KR101994483B1 - Pharmaceutical composition comprising the sprout extracts of zingiber officinale as an effective component for prevention or treatment of diabetes and health functional food comprising the same - Google Patents
Pharmaceutical composition comprising the sprout extracts of zingiber officinale as an effective component for prevention or treatment of diabetes and health functional food comprising the same Download PDFInfo
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- KR101994483B1 KR101994483B1 KR1020170166246A KR20170166246A KR101994483B1 KR 101994483 B1 KR101994483 B1 KR 101994483B1 KR 1020170166246 A KR1020170166246 A KR 1020170166246A KR 20170166246 A KR20170166246 A KR 20170166246A KR 101994483 B1 KR101994483 B1 KR 101994483B1
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- ginger
- extract
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- present
- ginger root
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Images
Classifications
-
- 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)
- A61K36/906—Zingiberaceae (Ginger family)
- A61K36/9068—Zingiber, e.g. garden ginger
-
- 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
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
-
- 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
- A23V2200/328—Foods, ingredients or supplements having a functional effect on health having effect on glycaemic control and diabetes
-
- Y10S514/866—
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- Health & Medical Sciences (AREA)
- Natural Medicines & Medicinal Plants (AREA)
- Life Sciences & Earth Sciences (AREA)
- Botany (AREA)
- Mycology (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Medical Informatics (AREA)
- Epidemiology (AREA)
- Food Science & Technology (AREA)
- Alternative & Traditional Medicine (AREA)
- Biotechnology (AREA)
- Nutrition Science (AREA)
- Medicinal Chemistry (AREA)
- Microbiology (AREA)
- Pharmacology & Pharmacy (AREA)
- Polymers & Plastics (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
- Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
본 발명은 생강싹(sprout of Zingiber officinale) 추출물을 유효성분으로 함유하는 항당뇨 활성의 조성물로서, 보다 상세하게는, 생강싹의 에탄올 추출물, 이의 헥센 분획물, 에틸아세테이트 분획물 또는 부탄올 분획물을 포함하는 전분분해효소 저해활성이 우수한 항당뇨 활성의 의약 및 기능성 식품 조성물에 관한 것이다. 본 발명의 생강싹 추출물은 우수한 전분분해효소 저해활성에 의한 항당뇨 활성을 나타내어, 당뇨병의 예방 또는 개선용 의약품 및 건강 기능 식품으로 사용할 수 있는 뛰어난 효과가 있다. 뿐만 아니라, 본 발명의 생강싹 추출물은 열 안정성이 우수하고, pH 2의 산성조건 및 혈장 내에서도 전분분해효소 저해활성의 손실이 나타나지 않아, 액상, 크림, 분말, 환, 정 등의 다양한 형태로 손쉽게 가공될 수 있어 제약 산업 및 식품 산업상 매우 유용하게 이용될 수 있다. 또한, 생강 저장 중에 생성되는 싹을 이용하므로 불용 자원의 재이용 및 환경 보존에 기여할 수 있는 장점이 있다. The present invention relates to an antidiabetic activity composition containing an extract of sprout of Zingiber officinale as an active ingredient, and more particularly to a composition containing an ethanol extract of ginger root, a hexane fraction thereof, an ethyl acetate fraction or a butanol fraction The present invention relates to a medicament and a functional food composition having an antidiabetic activity which is excellent in inhibitory activity against a decomposition enzyme. The ginger root extract of the present invention exhibits an antidiabetic activity due to its excellent inhibitory activity against starcholytic enzymes, and has an excellent effect that it can be used as a medicament for the prevention or improvement of diabetes and a health functional food. In addition, the ginger root extract of the present invention is excellent in thermal stability and does not exhibit the activity of inhibiting the activity of inhibiting the activity of starch hydrolyzing enzyme even in an acidic condition of pH 2 and plasma, and can be easily formulated into various forms such as liquid, cream, powder, And can be very usefully used in the pharmaceutical industry and the food industry. Also, since the sprouts generated during the storage of ginger are used, there is an advantage that it can contribute to reuse of insoluble resources and environmental preservation.
Description
본 발명은 생강싹(sprout of Zingiber officinale) 추출물을 유효성분으로 함유하는 항당뇨 활성의 조성물로서, 보다 상세하게는, 생강싹의 에탄올 추출물, 이의 헥센 분획물, 에틸아세테이트 분획물 또는 부탄올 분획물을 포함하는 전분분해효소 저해활성이 우수한 항당뇨 활성의 의약 및 기능성 식품 조성물에 관한 것이다.The present invention relates to a sprout of Zingiber officinale extract as an active ingredient, and more particularly to a composition of an anti-diabetic activity comprising an ethanol extract of ginger root, a hexane fraction thereof, an ethyl acetate fraction or a butanol fraction, ≪ / RTI >
현대인에게 가장 흔한 질병 중 하나인 당뇨병은 인슐린의 분비량이 부족하거나 정상적인 기능이 이루어지지 않는 등의 원인으로 발생하는 대사질환의 일종으로서 혈중 포도당 농도가 높은 것이 특징이며, 고혈당으로 인하여 여러 증상 및 징후를 일으킨다. 당뇨병은 제1형과 제2형으로 구분되는데, 제1형 당뇨병은 유전적 영향에 의해 인슐린을 전혀 생산하지 못하는 것이 원인이 되어 발생하며, 제2형 당뇨병은 인슐린 기능이 떨어져 인슐린 저항성(insulin resistance)이 원인으로 알려져 있다. 특히, 제2형 당뇨병은 식생활의 서구화에 따른 고열량, 고지방, 고단백의 식단, 운동부족, 스트레스 등 환경적인 요인이 크게 작용하는 것으로 알려져 있다.Diabetes mellitus, one of the most common diseases among modern people, is a type of metabolic disorder caused by lack of insulin secretion or lack of normal functions. It is characterized by a high blood glucose level, and due to hyperglycemia, various symptoms and signs Cause. Diabetes mellitus is divided into Type 1 and Type 2, Type 1 diabetes is caused by the inability to produce insulin at all due to genetic influences, Type 2 diabetes has insulin resistance, insulin resistance ) Is known to be the cause. In particular, type 2 diabetes is known to be affected by environmental factors such as high calorie, high fat, high protein diet, lack of exercise, and stress due to westernization of dietary habits.
당뇨병의 치료를 위해서는 제1형 당뇨병의 경우에는 인슐린 주사가 필수적이며, 제2형 당뇨병의 경우에는 생활습관 교정 및 약물치료가 필요하며, 대표적으로 인슐린 분비 촉진제(예, 레파글리나이드, 미티글이나이드) 및 소장에서의 탄수화물 흡수 지연제[글루코바이: 성분명-아카보즈(acarbose), 베이슨: 성분명-보글리보스 (voglibose)] 등이 이용되고 있다.For the treatment of diabetes, insulin injection is essential for type 1 diabetes, and lifestyle modification and medication are needed for type 2 diabetes. Typically, insulin secretagogues (eg, repaglinide, Carbohydrate absorption retarder [Glucobyme: ingredient name - acarbose, basin: ingredient name - voglibose], and the like.
