KR101303306B1 - Composition comprising an extract of Akebiae Caulis for preventing and treating obesity - Google Patents
Composition comprising an extract of Akebiae Caulis for preventing and treating obesity Download PDFInfo
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- KR101303306B1 KR101303306B1 KR1020110048433A KR20110048433A KR101303306B1 KR 101303306 B1 KR101303306 B1 KR 101303306B1 KR 1020110048433 A KR1020110048433 A KR 1020110048433A KR 20110048433 A KR20110048433 A KR 20110048433A KR 101303306 B1 KR101303306 B1 KR 101303306B1
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- 239000011718 vitamin C Substances 0.000 description 1
- 235000019165 vitamin E Nutrition 0.000 description 1
- 239000011709 vitamin E Substances 0.000 description 1
- 229940046009 vitamin E Drugs 0.000 description 1
- 229940011671 vitamin b6 Drugs 0.000 description 1
- 239000011787 zinc oxide Substances 0.000 description 1
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Abstract
본 발명은 으름덩굴 추출물을 유효성분으로 함유하는 비만증의 예방 및 치료용 조성물에 관한 것이다.The present invention relates to a composition for the prevention and treatment of obesity, which contains the slime extract as an active ingredient.
Description
본 발명은 으름덩굴 추출물을 유효성분으로 함유하는 비만증의 예방 및 치료용 조성물에 관한 것이다.
The present invention relates to a composition for the prevention and treatment of obesity, which contains the slime extract as an active ingredient.
[문헌 1] 이상인, 안덕균. 본초학. 영림사, 1992.[Document 1] More than, Andeokgyun. Herbology. Younglimsa, 1992.
[문헌 2] H.H. Lee, K.J. Kim, O.H. Lee, B.Y. Lee. Effect of Pycnogenol on Glucose Transport in Mature 3T3-L1 Adipocytes. Phytother. Res. 24: 12421249. 2010.Document 2 H.H. Lee, K.J. Kim, O. H. Lee, B.Y. Lee. Effect of Pycnogenol on Glucose Transport in Mature 3T3-L1 Adipocytes. Phytother. Res. 24: 12421249. 2010.
[문헌 3] Lin, J., Della-Fera, MA. AND Baile, C.A. Green tea polyphenol epigallocatechin gallate inhibits adipogenesis and induces apoptosis in 3T3-L1 adipocytes. Obes Res 13:982-990, 2005.[Reference 3] Lin, J., Della-Fera, MA. AND Baile, C. A. Green tea polyphenol epigallocatechin gallate inhibits adipogenesis and induces apoptosis in 3T3-L1 adipocytes. Obes Res 13: 982-990, 2005.
[문헌 4] Gao CL, Zhu JG et al. Mitochondrial dysfunction is induced by the overexpression of UCP4 in 3T3-L1 adipocytes. Int J Mol Med 25: 71-80, 2010.4 Gao CL, Zhu JG et al. Mitochondrial dysfunction is induced by the overexpression of UCP4 in 3T3-L1 adipocytes. Int J Mol Med 25: 71-80, 2010.
[문헌 5] Renu G.J, Lucy G.A. Ectopic Expression of Hel-N1, an RNA-Binding Protein, increases Glucose Transporter (GLUT1) Expression in 3T3-L1 Adipocytes. Molecular and cellular biololgy, p.95462, 1997.Renu G.J, Lucy G.A. Ectopic Expression of Hel-N1, an RNA-Binding Protein, increases Glucose Transporter (GLUT1) Expression in 3T3-L1 Adipocytes. Molecular and cellular biololgy, p. 95462, 1997.
[문헌 6] Nugent C, Prins JB, Whitehead JP. Potentiation of glucose uptake in 3T3-L1 adipocytes by PPAR gamma agonists is maintained in cells expressing a PPAR gamma dominant-negative mutant: evidence for selectivity in the downstream responses to PPAR gamma activation. Mol Endocrinol.15(10):1729-38, 2001.
6 Nugent C, Prins JB, Whitehead JP. Potentiation of glucose uptake in 3T3-L1 adipocytes by PPAR gamma agonists is maintained in cells expressing a PPAR gamma dominant-negative mutant: evidence for selectivity in the downstream responses to PPAR gamma activation. Mol Endocrinol. 15 (10): 1729-38, 2001.
최근 세계 각국에서 비만은 심각한 보건문제로 인식되고 있으며 우리나라 역시 세계화의 영향과 산업성장으로 인해 식생활을 포함한 생활습관의 불균형과 활동량 부족으로 비만 인구가 증가하는 것으로 보고되고 있다. 또한 당뇨병, 암, 고혈압, 동맥경화와 같은 순환기계 질환 등 비만과 관련된 질환의 발병률도 증가하고 있으며, 주요 사망원인들 중의 하나로서, 그 심각성은 점점 고조되고 있는 실정이다. 비만이 생기는 원인은 내분비계 및 인체 내 대사와 같은 유전적인 부분도 있지만, 직접적인 원인은 에너지 축적과 에너지 소비의 불균형 즉, 열량 섭취량 및 열량 소비량의 불균형 때문이다. 한의학에서는 비만의 원인으로 膏梁珍味, 氣虛, 濕滯 등으로 보고 있으며 치료법으로는 利水化濕, 化痰, 淸熱通腑, 活血祛瘀의 약물요법을 쓰고 있다.Recently, obesity is recognized as a serious health problem in various countries around the world, and Korea is also reported to increase obesity population due to the imbalance of lifestyle and lack of activity due to globalization and industrial growth. In addition, the incidence of obesity-related diseases, such as diabetes, cancer, high blood pressure, circulatory diseases such as arteriosclerosis is also increasing, and as one of the major causes of death, the severity is increasing. Obesity can also be caused by genetics such as the endocrine system and metabolism in the human body, but the direct cause is due to an imbalance between energy accumulation and energy consumption, i.e., calorie intake and calorie consumption. In oriental medicine, the causes of obesity are reported as 膏 梁 珍味, 氣虛, 濕 滯, and the treatment is using the drug therapy of 利 水 化濕, 化痰, 淸 熱 通腑, 活血 祛瘀.
