, 본 발명의 식품 조성물이 드링크제와 음료류로 제조되는 경우에는 본 발명의 대파의 엽초부, 엽신부 또는 이들 혼합물의 추출물 이외에 구연산, 액상과당, 설탕, 포도당, 초산, 사과산, 과즙, 및 각종 식물 추출액 등을 추가로 포함시킬 수 있다.The food composition of the present invention may contain, as an active ingredient, components that are commonly added in the manufacture of foods, as well as extracts of leafhopper, leafhopper, or mixture thereof, such as protein, carbohydrate, fat, , Seasoning agents, and flavoring agents. Examples of the above-mentioned carbohydrates are monosaccharides such as glucose, fructose, and the like; Disaccharides such as maltose, sucrose, oligosaccharides and the like; And polysaccharides such as dextrin, cyclodextrin and the like, and sugar alcohols such as xylitol, sorbitol and erythritol. Natural flavorings such as tau martin and stevia extract (e.g., rebaudioside A and glycyrrhizin) and synthetic flavorings (saccharine, aspartame, etc.) can be used as flavorings. For example, when the food composition of the present invention is prepared from a drink and a beverage, it may contain citric acid, liquid fructose, sugar, glucose, acetic acid, malic acid, juice, and various plants An extract, and the like.
본 발명은 상기 대파의 엽초부, 엽신부 또는 이들 혼합물의 추출물을 유효성분으로 포함하는 지방간 질환의 개선, 예방 또는 치료용 식품 조성물을 포함하는 건강기능식품을 제공한다. 건강기능식품이란, 대파의 엽초부, 엽신부 또는 이들 혼합물의 추출물을 음료, 차류, 향신료, 껌, 과자류 등의 식품소재에 첨가하거나, 캡슐화, 분말화, 현탁액 등으로 제조한 식품으로, 이를 섭취할 경우 건강상 특정한 효과를 가져오는 것을 의미하나, 일반 약품과는 달리 식품을 원료로 하여 약품의 장기 복용시 발생할 수 있는 부작용 등이 없는 장점이 있다. 이와 같이 하여 얻어지는 본 발명의 건강기능식품은, 일상적으로 섭취하는 것이 가능하기 때문에 매우 유용하다. 이와 같은 건강기능식품에 있어서의 대파의 엽초부, 엽신부 또는 이들의 혼합물 추출물의 첨가량은, 대상인 건강기능식품의 종류에 따라 달라 일률적으로 규정할 수 없지만, 식품 본래의 맛을 손상시키지 않는 범위에서 첨가하면 되며, 대상 식품에 대하여 통상 0.01 내지 50 중량%, 바람직하기로는 0.1 내지 20 중량%의 범위이다. 또한, 환제, 과립제, 정제 또는 캡슐제 형태의 건강기능식품의 경우에는 통상 0.1 내지 100 중량% 바람직하기로는 0.5 내지 80 중량%의 범위에서 첨가하면 된다. 한 구체예에서, 본 발명의 건강기능식품은 환제, 정제, 캡슐제 또는 음료의 형태일 수 있다.The present invention provides a health functional food comprising a food composition for improving, preventing or treating fatty liver disease comprising as an effective ingredient an extract of leafhopper, leaf extract or mixture thereof. The health functional food refers to a food prepared by adding leaf extract, leaf extract or mixture thereof to a food material such as beverage, tea, spice, gum or confection, or encapsulated, powdered or suspended, However, unlike general medicines, it has the advantage that there are no side effects that can occur when a drug is taken for a long time by using the food as a raw material. The health functional food of the present invention thus obtained is very useful because it can be ingested routinely. The amount of the leaf extract, leaf extract or mixture thereof extracted from the healthy functional food can not be uniformly determined depending on the type of the health functional food to be treated. However, in the range that does not impair the original taste of the food And it is usually in the range of 0.01 to 50% by weight, preferably 0.1 to 20% by weight based on the target food. In the case of health functional foods in the form of pills, granules, tablets or capsules, they may be added usually in the range of 0.1 to 100% by weight, preferably 0.5 to 80% by weight. In one embodiment, the health functional food of the present invention may be in the form of a pill, tablet, capsule or beverage.
또한, 본 발명은 지방간 질환의 개선, 예방 또는 치료용 의약 또는 식품의 제조를 위한 대파의 엽초부, 엽신부 또는 이들의 혼합물 추출물의 용도를 제공한다. 상기한 바와 같이 대파의 엽초부, 엽신부 또는 이들 혼합물의 추출물은 지방간 질환의 개선, 예방 또는 치료를 위한 용도로 이용될 수 있다.In addition, the present invention provides the use of a leaf herb, a leaf lobe or a mixture thereof for the production of a medicament or food for the improvement, prevention or treatment of fatty liver disease. As described above, the leaf herb, leaf bud or extract of the mixture can be used for the improvement, prevention or treatment of fatty liver disease.
또한, 본 발명은 포유동물에게 유효량의 대파의 엽초부, 엽신부 또는 이들 혼합물의 추출물을 투여하는 것을 포함하는 지방간 질환의 개선, 예방 또는 치료 방법을 제공한다.The present invention also provides a method of improving, preventing or treating a fatty liver disease comprising administering to a mammal an effective amount of a leaf sheath, a leaf lobe or an extract of the mixture.
여기에서 사용된 용어 "포유동물"은 치료, 관찰 또는 실험의 대상인 포유동물을 말하며, 바람직하게는 인간을 말한다.The term "mammal " as used herein refers to a mammal that is the subject of treatment, observation or experimentation, preferably a human.