한편, 생강(Zingiber officinale)은 생강과(Zingiberaceae)의 아열대, 열대산 다년생 초본 식물의 하나로, 지하부 근경은 특이한 맛과 향을 가지고 있어, 국내에서는 마늘, 고추와 함께 3대 향신료로 이용되고 있다. 생강은 그대로 식품으로 이용되기도 하며, 김치, 젓갈, 과자류 등의 다양한 식품 산업에 양념으로 이용되고 있다. 또한, 건조한 생강(건강 또는 건조분말), 올레오레진(생강 함유지), 생강 정유 등의 제품 형태로 가공되어 식의약 소재 및 화장품 소재로 이용되기도 한다(김선희 2013. 한양대학교 박사논문). 실제 생강은 본초강목에 백사(백가지 질병)를 방지한다고 기술될 만큼 다양한 생리 활성이 알려져 있으며, 한방에서는 건위약 및 소화 불량, 구토, 진통에 효과가 우수하다고 알려져 있다. 생강의 생약명은 [건강]이며, 일반인은 [새앙]으로도 많이 부른다. 고려시대에는 왕의 하사품으로 사용될 만큼 진기하였으며, 인도의 고대의학서적 [아유르데바]에는 "신이 내린 치료제"로 소개되어 있다. On the other hand, ginger (Zingiber officinale ) is one of the subtropical and tropical perennial herbaceous plants of the ginger (Zingiberaceae). Its rootstock has unusual taste and aroma, and it is used as three spices together with garlic and pepper in Korea. Ginger is used as food as it is, and it is used as seasoning in various food industries such as kimchi, salted fish, and confectionery. In addition, they are processed into product forms such as dried ginger (health or dried powder), oleoresin (preserved ginger), and ginger essential oil, and they are also used as food materials and cosmetic materials (Kim Sunhee 2013. Ph.D. Actual ginger is known to have a wide variety of physiological activities that are said to prevent white spots (100 kinds of diseases) in the main stem of the ginger, and it is known that it has excellent effects on the placebo and digestive disorders, vomiting and analgesia. The generic name of the ginger is [health], and the general person is also called [schan]. In the Goryeo Dynasty, it was used as a royal slave, and in ancient medicine books [Ayurveda] of India, it is introduced as "God's treatment".
생강의 주요 생산국은 인도이며, 전 세계 생산량의 50% 이상을 차지한다. 국내에서는 경북 안동, 영주 지역과 충남 서산, 당진 지역, 전북 완주 지역에서 주로 생산되고 있다. 생강의 품종은 재래종, 개량종으로, 또한, 조생종, 중생종, 만생종으로 분류하며, 크기에 따라, 소생강, 중생강, 대생강 등으로 구분한다. 농촌진흥청 식품분석 DB(2006)에 따르면, 생강은 단백질 1.5%, 지방 0.2%, 탄수화물 13.9%, 회분 1.1%, 수분 83.3%를 함유하며, 다양한 비타민과 미네랄, 필수 아미노산을 포함하고 있다. 특히, 생강은 100g당 344mg의 칼륨을 가지고 있어, 염분 배출에도 도움이 된다. 생강의 유효성분으로는 다양한 탄화수소(hydrocarbon), 케톤(ketone)류의 증류 성분과 진지베렌(zingiberene) 등의 향기 성분, 보르네올(borneol), 진지베롤(gingiberol) 등의 기름 성분, 매운맛의 6-진제롤(6-ginegerol), 6-샤가올(6-shagaol) 등이 알려져 있다. 이들 성분은 항산화 활성(이은주 외, 2011, 한국식품과학회지, 43: 469-474), 항균 작용(이명희 외, 2012. J. Fd Hyg. Safety 27: 109-116), 항 위염, 항 위궤양 작용(김신정 외, 2012, J Korean Soc Food Sci Nutr, 41: 1528-1533), 항암 작용(이예솔, 2013. 울산대학교 박사논문), 항진해 작용. 해독 진정 작용(한효선 2016, 건국대학교 박사논문), 판크레아틱 리파아제(pancreatic lipase) 저해 작용(Bae JS 등, 2011. Korean J. Food Preserv. 18: 390-396), 혈중 콜레스테롤 저하 작용(오수정, 2015. 이화여자대학교 석사논문), 간보호 작용(Koo SW 외, 2012, J Vet Clin 29: 441-446), 뇌세포 보호 작용(정길생 외, 2010, 생약학회지 41: 190-195) 등 다양한 생리 활성이 알려져, 현재 생강은 식품 부재료이면서 건강 기능성 식품 소재로 각광받고 있다. The main producer of ginger is India, accounting for more than 50% of global production. In Korea, it is mainly produced in Andong, Youngju, Chungnam, Seosan, Dangjin, and Jeonbuk Wanju. Ginger varieties are classified into native and improved varieties. Also, they are classified into early, late, and late-life varieties. Depending on their size, they are divided into small ginseng, regenerated ginseng, and large ginseng. Ginger contains 1.5% protein, 0.2% fat, 13.9% carbohydrate, 1.1% ash and 83.3% water content and contains various vitamins, minerals and essential amino acids according to the Food Analysis DB (2006) of the Rural Development Administration. In particular, ginger has 344 mg of potassium per 100 g, which is also beneficial for saline emissions. The effective components of ginger include distillation components of various hydrocarbons and ketones, aroma components such as zingiberene, oil components such as borneol and gingiberol, - 6-ginegerol, 6-shagaol, and the like are known. These components have antioxidant activity (Lee, EJ et al., 2011, The Korean Society of Food Science and Technology, 43: 469-474), antimicrobial activity (Lee, Myunghee et al., 2012. J. Fd Hyg. Safety 27: 109-116) (Kim, Shin Jung et al., 2012, J Korean Soc Food Sci Nutr, 41: 1528-1533), anti-cancer effect (Yae Yeol, 2013. doctoral dissertation, Ulsan University) Inhibitory effect of pancreatic lipase (Bae JS et al., 2011. Korean J. Food Preserv. 18: 390-396), blood cholesterol lowering effect (Koo SW et al., 2012, J Vet Clin 29: 441-446), brain-cell protective action (Jeong Gil-hyun et al., 2010, Journal of the Korean Pharmacopoeia 41: 190-195) Physiological activity is known, and now ginger is a food ingredient, but it is attracting attention as health functional food material.
생강의 부위 중에서 생강싹은 기온이 올라가는 4월 이후 생강의 지하부에서 지속적으로 불규칙적으로 발생하며(도 1), 생강 농가에서는 장기간의 저장기간 동안 싹이 나타나지 않아야만 상품성을 유지할 수 있으므로, 생강싹은 회피의 대상으로 인식되고 있다. 그러나, 생강싹은 부드러운 식감과 함께 생강 특유의 향미를 나타내므로, 이를 이용한 고부가가치 식의약품 소재 개발의 필요성이 대두되나, 현재까지는 이에 대한 연구는 전무한 상태이며, 특별한 용도 없이 버려지고 있는 생강싹의 전분분해효소 저해에 의한 강력한 항당뇨 활성은 알려진 바 없다. Ginger sprouts in the ginger area continuously irregularly occur at the bottom of the ginger after April when the temperature rises (Fig. 1). Since the ginger sprout can keep its merchantability only when the ginger root does not show shoots for a long period of storage, It is recognized as an object of avoidance. However, since ginger sprouts exhibit a unique taste of ginger with a soft texture, there is a need to develop a high value-added pharmaceutical material using such ginger sprouts. However, no study has been made so far, and ginger sprouts No strong antidiabetic activity due to inhibition of starch degrading enzymes is known.