지방세포는 비만과 관련이 있는 생리적 요소로서 지방 세포내의 중성지방 양이 늘어나거나 지방세포의 수가 증가하여 비만을 유도하게 된다. 세포내 중성지방이 증가하게 되면 운동이나 식이조절을 통해 축적된 중성지방을 분해 할 수 있지만 지방세포 수의 증가로 비만이 유도된 경우에는 생성된 지방세포를 파괴하거나 제거해야 하므로 비만을 치료하는데 많은 어려움이 있다. 전구 지방세포에서 지방세포로의 분화는 성장과정 중 특정 호르몬의 급작스러운 양적변화와 과잉 영양 공급에 의해 유발될 수 있으며 지방세포 분화 유도인자에 의해 신체 내 특정부위에서 지방세포 분화가 일어날 수 있다. 또한 지방세포의 크기는 한계가 있으므로 과잉에너지를 섭취 할 때 에너지를 신속히 저장하기 위해서 지방세포수의 증가가 일어나게 된다. 3T3-L1 세포주는 3T3 쥐 배아의 섬유아세포로부터 유래 된 전구 지방세포주로써 비만이나 대사성 질환 연구에 가장 많이 이용되고 있다. Fat cells are a physiological factor associated with obesity, which leads to an increase in the amount of triglycerides in fat cells or an increase in the number of fat cells. Increasing intracellular triglycerides can degrade triglycerides accumulated through exercise or dietary control, but when obesity is induced by an increase in the number of adipocytes, fat cells produced must be destroyed or removed. There is difficulty. Differentiation of progenitor cells into adipocytes may be caused by sudden quantitative changes and excess nutrition of certain hormones during the growth process, and adipocyte differentiation may occur at specific sites in the body by adipocyte differentiation inducing factors. In addition, since the size of fat cells is limited, an increase in the number of fat cells occurs in order to store energy quickly when excess energy is ingested. The 3T3-L1 cell line is a progenitor cell line derived from fibroblasts of 3T3 mouse embryos and is most used for the study of obesity and metabolic diseases.
전 세계적으로 급증하고 있는 비만 인구와 그로 인한 대사성 질환 발병률은 심각한 보건문제로 떠오르고 있다. 최근 WHO(세계보건기구)의 보고에 의하면 약 10억의 인구가 과체중이며, 그중 한국인의 경우 서구인에 비해 BMI(체질량지수)를 기준으로 한 비만도는 낮지만 당뇨, 대사질환 발병률은 상대적으로 높은 수준으로 나타나 비만 및 대사질환의 관심이 높아지고 있다. 현재 시판되고 있는 많은 비만 치료제의 대부분은 중추 신경계에 작용하여 식욕을 억제하거나 지방의 소화를 막아 체외로 배출하는 역할을 하고 있다. 하지만 부작용으로 두통과 불면증, 혈압상승, 식욕부진 등의 부작용이 나타나는 것으로 보고되어 있고 향정신성 의약품으로 분류되어 있는 약도 있어 위험성이 크다. 이러한 상황에서 많은 연구자들은 식품 소재와 한약재로부터 비만 치료제를 찾는 연구들을 진행하고 있다.The world's rapidly growing obese population and the resulting incidence of metabolic diseases are a serious health problem. According to a recent report by the World Health Organization (WHO), about 1 billion people are overweight. Among Koreans, obesity is lower based on BMI (body mass index), but the incidence of diabetes and metabolic diseases is relatively high. Appears as the interest of obesity and metabolic diseases are increasing. Many of the current anti-obesity drugs on the market act on the central nervous system to suppress appetite or prevent the digestion of fat, which acts to release it in vitro. However, side effects such as headache, insomnia, increased blood pressure, and loss of appetite have been reported as side effects, and there are also drugs classified as psychotropic drugs, which is very dangerous. In this situation, many researchers are working on finding obesity drugs from food ingredients and herbal medicines.
으름덩굴 잎은 으름덩굴과(Lardizabalaceae)에 속하는 으름덩굴의 잎으로 전국 산간지에 산재되어 있으며, 한방에서는 으름덩굴의 줄기를 목통(木通, Akebiae Caulis)으로 사용하고 있으며 열매와 잎은 무독성으로 식용하고 있다(이상인, 안덕균. 본초학. 영림사, 1992). The vine leaves are the leaves of the vines belonging to the genus Lardizabalaceae and are scattered in mountainous areas nationwide. (Lee Sang-in, Ahn Deok-kyun, Herbology, Younglimsa, 1992).
항비만 소재를 탐색하기 위한 in vitro 연구에서는 3T3-L1 cell을 이용하여 지방세포의 분화를 억제하고 지방생성을 저해하는 소재의 효능 및 기전을 연구한 바, 으름덩굴 추출물이 지방세포 분화 및 지방생성을 억제함을 확인하여 본 발명을 완성하게 되었다.
In to search for an anti-obesity material In vitro studies were conducted using 3T3-L1 cells to study the efficacy and mechanism of inhibiting the differentiation of adipocytes and inhibiting adipogenesis, confirming that the scabbard extract inhibits adipocyte differentiation and adipogenesis. To complete.
