KR20020035230A - Plant extract from Adenophora remotiflora and composition containing the same with hypoglycemic activity - Google Patents

Plant extract from Adenophora remotiflora and composition containing the same with hypoglycemic activity Download PDF

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KR20020035230A
KR20020035230A KR1020000065428A KR20000065428A KR20020035230A KR 20020035230 A KR20020035230 A KR 20020035230A KR 1020000065428 A KR1020000065428 A KR 1020000065428A KR 20000065428 A KR20000065428 A KR 20000065428A KR 20020035230 A KR20020035230 A KR 20020035230A
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김정상
장동재
권정숙
손건호
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김정상
손건호
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    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18Magnoliophyta (angiosperms)
    • A61K36/185Magnoliopsida (dicotyledons)
    • A61K36/34Campanulaceae (Bellflower family)
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    • A61P3/08Drugs for disorders of the metabolism for glucose homeostasis
    • A61P3/10Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A61K2236/30Extraction of the material
    • A61K2236/33Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones

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Abstract

PURPOSE: A composition containing an Adenophora remotiflora extract capable of inhibiting a sudden rise of a glucose concentration in blood after a meal by reducing alpha-glucosidase and alpha-amylase enzyme activity is provided which has an excellent effect on prevention and/or treatment of diabetics in human and animal. CONSTITUTION: Adenophora remotiflora is extracted in water or an organic solvent and a composition for reducing a blood sugar concentration contains the Adenophora remotiflora extract as an effective component. For an example, leaves, roots and stems of Adenophora remotiflora are extracted 10 times in water for 12 hr, concentrated and fractionated. The extract is orally administered to a Sprague-Dawley rat(150-200g) at a dose of 1, 5 and 10mg/kg weight and the dose of 5mg/kg weight exhibits an excellent effect on inhibition of blood sugar.

Description

모시대 추출물과 그를 함유한 혈당강하용 조성물{Plant extract from Adenophora remotiflora and composition containing the same with hypoglycemic activity}Plant extract from Adenophora remotiflora and composition containing the same with hypoglycemic activity

본 발명은 모시대 추출물과 그를 함유한 혈당강하용 조성물에 관한 것으로, 보다 상세하게는 식후 혈중 포도당 농도의 상승을 조절하는 혈당강하기능을 가져 당요병이나 심혈관계 질환과 같은 합병증 조절에 유효한 약리학적 효과를 갖는 모시대 추출물, 그의 제조방법 및 이를 유효성분으로 포함하는 조성물에 관한 것이다.The present invention relates to a seedling extract and a composition for lowering blood glucose containing the same, and more particularly, to have a hypoglycemic effect of controlling the rise of blood glucose concentration after a meal, which is effective in controlling complications such as diabetes and cardiovascular diseases. It relates to a seedling extract having a, a method for preparing the same and a composition comprising the same as an active ingredient.

최근 산업사회의 발달과 생활수준의 향상에 따른 식생활 양상의 변화로 고지혈증, 동맥경화, 고혈압, 당뇨병과 같은 심혈관계 만성 성인병 환자가 급증하고 있는 추세이다. 그 중 당뇨병 환자는 전세계적으로 2억 5천만명이 있는 것으로 추정되고 있어 당뇨병 환자의 치료 및 질환조절에 있어서 혈액 내의 포도당 농도를 조절하는 것이 매우 중요한 일이 되고 있다. 당뇨병은 그 자체가 질환이라기 보다는 장기간 이 질환에 걸리면 당뇨병성 신경병증, 망막병증, 백내장, 신증 등의 합병증이 나타나 환자의 정상적인 삶을 영위할 수 없을뿐만 아니라 치명적인 결과까지 초래할 수 있는데, 심한 경우 당뇨병은 실명, 심부전, 다리절단 등의 주요원인이 되는 만성질환이다(Alper, J.: New insights into type 2 diabetes. Science, 289, 37(2000)).Recently, due to the change of dietary habits according to the development of industrial society and the improvement of living standard, the cardiovascular chronic geriatric patients such as hyperlipidemia, arteriosclerosis, hypertension and diabetes are increasing rapidly. It is estimated that there are 250 million people with diabetes worldwide, so controlling the concentration of glucose in the blood is very important in the treatment and control of diseases. Diabetes is not a disease in itself, but a long-term disease can lead to complications such as diabetic neuropathy, retinopathy, cataracts and nephropathy, which can lead to fatal consequences as well as to the normal life of the patient. Is a chronic disease that is a major cause of blindness, heart failure, and amputation (Alper, J .: New insights into type 2 diabetes. Science, 289, 37 (2000)).