생강과 관련된 특허로는 대한민국 등록특허 제10-1476092호에 [숙취 해소 음료 및 그의 제조 방법], 등록특허 제10-1195527호에 [생강 추출물의 제조방법, 이로부터 제조된 생강 추출물을 포함하는 구강용 조성물 및 그 제조방법], 등록특허 제10-1145073호에 [증숙된 생강 추출물 함유 미백용 화장료 조성물 및 그 제조방법], 등록특허 제10-1384633호에 [생강 추출물 또는 이의 분획물을 유효성분으로 포함하는 톡소플라즈마증의 예방 및 치료용 조성물] 등 다양한 효능의 생강 조성물이 알려져 있으나, 유용 생리 활성을 나타내는 생강싹에 대한 특허는 없으며, 특히, 생강싹의 강력한 항당뇨 활성에 관한 특허는 알려진 바 없다.Patents related to ginger include Korea Patent No. 10-1476092 [hangover drink and its preparation method], and Korean Patent No. 10-1195527 [method of producing ginger extract, oral cavity containing ginger extract manufactured therefrom And a method for producing the same], and a method for producing a whitening cosmetic composition containing the ginger extract and a method for producing the same, which are disclosed in Patent No. 10-1145073 A composition for prevention and treatment of toxoplasmosis which is included in the ginger composition of the present invention]. However, there is no patent for ginger sprouts exhibiting useful physiological activity, and in particular, a patent for a strong antidiabetic activity of ginger sprouts is known none.
본 발명은 상기와 같은 종래 기술의 문제점을 해결하기 위하여 안출된 것으로서, 본 발명에서 해결하고자 하는 과제는, 생강 농가에서 저장기간 동안 발아하여 상품성을 떨어뜨리는 것으로 알려진 생강싹의 추출물을 조제하고, 이를 유효성분으로 함유하는 항당뇨 활성을 갖는 의약 및 기능성 식품 조성물을 제공하고자 하는 것이다.Disclosure of Invention Technical Problem [8] Accordingly, the present invention has been made to solve the above-mentioned problems occurring in the prior art, and an object of the present invention is to provide an extract of ginger root which is known to lower germination and germination during storage in a ginger farm, And to provide a medicament and a functional food composition having antidiabetic activity as an active ingredient.
상기와 같은 과제를 해결하기 위하여, 본 발명은 생강싹(Sprout of Zingiber officinale) 추출물을 유효성분으로 함유하는 당뇨병의 예방 또는 치료용 약학적 조성물을 제공한다.In order to solve the above problems, the present invention provides a pharmaceutical composition for preventing or treating diabetes comprising an extract of Sprout of Zingiber officinale as an active ingredient.
상기 생강싹 추출물은 생강싹 에탄올 추출물인 것이 바람직하다.The ginger root extract is preferably an ethanol extract of ginger root.
상기 생강싹 추출물은 생강싹 에탄올 추출물의 헥센 분획물, 에틸아세테이트 분획물 또는 부탄올 분획물인 것이 바람직하다.Preferably, the ginger root extract is a hexane fraction, an ethyl acetate fraction or a butanol fraction of an ethanol extract of ginger root.
또한, 본 발명은 상기 기재된 생강싹 추출물을 유효성분으로 함유하는 당뇨병의 예방 또는 개선용 건강 기능 식품을 제공한다.The present invention also provides a health functional food for preventing or ameliorating diabetes containing the ginger root extract described above as an active ingredient.
본 발명의 생강싹 추출물은 우수한 전분분해효소 저해활성에 의한 항당뇨 활성을 나타내어, 당뇨병의 예방 또는 개선용 의약품 및 건강 기능 식품으로 사용할 수 있는 뛰어난 효과가 있다. 뿐만 아니라, 본 발명의 생강싹 추출물은 열 안정성이 우수하고, pH 2의 산성조건 및 혈장 내에서도 전분분해효소 저해활성의 손실이 나타나지 않아, 액상, 크림, 분말, 환, 정 등의 다양한 형태로 손쉽게 가공될 수 있어 제약 산업 및 식품 산업상 매우 유용하게 이용될 수 있다. 또한, 생강 저장 중에 생성되는 싹을 이용하므로 불용 자원의 재이용 및 환경 보존에 기여할 수 있는 장점이 있다. The ginger root extract of the present invention exhibits an antidiabetic activity due to its excellent inhibitory activity against starcholytic enzymes, and has an excellent effect that it can be used as a medicament for the prevention or improvement of diabetes and a health functional food. In addition, the ginger root extract of the present invention is excellent in thermal stability and does not exhibit the activity of inhibiting the activity of inhibiting the activity of starch hydrolyzing enzyme even in an acidic condition of pH 2 and plasma, and can be easily formulated into various forms such as liquid, cream, powder, And can be very usefully used in the pharmaceutical industry and the food industry. Also, since the sprouts generated during the storage of ginger are used, there is an advantage that it can contribute to reuse of insoluble resources and environmental preservation.
도 1은 저장 중 발아한 생강싹을 나타내고,
도 2는 생강 지하부로부터 회수된 생강싹을 나타내며,
도 3은 생강싹이 성장하여 생강 줄기가 생성된 것을 보여준다.Figure 1 shows ginger buds germinated during storage,
2 shows the ginger shoot recovered from the root of ginger,
Fig. 3 shows that ginger shoots were grown to produce ginger stems.
이하, 본 발명을 상세하게 설명한다.Hereinafter, the present invention will be described in detail.
본 발명은 생강싹 추출물을 함유하는 강력한 항당뇨 활성을 나타내는 의약 및 기능성 식품 조성물에 관한 것이다. 본 발명의 발명자는 생강싹으로부터 에탄올 추출물 및 이의 헥센, 에틸아세테이트, 부탄올로 순차적 유기용매 분획하여 얻어지는 각 분획물들과 물 잔류물을 조제하고, 이중 에탄올 추출물과 이의 헥센 분획물, 에틸아세테이트 분획물 및 부탄올 분획물에서 강력한 전분분해효소 저해활성을 확인하였다.The present invention relates to a medicament and a functional food composition exhibiting a strong antidiabetic activity containing ginger root extract. The inventors of the present invention prepared each fraction and a water residue obtained by fractionating the ethanol extract from ginger sprouts with hexane, ethyl acetate and butanol sequentially, and a water residue. The extract of the double ethanol and its hexane fraction, ethyl acetate fraction and butanol fraction And the activity of the enzyme was inhibited.
상기와 같은 본 발명의 생강싹 추출물 및 이의 활성 분획물들의 전분분해효소 저해활성은 당뇨병의 예방, 치료, 개선을 위한 의약 및 기능성 식품의 용도로서 적용 가능하며, 따라서, 본 발명은 생강싹(Sprout of Zingiber officinale) 추출물을 유효성분으로 함유하는 항당뇨 활성의 의약 및 기능성 식품 조성물을 제공한다.The activity of the ginger extract of the present invention and its active fractions can be applied to the use of medicines and functional foods for the prevention, treatment and improvement of diabetes. Thus, the present invention relates to a ginger sprout Zingiber officinale ) as an active ingredient, and to provide a pharmaceutical and functional food composition having antidiabetic activity.