상기 목적을 수행하기 위하여, 본 발명은 으름덩굴 추출물을 유효성분으로 함유하는 비만증의 예방 및 치료용 약학조성물을 제공한다. In order to carry out the above object, the present invention provides a pharmaceutical composition for the prevention and treatment of obesity, containing the snail extract as an active ingredient.
또한, 본 발명은 으름덩굴 추출물을 유효성분으로 함유하는 비만증의 예방 및 개선용 건강기능식품을 제공한다.In another aspect, the present invention provides a health functional food for the prevention and improvement of obesity containing the slime extract as an active ingredient.
또한, 본원에서 정의되는 추출물은 으름덩굴의 잎, 뿌리, 줄기 또는 전초, 바람직하게는 잎의 정제수를 포함한 물, 탄소수 1 내지 4의 저급알코올 또는 이들의 혼합용매로부터 선택된 용매, 바람직하게는 물 또는 물 및 에탄올 혼합용매, 보다 바람직하게는 물 또는 50-99% 물 및 에탄올 혼합용매에 가용한 추출물을 포함한다. In addition, the extract as defined herein is a solvent, preferably water, selected from the leaves, roots, stems or starches of the vines, preferably water containing purified water of the leaves, lower alcohols having 1 to 4 carbon atoms or mixed solvents thereof. Water and ethanol mixed solvents, more preferably water or extracts soluble in 50-99% water and ethanol mixed solvents.
이하, 본 발명의 추출물을 수득하는 방법을 상세히 설명한다.Hereinafter, the method for obtaining the extract of the present invention will be described in detail.
본 발명의 추출물은 으름덩굴 잎을 세척, 건조 및 세절하고, 상기 시료 총 중량의 약 1배 내지 200배(w/w), 바람직하게는 10배 내지 100배(w/w)의 정제수를 포함한 물, 주정, 탄소수 1 내지 4의 저급 알콜, 또는 이들의 혼합용매, 바람직하게는 물 또는 물 및 에탄올 혼합용매, 보다 바람직하게는 물 또는 50-99% 물 및 에탄올 혼합용매를 가하여 1시간 내지 1주일, 바람직하게는 2시간 내지 12시간 동안, 10℃ 내지 150℃, 바람직하게는 20℃ 내지 100℃, 보다 바람직하게는 60℃ 내지 90℃에서 냉침추출, 열수추출, 초음파 추출, 환류냉각 추출 등의 추출방법, 바람직하게는 열수추출법을 수행하여 추출물을 수득하는 제 1단계; 상기에서 얻은 추출물을 여과하고 여과물을 동결 건조시키는 제 2단계의 제조공정을 통하여 본 발명의 추출물을 수득할 수 있다.The extract of the present invention washes, dries, and shreds stalk leaves, and includes about 1 to 200 times (w / w), preferably 10 to 100 times (w / w) purified water, of the total weight of the sample. Water, alcohol, lower alcohols having 1 to 4 carbon atoms, or mixed solvents thereof, preferably water or water and ethanol mixed solvent, more preferably water or 50-99% water and ethanol mixed solvent, for 1 hour to 1 Cold extraction, hot water extraction, ultrasonic extraction, reflux cooling extraction, etc. at 10 ° C. to 150 ° C., preferably 20 ° C. to 100 ° C., more preferably 60 ° C. to 90 ° C. for a week, preferably 2 to 12 hours. Extraction method of, preferably, a first step of obtaining an extract by performing a hot water extraction method; The extract of the present invention may be obtained through the second step of manufacturing the filter obtained by filtration and freeze-drying the filtrate.
따라서, 본 발명은 상기의 제조방법으로 얻어진 으름덩굴 추출물을 유효성분으로 함유하는 비만증의 예방 및 치료용 약학 조성물을 제공한다.Accordingly, the present invention provides a pharmaceutical composition for the prevention and treatment of obesity, containing the scab extract obtained by the above production method as an active ingredient.
상기에서 제조된 조합 추출물은 3T3-L1 adipocyte를 이용한 세포생존율 실험, 세포내 지질축적실험, 지방세포의 지방분해에 미치는 영향실험, 지방생성억제실험, 세포사멸에 미치는 영향실험 등을 통하여 지방세포 분화 억제 및 지방생성 억제 효능과 세포사멸 유도능이 탁월함을 확인하였다.The combination extract prepared above was characterized by differentiation of adipocytes through cell viability test using 3T3-L1 adipocyte, intracellular lipid accumulation test, effect test on adipocyte lipolysis, fat production suppression test, and effect test on cell death. It was confirmed that the inhibitory and adipogenic inhibitory effect and the ability to induce apoptosis were excellent.
본 발명의 약학 조성물은 조성물 총 중량에 대하여 상기 추출물을 0.1 내지 50 중량%로 포함한다.The pharmaceutical composition of the present invention comprises 0.1 to 50% by weight of the above extract, based on the total weight of the composition.
그러나 상기와 같은 조성은 반드시 이에 한정되는 것은 아니고, 환자의 상태 및 질환의 종류 및 진행 정도에 따라 변할 수 있다.However, the composition is not limited thereto, and may vary depending on the condition of the patient, the type of disease, and the progress of the disease.
본 발명의 추출물 자체는 독성 및 부작용이 거의 없으므로 예방 목적으로 장기간 복용 시에도 안심하고 사용할 수 있는 약제이다. Since the extract of the present invention has little toxicity and side effects, it can be safely used even for long-term administration for the purpose of prevention.