음식물 중에 포함된 탄수화물은 소장에서 탄수화물 분해효소의 작용을 받아 포도당으로 소화되어 혈액내로 흡수된다. 음식물을 섭취하면 혈액내 포도당 농도는 급격히 상승하는데, 당뇨병과 같은 질환의 경우 혈중 인슐린 농도가 부족하거나 인슐린 작용에 결함이 있어 상승된 포도당 농도는 계속 유지되어 위험한 상태를 초래할 수 있다. 즉, 인슐린 부족시(제1형 당뇨병)나 정상적인 인슐린 분비가 이뤄지지만 인슐린 수용체에 이상이 있는(제2형 당뇨병) 경우, 포도당이 세포내로 들어가지 못하고 에너지원으로 이용되지 못하게 되면, 혈당이 높아지고 근육내의 단백질 및 지방세포가 분해되며 포도당 신생을 통해 혈중 포도당 농도가 더욱 상승하게 된다. 간장에서는 포도당신생과 단백질 분해가 증가하게 되고 지방세포의 분해도 증가되어 지방산과 글리세롤 유리가 일어나는데, 지방산은 주요 에너지원이기도 하나 활성아세트산의 생성을 야기하여 케톤체의 축적을 초래하고 유리된 지방산이 간장 내에서 중성지방으로 전환되기 때문에 합병증으로 지방간이 발생하기도 한다.Carbohydrates contained in food are digested into glucose by the action of carbohydrate degrading enzymes in the small intestine and absorbed into the blood. Ingestion of food rapidly increases blood glucose levels. In diseases such as diabetes, elevated insulin levels may be maintained due to insufficient insulin levels in the blood or impaired insulin action, leading to dangerous conditions. That is, when insulin deficiency (type 1 diabetes) or normal insulin secretion occurs, but abnormalities in the insulin receptor (type 2 diabetes), when glucose cannot enter the cell and cannot be used as an energy source, Protein and fat cells in the muscle are broken down and the blood glucose concentration is increased further through the formation of glucose. Fatty acid and glycerol release occur in the liver, resulting in increased cell viability and proteolysis, and fatty acid and glycerol release. Fatty acids, which are also a major energy source, cause the production of active acetic acid, leading to the accumulation of ketone bodies. Since it is converted to triglycerides, fatty liver can occur as a complication.

이와같이 고혈당은 당뇨와 관련된 여러가지 합병증의 주요 원인이 되므로 가능한 정상 수준으로 유지하는 것이 매우 중요하다. 그러므로 이와같은 인슐린 부족으로 인한 당뇨병을 치료하기 위해 인슐린 투여, 설포닐우레아 제제투여, 식이요법 등의 국한적인 치료를 하고 있지만, 임상적으로 혈당을 정상으로 유지한다는 것은 쉬운 일이 아니다. 특히 식후 혈당 상승을 조절하는 것은 더 어려운 것으로 여겨진다(Baron, A.D.: Diabetes Res. Clin. Pract., 40, 51S(1998)). 식후 혈당 조절에 사용하는 대표적인 약물이 알파글루코시다제 저해제이다. 알파글루코시다제는 탄수화물 소화과정이 마지막 단계인 말토스의 포도당으로 전환을 촉매하는 효소이다. 따라서, 이 효소의 저해제는 식이성 탄수화물의 소화를 저하시킴으로써 식후 급격한 혈당상승을 억제하는 것으로 알려져있다(Mooradian, A.D. and Thurman, J.E. Drugs , 57, 19(1999)).As such, hyperglycemia is a major cause of various complications related to diabetes, so it is very important to keep it as normal as possible. Therefore, in order to treat diabetes due to insulin deficiency, there are limited treatment such as insulin administration, sulfonylurea preparation, and diet, but it is not easy to keep blood sugar level clinically. In particular, controlling post-prandial blood sugar elevations is considered more difficult (Baron, A.D .: Diabetes Res. Clin. Pract., 40, 51S (1998)). Representative drugs used for post-prandial blood sugar control are alphaglucosidase inhibitors. Alphaglucosidase is an enzyme that catalyzes the conversion of maltose to glucose, the last step in carbohydrate digestion. Thus, inhibitors of this enzyme are known to inhibit rapid blood glucose elevation after meals by reducing the digestion of dietary carbohydrates (Mooradian, A.D. and Thurman, J.E.Drugs, 57, 19 (1999)).