상기 생강싹 추출물은 생강싹 에탄올 추출물인 것이 바람직하다. 또한, 상기 생강싹 추출물은 생강싹 에탄올 추출물로부터 헥센, 에틸아세테이트 및 부탄올의 유기용매로 순차 분획된 헥센 분획물, 에틸아세테이트 분획물 또는 부탄올 분획물인 것이 바람직하다.The ginger root extract is preferably an ethanol extract of ginger root. Preferably, the ginger root extract is a hexane fraction, an ethyl acetate fraction, or a butanol fraction, which is sequentially fractionated from an ethanolic extract of ginger root by an organic solvent of hexene, ethyl acetate and butanol.
바람직한 구체예로서, 본 발명은 생강 지하부로부터 생강싹을 회수하는 단계; 회수된 생강싹으로부터 활성 물질을 용매로 추출하는 단계; 생강싹 추출물의 순차적 유기 용매 분획을 통한 활성 성분을 정제하는 단계; 상기 물질의 안정성 평가 단계로 구성된다.As a preferred embodiment, the present invention relates to a method for producing ginger root, Extracting the active substance from the recovered ginger root with a solvent; Purifying the active ingredient through a sequential organic solvent fraction of ginger root extract; And a stability evaluation step of the material.
본 발명에서 언급되는 "생강싹(Sprout of Zingiber officinale)"은 생강의 지하부를 수확한 이후, 예를 들어, 기온이 18℃ 이상으로 상승하는 경우에 생강 지하부에서 자라나는 것을 말하며, 흰색~붉은색을 나타낸다. 다만, 시간이 더 경과하여 줄기와 잎이 나타나는 경우는 본 발명에서 언급되는 "생강싹"에 포함되지 않는다. The term "ginger bud (Sprout of Zingiber officinale "refers to growing in the root of ginger when the temperature is raised above 18 ℃ after harvesting the root of ginger, for example, it shows white to red color. Is not included in the "ginger shoot" mentioned in the present invention.
본 발명의 항당뇨 조성물에 포함되는 "생강싹 추출물"은 생강싹을 추출하여 얻은 것을 의미하며, 생강싹은 생강 지하부로부터 3cm 이하의 크기로 무작위로 생성되며, 이를 손으로 간단히 뜯어 수확한 후, 이를 수세하여 사용하거나, 분쇄된 분말을 사용할 수 있으며, 바람직하게는 생강싹을 마쇄하여 사용하는 것이다. The "ginger root extract" contained in the antidiabetic composition of the present invention refers to a ginger root obtained by extracting ginger buds. The ginger buds are randomly generated at a size of 3 cm or less from the root of the ginger. The ginger buds are harvested by hand, It may be washed with water or pulverized powder may be used. Preferably, ginger sprouts are ground and used.
본 발명에서 "생강싹 추출물"은 다양한 추출 용매를 이용하여 제조할 수 있다. 예를 들어, 물(냉수, 열수), 주정, 탄소수 1~4의 무수 또는 함수 저급 알코올(메탄올, 에탄올, 주정, 프로판올, 부탄올 등), 상기 저급알코올과 물과의 혼합용매 등을 이용하며 제조할 수 있다. 바람직하게는, 에탄올 또는 주정을 사용하여 추출하는 경우 최대의 활성물질의 수율을 나타낸다. In the present invention, "ginger root extract" can be prepared using various extraction solvents. For example, water (cold water, hot water), alcohol, water, or a lower alcohol having 1 to 4 carbon atoms (methanol, ethanol, alcohol, propanol, butanol, etc.) can do. Preferably, it represents the maximum yield of active material when extracted with ethanol or alcohol.
또한, 본 발명의 "생강싹 추출물"은 상기 추출 용매를 사용한 추출 과정 이후, 추가적인 유기 용매 분획에 의한 활성 분획물로서도 존재할 수 있다. 바람직한 구체예로서, 본 발명의 생강싹 에탄올 추출물은 헥센, 에틸아세테이트 및 부탄올의 순차적 유기 용매 분획을 수행하여 헥센 분획물, 에틸아세테이트 분획물, 부탄올 분획물 및 유기 용매 분획 후의 물 잔류물로서 분획될 수 있다. In addition, the "ginger root extract" of the present invention may exist as an active fraction by an additional organic solvent fraction after the extraction using the above-mentioned extraction solvent. As a preferred embodiment, the ginger sprout ethanol extract of the present invention can be fractionated as a hexane fraction, an ethyl acetate fraction, a butanol fraction and a water residue after an organic solvent fraction by performing a sequential organic solvent fraction of hexene, ethyl acetate and butanol.
본 발명의 생강싹 추출물 및 이의 유기 용매 분획물은 감압증류 및 동결건조, 또는 분무건조 등과 같은 통상적인 분말화 과정을 거쳐 분말로 제조될 수 있다. 이들은 100℃의 열처리와 pH 2의 인체 위 내의 pH에서도 활성을 유지한다.The ginger root extract of the present invention and its organic solvent fraction may be powdered by a conventional powdering process such as vacuum distillation and freeze drying or spray drying. They maintain their activity even at 100 ° C heat treatment and pH 2 in human body.
본 발명의 생강싹 에탄올 추출물과 이들의 활성 분획물은 강력한 항당뇨 활성을 나타내는 것으로 확인되었으며, 특히, 강력한 알파-아밀라제 및 알파-글루코시다제에 대한 전분분해효소 저해활성을 나타내어, 제1형 당뇨병, 제2형 당뇨병 또는 당뇨병성 망막증과 당뇨병성 신증 등과 같은 당뇨 합병증의 예방 및 치료/개선과 관련되는 약학적 조성물 및 건강 기능 식품의 소재로 사용될 수 있다.The ginger sprout ethanol extract of the present invention and the active fractions thereof showed strong antidiabetic activity and exhibited a starcholytic enzyme inhibitory activity against potent alpha-amylase and alpha-glucosidase, Type 2 diabetes or diabetic nephropathy such as diabetic retinopathy and diabetic nephropathy, and a health functional food.
바람직한 구체예로서, 본 발명의 항당뇨 조성물은 약학적 조성물의 용도로서 적용될 수 있다. In a preferred embodiment, the antidiabetic composition of the present invention can be applied as a pharmaceutical composition.
상기 약학적 조성물은 각각의 사용 목적에 맞게 통상의 방법에 따라 산제, 과립제, 정제, 캡슐제, 현탁제, 에멀젼, 시럽, 에어로졸 등의 경구 제형, 멸균 주사용액의 주사제 등 다양한 형태로 제형화하여 사용할 수 있으며, 경구 투여하거나 정맥 내, 복강 내, 피하, 직장, 국소 투여 등을 포함한 다양한 경로를 통해 투여될 수 있다. The pharmaceutical composition may be formulated into various forms such as powders, granules, tablets, capsules, oral formulations such as suspensions, emulsions, syrups and aerosols, injections of sterilized injection solutions, etc. And can be administered via various routes including oral administration or intravenous, intraperitoneal, subcutaneous, rectal, topical administration, and the like.