본 발명의 조성물은 약학적 조성물의 제조에 통상적으로 사용하는 적절한 담체, 부형제 및 희석제를 더 포함할 수 있다.The compositions of the present invention may further comprise suitable carriers, excipients and diluents conventionally used in the manufacture of pharmaceutical compositions.
본 발명의 조성물은 각각 통상의 방법에 따라 산제, 과립제, 정제, 캡슐제, 현탁액, 에멀젼, 시럽, 에어로졸 등의 경구형 제형, 외용제, 좌제 및 멸균 주사용액의 형태로 제형화하여 사용될 수 있으며, 추출물을 포함하는 조성물에 포함될 수 있는 담체, 부형제 및 희석제로는 락토즈, 덱스트로즈, 수크로스, 솔비톨, 만니톨, 자일리톨, 에리스리톨, 말티톨, 전분, 아카시아 고무, 알지네이트, 젤라틴, 칼슘 포스페이트, 칼슘 실리케이트, 셀룰로즈, 메틸 셀룰로즈, 미정질 셀룰로스, 폴리비닐 피롤리돈, 물, 메틸히드록시벤조에이트, 프로필히드록시벤조에이트, 탈크, 마그네슘 스테아레이트 및 광물유를 들 수 있다. 제제화할 경우에는 보통 사용하는 충진제, 증량제, 결합제, 습윤제, 붕해제, 계면활성제 등의 희석제 또는 부형제를 사용하여 조제된다. 경구투여를 위한 고형제제에는 정제, 환제, 산제, 과립제, 캡슐제 등이 포함되며, 이러한 고형제제는 상기 추출물에 적어도 하나 이상의 부형제 예를 들면, 전분, 칼슘카보네이트(calcium carbonate), 수크로스(sucrose) 또는 락토오스(lactose), 젤라틴 등을 섞어 조제된다. 또한 단순한 부형제 이외에 마그네슘 스테아레이트 탈크 같은 윤활제들도 사용된다. 경구를 위한 액상제제로는 현탁제, 내용액제, 유제, 시럽제 등이 해당되는데 흔히 사용되는 단순희석제인 물, 리퀴드 파라핀 이외에 여러 가지 부형제, 예를 들면 습윤제, 감미제, 방향제, 보존제 등이 포함될 수 있다. 비경구 투여를 위한 제제에는 멸균된 수용액, 비수성용제, 현탁제, 유제, 동결건조제제, 좌제가 포함된다. 비수성용제, 현탁제로는 프로필렌글리콜 (propylene glycol), 폴리에틸렌 글리콜, 올리브 오일과 같은 식물성 기름, 에틸올레이트와 같은 주사 가능한 에스테르 등이 사용될 수 있다. 좌제의 기제로는 위텝솔(witepsol), 마크로골, 트윈(tween) 61, 카카오지, 라우린지, 글리세롤젤라틴 등이 사용될 수 있다.The composition of the present invention may be formulated in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols and the like, oral preparations, suppositories and sterilized injection solutions, Examples of carriers, excipients and diluents that can be included in the composition containing the extract include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate , Cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. In the case of formulation, a diluent or excipient such as a filler, an extender, a binder, a wetting agent, a disintegrant, or a surfactant is usually used. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, and the solid preparations may include at least one excipient such as starch, calcium carbonate, sucrose in the extract. ) Or lactose, gelatin and the like are mixed. In addition to simple excipients, lubricants such as magnesium stearate talc are also used. Examples of the liquid preparation for oral use include suspensions, solutions, emulsions, and syrups. In addition to water and liquid paraffin, simple diluents commonly used, various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like may be included . Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Examples of the suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like. As a suppository base, witepsol, macrogol, tween 61, cacao paper, laurin, glycerol gelatin and the like can be used.
본 발명의 조성물의 바람직한 투여량은 환자의 상태 및 체중, 질병의 정도, 약물형태, 투여경로 및 기간에 따라 다르지만, 당업자에 의해 적절하게 선택될 수 있다. 그러나 바람직한 효과를 위해서, 본 발명의 추출물은 1일 0.5g/kg 내지 5g/kg으로, 바람직하게는 1g/kg 내지 3g/kg으로 투여하는 것이 좋다. 투여는 하루에 한번 투여할 수도 있고, 수회 나누어 투여할 수 있다. 따라서, 상기 투여량은 어떠한 면으로든 본 발명의 범위를 한정하는 것은 아니다.The preferred dosage of the composition of the present invention varies depending on the condition and the weight of the patient, the degree of disease, the type of drug, the route of administration and the period of time, but can be appropriately selected by those skilled in the art. However, for the desired effect, the extract of the present invention is preferably administered at 0.5 g / kg to 5 g / kg per day, preferably 1 g / kg to 3 g / kg per day. The administration may be carried out once a day or divided into several doses. Thus, the dosage amounts are not intended to limit the scope of the invention in any manner.
본 발명의 조성물은 쥐, 생쥐, 가축, 인간 등의 포유동물에 다양한 경로로 투여될 수 있다. 투여의 모든 방식은 예상될 수 있는데, 예를 들면, 경구, 직장 또는 정맥, 근육, 피하, 자궁내 경막 또는 뇌혈관내(intracerebroventricular) 주사에 의해 투여될 수 있다.The composition of the present invention may be administered to mammals such as rats, mice, livestock, humans, and the like in various routes. All modes of administration may be expected, for example, by oral, rectal or intravenous, intramuscular, subcutaneous, intra-uterine or intracerebroventricular injections.
또한, 본 발명은 으름덩굴 추출물을 유효성분으로 함유하는 비만증의 예방 및 개선용 건강기능식품을 제공한다. In another aspect, the present invention provides a health functional food for the prevention and improvement of obesity containing the slime extract as an active ingredient.