여기에서 사용된 용어 "유효량"은 연구자, 수의사, 의사 또는 기타 임상의에 의해 생각되는 조직계, 동물 또는 인간에서 생물학적 또는 의학적 반응을 유도하는 유효 성분 또는 약학적 조성물의 양을 의미하는 것으로, 이는 해당 질환 또는 장애의 증상의 완화를 유도하는 양을 포함한다. 본 발명의 유효 성분에 대한 유효량 및 투여횟수는 원하는 효과에 따라 변화될 수 있다. 그러므로, 투여될 최적의 투여량은 당업자에 의해 쉽게 결정될 수 있으며, 질환의 종류, 질환의 중증도, 조성물에 함유된 유효성분 및 다른 성분의 함량, 제형의 종류, 및 환자의 연령, 체중, 일반 건강 상태, 성별 및 식이, 투여 시간, 투여 경로 및 조성물의 분비율, 치료기간, 동시 사용되는 약물을 비롯한 다양한 인자에 따라 조절될 수 있다. 본 발명의 예방, 치료 또는 개선 방법에 있어서, 성인의 경우, 대파의 엽초부, 엽신부 또는 이들 혼합물의 추출물을 1일 1회 내지 수회 투여시, 0.001 g/kg 내지 10 g/kg의 용량으로 투여하는 것이 바람직하다.As used herein, the term "effective amount" refers to the amount of active ingredient or pharmaceutical composition that elicits a biological or medical response in a tissue system, animal, or human, as contemplated by a researcher, veterinarian, physician or other clinician, ≪ / RTI > inducing a reduction of the symptoms of the disease or disorder. The effective amount and the administration frequency for the active ingredient of the present invention can be changed according to the desired effect. Thus, the optimal dosage to be administered can be readily determined by those skilled in the art and will vary with the nature of the disease, the severity of the disease, the amount of active and other ingredients contained in the composition, the type of formulation, and the age, The age, body weight, sex, diet, time of administration, route of administration and fraction of the composition, duration of treatment, concurrent medication, and the like. In the prevention, treatment or improvement method of the present invention, in the case of an adult, a dose of 0.001 g / kg to 10 g / kg is administered once to several times a day, Administration.
본 발명의 치료방법에서 대파의 엽초부, 엽신부 또는 이들 혼합물의 추출물을 유효 성분으로 포함하는 조성물은 경구, 직장, 정맥내, 동맥내, 복강내, 근육내, 흉골내, 경피, 국소, 안구내 또는 피내 경로를 통해 통상적인 방식으로 투여할 수 있다.
In the treatment method of the present invention, the composition comprising the extract of leafhopper, leaf extract or mixture thereof as an active ingredient can be administered orally, rectally, intravenously, intraarterially, intraperitoneally, intramuscularly, intrasternally, transdermally, Lt; RTI ID = 0.0 > and / or < / RTI > intradermal route.
이하, 본 발명의 이해를 돕기 위하여 바람직한 실시예를 제시하나, 하기 실시예는 본 발명을 예시하는 것일 뿐 본 발명의 범주 및 기술사상 범위 내에서 다양한 변경 및 수정이 가능함은 당업자에게 있어서 명백한 것이며, 이러한 변형 및 수정이 첨부된 특허청구범위에 속하는 것도 당연한 것이다.It will be apparent to those skilled in the art that various modifications and variations can be made in the present invention without departing from the spirit or scope of the present invention. Such variations and modifications are intended to be within the scope of the appended claims.
실시예 1. 70 부피% 에탄올_대파의 엽초부EXAMPLES Example 1: Sheep section of 70 vol.% Ethanol-
진도에서 채취한 대파의 엽초부를 70 부피% 에탄올 수용액과 1 : 10의 중량비로 혼합하여 25 ℃에서 24시간 동안 추출한 후 8000xg로 30분 동안 원심분리한 후 침전물을 제거하고 상등액만 수득하여 대파 엽초부 추출물을 제조하였다.
The leaf sheaths of the larvae collected from the jinhyo were mixed with a 70% by volume ethanol aqueous solution at a weight ratio of 1:10, and the mixture was extracted at 25 ° C for 24 hours. After centrifugation at 8000 xg for 30 minutes, the precipitate was removed and only the supernatant was obtained. The extract was prepared.
실시예 2. 70 부피% 에탄올_대파의 엽신부Example 2. 70 vol.% Ethanol-lobe wax
상기 실시예 1과 동일하게 실시하되, 대파의 엽초부 대신 대파의 엽신부를 사용하여 대파 엽신부 추출물을 제조하였다.
The extract was prepared in the same manner as in Example 1 except that the leaves were used instead of the leaves.
실시예 3. 70 부피% 에탄올_대파의 엽초부 : 엽신부 = 1 : 0.6 중량비Example 3: 70 vol.% Ethanol-leaf sheath: lint = 1: 0.6 weight ratio
상기 실시예 1과 동일하게 실시하되, 대파의 엽초부 대신 대파의 엽초부와 엽신부를 1 : 0.6의 중량비로 혼합한 혼합물을 사용하여 추출물을 제조하였다.
The extract was prepared in the same manner as in Example 1 except that a mixture of a leaf sheath part and a leaf part was used at a weight ratio of 1: 0.6 in place of the leaf sheath part of the green onion.
실시예Example 4. 20 부피% 에탄올_대파의 4. 20 vol% ethanol-laden 엽초부Sheath part
상기 실시예 1과 동일하게 실시하되, 추출용매를 70 부피% 에탄올 수용액 대신 20 부피% 에탄올 수용액을 이용하여 대파 엽초부 추출물을 제조하였다.
The extract was prepared in the same manner as in Example 1 except that a 20% by volume ethanol aqueous solution was used instead of the 70% ethanol aqueous solution.
실시예 5. 물_대파의 엽초부Example 5: Sheath portion of water-
상기 실시예 1과 동일하게 실시하되, 추출용매를 70 부피% 에탄올 수용액 대신 물을 이용하여 대파 엽초부 추출물을 제조하였다.