현재, 아카보스(Acarbos), 보글리보스(Voglibose), 미글리톨(Miglitol) 등 다양한 알파글루코시다제 저해제가 개발되어 임상적으로 이용되고 있으나, 이들 약제에 내성을 대비하여 다양한 약제를 확보하는 것이 필요하고, 특히 당뇨병 환자용 식품에 사용하기 위한 소재의 개발이 필요하다.Currently, various alpha glucosidase inhibitors such as Acarbos, Boglibose, and Miglitol have been developed and used clinically, but it is necessary to secure various drugs in preparation for resistance to these drugs. In particular, the development of materials for use in foods for diabetics is needed.

최근에는 당뇨병의 치료를 위해 합성된 약물보다는 천연물에서의 약리효과가 있는 물질을 탐색하는 연구가 이루어지고 있으며, 약 400여종의 야생식용식물이 당뇨병 치료에 효과가 있다고 보고된 바 있다(Bailey CJ, Day C. 1989. Diabetes Care 12:553-564). 약용작물 또는 식용작물이 혈당강하에 효과를 나타내었다는 연구가 계속 이루어지고 있으나, 과학적인 증명이 시작된 것은 최근의 일이며 그 대부분은 아직 약리효능과 생리적 기전 및 유효성분의 분석에 대해 임상 및 과학적으로 입증되지 못하는 실정이며, 야생식용식물은 그 성분 및 약리효과에 대하여 국내외적으로 많은 관심이 집중되고 있다.Recently, research has been conducted to search for substances that have pharmacological effects in natural products, rather than drugs synthesized for the treatment of diabetes, and about 400 wild edible plants have been reported to be effective in treating diabetes (Bailey CJ, Day C. 1989. Diabetes Care 12: 553-564. Although studies on medicinal crops or edible crops have been shown to have an effect on hypoglycemic activity, scientific proofs have recently begun, and most of them have been clinically and scientifically analyzed for the analysis of pharmacological efficacy, physiological mechanisms and active ingredients. It is unproven, and wild edible plants have attracted much attention both at home and abroad about its components and pharmacological effects.

특히, 우리의 고유식품이며 전통적으로 질병치료에 이용되어오던 야색식용식 물의효능에 대해 적극적으로 과학적 연구가 이루어져, 씀바귀(Ixeris dentata Nakai), 산복사(Prunus davidiana Fr.) 및 갈대(Phragmites communis Trin)의 메탄올 추출물이 alloxan 당뇨유발 흰쥐에서 유의적으로 혈당을 낮춘다고 보고(Choi JS et al., 1990, Kor. J. Pharmacogn 21(2):153-157)된 바 있으며,스트렙토조토신 당뇨 유발 흰쥐에서 닭의 장풀(Commeinae herba) 물추출물과 메탄올 추출물이 혈당강하효과를 보였다고 보고(박수영, 1992, 석사학위논문, 덕성여자대학교대학원)하였다. 그 외 두릅나무를 비롯한 여러가지 국내 자생 야생식용식물의 혈당강하효과에 대한 연구가 이루어져 왔다.In particular, scientific researches have been conducted on the efficacy of the non-edible edible water, which is our own food and has been traditionally used to treat diseases. Therefore, Ixeris dentata Nakai, Prunus davidiana Fr. and Phragmites communis Trin Methanol extract has been reported to significantly lower blood sugar levels in alloxan diabetic rats (Choi JS et al., 1990, Kor. J. Pharmacogn 21 (2): 153-157), and induced streptozotocin diabetes. In rats, water extract and methanol extract of commeinae herba were reported to have a hypoglycemic effect (Soo Young Park, 1992, Master Thesis, Duksung Women's University). In addition, studies on the hypoglycemic effect of various domestic wild edible plants including elm have been conducted.