본원에서 정의되는 "건강기능식품"은 건강기능식품에 관한 법률 제6727호에 따른 인체에 유용한 기능성을 가진 원료나 성분을 사용하여 제조 및 가공한 식품을 의미하며, "기능성"이라 함은 인체의 구조 및 기능에 대하여 영양소를 조절하거나 생리학적 작용 등과 같은 보건 용도에 유용한 효과를 얻을 목적으로 섭취하는 것을 의미한다.&Quot; Health functional food "as defined herein means food prepared and processed using raw materials or ingredients having functionality useful to the human body in accordance with Law No. 6727 on Health Functional Foods." Functional " Structure and function of the nutrient to control or physiological effects, such as to obtain a beneficial effect for health is intended to eat.
본 발명의 비만증 예방을 위한 건강기능식품은, 조성물 총 중량에 대하여 상기 추출물을 0.01 내지 95%, 바람직하게는 1 내지 80%중량 백분율로 포함한다.The dietary supplement for preventing obesity of the present invention comprises the extract in an amount of 0.01 to 95%, preferably 1 to 80% by weight based on the total weight of the composition.
또한, 비만증 예방을 위한 목적으로 정제, 캅셀, 분말, 과립, 액상, 환 등의 형태인 건강기능식품으로 제조 및 가공이 가능하다.In addition, it is possible to manufacture and process as a dietary supplement in the form of tablets, capsules, powders, granules, liquid, pills for the purpose of preventing obesity.
본 발명은 비만증의 예방 및 치료의 효과를 나타내는 상기 추출물을 포함하는 건강보조식품을 제공한다. 상기 추출물을 첨가할 수 있는 식품으로는, 예를 들어, 각종 식품류, 음료, 껌, 차, 비타민 복합제, 건강 기능성 식품류 등이 있다.The present invention provides a dietary supplement comprising the extract showing the effect of preventing and treating obesity. Examples of foods to which the extract can be added include various foods, beverages, gums, tea, vitamin complexes, and health functional foods.
본 발명의 추출물을 포함하는 조성물은 비만증의 예방 및 개선을 위한 약제, 식품 및 음료 등에 다양하게 이용될 수 있다. 본 발명의 추출물을 첨가할 수 있는 식품으로는, 예를 들어, 각종 식품류, 음료, 껌, 차, 비타민 복합제, 건강보조 식품류 등이 있고, 분말, 과립, 정제, 캡슐 또는 음료인 형태로 사용할 수 있다.The composition comprising the extract of the present invention can be used in various ways, such as drugs, foods and drinks for the prevention and improvement of obesity. Examples of the foods to which the extract of the present invention can be added include various foods, beverages, gums, tea, vitamin complexes, health supplements and the like, and they can be used as powders, granules, tablets, capsules or beverages have.
또한, 본 발명은 비만증의 예방 및 개선효과를 갖는 으름덩굴 추출물을 주성분으로 함유하는 식품첨가제를 제공한다. 본 발명의 추출물은 비만증의 예방 및 개선을 목적으로 식품 또는 음료에 첨가될 수 있다. 이때, 식품 또는 음료 중의 상기 추출물의 양은 일반적으로 본 발명의 건강식품 조성물은 전체 식품 중량의 1 내지 5 중량%로 가할 수 있으며, 건강 음료 조성물은 100㎖를 기준으로 0.02 내지 10g, 바람직하게는 0.3 내지 1g의 비율로 가할 수 있다. In addition, the present invention provides a food additive containing as a main component of the extract of the creeper having the effect of preventing and improving obesity. The extract of the present invention can be added to food or beverage for the purpose of prevention and improvement of obesity. At this time, the amount of the extract in the food or beverage is generally added to the health food composition of the present invention 1 to 5% by weight of the total food weight, the health beverage composition is 0.02 to 10g, preferably 0.3 based on 100ml To 1 g.
본 발명의 건강 음료 조성물은 지시된 비율로 필수 성분으로서 으름덩굴 추출물을 함유하는 것 외에 액체성분에는 특별한 제한점은 없으며 통상의 음료와 같이 여러 가지 향미제 또는 천연 탄수화물 등을 추가 성분으로서 함유할 수 있다. 상술한 천연 탄수화물의 예는 모노사카라이드, 예를 들어, 포도당, 과당 등의 디사카라이드, 예를 들어 말토스, 수크로스 등의 및 폴리사카라이드, 예를 들어 덱스트린, 시클로덱스트린 등과 같은 통상적인 당 및 자일리톨, 소르비톨, 에리트리톨 등의 당알콜이다. 상술한 것 이외의 향미제로서 천연 향미제(타우마틴, 스테비아 추출물(예를 들어 레바우디오시드 A, 글리시르히진 등)) 및 합성 향미제(사카린, 아스파르탐 등)를 유리하게 사용할 수 있다. 상기 천연 탄수화물의 비율은 본 발명의 조성물 100㎖당 일반적으로 약 1 내지 20g, 바람직하게는 약 5 내지 12g이다.The health beverage composition of the present invention, in addition to containing the scab extract as an essential ingredient in the indicated ratio, there is no particular limitation on the liquid component and may contain various flavors or natural carbohydrates as additional ingredients, such as ordinary drinks. . Examples of the above-mentioned natural carbohydrates are conventional monosaccharides such as disaccharides such as glucose and fructose, such as maltose, sucrose and the like, and polysaccharides such as dextrin, cyclodextrin and the like. Sugars and sugar alcohols such as xylitol, sorbitol, and erythritol. As natural flavors other than those described above, natural flavors (such as tau martin, stevia extract (e.g., rebaudioside A, glycyrrhizin)) and synthetic flavors (saccharin, aspartame, etc.) have. The ratio of the natural carbohydrate is generally about 1 to 20 g, preferably about 5 to 12 g per 100 ml of the composition of the present invention.