이러한 약학적 조성물에는 추가적으로 담체, 부형제 또는 희석제 등이 더 포함될 수 있으며, 포함될 수 있는 적합한 담체, 부형제 또는 희석제의 예로는 락토오스, 덱스트로오스, 수크로오스, 솔비톨, 만니톨, 자일리톨, 에리쓰리톨, 말티톨, 전분, 아카시아 고무, 알지네이트, 젤라틴, 칼슘 포스페이트, 칼슘 실리케이트, 셀룰로스, 메틸 셀룰로스, 비정질 셀룰로스, 폴리비닐 피롤리돈, 물, 메틸하이드록시벤조에이트, 프로필하이드록시벤조에이트, 탈크, 마그네슘 스테아레이트 및 광물유 등을 들 수 있다. 또한, 본 발명의 약학적 조성물은 충전제, 항응집제, 윤활제, 습윤제, 향료, 유화제, 방부제 등을 추가로 더 포함할 수도 있다. Such pharmaceutical compositions may further comprise carriers, excipients or diluents, and examples of suitable carriers, excipients or diluents that may be included include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, But are not limited to, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, amorphous cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, And the like. In addition, the pharmaceutical composition of the present invention may further include a filler, an anti-coagulant, a lubricant, a wetting agent, a flavoring agent, an emulsifying agent, an antiseptic, and the like.
본 발명의 약학적 조성물은 약제학적으로 유효한 양으로 투여한다. 본 발명에서, "약제학적으로 유효한 양"은 의학적 치료에 적용 가능한 합리적인 수혜/위험 비율로 질환을 치료하기에 충분한 양을 의미하며, 유효 용량 수준은 환자의 질환의 종류, 중증도, 약물의 활성, 약물에 대한 민감도, 투여 시간, 투여 경로 및 배출 비율, 치료 기간, 동시 사용되는 약물을 포함한 요소 및 기타 의학 분야에 잘 알려진 요소에 따라 결정될 수 있다. The pharmaceutical composition of the present invention is administered in a pharmaceutically effective amount. In the present invention, "pharmaceutically effective amount" means an amount sufficient to treat a disease at a reasonable benefit / risk ratio applicable to medical treatment, and the effective dose level will depend on the type of disease, severity, The sensitivity to the drug, the time of administration, the route of administration and the rate of release, the duration of the treatment, factors including co-administered drugs, and other factors well known in the medical arts.
본 발명의 약학적 조성물은 개별 치료제로 투여하거나 다른 치료제와 병용하여 투여될 수 있고, 종래의 치료제와 순차적으로 또는 동시에 투여될 수 있으며, 단일 또는 다중 투여될 수 있다. 상기한 요소들을 모두 고려하여 부작용 없이 최소한의 양으로 최대 효과를 얻을 수 있는 양을 투여하는 것이 중요하며, 이는 당업자에 의해 용이하게 결정될 수 있다. The pharmaceutical composition of the present invention may be administered as an individual therapeutic agent or in combination with other therapeutic agents, and may be administered sequentially or simultaneously with conventional therapeutic agents, and may be administered singly or multiply. It is important to take into account all of the above factors and to administer the amount in which the maximum effect can be obtained in a minimal amount without side effects, which can be easily determined by those skilled in the art.
본 발명의 약학적 조성물에서 항당뇨 활성을 갖는 성분의 유효량은 환자의 나이, 성별, 체중에 따라 달라질 수 있으며, 일반적으로는 체중 당 1 내지 5,000mg, 바람직하게는 100 내지 3,000mg을 매일 또는 격일 투여하거나 1일 1 내지 3회로 나누어 투여할 수 있다. 그러나, 투여 경로, 질병의 중증도, 성별, 체중, 연령 등에 따라서 증감될 수 있으므로 상기 투여량이 어떠한 방법으로도 본 발명의 범위를 한정하는 것은 아니다. The effective amount of the component having an anti-diabetic activity in the pharmaceutical composition of the present invention may vary depending on the age, sex and body weight of the patient, and is generally 1 to 5,000 mg, preferably 100 to 3,000 mg per body weight, Or one to three divided doses per day. However, the dosage may not be limited in any way because it may be increased or decreased depending on route of administration, severity of disease, sex, weight, age, and the like.
본 발명의 약학적 조성물은 다양한 경로를 통하여 대상에 투여될 수 있다. 투여의 모든 방식은 예상될 수 있는데, 예를 들면, 경구, 직장 또는 정맥, 근육, 피하, 자궁내 경막 또는 뇌혈관 내(intra-cerebroventricular) 주사에 의해 투여될 수 있다. 본 발명에서 "투여"는 임의의 적절한 방법으로 환자에게 소정의 물질을 제공하는 것을 의미하며, 본 발명의 약학적 조성물의 투여 경로는 목적 조직에 도달할 수 있는 한 일반적인 모든 경로를 통하여 경구 또는 비경구 투여될 수 있다. 또한, 본 발명의 조성물은 유효성분을 표적 세포로 전달할 수 있는 임의의 장치를 이용해 투여될 수도 있다. The pharmaceutical composition of the present invention can be administered to a subject through various routes. All modes of administration may be expected, for example, by oral, rectal or intravenous, intramuscular, subcutaneous, intra-uterine or intra-cerebroventricular injections. In the present invention, "administration" means providing a predetermined substance to a patient by any suitable method, and the administration route of the pharmaceutical composition of the present invention is either oral or non-oral May be administered orally. The composition of the present invention may also be administered using any device capable of delivering an effective ingredient to a target cell.
본 발명에서 "대상"은, 특별히 한정되는 것은 아니지만, 예를 들어, 인간, 원숭이, 소, 말, 양, 돼지, 닭, 칠면조, 메추라기, 고양이, 개, 마우스, 쥐, 토끼 또는 기니아 피그를 포함하고, 바람직하게는 포유류, 보다 바람직하게는 인간을 의미한다. In the present invention, the term "object" includes, but is not limited to, human, monkey, cow, horse, sheep, pig, chicken, turkey, quail, cat, dog, mouse, rat, rabbit or guinea pig , Preferably a mammal, more preferably a human.
바람직한 구체예로서, 본 발명의 항당뇨 조성물은 건강 기능 식품의 용도로서 적용될 수 있다. In a preferred embodiment, the antidiabetic composition of the present invention can be applied as a health functional food.
본 발명의 항당뇨 활성이 우수한 유효성분을 포함하는 식품으로는, 예를 들어, 각종 식품류, 음료, 껌, 차, 비타민 복합제, 건강보조 식품류 등이 있고, 분말, 과립, 정제, 캡슐 또는 음료인 형태로 사용할 수 있다. Examples of the foods containing the active ingredient having excellent antidiabetic activity of the present invention include various foods, beverages, gums, tea, vitamin complexes, health supplement foods and the like, and may be powder, granule, tablet, capsule or beverage Can be used.
본 발명의 활성 성분은 일반적으로 전체 식품 중량의 0.01 내지 15중량%로 가할 수 있으며, 건강음료 조성물은 100 ml를 기준으로 0.02 내지 10g, 바람직하게는 0.3 내지 1g의 비율로 가할 수 있다. The active ingredient of the present invention may generally be added in an amount of 0.01 to 15% by weight of the total food, and the health beverage composition may be added in a proportion of 0.02 to 10 g, preferably 0.3 to 1 g, based on 100 ml.