본 발명에서는 식후 혈당 상승을 억제하는 식품소재를 탐색하기 위하여 한국산 산채류 67종으로부터 물 또는 유기용매 추출물을 제조하고, 이들의 알파글루코시다제 및 알파아밀라제 효소 저해정도를 평가하였다.In the present invention, water or organic solvent extracts were prepared from 67 Korean wild vegetables to search for foodstuffs to suppress the post-prandial blood sugar rise, and their alphaglucosidase and alpha amylase enzyme inhibition levels were evaluated.

본 발명자들은 생체에 부작용이 없으면서 혈당강하작용이 우수한 물질을 제공하고자 산채류로부터 다양한 용매로 추출하여 수득되는 각종 추출물에 대해 약리학적 효과를 시험한 결과, 모시대의 물 또는 알콜 추출물이 알파글루코시다제와 알파아밀라제 저해활성 및 흰쥐의 혈당상승억제 효과가 가장 높음을 확인하고 본 발명을 완성하게 되었다.The present inventors have tested the pharmacological effects of various extracts obtained by extracting various solvents from wild vegetables to provide a substance with excellent hypoglycemic activity without adverse effects on the living body. It was confirmed that the alpha-amylase inhibitory activity and the blood glucose increase inhibitory effect of the rat is the highest and completed the present invention.

즉, 본 발명 방법에 의한 모시대 추출물은 알파글루코시다제 및 알파아밀라제 효소의 활성을 억제하여 혈중 포도당 농도를 현저히 감소시키는 효과가 있음을 확인할 수 있었다.That is, the seedling extract according to the method of the present invention was found to have an effect of significantly reducing blood glucose concentration by inhibiting the activity of alpha glucosidase and alpha amylase enzymes.

따라서, 본 발명의 목적은 혈당강하작용을 나타내는 모시대 추출물 및 그 제조방법을 제공하는 것이다.Accordingly, it is an object of the present invention to provide a seedling extract and a method for producing the same, which exhibit a hypoglycemic action.

본 발명의 다른 목적은 상기 모시대 추출물을 유효성분으로 함유하는 조성물을 제공하는 것이다.Another object of the present invention is to provide a composition containing the seedling extract as an active ingredient.

이하, 본 발명의 구성을 구체적으로 설명한다.EMBODIMENT OF THE INVENTION Hereinafter, the structure of this invention is demonstrated concretely.

도 1은 모시대 물분획의 흰쥐에서의 전분투여에 대한 혈당상승 억제효과를 나타내는 그림이다.1 is a diagram showing the effect of inhibiting the blood glucose increase on starch administration in rats of the water-time fraction.

본 발명은 인체와 동물에서 탄수화물 대사를 조절함으로써 식후 혈중 포도당농도의 급격한 상승을 조절하여 당뇨병과 같은 심혈관계 질환의 치료 및 합병증 조절에 유효한 모시대 추출물 및 이의 제조방법 그리고 상기 추출물을 유효성분으로 하여 약학적으로 허용가능한 담체를 포함하는 약학적 조성물 및 혈당강하 기능성 식품에 관한 것이다.The present invention regulates carbohydrate metabolism in humans and animals to control the rapid rise in blood glucose levels after meals to treat cardiovascular diseases such as diabetes and control complications. It relates to a pharmaceutical composition and a hypoglycemic functional food comprising a generally acceptable carrier.

본 발명의 혈당강하용 조성물 또는 기능성(건강보조)식품에서 주성분으로 사용되는 모시대는(Adenophora remotiflora)는 잔대와 같이 초롱꽃과에 속하는 여러해살이 풀이다. 산지에 절로 나고 우리나라 각지에서 자란다. 높이 40-100cm 정도로, 뿌리가 굵으며 잎은 타원 모양에 예리한 톱니가 있다. 8-9월에 자주색 꽃이 피는데 원줄기 끝에서 밑을 향해 달려있다. 흰색 꽃이 피는 것을 흰 모싯대라하여 구별한다. 어린 순은 먹고 뿌리는 약으로 쓰며 관상초로 심는다(안덕균, 원색한국 본초도감. 교학사(1998)).Adenophora remotiflora, which is used as a main ingredient in the blood sugar-lowering composition or functional (health supplement) food of the present invention, is a perennial herb belonging to a campanula, such as a twig. It is a temple in the mountains and grows all over Korea. The height is 40-100cm, the roots are thick, and the leaves are oval-shaped and have sharp teeth. Purple flowers bloom in August-September, hanging downward from the end of the main stem. White flowers are distinguished by white fleet. Young Shun eats and roots as a medicine and is planted with Kwanchocho (Andeokgyun, primary color book of Korea. Kyohaksa (1998)).