상기 외에 본 발명의 조성물은 여러 가지 영양제, 비타민, 광물(전해질), 합성 풍미제 및 천연 풍미제 등의 풍미제, 착색제 및 중진제(치즈, 초콜릿 등), 펙트산 및 그의 염, 알긴산 및 그의 염, 유기산, 보호성 콜로이드 증점제, pH 조절제, 안정화제, 방부제, 글리세린, 알콜, 탄산음료에 사용되는 탄산화제 등을 함유할 수 있다. 그밖에 본 발명의 조성물들은 천연 과일 쥬스 및 야채 음료의 제조를 위한 과육을 함유할 수 있다. 이러한 성분은 독립적으로 또는 조합하여 사용할 수 있다. 이러한 첨가제의 비율은 그렇게 중요하진 않지만 본 발명의 조성물 100 중량부 당 0 내지 약 20 중량부의 범위에서 선택되는 것이 일반적이다.
In addition to the above, the composition of the present invention includes various nutrients, vitamins, minerals (electrolytes), flavors such as synthetic flavors and natural flavors, coloring and neutralizing agents (such as cheese and chocolate), pectic acid and salts thereof, alginic acid and its Salts, organic acids, protective colloidal thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated drinks, and the like. In addition, the compositions of the present invention may contain flesh for the production of natural fruit juices and vegetable beverages. These components may be used independently or in combination. The proportion of such additives is not so critical, but is generally selected in the range of 0 to about 20 parts by weight per 100 parts by weight of the composition of the present invention.
본 발명의 조합 추출물은 3T3-L1 adipocyte를 이용한 세포생존율실험, 세포 분화 및 지방생성 억제실험, 세포사멸에 미치는 영향실험을 통하여 지방세포 분화 억제 및 지방 생성 억제 효능과 세포사멸 유도능이 탁월함을 확인한 바, 비만증의 예방 및 치료에 유용한 약학조성물 및 건강기능식품에 이용될 수 있다.
Combination extract of the present invention was found to be excellent in inhibiting fat cell differentiation and inhibiting fat production and inducing apoptosis through cell viability test, cell differentiation and adipogenesis inhibition test, and cell death test using 3T3-L1 adipocyte. It can be used in pharmaceutical compositions and dietary supplements useful for the prevention and treatment of obesity.
도 1은 성숙된 지방분화세포에 대한 세포독성 효과를 나타낸 도이고(데이타는 평균(mean)±S.D으로 표기되고 별표는(*) 대조군과 유의적으로 상이함, p<0.05.),
도 2는 성숙된 지방분화세포에 대한 시료의 지방 축적 효과를 나타낸 도이고 (데이타는 평균(mean)±S.D으로 표기되고 별표는(*) 대조군과 유의적으로 상이함, p<0.05.),
도 3은 성숙된 지방분화세포 내 triacylglycerol 생성 억제 활성을 나타낸 도이고 (데이타는 평균(mean)±S.D으로 표기되고 별표는(*) 대조군과 유의적으로 상이함, p<0.05.)
도 4는 성숙된 지방분화 세포내의 유리 글리세롤 수준에 미치는 시료의 효과를 나타낸 도이고(데이타는 평균(mean)±S.D으로 표기되고 별표는(*) 대조군과 유의적으로 상이함, p<0.05.),
도 5는 지방분화세포에서의 포도당 흡수 수준에 미치는 시료의 효과를 나타낸 도이다(세포 사멸된 세포는 화살표로 표시하고 (데이타는 평균(mean)±S.D으로 표기되고 별표는(*) 대조군과 유의적으로 상이함, p<0.05.).1 is a diagram showing the cytotoxic effect on mature adipocytes (data are expressed as mean ± SD and asterisks are significantly different from the control ( p <0.05 )).
2 is a diagram showing the fat accumulation effect of the sample on the mature adipocytes (data is expressed as mean ± SD and asterisk (*) significantly different from the control, p <0.05 ),
3 is a diagram showing the triacylglycerol production inhibitory activity in mature adipocytes (data is expressed as mean ± SD and asterisk (*) significantly different from the control, p <0.05 ).
4 is a diagram showing the effect of a sample on free glycerol levels in mature adifferentiated cells (data is expressed as mean ± SD and asterisks are significantly different from the control, p <0.05) . ),
5 is a diagram showing the effect of the sample on the glucose uptake level in adipose cells (apoptotic cells are indicated by an arrow (data is expressed as mean ± SD and asterisk (*)) Differently, p <0.05 ).
이하, 본 발명을 참고예, 실시예 및 실험예에 의해 상세히 설명한다. Hereinafter, the present invention will be described in detail by reference examples, examples and experimental examples.
단, 하기 참고예, 실시예 및 실험예는 본 발명을 예시하는 것일 뿐, 본 발명의 내용이 하기 참고예, 실시예 및 실험예에 한정되는 것은 아니다.
However, the following Reference Examples, Examples and Experimental Examples are merely illustrative of the present invention, and the content of the present invention is not limited to the following Reference Examples, Examples and Experimental Examples.