본 발명의 건강 기능 식품은 지시된 비율로 필수 성분으로서 상기 화합물을 함유하는 것 외에 식품학적으로 허용 가능한 식품보조 첨가제, 예컨대, 천연 탄수화물 및 다양한 향미제 등을 추가 성분으로서 함유할 수 있다. 상기 천연 탄수화물의 예로는 포도당, 과당 등의 단당류, 말토오스, 수크로오스 등의 이당류 및 덱스트린, 시클로덱스트린 등의 다당류와 같은 통상적인 당 및 자일리톨, 소르비톨, 에리쓰리톨 등의 당알코올이 있다. 상기 향미제로는 타우마틴, 레바우디오시드 A, 글리시르히진, 사카린, 아스파르탐 등을 사용할 수 있다. 상기 향미제의 비율은 본 발명의 건강 기능 식품 100ml당 일반적으로 약 1 내지 20g, 바람직하게는 약 5 내지 12g을 사용한다. 상기 외에 본 발명의 건강 기능 식품은 여러 가지 영양제, 비타민, 광물, 합성 풍미제 및 천연 풍미제 등의 풍미제, 착색제 및 중진제, 펙트산 및 그의 염, 알긴산 및 그의 염, 유기산, 보호성 콜로이드 증점제, pH 조절제, 안정화제, 방부제, 글리세린, 알코올, 탄산음료에 사용되는 탄산화제 등을 함유할 수 있다. The health functional food of the present invention may contain, as an additional ingredient, a food-acceptable food-aid additive such as natural carbohydrates and various flavors, in addition to containing the above-mentioned compound as an essential ingredient in the indicated ratio. Examples of the natural carbohydrate include sugar sugars such as glucose, monosaccharides such as fructose, disaccharides such as maltose and sucrose, polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol and erythritol. Examples of the flavor agent include tau martin, rebaudioside A, glycyrrhizin, saccharin, and aspartame. The proportion of the above-mentioned flavoring agent is generally about 1 to 20 g, preferably about 5 to 12 g per 100 ml of the health functional food of the present invention. In addition to the above, the health functional food of the present invention may contain various kinds of nutrients, vitamins, minerals, flavors such as synthetic flavors and natural flavors, colorants and heavy stabilizers, pectic acid and its salts, alginic acid and its salts, Thickening agents, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonating agents used in carbonated drinks, and the like.
그 밖에 본 발명의 건강 기능 식품은 천연 과일 주스 및 과일 주스 음료 및 야채 음료 등의 제조를 위한 과육을 함유할 수도 있다. 이러한 성분은 독립적으로 또는 조합하여 사용할 수 있다. 이러한 첨가제의 비율은 본 발명의 상기 활성 분획물 100 중량부 당 0.01 내지 약 20중량부의 범위에서 선택되는 것이 일반적이다.In addition, the health functional food of the present invention may contain flesh for producing natural fruit juice, fruit juice drink, vegetable drink and the like. These components may be used independently or in combination. The ratio of such additives is generally selected in the range of 0.01 to about 20 parts by weight per 100 parts by weight of the active fraction of the present invention.
이하에서는 실시예를 통하여 본 발명을 더욱 상세하게 설명한다. 하기 실시예는 본 발명의 바람직한 일 구체예일 뿐이며, 본 발명의 권리범위가 하기 실시예의 범위로 한정되는 것은 아니다. Hereinafter, the present invention will be described in more detail by way of examples. The following examples are only exemplary embodiments of the present invention, and the scope of the present invention is not limited to the scope of the following examples.
[실시예][Example]
실시예Example 1: 생강 부위별 추출물의 조제 1: Preparation of ginger extract
2016년 경북 안동에서 재배한 생강 지하부 및 지상부를 구입한 후, 이물질을 제거하고, 껍질을 거피한 생강 지하부, 생강 지하부의 껍질, 생강싹, 생강잎으로 각각 구분하고, 이를 에탄올 추출물 제조에 사용하였다. 특히, 생강싹의 경우, 2017년 4월 이후 지하부에서 불규칙하게 생성된 생강싹을 손으로 뜯어 회수하였다. 이 경우 지하부와 싹은 경계는 분명하였고, 손쉽게 회수할 수 있었다. 생강싹이 난 생강 지하부와 이로부터 분리된 생강싹은 도 1 및 도 2에 나타내었다. 제때 회수하지 않은 생강싹은 계속 성장하게 되며, 이 경우 줄기로 성장한 생강 지하부는 도 3에 나타내었다. After purchasing the ginger underground and the ground part in Gyeongbuk Province, Gyeongbuk Province in 2016, the foreign matter was removed and the ginger root, ginger root, ginger root and ginger leaf were separated from each other and used for the production of ethanol extract . Especially, in case of ginger shoots, irregularly formed ginger shoots were recovered by hand after April, 2017. In this case, the boundaries between the underground and shoots were clear and could be easily recovered. The ginger root and the ginger shoot separated therefrom are shown in FIGS. 1 and 2. The ginger shoots that have not been recovered in time will continue to grow, and the ginger basement grown in the stem in this case is shown in Fig.
한편, 각각 생강 부위별 에탄올 추출물 조제시에는 각각의 시료에 대해 10배의 에탄올(95%, 덕산, 한국)을 가하고, 상온에서 3회 추출한 후 추출액을 모아 필터링한 후, 감압 농축하여 분말로 제조하였다. 조제된 추출물의 성분 분석으로 총 폴리페놀, 총 플라보노이드, 총 당 및 환원당 함량을 측정하였다. 총 폴리페놀 함량은 추출 검액 400μl에 50μl의 Folin-ciocalteau, 100μl의 Na2CO3 포화용액을 넣고 실온에서 1시간 방치한 후 725nm에서 흡광도를 측정하였다. 표준시약으로는 tannic acid를 사용하였다. 총 플라보노이드 함량은 각각의 시료를 18시간 메탄올 교반 추출하고, 여과한 추출 검액 400μl에 90% diethylene glycol 4ml를 첨가하고 다시 1N NaOH 40μl를 넣고 37℃에서 1시간 반응 후 420nm에서 흡광도를 측정하였다. 표준시약으로는 rutin을 사용하였다. 환원당은 DNS법으로, 총 당은 phenol-sulfuric acid법을 이용하여 정량하였다.For the preparation of ethanol extracts for each ginger part, 10 times ethanol (95%, Duksan, Korea) was added to each sample, and the extracts were collected three times at room temperature. The extracts were collected by filtration and concentrated under reduced pressure to give powder Respectively. Total polyphenols, total flavonoids, total sugars and reducing sugar content were determined by analysis of the constituents of the prepared extracts. Total polyphenol content was determined by adding 50 μl of Folin-ciocalteau and 100 μl of saturated Na 2 CO 3 solution to 400 μl of the extract solution, leaving it at room temperature for 1 hour and measuring the absorbance at 725 nm. Tannic acid was used as a standard reagent. The total flavonoid content of each sample was measured by stirring for 18 hours in methanol. To the 400 μl of the filtered extract, 4 ml of 90% diethylene glycol was added and 40 μl of 1N NaOH was added. The reaction was carried out at 37 ° C. for 1 hour and then absorbance was measured at 420 nm. As a standard reagent, rutin was used. Reducing sugar was quantified by DNS method and total sugar was quantified by phenol-sulfuric acid method.