본 발명 모시대 추출물의 제조방법으로는 공지방법을 사용하는 것이 가능하다, 예컨대 모시대의 잎, 줄기, 뿌리 등을 채취하여 생 또는 건조된 상태의 것을 잘게 자른 후 물, 메탄올, 에탄올 등의 극성 용매 및 에테르, 헥산, 클로로포름, 에틸아세테이트 등의 비극성 용매들을 가한 후 실온에서 침적추출하거나 가온추출할 수 있다.It is possible to use a known method as a method of preparing the present invention, such as leaves, stems, roots, etc. of fine seedlings, finely chopped raw or dried ones, and then polar solvents such as water, methanol, ethanol, and the like. Non-polar solvents such as ether, hexane, chloroform, ethyl acetate and the like may be added, followed by deposition or warm extraction at room temperature.

본 발명의 바람직한 한 실시예에 있어서 모시대를 물 또는 메탄올 용매하에서 얻은 추출물이 혈당강하효과가 특히 뛰어났다.In one preferred embodiment of the present invention, the extract obtained from the hairy time under water or methanol solvent was particularly excellent in hypoglycemic effect.

본 발명 모시대 추출물은 흰쥐에 아주 많은 양을 경구투여하여도 전혀 급성독성을 나타내지 않았다. 이와같이 본 발명 모시대 추출물은 부작용이나 독성 등의 문제가 없어 스트레스, 지방과다섭취, 당(또는 탄수화물) 과다섭취 및 운동부족 등으로 인한 당뇨병 유발을 임상적으로 예방 또는 치료할 수 있는 치료제로 이용하거나 기능성 식품의 유효성분으로도 이용할 수 있는 것으로 판단되었다.The seedling extract of the present invention did not show any acute toxicity even when administered orally in large amounts in rats. As described above, the seedling extract of the present invention does not have problems such as side effects or toxicity, and thus is used as a therapeutic food or a functional food that can clinically prevent or treat diabetes caused by stress, excessive intake of fat, excessive intake of sugar (or carbohydrates) and lack of exercise. It could be used as an effective ingredient of.

본 발명에 모시대 추출물은 당뇨병을 예방 또는 치료하기 위하여 임상적으로 이용시 투여량은 당뇨병의 중증도, 환자의 상태, 연령, 합병증 등의 다양한 요인에 따라 전문가에 의해 결정될 수 있으며, 효과적인 약물요법이 이행되기 위해서는 환자의 생활스타일과 같은 환자의 요소와 약리학적 요소 등을 분석 종합하여 환자에게 제공하여야 한다.In the present invention, the seedling extract may be determined by a specialist according to various factors such as the severity of the diabetes mellitus, the patient's condition, age, and complications when clinically used to prevent or treat diabetes mellitus. In order to analyze the patient's factors such as the patient's lifestyle and pharmacological factors and provide them to the patient.

본 발명에 따른 모시대 추출물은 실질적으로 부작용이나 독성없이 안전하게 투여할 수 있는 약제로 판단되며, 약학적 조성물을 임상적으로 이용시 약제학적 분야에서 통상적인 방법에 따라 통상적인 약제학적 제제로 제형화시켜 이용할 수 있다.The seedling extract according to the present invention is judged to be a drug that can be safely administered without substantially any side effects or toxicity, and when used in a clinical use, it is formulated into a conventional pharmaceutical preparation according to a conventional method in the pharmaceutical field. Can be.