실시예Example 1. 추출물의 제조 1. Preparation of Extract
으름덩굴 잎을 2009년 6월 전북 임실군 강천산에서 채취하여 건조한 후 으름덩굴 잎 35kg을 80% 에탄올 560L를 가하여 75℃에서 5시간 동안 가열추출하고 여액을 농축기[(주)명일이엔지]로 감압 농축하여 동결건조기(LYOPH-PRIDE 100R, (주)영일이엔지)를 이용하여 -40℃에서 48시간 동안 동결건조시켜 본 발명의 으름덩굴 잎 80% 에탄올 추출물 4.1kg(수득률: 11.7%, 이하 “AQE"라 함)을 수득하여 하기 실험예의 시료로 사용하였다.
After harvesting the leaves of the vines in Gangcheonsan, Imsil-gun, Jeonbuk, in June 2009, 35kg of vine leaves were added to 560L of 80% ethanol, and extracted by heating at 75 ℃ for 5 hours, and the filtrate was concentrated under reduced pressure with a concentrator [Myeongil ENG]. Freeze-dried for 48 hours at -40 ℃ using a freeze dryer (LYOPH-PRIDE 100R, Co., Ltd. Yeongil ENG) 4.1kg (80% ethanol extract of the mash leaf of the present invention yield: 11.7%, below "AQE" ) Was used as a sample of the following experimental example.
참조예Reference Example 1 . 세포 배양과 분화 One . Cell Culture and Differentiation
3T3-L1 전지방세포는 10% Bovine calf serum(BCS)과 항생제가 포함된 Dulbeccos Modified Eagles Medium(DMEM) 배지를 사용하였으며, 5%의 이산화탄소의 공급과 37℃가 유지되는 배양기에서 배양하였다. 마우스의 전지방 세포주 3T3-L1은 ATCC(American Type Culture Collection, CL-173, USA)에서 구입하였고, DMEM 배양액, 트립신, FBS(Fetal bovine serum)와 BCS(Bovine calf serum)는 하이클론(Hyclone, USA)에서 구입하였다.3T3-L1 cells were cultured in a Dulbeccos Modified Eagles Medium (DMEM) medium containing 10% Bovine calf serum (BCS) and antibiotics. The mouse cell line 3T3-L1 was purchased from the American Type Culture Collection (ATCC), CL-173, USA, and DMEM culture, trypsin, FBS (Fetal bovine serum) and BCS (Bovine calf serum) were selected from Hyclone, USA).
세포는 6-웰 플레이트(well plate)에 웰 당 1×105 cells/well의 3T3-L1 세포를 분주해 세포가 100%로 증식될 때까지 10% BCS를 포함한 DMEM 배지에서 배양하였으며, 지방세포로 분화시키기 위하여 10% FBS와 MDI solution(0.5mM 3-isobutyl-1-methyxanthine, 1μM dexamethason, 10μg/ml insulin, Sigma)을 포함한 DMEM 배지를 3일 처리하였고 다시 10% FBS와 10μg/ml 인슐린(insulin, Sigma I6634)을 포함한 DMEM 배지를 3일 동안 처리하였고 그 이후는 10% FBS를 포함한 DMEM 배지로 배양하여 세포 내 지방구의 형성을 근거하여 지방세포로 분화시켰다.
Cells were cultured in DMEM medium containing 10% BCS until a 6% well plate was dispensed with 1 × 10 5 cells / well of 3T3-L1 cells per well into 100% cells. DMEM medium containing 10% FBS and MDI solution (0.5 mM 3-isobutyl-1-methyxanthine, 1 μM dexamethason, 10 μg / ml insulin, Sigma) was treated for 3 days to differentiate into 10% FBS and 10 μg / ml insulin ( DMEM medium containing insulin, Sigma I6634) was treated for 3 days, and then cultured in DMEM medium containing 10% FBS to differentiate into adipocytes based on the formation of intracellular fat globules.
실험예Experimental Example 1. 세포독성 실험 1. Cytotoxicity Test
상기 실시예에서 얻은 AQE에 대한 세포독성을 확인하기 위하여 문헌에 기재된 방법을 응용하여 하기와 같이 실험을 수행하였다(H.H. Lee, K.J. Kim, O.H. Lee, B.Y. Lee. Effect of Pycnogenol on Glucose Transport in Mature 3T3-L1 Adipocytes. Phytother. Res. 24: 12421249. 2010.).In order to confirm the cytotoxicity to AQE obtained in the above example, the experiment was performed by applying the method described in the literature (HH Lee, KJ Kim, OH Lee, BY Lee.Effect of Pycnogenol on Glucose Transport in Mature 3T3) -L1 Adipocytes.Phytother.Res. 24: 12421249. 2010.).
상기 실시예에서 얻은 AQE에 대한 세포독성을 측정하기 위해 24 well plate에 2×105 cells/well의 3T3-L1 세포를 분주한 후 10% BCS를 포함한 DMEM 배지(Dulbeccos Modified Eagles Medium, Hyclone, SH30243, 10% Fetal bovine serum (BCS), Hyclone , SH30401)를 이용하여 24시간 동안 배양한 후 3일 동안 매일 1회 배지를 교체하면서 AQE 시료를 도 1과 같이 농도별로 처리하였다. In order to measure cytotoxicity against AQE obtained in the above example, 2 × 10 5 cells / well of 3T3-L1 cells were dispensed on a 24 well plate and then DMEM medium containing 10% BCS (Dulbeccos Modified Eagles Medium, Hyclone, SH30243). , 10% Fetal bovine serum (BCS), Hyclone, SH30401) was incubated for 24 hours and then AQE samples were treated by concentration as shown in Figure 1 while replacing the medium once daily for 3 days.