The extract was prepared in the same manner as in Example 1 except that the extraction solvent was replaced with 70 vol.% Ethanol aqueous solution and water was used.
실시예 6. 70 부피% 헥산_대파의 엽초부Example 6: Sheep section of 70 vol.% Hexane-
상기 실시예 1과 동일하게 실시하되, 추출용매를 70 부피% 에탄올 수용액 대신 70 부피% 헥산 수용액을 이용하여 대파 엽초부 추출물을 제조하였다.
The extract was prepared in the same manner as in Example 1 except that a 70% by volume hexane aqueous solution was used instead of the 70% by volume aqueous ethanol solution to prepare a sheath extract.
비교예 1.Comparative Example 1 70 부피% 에탄올_대파의 뿌리70 vol.% Ethanol _ root of roots
상기 실시예 1과 동일하게 실시하되, 대파의 엽초부 대신 대파의 뿌리를 사용하여 대파 뿌리 추출물을 제조하였다.
The same procedure as in Example 1 was carried out except that the roots of Robinia pseudo-acacia var.
<시험예><Test Example>
HepG2 세포는 American Type Culture Collection (ATCC, Manassa, Virginia, USA)에서 구입하였으며, DMEM(Dulbecco's Modified Eagle's Media)과 bovine serum albumin(BSA)은 Daegu(한국)에서 구입하였다. HepG2 cells were purchased from the American Type Culture Collection (ATCC, Manassas, Va., USA) and DMEM (Dulbecco's Modified Eagle's Media) and bovine serum albumin (BSA) were purchased from Daegu (Korea).
또한, 오일 레드 O(Oil Red O) 용액 및 MTT(3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide)는 Sigma-Aldrich(USA)에서 구입하였다. An oil red O solution and MTT (3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyltetrazolium bromide) were also purchased from Sigma-Aldrich (USA).
본 발명에서의 용어 "오일 레드 오 염색"이란 오일 레드 오를 사용하여 세포 내 지방을 붉게 염색하는 염색 방법의 일종을 말한다. 또한, 본 발명에서의 용어, "오일 레드 오 용액"이란 오일 레드 오를 이소프로필 알코올, 에탄올, 메탄올, 프로필렌글라이콜 (propylene glycol) 등의 용매에 용해시킨 용액을 말하며, 상기 (용매)에는 바람직하게는 이소프로필 알코올(isopropyl alchol)이 포함되고 용해시키는 양은 용매의 종류 및 염색의 조건에 따라 달라질 수 있으나 염색효율을 극대화하면서 경제성을 고려하여 포화되는 지점까지 용해시키는 것이 바람직하다.The term "oil red dyed" in the present invention refers to a kind of dyeing method in which oil fat red is used to stain the intracellular fat red. The term "oil redo solution" in the present invention refers to a solution obtained by dissolving an oil red oil in a solvent such as isopropyl alcohol, ethanol, methanol, propylene glycol or the like, The amount of isopropyl alcohol to dissolve depends on the kind of solvent and the condition of dyeing, but it is preferable to dissolve to the point where saturation is taken into account in consideration of economical efficiency while maximizing dyeing efficiency.
시험예Test Example 1. 지방축적 억제 확인_정량적 확인 1. Identification of fat accumulation inhibition _ Quantitative confirmation
도 1은 오일 레드 O 용액으로 염색된 대조군, OA군, OA+실시예 1 내지 OA+실시예 6 및 OA+비교예 1의 HepG2 세포를 용해한 후 측정한 흡광도 그래프이다.FIG. 1 is a graph of absorbance of a control, OA group, OA + Example 1 to OA + Example 6, and OA + Comparative Example 1 HepG2 cells stained with Oil Red O solution.
구체적으로, 상기 대조군, OA군, OA+실시예 1 내지 OA+실시예 6 및 OA+비교예 1의 세포내 축적 지방을 정량적으로 확인하기 위하여 다음과 같은 시험을 실시하였다.Specifically, the following tests were performed to quantitatively determine intracellular accumulated fats of the control group, OA group, OA + Example 1 to OA + Example 6 and OA + Comparative Example 1.
대조군은 HepG2 세포를 24-well 세포배양 플레이트에 접종하고 1% BSA이 첨가된 낮은 글루코스 DMEM 배지 37 ℃, 5% CO2 조건 하에 24시간 배양한 후 실온(23 내지 26 ℃)에서 15분 동안 4% 포름알데히드 200 uL로 세포를 고정한 후 인산 완충 식염수(PBS)로 3회 세척한 다음 5분 동안 60% 이소프로판올 200 uL와 함께 배양하고, 마지막으로 0.1% 오일 레드 O 용액 200 uL로 실온에서 60분 동안 염색하였다.In the control group, HepG2 cells were inoculated on a 24-well cell culture plate and cultured in a low glucose DMEM medium supplemented with 1% BSA at 37 ° C and 5% CO 2 for 24 hours, followed by incubation at room temperature (23-26 ° C) The cells were washed with phosphate buffered saline (PBS) three times and then incubated with 200 μL of 60% isopropanol for 5 minutes. Finally, the cells were incubated with 200 μL of 0.1% Oil Red O solution for 60 minutes at room temperature Lt; / RTI >
또한, OA군(유도군)은 1% BSA이 첨가된 낮은 글루코스 DMEM 배지에 0.5 mM 올레인산(oleic acid)을 첨가한 후 HepG2 세포를 37 ℃, 5% CO2 조건 하에 24시간 배양한 다음 상기 대조군과 동일한 방법으로 염색하였다.OA group (induction group) was prepared by adding 0.5 mM oleic acid to low glucose DMEM medium supplemented with 1% BSA and then incubating HepG2 cells for 24 hours at 37 ° C and 5% CO 2, Lt; / RTI >
또한, 처리군(OA+실시예 1 내지 OA+실시예 5 및 OA+비교예 1)은 상기 OA군의 배지를 실시예 1 내지 5 및 비교예 1에서 제조된 200 ㎍/㎖의 대파 추출물로 각각 처리한 후 HepG2 세포를 37 ℃, 5% CO2 조건 하에 24시간 배양한 다음 상기 대조군과 동일한 방법으로 염색하였다. In the treatment group (OA + Example 1 to OA + Example 5 and OA + Comparative Example 1), the OA group medium was treated with the 200 占 퐂 / ml of the Robiniae extract prepared in Examples 1 to 5 and Comparative Example 1 , HepG2 cells were cultured for 24 hours at 37 ° C and 5% CO 2, and then stained in the same manner as the control group.