본 발명 모시대 추출물은 음료수, 아이스크림, 양념, 인스턴트 식품 등의 식품이나 가축사료 등에 혼합하여 고상, 액상, 호상 또는 젤리 형태로 혈당을 강하시킬 수 있는 기능성 식품의 제조에 이용될 수도 있으며, 필요에 따라 식용색소, 식용향료, 생약초, 과일, 올리고당, 키토산, 구연산 등의 통상적인 성분을 적량 가하여 좋은 맛과 향을 나타내게 할 수 있다.The seedling extract of the present invention may be mixed with foods such as beverages, ice cream, condiments, instant foods, or livestock feeds, or the like, and may be used to prepare functional foods capable of lowering blood sugar in the form of solid, liquid, lake or jelly. Food coloring, food flavorings, herbal medicines, fruits, oligosaccharides, chitosan, citric acid and the like can be appropriately added to make a good taste and aroma.

본 발명 조성물 또는 기능성 식품에는 필요에 따라 모시대 추출물 이외에 인체의 당흡수를 억제하여 당뇨병을 예방 및/또는 치료하는데 사용되는 공지 물질 또는 새로운 물질을 추가로 포함하여 상승작용을 발휘할 수도 있다.The composition or functional food of the present invention may exert synergism by additionally including a known substance or a new substance used to prevent and / or treat diabetes by inhibiting glucose absorption of the human body in addition to the extract of the seedlings as needed.

이하, 본 발명의 구성 및 작용효과를 실시예를 통하여 보다 상세히 설명한다. 단, 하기 실시예는 본 발명을 예시하기 위한 목적일 뿐 본 발명의 권리범위는 이에 한정되지 않는다.Hereinafter, the configuration and operation of the present invention will be described in more detail with reference to Examples. However, the following examples are only for the purpose of illustrating the present invention is not limited to the scope of the present invention.

실시예 1 : 모시대 추출물의 제조Example 1: Preparation of seedling extract

모시대의 잎, 줄기 및 뿌리부분을 풍건하여 분쇄한 다음, 10배의 메탄올을 가하여 12시간씩 2회 추출한 다음, 여과액을 모아 회전증발농축기에서 용매를 증발시키고, 남은 잔사를 일부 취하여 동결시키고(-70℃), 나머지는 순차 용매분획에 사용하였다. 즉, 헥산, 에틸아세테이트, 그리고 증류수(물)로 차례로 용매 분획을 수행한 다음, 용매를 회전증발농축기에서 증발시키고 건조시켜 시료로 사용하였다.The leaves, stems and roots of the seedlings were air-dried and pulverized, and then extracted twice with 12 times of methanol for 10 hours. The filtrates were collected, the solvent was evaporated in a rotary evaporator, and the remaining residue was taken and frozen ( -70 DEG C), the rest was used in sequential solvent fractions. That is, the solvent fractions were sequentially performed with hexane, ethyl acetate, and distilled water (water), and then the solvent was evaporated and dried in a rotary evaporator to use as a sample.

실시예 2 : 모시대 추출물 및 용매 분획의 알파글루코시다제 저해 활성 측정Example 2: Determination of alphaglucosidase inhibitory activity of seedling extract and solvent fraction

본 실험에 사용한 알파글루코시다제는 쥐의 소장에서 추출하여 -70℃에서 보관하면서 사용하였으며, 돼지 췌장 알파아밀라제는 시그마사(St Louis, MO, USA)에서 구매하였다. 알파글루코시다제 효소기질은 파라-니트로페닐알파-디-글루코피라노사이드(para-nitrophenyl-alpha-D-glucopyranoside)를 사용하였으며, 알파아밀라제 기질로서는 파라-니트로페닐-알파-디-말토페타오사이드(para-nitrophenyl- alpha-D-maltopentaoside)를 사용하였다. 한편, 양성대조구로는 혈당상승억제제로 임상적으로 사용되고 있는 아카보스(Bayer-Korea)를 사용하였다.The alpha glucosidase used in this experiment was extracted from the small intestine of the rat and stored at -70 ° C. Porcine pancreatic alpha amylase was purchased from Sigma (St Louis, MO, USA). Para-nitrophenyl-alpha-D-glucopyranoside was used as an alphaglucosidase enzyme substrate, and para-nitrophenyl-alpha-di-maltopetao as an alpha amylase substrate. Para-nitrophenyl-alpha-D-maltopentaoside was used. On the other hand, as a positive control was used akabose (Bayer-Korea) which is clinically used as a blood glucose increase inhibitor.