그 결과, 표 1에 나타낸 바와 같이, 생강의 부위별 추출 수율은 생강잎 > 생강싹 > 거피 생강 지하부 > 생강 지하부 껍질의 순으로 나타났으며, 생강싹은 3.9%의 높은 수율을 나타내었다. 총 폴리페놀 함량 분석 결과, 거피 생강 지하부에서 가장 높은 총 폴리페놀 함량(53.3mg/g)을 나타내었으며, 생강잎에서는 상대적으로 가장 낮은 31.9mg/g의 함량을 나타내었다. 생강싹의 경우, 생강 지하부와 생강잎의 중간 정도의 함량을 나타내었다. 한편, 총 플라보노이드 함량 측정 결과, 생강잎에서 가장 높은 16.2mg/g을 나타내었으며, 생각싹에서 가장 낮은 4.2mg/g 함량을 나타내었다. 총 당 및 환원당 함량의 경우, 생강잎에서 가장 높았으며, 생강싹, 거피 생강 지하부 및 생강 지하부 껍질 순으로 나타났다. As a result, as shown in Table 1, the extraction yields of the ginger root were in the order of ginger leaf> ginger root> ginger root, ginger root, and ginger root were 3.9%. The total polyphenol content of the ginger root showed the highest total polyphenol content (53.3 mg / g) in the bottom part of the ginger, and the lowest content of 31.9 mg / g in the ginger root. In the case of ginger shoots, the content of ginger root was about the middle of ginger root. On the other hand, total flavonoid content was 16.2 mg / g in ginger leaf and 4.2 mg / g in thought shoot. Total sugar and reducing sugar contents were highest in ginger leaf, ginger root, ginger root, and ginger root.
실시예Example 2: 생강 부위별 추출물의 2: Extract of ginger 항당뇨Anti-diabetic 활성 평가 Activity evaluation
실시예 1에서 얻어진 생강의 다양한 부위별 에탄올 추출물의 항당뇨 활성을 in-vitro α-amylase 저해활성 및 α-glucosidase 저해 활성으로 평가하여 나타내었다. 먼저, α-amylase 저해활성은 Lim 등이 보고한 방법(Li, CS 등, 2005. J. Kor. Soc. Appl. Biol. Chem. 48: 103-108)을 수정하여 사용하였다. 시료 2.5μL와 50mM phosphate buffer(pH 6.8)로 희석한 α-amylase(0.25U/mL) 25μL를 혼합하여 37℃에서 10분간 preincubation한 후, 0.5% soluble starch(Samchun Chemicals Co., Korea) 25μL를 가하여 37℃에서 10분간 반응하였다. 이후, 2N acetic acid를 25μL를 가하여 반응을 정지시켰으며, 반응액에 0.75% KI/I2 용액 10μL를 가하여 발색하였다. 발색액은 540nm에서 흡광도를 측정하였으며, 각각의 실험은 3반복한 후 평균값을 구하여 다음의 식으로 저해율을 계산하였다.The anti-diabetic activity of ethanol extracts of various parts of ginger obtained in Example 1 was evaluated by in-vitro α-amylase inhibitory activity and α-glucosidase inhibitory activity. First, the α-amylase inhibitory activity was modified by the method reported by Lim et al. (Li, CS et al., 2005. J. Kor. Soc. Appl. Biol. Chem. 25 μL of α-amylase (0.25 U / mL) diluted in 50 mM phosphate buffer (pH 6.8) was preincubated at 37 ° C. for 10 minutes and then 25 μL of a 0.5% soluble starch (Samchun Chemicals Co., Korea) Followed by reaction at 37 占 폚 for 10 minutes. Then, 25 μL of 2N acetic acid was added to stop the reaction, and 10 μL of 0.75% KI / I 2 solution was added to the reaction solution to develop color. The absorbance at 540 nm was measured, and each experiment was repeated three times and the average value was obtained. The inhibition rate was calculated by the following equation.
저해율 (Inhibition rate %% ) = 시료 ) = Sample 첨가구Shu Furniture 효소활성/ Enzyme activity / 대조구Control 첨가구Shu Furniture 효소활성 x 100 Enzyme activity x 100
한편, α-glucosidase 저해활성은 pNPG(p-nitrophenol glucoside; Sigma Co., USA)를 이용하여 평가하였으며, 생강 추출물 시료 2.5μl와 50mM Sodium acetate buffer(pH 5.6)로 희석한 α-glucosidase(0.25U/ml) 25μl를 혼합하여 37℃에서 10분간 preincubation 하고, 1mM pNPG 용액 25μl를 가하여 60℃에서 10분간 반응하였다. 이후, 1M NaOH 25μl를 가하여 반응을 정지시키고, 405nm에서 흡광도를 측정하여 저해율을 계산하였다. The α-glucosidase inhibitory activity was evaluated using pNPG (p-nitrophenol glucoside; Sigma Co., USA), and α-glucosidase (0.25 U) diluted with 2.5 μl of ginger extract sample and 50 mM sodium acetate buffer / ml) were preincubated at 37 ° C for 10 minutes, 25 μl of 1 mM pNPG solution was added, and reacted at 60 ° C for 10 minutes. Thereafter, the reaction was stopped by adding 25 μl of 1M NaOH, and the inhibition rate was calculated by measuring the absorbance at 405 nm.
저해율 (Inhibition rate %% ) = [1-(시료 ) = [1- (sample 첨가구Shu Furniture 효소활성/ Enzyme activity / 대조구Control 첨가구Shu Furniture 효소활성)] x 100 Enzyme activity)] x 100
그 결과, 표 2에 나타낸 바와 같이, 임상에서 당뇨병 치료제로 사용하고 있는 acarbose는 농도 의존적으로 강력한 α-amylase 저해활성 및 α-glucosidase 저해활성을 나타내었다. 생강은 생강피, 생강잎, 거피 생강, 생강싹 모두 0.5mg/ml 농도에서 4.5~7.9%의 미약한 α-amylase 저해활성을 나타내었으며, 동일 농도의 acarbose에 비해 미미한 저해활성을 나타내었다. 그러나, α-glucosidase 저해활성의 경우, 생강싹 추출물(0.5mg/ml)이 17.4%의 저해를 나타내어, acarbose 62.5μg/ml 농도의 저해활성의 약 1/2의 α-glucosidase 저해활성을 나타내었다. 이러한 저해활성은 고농도 처리시에 보다 강력하게 나타났으며, 농도의존적인 저해 활성을 나타내었다. 따라서, 생강싹 추출물은 제2형 당뇨병의 예방 및 치료에 이용 가능함을 확인하였다.As a result, as shown in Table 2, acarbose used as a therapeutic agent for diabetes in the clinic showed strong α-amylase inhibitory activity and α-glucosidase inhibitory activity in a concentration-dependent manner. The ginger showed 4.5 ~ 7.9% weak α-amylase inhibitory activity at 0.5mg / ml concentration of ginger, ginger leaf, ginger ginger and ginger sprout and showed a slight inhibitory activity compared with acarbose at the same concentration. However, in the case of α-glucosidase inhibitory activity, ginger shoot extract (0.5 mg / ml) showed inhibition of 17.4% and showed about ½ of the inhibitory activity of acarbose at 62.5 μg / ml of α-glucosidase inhibitory activity . These inhibitory activities were more strongly exhibited at the high concentration treatment and showed a concentration dependent inhibitory activity. Therefore, it was confirmed that ginger root extract can be used for the prevention and treatment of type 2 diabetes.