상기 오일 레드 O 용액으로 염색된 각 군의 세포를 10분 동안 이소프로판올에 용해하고, 용해된 용액 100 ㎕를 96-well 플레이트에 옮겨 510 nm의 파장에서 흡광도를 측정하였다. Cells of each group stained with the oil red O solution were dissolved in isopropanol for 10 minutes, and 100 μl of the dissolved solution was transferred to a 96-well plate and absorbance was measured at a wavelength of 510 nm.
도 1에 도시된 바와 같이, OA군은 올레인산에 의해 지방의 축적량이 대조군에 비하여 60% 가량 증가하였으나, 상기 OA군에 실시예 1 내지 3의 추출물을 처리(OA+실시예 1 내지 OA+실시예 3)하면 지방 축적량이 대조군에 비하여 각각 20%, 25% 및 14%정도 밖에 증가하지 않는 것을 확인하였다. 이는 약 30 내지 34%의 지방 축적 억제효과를 나타내는 것으로서, 실시예 1 내지 3에 따라 제조된 추출물이 우수한 지방축적 억제 효과를 나타낸다는 것을 의미한다.As shown in FIG. 1, in the OA group, the accumulation amount of fat was increased by 60% as compared with the control group by oleic acid. However, the OA group was treated with the extracts of Examples 1 to 3 (OA + Example 1 to OA + ) Showed that the accumulation of fat was only 20%, 25% and 14%, respectively, as compared with the control group. This indicates a fat accumulation inhibitory effect of about 30 to 34%, which means that the extract prepared according to Examples 1 to 3 exhibits an excellent fat accumulation inhibitory effect.
한편, OA+실시예 4 내지 OA+실시예 6 및 OA+비교예 1은 지방의 축적량이 대조군에 비하여 각각 38%, 43%, 49% 및 60% 증가하므로, 실시예 4 내지 6 및 비교예 1은 실시예 1 내지 실시예 3에 비하여 지방축적 억제 효과가 우수하지 못한 것을 확인하였다.
On the other hand, in Examples 4 to 4, OA + Example 6 and OA + Comparative Example 1, the accumulation amount of fat was increased by 38%, 43%, 49% and 60%, respectively, It was confirmed that the fat accumulation inhibitory effect was not superior to that of Examples 1 to 3.
시험예Test Example 2. 지방축적 억제 확인_사진 확인 2. Confirmation of fat accumulation _ Check photos
상기 시험예 1의 방법에 따라 염색된 각 군의 세포 중 대조군, OA군 및 OA+실시예 1의 세포만 1 ㎖의 물로 세척한 후 적색가시화상현미경(올림푸스, 도쿄, 일본)으로 확인하였다.Of the cells of each group stained according to the method of Test Example 1, only the control group, OA group, and OA + cells of Example 1 were washed with 1 ml of water and identified by a red visible-light microscope (Olympus, Tokyo, Japan).
도 2는 대조군, OA군 및 상기 OA군에 실시예 1에 따라 제조된 추출물로 처리한 군(OA+실시예 1)의 HepG2 세포에 축적된 지방을 적색가시화상현미경으로 촬영한 사진이다.FIG. 2 is a photograph of the fat accumulated in HepG2 cells of the group treated with the extract prepared according to Example 1 (OA + Example 1) in the control group, the OA group and the OA group by a red visible-light microscope.
도 2에 도시된 바와 같이, OA+실시예 1이 OA군에 비하여 세포 내 지방 축적량이 월등히 낮은 것을 확인하였다. 이러한 결과는 본 발명의 실시예 1에 따라 제조된 대파 엽초부 추출물이 지방축적 억제 효과가 우수하다는 것을 의미한다.
As shown in FIG. 2, it was confirmed that OA + Example 1 had significantly lower intracellular fat accumulation than OA group. These results indicate that the extract of rapeseed leaf herb prepared according to Example 1 of the present invention is excellent in inhibiting fat accumulation.
시험예Test Example 3. 세포독성 확인 3. Cytotoxicity check
OA군(유도군)은 HepG2 세포를 24-well 플레이트에 접종하고 1% BSA이 첨가된 낮은 글루코스 DMEM 배지에 0.5 mM 올레인산(oleic acid)을 첨가한 후 37 ℃, 5% CO2 조건 하에 24시간 배양한 다음 5 mg/ml MTT 스톡을 10 ㎕ 넣고 3시간 후 배지를 제거하여 보라색으로 변화된 세포를 확인한 다음 DMSO에 용해시켜 96-well 플레이트로 100 ㎕씩 옮겨 540 nm에서 흡광도를 측정하여 세포독성을 확인하였다. OA group (derived group) is 24 hours under and then inoculated into HepG2 cells in 24-well plate was added to 0.5 mM oleic acid (oleic acid) in low-glucose DMEM medium supplemented with a 1% BSA was added 37 ℃, 5% CO 2 conditions After incubation, 10 μl of 5 mg / ml MTT stock was added. After 3 hours, the medium was removed and the cells changed to purple. The cells were then dissolved in DMSO and transferred to a 96-well plate in an amount of 100 μl. The absorbance at 540 nm was measured. Respectively.