모시대의 알콜(메탄올) 추출물 및 그 용매 분획물(물층, 헥산층, 에틸아세테이트층)을 건조시킨 후, 디메틸설폭사이드 또는 물에 일정 농도로 용해시켜 용액을 제조하고, 이를 쥐의 소장에서 추출한 알파글루코시다제 함유 효소반응액에 넣어 알파글루코시다제 저해정도를 살펴본 결과, 1mg/ml에서 알콜추출물과 물층은 효소활성을 각각 37.7, 50.7% 저해하는 것으로 나타났다. 물층은 0.1mg/ml의 농도에서도 효소활성을 22.8% 저해하는 것으로 나타났으며, 이는 임상적으로 사용하고 있는 아카보스보다는 약간 낮은 수준이었다(표1).The alcoholic (methanol) extract and its solvent fractions (water layer, hexane layer, ethyl acetate layer) of the era were dried and dissolved in dimethyl sulfoxide or water at a constant concentration to prepare a solution, which was extracted from the small intestine of the mouse. As a result of examining the degree of alpha glucosidase inhibition in the enzyme-containing enzyme reaction solution, the alcohol extract and the water layer inhibited the enzyme activity by 37.7 and 50.7%, respectively, at 1 mg / ml. The water layer was found to inhibit 22.8% of enzyme activity even at a concentration of 0.1mg / ml, which was slightly lower than the acarbose used clinically (Table 1).

한편, 헥산층은 1mg/ml농도에서 효소활성을 28.1% 저해하였으며, 에틸아세테이트층은 효소활성을 전혀 저해하지 않았다.On the other hand, the hexane layer inhibited the enzyme activity by 28.1% at a concentration of 1mg / ml, the ethyl acetate layer did not inhibit the enzyme activity at all.

실시예 3 : 모시대 추출물의 알파아밀라제 효소 저해활성Example 3: Alpha amylase enzyme inhibitory activity

모시대의 알콜추출물 및 그 용매분획을 디메틸설폭사이드 또는 증류수에 5mg/ml 정도로 용해시킨 다음, 돼지 췌장에서 분리한 알파아밀라제 용액에 첨가하여 효소활성 저해정도를 측정한 결과,메탄올 추출물, 물층, 에틸아세테이트층, 헥산층은 효소활성을 각각 61.0, 62.4, 100, 42.5% 저해하였다. 이렇듯 모시대의 알콜추출물이나 그것의 물과 에틸아세테이트 분획은 알파아밀라제 저해활성이 매우 높은 것으로 나타났다(표 2).The alcoholic extract and its solvent fractions of the Moshi era were dissolved in dimethyl sulfoxide or distilled water at about 5 mg / ml, and then added to the alpha amylase solution isolated from the pig pancreas to measure the inhibition of enzyme activity. The layer and hexane layer inhibited enzymatic activity 61.0, 62.4, 100 and 42.5%, respectively. As such, the alcoholic extract of the era, or its water and ethyl acetate fractions, showed very high alpha amylase inhibitory activity (Table 2).

*. 모든 시료의 농도는 5mg/ml임.*. The concentration of all samples is 5 mg / ml.

** : 측정은 3회 반복하였으며, 결과는 평균값으로 나타냄.**: The measurement was repeated three times, and the result is expressed as an average value.

알파아밀라제는 인체의 췌장에서 분비되는 효소로서 전분의 초기 소화에 관여한다. 즉, 전분을 분해하여 주로 맥아당(maltose) 단위로 전환하는데 관여하는데, 이를 저해할 경우 혈당의 상승억제효과가 있는 것으로 알려져 있다. 임상적으로 사용되고 있는 아카보스는 아밀라제저해활성이 매우 높은 것으로 보고되고 있다.Alpha amylase is an enzyme secreted by the human pancreas and is involved in the early digestion of starch. In other words, starch is mainly involved in the conversion to maltose units by breaking down starch, which is known to have a synergistic inhibitory effect on blood glucose. Clinically used acarbose has been reported to have a very high amylase inhibitory activity.