실시예Example 3: 생강싹 추출물로부터 순차적 3: Sequential from ginger root extract 유기용매Organic solvent 분획물의Fraction 제조 및 이들의 유용성분 분석 And analysis of their useful components
실시예 1에서 제조한 생강싹 에탄올 추출물을 대상으로 헥센, 에틸아세테이트, 부탄올로 순차적 유기용매 분획하였으며, 최종적으로 물 잔류물을 회수하였다. 유기용매 분획 효율 및 성분 분석 결과는 표 3에 나타내었다. 성분 분석은 실시예 1의 방법과 동일하게 총 폴리페놀, 총 플라보노이드, 총 당 및 환원당 함량을 측정하였다. The ginger sprout ethanol extract prepared in Example 1 was fractionated with hexane, ethyl acetate and butanol sequentially, and finally the water residue was recovered. The organic solvent fraction efficiency and component analysis results are shown in Table 3. The total polyphenols, total flavonoids, total sugars and reducing sugar content were measured in the same manner as in Example 1.
표 3에 나타낸 바와 같이, 생강싹 추출물의 분획물 중 물 잔류물이 50.1%로 분획 효율이 가장 높았으며, 에틸아세테이트 분획물과 부탄올 분획물은 각각 추출물의 25.4% 및 26.6%를 차지하였다. 반면, 헥센 분획물은 추출물의 4.2%이 차지하여 생강싹 추출물이 생강 추출물과는 달리 지용성 오일 성분은 매우 낮은 상태임을 알 수 있었다. As shown in Table 3, the fraction of ginger root extract had the highest fractionation efficiency of 50.1%, while the ethyl acetate fraction and the butanol fraction accounted for 25.4% and 26.6% of the extract, respectively. On the other hand, the fraction of hexane occupied 4.2% of the extract, and the extract of ginger root was very low in oil - soluble oil component, unlike ginger extract.
분획물의 성분 분석 결과, 총 폴리페놀 함량은 특이하게도 헥센 분획물에서 가장 높았으며, 에틸아세테이트 분획물, 부탄올 분획물 및 물 잔류물 순으로 나타났다. 또한, 총 플라보노이드 함량 역시 헥센 분획물에서 가장 높았으며, 특이하게 에틸아세테이트 분획물에서는 거의 검출되지 않았다. 반면, 총 당 및 환원당 함량은 각각 부탄올 분획물 및 에틸아세테이트 분획물에서 가장 높은 함량을 나타내었다. 따라서, 생강싹 추출물의 헥센 분획물 및 에틸아세테이트 분획물은 페놀성 배당체가 주요 성분으로 예상되며, 다양한 생리 활성을 나타내리라 판단되었다. As a result of the analysis of the fractions, total polyphenol contents were highest in hexane fraction, ethyl acetate fraction, butanol fraction and water residue. Total flavonoid content was also highest in the hexane fraction, and was not detected in the ethyl acetate fraction. On the other hand, total sugar and reducing sugar contents were the highest in butanol fraction and ethyl acetate fraction, respectively. Therefore, the hexane fraction and the ethyl acetate fraction of the ginger root extract were expected to exhibit various physiological activities.
실시예Example 4: 생강싹 추출물의 순차적 4: Sequential of ginger root extract 유기용매Organic solvent 분획물의Fraction 항당뇨Anti-diabetic 활성 평가 Activity evaluation
상기 실시예 3에서 조제한 생강싹 에탄올 추출물의 순차적 유기 용매 분획물들을 대상으로 항당뇨 활성을 평가하였으며, 그 결과는 표 4에 나타내었다.The anti-diabetic activity of the organic solvent fractions of the ethanol extract of ginger root prepared in Example 3 was evaluated. The results are shown in Table 4.
그 결과, 표 4에 나타낸 바와 같이, 생강싹 추출물의 헥센 분획물 및 에틸아세테이트 분획물은 매우 강력한 α-amylase 저해활성을 나타내었으며, 이는 임상에서 치료제로 사용되고 있는 acarbose의 동일농도에서의 저해활성의 1/2에 해당되는 활성이었다. 또한 α-glucosidase 저해활성은 생강싹 추출물의 부탄올 분획에서만 강력하게 나타났다. 즉, 생강싹 추출물의 분획물 중 헥센 및 에탈아세테이트 분획물은 우수한 α-amylase 저해활성, 부탄올 분획물은 우수한 α-glucosidase 저해활성을 나타냄을 의미하고 있다. 이러한 저해활성은 고농도 처리시에 보다 강력하게 나타났으며, 농도의존적인 저해 활성을 나타내었다. 따라서, 생강싹 추출물 및 이의 활성 분획물들은 제2형 당뇨병의 예방 및 치료에 이용 가능함을 확인하였다. As a result, as shown in Table 4, the hexane fraction and the ethyl acetate fraction of the extract of ginger root showed very strong? -Amylase inhibitory activity, and the inhibitory activity of acarbose at the same concentration of 1 / 2, respectively. In addition, α-glucosidase inhibitory activity was strong only in the butanol fraction of ginger root extract. That is, the hexane and ethalacetate fractions of the ginger root extract fractions exhibit excellent .alpha.-amylase inhibitory activity and the butanol fractions exhibit excellent .alpha.-glucosidase inhibitory activity. These inhibitory activities were more strongly exhibited at the high concentration treatment and showed a concentration dependent inhibitory activity. Therefore, it was confirmed that ginger root extract and its active fractions can be used for the prevention and treatment of type 2 diabetes.
실시예Example 5: 생강싹 추출물 및 이의 활성 5: Ginger root extract and its activity 분획물의Fraction 혈장, 산 및 열 안정성 평가 Plasma, acid and thermal stability assessment
상기 실시예 1 및 실시예 3에서 얻은 생강싹 추출물과 이의 헥센 분획물, 에틸아세테이트 분획물 및 부탄올 분획물을 대상으로 각각 α-amylase 저해활성과 α-glucosidase 저해활성에 대한 열 안정성 및 산 안정성을 확인하였다. 상기 추출물과 분획물들은 100℃에서 1시간 열 처리, pH 2(0.01M HCl)에서의 1시간 처리시 α-amylase 저해활성과 α-glucosidase 저해활성의 심각한 감소가 나타나지 않았다. 따라서, 본 발명의 생강싹 추출물 및 이의 활성 분획물들은 일반적인 식품 가공 공정 및 소화 흡수 단계에서도 항당뇨 활성을 유지할 것으로 판단된다. The ginger extracts obtained in Examples 1 and 3, their hexane fractions, ethyl acetate fractions and butanol fractions were examined for their α-amylase inhibitory activity and α-glucosidase inhibitory activity, respectively. The extracts and fractions did not show a significant decrease in the? -Amylase inhibitory activity and the? -Glucosidase inhibitory activity after 1 hour heat treatment at 100 ° C and 1 hour treatment at pH 2 (0.01M HCl). Therefore, the ginger root extract of the present invention and its active fractions are expected to maintain antidiabetic activity even in the general food processing step and digestion and absorption step.
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