또한, 7개의 처리군은 상기 OA군의 DMEM 배지를 실시예 1 내지 6 및 비교예 1에서 제조된 200 ㎍/㎖의 추출물로 각각 처리한 후 37 ℃, 5% CO2 조건 하에 24시간 배양한 다음 5 mg/ml MTT 스톡을 10 ㎕ 넣고 3시간 후 배지를 제거하여 보라색으로 변화된 세포를 확인한 다음 DMSO에 용해시켜 96-well 플레이트로 100 ㎕씩 옮겨 540 nm에서 흡광도를 측정하여 세포독성을 확인하였다. In the 7 treatment groups, the DMEM medium of the OA group was treated with the extracts of 200 / / ml prepared in Examples 1 to 6 and Comparative Example 1, respectively, and cultured for 24 hours at 37 캜 and 5% CO 2 Next, 10 μl of the 5 mg / ml MTT stock was added, and after 3 hours, the cells were changed to purple. The cells were then dissolved in DMSO and transferred to a 96-well plate at 100 μl. The absorbance at 540 nm was measured to confirm cytotoxicity .
(%)Cell viability
(%)
위 표 1에 나타낸 바와 같이, 본 발명의 실시예 1 내지 6 및 비교예 1에 따라 제조된 추출물은 독성을 보이지 않는 것을 확인하였다.
As shown in Table 1, it was confirmed that the extract prepared according to Examples 1 to 6 and Comparative Example 1 of the present invention showed no toxicity.
시험예 4. Test Example 4. FASNFASN 및 And SREBP1c SREBP1c 발현 측정Expression measurement
지방산 합성과 관련된 유전자들의 발현 감소에 의한 지방산 합성 감소를 확인하기 위하여 지방산 합성과 관련된 대표 유전자인 FASN 및 SREBP1c 유전자들 발현 변화를 측정하였다.Genes involved in fatty acid synthesis A representative gene FASN and SREBP1c the gene expression changes associated with fatty acid synthesis was measured in order to confirm the fatty acid synthesis decreases due to decrease expression.
구체적으로, 대조군은 HepG2 세포를 24-well 세포배양 플레이트에 접종하고 1% BSA이 첨가된 낮은 글루코스 DMEM 배지 37 ℃, 5% CO2 조건 하에 24시간 배양하였다.Specifically, HepG2 cells were inoculated into 24-well cell culture plates and cultured in a low glucose DMEM medium supplemented with 1% BSA at 37 ° C and 5% CO 2 for 24 hours.
또한, OA군(유도군)은 HepG2 세포를 24-well 세포배양 플레이트에 접종하고 1% BSA이 첨가된 낮은 글루코스 DMEM 배지에 0.5 mM 올레인산(oleic acid)을 첨가한 후 37 ℃, 5% CO2 조건 하에 24시간 배양하였다.In the OA group (induction group), HepG2 cells were inoculated on a 24-well cell culture plate, and 0.5 mM oleic acid was added to low glucose DMEM medium supplemented with 1% BSA, followed by addition of 5% CO 2 Lt; / RTI > for 24 hours.
또한, 처리군(OA+실시예 1 내지 OA+실시예 5 및 OA+비교예 1)은 상기 OA군의 배지를 실시예 1 내지 5 및 비교예 1에서 제조된 200 ㎍/㎖의 대파 추출물로 각각 처리한 후 HepG2 세포를 37 ℃, 5% CO2 조건 하에 24시간 배양하였다.In the treatment group (OA + Example 1 to OA + Example 5 and OA + Comparative Example 1), the OA group medium was treated with the 200 占 퐂 / ml of the Robiniae extract prepared in Examples 1 to 5 and Comparative Example 1 HepG2 cells were cultured for 24 hours at 37 ° C and 5% CO 2 .
상기 각 배양한 세포를 수거하여 RNA를 추출하고 상기 추출된 RNA 2 ug을 주형으로 역전사효소를 이용하여 cDNA를 합성한 후 다시 이를 주형으로 하여 FASN 및 SREBP1c 유전자들의 발현 변화를 real-time qPCR(quantitative polymerase chain reaction)을 이용하여 관찰하였다.Each of the return of the cultured cells to extract RNA, and wherein the extracted RNA 2 ug of after synthesizing cDNA using a reverse transcriptase as a template FASN and expression changes in SREBP1c gene as a template back to real-time qPCR (quantitative polymerase chain reaction).
유도 배수 FASN
Induction drainage
유도 배수 SPEBP1C
Induction drainage
위 표 2에 나타낸 바와 같이, OA군으로 처리시 증가되었던 FASN 및 SREBP1c 유전자 전사가 실시예 1 내지 6의 추출물로 처리시 감소되는 것을 확인하였다.As shown in the above Table 2, the increase in processing the OA group was FASN and SREBP1c It was confirmed that gene transcription was reduced upon treatment with the extracts of Examples 1 to 6.
특히, 실시예 1 내지 3의 추출물로 처리한 군이 실시예 4 내지 6의 추출물로 처리한 군에 비하여 FASN 및 SREBP1c 유전자 전사가 더욱 감소하는 것을 확인하였다.In particular, Examples 1 to the group treated with the extract of Example 3 from 4 to FASN and SREBP1c compared with the group treated with the extract of the 6 Gene transcription was further reduced.
반면, 비교예 1의 추출물은 OA군과 유사한 FASN 및 SREBP1c 유전자 전사를 보이므로 효과가 미미하다는 것을 보여준다.