실시예 4 : 흰쥐를 이용한 모시대 추출물의 혈당강하 효과 측정Example 4 Measurement of Hypoglycemic Effect of Hairy Age Extract Using Rats

본 실험에서는 스프레그-도올리(Sprague-Dawley)계 흰쥐(150-200g)를 사용하였으며, 흰쥐 체중 1kg당 가용성 전분과 모시대 또는 잔대 추출물을 1, 5, 10mg/kg체중을 혼합하여 경구투여하기 전(0분), 투여후 30, 60, 120분에 각각 쥐의 꼬리부분에서 채취하여 혈당 측정계를 이용하여 혈당을 측정하였다.In this experiment, Sprague-Dawley rats (150-200 g) were used. Oral administration of 1, 5, and 10 mg / kg body weight of soluble starch and seedling or stalk extract per 1 kg of rat body weight Blood glucose was measured using a blood glucose meter before and after (0 minutes) and 30, 60 and 120 minutes after administration, respectively.

하룻밤동안 절식시킨 흰쥐에게 가용성 전분 1g/kg 수준으로 투여하기 전에 혈당을 측정하고, 전분을 각기 다른 함량의 모시대추출물과 함께 경구투여한 후 30분 간격으로 꼬리정맥에서 혈액을 채취하여 혈당농도를 측정한 결과, 모시대추출물이 포함되지 않은 대조군에서는 전분투여 30분 후 급격한 혈당상승을 보인 반면, 모시대 추출물을 체중 1kg당 1mg이상으로 투여한 흰쥐의 경우는 전분부하에 의한 혈당상승이 대조군에 비하여 완만하게 진행되는 것으로 나타났다. 그러나, 체중 1kg당 1mg 농도로 투여한 모시대 추출물에서는 혈당저하효과가 관찰되지 않았다. 한편, 모시대 추출물을 체중 kg당 10mg 투여한 경우는 5mg 투여한 흰쥐에서보다 혈당상승억제 효과가 큰 것으로 나타났다(도 1). 모든 실험은 3회 반복하여 수행하였으며 결과는 평균값이다.Glucose was measured before administering 1g / kg of soluble starch to overnight fasted rats, and blood glucose was collected from tail vein at 30 minute intervals after oral administration of starch with different amounts of seedling extracts. As a result, the control group containing no seedling extract showed a rapid blood sugar increase 30 minutes after starch administration, whereas in rats treated with the seedling extract more than 1 mg / kg body weight, the increase in blood sugar level by starch load was slower than that of the control group. It was shown to proceed. However, no hypoglycemic effect was observed in the seedling extract administered at a concentration of 1 mg / kg body weight. On the other hand, administration of 10mg per kg body weight extract was shown to have a higher blood sugar suppression effect than in rats administered 5mg (Fig. 1). All experiments were performed three times and the results are average.

이상 살펴본 바와 같이 본 발명 모시대 추출물은 알파글루코시다제 및 알파아밀라제 효소활성을 억제하여 식후 혈중 포도당 농도의 급격한 상승을 억제하여 인체나 동물의 당뇨병 예방 및/또는 치료에 이용할 수 있는 매우 뛰어난 효과가 있어 본 발명은 의약식품산업상 매우 유용한 발명인 것이다.As described above, the seedling extract of the present invention has a very excellent effect that can be used to prevent and / or treat diabetes in humans or animals by inhibiting the rapid increase in blood glucose levels after meals by inhibiting alpha glucosidase and alpha amylase enzyme activities. The present invention is a very useful invention in the pharmaceutical food industry.

Claims (4)

모시대의 잎, 뿌리, 줄기로부터 물 또는 유기용매로 추출하여 혈당강하 효과를 갖는 모시대 추출물을 제조하는 것을 특징으로 하는 혈당강하용 모시대추출물의 제조방법.A method for producing a hypoglycemic extract, characterized in that the extract from the leaves, roots, stems of the seedlings with water or an organic solvent to prepare a seedling extract having a blood sugar-lowering effect. 상기 제1항 방법에 의해 제조된 혈당강하 효과를 갖는 모시대 추출물.A seedling extract having a hypoglycemic effect prepared by the method of claim 1. 상기 제2항의 모시대 추출물을 유효성분으로 포함하는 것을 특징으로 하는 혈당강하용 조성물.The composition for lowering blood sugar, comprising the extract of claim 2 as an active ingredient. 상기 제2항의 모시대 추출물을 유효성분으로 포함하는 것을 특징으로 하는 혈당강하용 식품.Food for hypoglycemic, comprising the extract of claim 2 as an active ingredient.
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