On the other hand, the extract of Comparative Example 1 is similar to the OA group FASN and SREBP1c It shows that the effect is insignificant because it shows gene transcription.
동물실험Animal experiment
동물은 7주령의 숫컷 18-22 g의 C57BL/6 마우스를 샘타코 바이오코리아(Korea)로부터 공급받아 1주일간 실험실 환경에 적응시킨 후 실험에 사용하였으며 고형사료와 물은 자유로이 공급하였다. 실험실은 일정한 온도(22±2 ℃)와 습도(50±10%) 및 12시간 낮과 밤의 주기를 유지하였다. Animals were fed with 18-22 g of C57BL / 6 mice, 7 weeks of age, from Sam Taco Bio Korea (Korea), adapted to the laboratory environment for 1 week, and used for the experiment. The solid feed and water were supplied freely. The laboratory maintained a constant temperature (22 ± 2 ° C), humidity (50 ± 10%) and a 12-hour day and night cycle.
시험예Test Example 5. 부검사진 5. Autopsy photos
실험동물은 체중변화가 일정하고 건강한 동물만을 선별하여 임의 배치법에 의해 정상군, 대조군 및 시료 투여군으로 나누었으며 실험군마다 10마리씩 사용하였다. 정상 동물모델 실험을 위하여 일반 식이를 급여한 정상군(normal), High fat-High sucrose diet를 급여한 대조군(control) 및 High fat-High sucrose diet와 실시예 3의 추출물 1%를 투여한 시료 투여군(실시예 3)으로 나누었다.The experimental animals were divided into normal group, control group and sample group by randomly selecting healthy animals with constant weight change, and 10 animals were used in each experimental group. High fat-high sucrose diet (control) and high fat-high sucrose diet (control) and 1% of the extract of Example 3 (normal) (Example 3).
투여는 연속 12주간 매일 1회 경구로 하였으며 실험 종료일에 실험동물을 희생시켰다.The doses were given once a day for 12 consecutive weeks and the animals were sacrificed at the end of the experiment.
도 3은 실시예 3의 추출물을 투여한 시료 투여군, 정상군 및 대조군의 마우스를 희생시켜 간을 촬영한 사진이다.FIG. 3 is a photograph of a liver taken at the sacrifice of a mouse administered with a sample administered with the extract of Example 3, a normal group, and a control group.
도 3에 도시된 바와 같이, 실시예 3의 추출물을 투여한 시료 투여군은 대조군에 비하여 지방축적으로 의한 간조직의 변형이 낮은 것을 확인하였다.
As shown in FIG. 3, the group administered with the extract of Example 3 showed less deformation of liver tissue due to fat accumulation than the control group.
시험예Test Example 6. 체중 및 간 조직과 6. Weight and liver tissue 부고환지방의Epididymal 무게 측정 Weighing
12주간 마우스의 체중을 측정하였으며, 실험 종료일에 마우스를 희생시킨 후 간 조직 및 부고환지방을 적출하여 중량을 측정하고 사진을 촬영하였다.The mice were weighed for 12 weeks. After the mice were sacrificed at the end of the experiment, liver and epididymal fat were extracted, weighed and photographed.
도 4는 12주 동안 실시예 3의 추출물을 투여한 시료 투여군, 정상군 및 대조군 마우스의 체중을 측정한 그래프이다.FIG. 4 is a graph showing body weight of a sample-administered group, a normal group, and a control mouse in which the extract of Example 3 was administered for 12 weeks.
도 4에 도시된 바와 같이, 고지방식이만 급여한 대조군의 체중이 가장 많이 증가하였으며, 고지방식이와 실시예 3의 추출물을 함께 섭취한 시료 투여군은 상기 대조군에 비하여 체중이 적게 증가하는 것을 확인하였다.
As shown in FIG. 4, the body weight of the control group fed only with the high fat diet increased the most, and the weight of the high fat diet and the extract group of Example 3 were lowered Respectively.
도 5a는 실시예 3의 추출물을 투여한 시료 투여군, 정상군 및 대조군 마우스를 희생시킨 후 추출된 간 조직의 무게를 나타낸 그래프이며, 도 5b는 상기 각 군에서 추출된 간 조직을 촬영한 사진이다. 또한, 도 6a는 실시예 3의 추출물을 투여한 시료 투여군, 정상군 및 대조군 마우스를 희생시킨 후 추출된 부고환지방의 무게를 나타낸 그래프이며, 도 5b는 상기 각 군에서 추출된 부고환지방을 촬영한 사진이다. FIG. 5A is a graph showing the weights of liver tissues extracted after sacrifice of a sample administration group, a normal group and a control mouse to which the extract of Example 3 was administered, and FIG. 5B is a photograph of liver tissues extracted from the above groups . 6A is a graph showing the weight of epididymal fat extracted after sacrifice of the sample administration group, the normal group and the control mouse to which the extract of Example 3 was administered. FIG. 5B is a graph showing the epididymal fat extracted from each group It is a photograph.
도 5 및 6에 도시된 바와 같이, 실시예 3의 추출물을 투여한 시료 투여군의 간 조직 무게는 정상군에 비해서 많이 나갔지만, 대조군에 비해서 적게 나가는 것을 확인하였다. 또한, 부고환지방 역시 실시예 3의 추출물을 투여한 시료 투여군의 부고환지방 무게가 정상군에 비해서 많이 나갔지만, 대조군에 비해서 적게 나가는 것을 확인하였다.
As shown in FIGS. 5 and 6, the weight of the liver of the sample administered with the extract of Example 3 was much higher than that of the normal group, but it was less than that of the control group. In addition, the epididymal fat weight of the sample treated with the extract of Example 3 was higher than that of the normal group, but it was lower than that of the control group.
시험예Test Example 7. 혈액 분석 7. Blood analysis
상기 희생된 마우스에서 채혈한 혈액을 상온에서 30분 응고시킨 후 3000 rpm에서 10분간 원심 분리하여 혈청을 회수하였다. 회수한 혈청을 Total cholesterol (TC), Low density lipoprotein-cholesterol (LDL-C), High density lipoprotein-cholesterol (HDL-C), Total protein, Albumin, Alkaline phosphatase (ALP), Aaspartate aminotransferase (AST), Alanine aminotransaminase (ALT)의 분석에 사용하였으며 녹십자의료재단(GC Labs, 한국)에 분석을 의뢰하였다.Blood collected from the sacrificed mice was coagulated at room temperature for 30 minutes, and centrifuged at 3000 rpm for 10 minutes to recover the serum. (TC), low density lipoprotein-cholesterol (LDL-C), high density lipoprotein-cholesterol (HDL-C), total protein, albumin, alkaline phosphatase (ALP), Aaspartate aminotransferase aminotransaminase (ALT), and was requested to be analyzed by the Green Cross Medical Foundation (GC Labs, Korea).
도 7a 내지 7f는 실시예 3의 추출물을 투여한 시료 투여군, 정상군(N) 및 대조군(C) 마우스에서 채혈한 혈액의 Alkaline phosphatase (ALP), Alanine aminotransaminase (ALT), Aaspartate aminotransferase (AST), Total cholesterol (TC), Low density lipoprotein-cholesterol (LDL-C), High density lipoprotein-cholesterol (HDL-C) 및 Total protein 수치를 나타낸 그래프이다.FIGS. 7A to 7F are graphs showing changes in the concentrations of Alkaline phosphatase (ALP), alanine aminotransaminase (ALT), and Aaspartate aminotransferase (AST) in the blood collected from the sample administration group, the normal group (N) (TC), low density lipoprotein-cholesterol (LDL-C), high density lipoprotein-cholesterol (HDL-C) and total protein levels.
도 7에 나타낸 바와 같이, ALP, ALT, AST, TC 및 LDL-C에서는 실시예 3의 추출물을 투여한 시료 투여군이 대조군(C)에 비해서는 수치가 낮고 정상군(N)dp 비해서는 수치가 높은 것을 확인하였다. 상기 ALP, ALT 및 AST는 간 독성 여부를 판단하는 대표적인 지표로서, 실시예 3의 추출물을 투여한 시료 투여군이 대조군에 비하여 감소되는 것으로 보아 대표 추출물에 의한 간독성의 개선 효과가 있는 것으로 확인하였다.As shown in FIG. 7, in the ALP, ALT, AST, TC and LDL-C, the sample to which the extract of Example 3 was administered had a lower value than the control (C) Respectively. The ALP, ALT, and AST are representative indexes for determining liver toxicity. As a result, the extract administered with the extract of Example 3 was found to be reduced compared to the control group. Thus, it was confirmed that hepatic toxicity was improved by the representative extract.
또한, LDL-C의 생성억제로 인해 지방간 개선에 효과가 있다는 것을 확인하였다.
In addition, it was confirmed that the inhibition of the production of LDL-C was effective in the improvement of fatty liver.
시험예Test Example 8. 동맥경화 지수 8. Atherosclerotic index
동맥경화 지수(Atherogenic Index, AI)는 Haglund의 계산법(1991)에 따라 아래와 같이 계산하여 구하였다.The atherogenic index (AI) was calculated according to Haglund's calculation (1991) as follows.
[수학식 1][Equation 1]
AI={(총 콜레스테롤량- HDL 콜레스테롤량)/HDL 콜레스테롤량}AI = {(total cholesterol level - HDL cholesterol level) / HDL cholesterol level}
도 8은 실시예 3의 추출물을 투여한 시료 투여군, 정상군 및 대조군 마우스에서 측정한 동맥경화 지수를 나타낸 그래프이다.8 is a graph showing the arteriosclerosis index measured in the sample administration group, the normal group and the control mouse to which the extract of Example 3 was administered.
도 8에 도시된 바와 같이, 실시예 3의 추출물을 투여한 시료 투여군의 지수는 정상군(normal)에 비해서는 높았으나, 대조군(control)에 비해서는 낮은 것을 확인하였다.
As shown in FIG. 8, the index of the group administered with the extract of Example 3 was higher than that of the normal group but lower than that of the control group.
하기에 본 발명의 분말을 함유하는 조성물의 제제예를 설명하나, 본 발명은 이를 한정하고자 함이 아닌 단지 구체적으로 설명하고자 함이다.Hereinafter, formulation examples of the composition containing the powder of the present invention will be described, but the present invention is not intended to be limited thereto but is specifically described.
제제예Formulation example 1. One. 산제의Sanje 제조 Produce
실시예 1에서 얻은 추출물 분말 500 mg500 mg of the extract powder obtained in Example 1
유당 100 mg
탈크 10 mgTalc 10 mg
상기의 성분들을 혼합하고 기밀포에 충진하여 산제를 제조한다.
The above components are mixed and filled in airtight bags to prepare powders.
제제예Formulation example 2. 정제의 제조 2. Preparation of tablets
실시예 1에서 얻은 추출물 분말 300 mg300 mg of the extract powder obtained in Example 1
옥수수전분 100 mg
유당 100 mg
스테아린산 마그네슘 2 mgMagnesium stearate 2 mg
상기의 성분들을 혼합한 후 통상의 정제의 제조방법에 따라서 타정하여 정제를 제조한다.
After mixing the above components, tablets are prepared by tableting according to the usual preparation method of tablets.
제제예Formulation example 3. 캅셀제의 제조 3. Preparation of capsules
실시예 1에서 얻은 추출물 분말 200 mg200 mg of the extract powder obtained in Example 1
결정성 셀룰로오스 3 mgCrystalline cellulose 3 mg
락토오스 14.8 mgLactose 14.8 mg
마그네슘 스테아레이트 0.2 mgMagnesium stearate 0.2 mg
통상의 캡슐제 제조방법에 따라 상기의 성분을 혼합하고 젤라틴 캡슐에 충전하여 캡슐제를 제조한다.
The above components are mixed according to a conventional capsule preparation method and filled in gelatin capsules to prepare capsules.
제제예Formulation example 4. 주사제의 제조 4. Preparation of injections
실시예 1에서 얻은 추출물 분말 600 mg600 mg of the extract powder obtained in Example 1
만니톨 180 mg180 mg mannitol
주사용 멸균 증류수 2974 mgSterile sterilized water for injection 2974 mg
Na2HPO4 ,12H2O 26 mgNa 2 HPO 4 , 12H 2 O 26 mg
통상의 주사제의 제조방법에 따라 1 앰플 당 상기의 성분 함량으로 제조한다.
It is prepared by the above-mentioned component content per ampoule according to the usual injection preparation method.
제제예Formulation example 5. 5. 액제의Liquid 제조 Produce
실시예 1에서 얻은 추출물 분말 4 g4 g of the extract powder obtained in Example 1
이성화당 10 g10 g per isomer
만니톨 5 g5 g mannitol
정제수 적량Purified water quantity
통상의 액제의 제조방법에 따라 정제수에 각각의 성분을 가하여 용해시키고 레몬향을 적량 가한 다음 상기의 성분을 혼합한 다음 정제수를 가하여 전체를 정제수를 가하여 전체 100g으로 조절한 후 갈색병에 충진하여 멸균시켜 액제를 제조한다.
Each component was added and dissolved in purified water according to the usual liquid preparation method, and the lemon flavor was added in an appropriate amount. Then, the above components were mixed, and purified water was added thereto. The whole was adjusted to 100 g with purified water, To prepare a liquid agent.
제제예Formulation example 6. 과립제의 제조 6. Preparation of granules
실시예 1에서 얻은 추출물 분말 1,000 mg1,000 mg of the extract powder obtained in Example 1
비타민 혼합물 적량Vitamin mixture quantity
비타민 A 아세테이트 70 Vitamin A Acetate 70
비타민 E 1.0 mgVitamin E 1.0 mg
비타민 B1 0.13 mgVitamin B1 0.13 mg
비타민 B2 0.15 mg0.15 mg of vitamin B2
비타민 B6 0.5 mgVitamin B6 0.5 mg
비타민 B12 0.2 Vitamin B12 0.2
비타민 C 10 mgVitamin C 10 mg
비오틴 10 Biotin 10
니코틴산아미드 1.7 mgNicotinic acid amide 1.7 mg
엽산 50
판토텐산 칼슘 0.5 mgCalcium pantothenate 0.5 mg
무기질 혼합물 적량Mineral mixture quantity
황산제1철 1.75 mg1.75 mg of ferrous sulfate
산화아연 0.82 mg0.82 mg of zinc oxide
탄산마그네슘 25.3 mgMagnesium carbonate 25.3 mg
제1인산칼륨 15 mgPotassium monophosphate 15 mg
제2인산칼슘 55 mgSecondary calcium phosphate 55 mg
구연산칼륨 90 mgPotassium citrate 90 mg
탄산칼슘 100 mg
염화마그네슘 24.8 mgMagnesium chloride 24.8 mg
상기의 비타민 및 미네랄 혼합물의 조성비는 비교적 과립제에 적합한 성분을 바람직한 실시예로 혼합 조성하였지만, 그 배합비를 임의로 변형 실시하여도 무방하며, 통상의 과립제 제조방법에 따라 상기의 성분을 혼합한 다음, 과립을 제조하고, 통상의 방법에 따라 건강기능식품 조성물 제조에 사용할 수 있다.
The composition ratio of the above-mentioned vitamins and minerals is comparatively comparatively mixed with the granules according to the preferred embodiment. However, the blending ratio may be arbitrarily changed, and the above components are mixed according to the ordinary granule preparation method, Can be prepared and used in the manufacture of a health functional food composition according to a conventional method.
제제예Formulation example 7. 기능성 음료의 제조 7. Manufacture of functional beverages
실시예 1에서 얻은 추출물 분말 1,000 mg 1,000 mg of the extract powder obtained in Example 1
구연산 1,000 mgCitric acid 1,000 mg
올리고당 100 g100 g of oligosaccharide
매실농축액 2 gPlum concentrate 2 g
타우린 1 gTaurine 1 g
정제수를 가하여 전체 900 mLPurified water was added to the flask to obtain a total of 900 mL
통상의 건강음료 제조방법에 따라 상기의 성분을 혼합한 다음, 약 1 시간 동안 85 ℃에서 교반 가열한 후, 만들어진 용액을 여과하여 멸균된 2 L 용기에 취득하여 밀봉 멸균한 뒤 냉장 보관한 다음 본 발명의 기능성 음료 조성물 제조에 사용한다. The above components were mixed according to a conventional health drink manufacturing method, and the mixture was stirred and heated at 85 DEG C for about 1 hour. The solution thus prepared was filtered and sterilized in a sterilized 2 L container, It is used in the production of the functional beverage composition of the invention.
상기 조성비는 비교적 기호음료에 적합한 성분을 바람직한 실시예로 혼합 조성하였지만, 수요계층, 수요국가, 사용용도 등 지역적, 민족적 기호도에 따라서 그 배합비를 임의로 변형 실시하여도 무방하다.Although the composition ratio is a mixture of the components suitable for the preferred beverage as a preferred embodiment, the compounding ratio may be arbitrarily varied depending on the regional and national preferences such as the demand level, the demanding country, and the intended use.
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