배양 후, 4일째 되는 날에 각 웰(well)에 0.25㎖의 XTT-PMS 용액(1mg XTT(XTT Sodium Cell Culture Test, Sigma, X4626) and 10g PMS(phenazine methosulfate, Sigma, P9625)/mL of DMEM(Dulbeccos Modified Eagles Medium (DMEM), Hyclone, SH30406)without phenol red)을 첨가하고 다시 2시간 배양하였다. 세포에 대한 세포독성도는 포르마잔(formazan)의 형성 정도를 microplate reader(BIO-TEK, 1010-1,USA)를 이용하여 450nm에서 흡광도 기기를 이용하여 측정하였다.On day 4 after incubation, 0.25 ml of XTT-PMS solution (1 mg XTT (XTT Sodium Cell Culture Test, Sigma, X4626) and 10 g PMS (phenazine methosulfate, Sigma, P9625) / mL of DMEM in each well) (Dulbeccos Modified Eagles Medium (DMEM), Hyclone, SH30406) without phenol red) was added and incubated for another 2 hours. The cytotoxicity of the cells was measured by using an absorbance device at 450 nm using a microplate reader (BIO-TEK, 1010-1, USA) to form formazan.
본 실험 결과, 도 1에 나타난 바와 같이, 본 발명의 AQE가 10ug/mL의 농도에서 최대안전 범위를 나타내고 있음을 확인하였다. AQE 시료의 세포독성을 평가한 결과 5ug/mL 이하의 농도에서 세포독성이 나타나지 않았다(도 1).
As a result of the experiment, as shown in Figure 1, it was confirmed that the AQE of the present invention shows a maximum safety range at a concentration of 10ug / mL. Evaluation of the cytotoxicity of the AQE sample showed no cytotoxicity at concentrations of 5 ug / mL or less (Fig. 1).
실험예Experimental Example 2. 지방세포 분화 및 지방 축적에 미치는 영향 실험 2. Experiment on the effect on adipocyte differentiation and fat accumulation
상기 실시예에서 얻은 AQE에 대한 지방세포 분화 및 지방 축적에 미치는 영향을 확인하기 위하여 문헌에 기재된 Oil Red O 염색방법을 응용하여 하기와 같이 실험을 수행하였다(Lin, J., Della-Fera, MA. AND Baile, C.A. Green tea polyphenol epigallocatechin gallate inhibits adipogenesis and induces apoptosis in 3T3-L1 adipocytes. Obes Res 13:982-990, 2005).In order to confirm the effects on the adipocyte differentiation and fat accumulation on the AQE obtained in the above example, the experiment was performed by applying the Oil Red O staining method described in the literature (Lin, J., Della-Fera, MA). AND Baile, CA Green tea polyphenol epigallocatechin gallate inhibits adipogenesis and induces apoptosis in 3T3-L1 adipocytes.Obes Res 13: 982-990, 2005).
분화시킨 3T3-L1 지방세포에서 지방 축적에 미치는 AQE의 효과를 알아보기 위해 Lin J 등의 방법에 따라 Oil Red O 염색을 실시하였다. 96시간동안 AQE를 처리한 세포에 Phosphate buffer saline(PBS)으로 2회 세척한 다음 10% formaldehyde로 10분간 상온에서 고정시켰다. 고정된 세포를 건조시킨 후 0.4μm filter로 여과한 Oil Red O (O5625, SIGMA, USA) 염색용액으로 20분간 염색한 후 증류수로 세척한 다음 isopropanol로 염색된 지방구를 용출시킨 후 microplate reader(BIO-TEK, 1010-1,USA)를 사용하여 510nm에서 흡광도를 측정하였다.To investigate the effect of AQE on fat accumulation in differentiated 3T3-L1 adipocytes, Oil Red O staining was performed according to Lin J et al. AQE-treated cells were washed twice with Phosphate buffer saline (PBS) for 96 hours and then fixed at room temperature for 10 minutes with 10% formaldehyde. After the fixed cells were dried, they were dyed with Oil Red O (O5625, SIGMA, USA) staining solution filtered with a 0.4μm filter for 20 minutes, washed with distilled water, and eluted fat spheres stained with isopropanol. -TEK, 1010-1, USA) was used to measure absorbance at 510 nm.
본 실험 결과, 3T3-L1 세포 분화 에서 지방 축척 정도를 평가한 결과는 도 2에 제시된 바와 같이 AQE 에서 지방축적 억제 활성이 나타났다.
As a result of evaluating the degree of fat accumulation in 3T3-L1 cell differentiation, it was shown that the fat accumulation inhibitory activity in AQE as shown in FIG.
실험예Experimental Example 3. 세포 내 지질 축적 분석( 3. Analysis of intracellular lipid accumulation IntracellularIntracellular TGTG AssayAssay ))
상기 실시예에서 얻은 AQE에 대한 세포 내 지질 축적을 분석하기 위하여 문헌에 기재된 방법을 응용하여 하기와 같이 실험을 수행하였다(Gao CL, Zhu JG et al. Mitochondrial dysfunction is induced by the overexpression of UCP4 in 3T3-L1 adipocytes. Int J Mol Med 25: 71-80, 2010).In order to analyze intracellular lipid accumulation for AQE obtained in the above example, the experiment was performed by applying the method described in the literature (Gao CL, Zhu JG et al. Mitochondrial dysfunction is induced by the overexpression of UCP4 in 3T3). -L1 adipocytes.Int J Mol Med 25: 71-80, 2010).
분화시킨 3T3-L1 지방세포에서 지질 축적(lipid accumulation)에 미치는 AQE의 효과를 알아보기 위해 현미경으로 충분한 지질의 축적 여부를 확인하고 기존의 배지를 제거한 후, 세포내 지질만을 염색하여 특정 파장의 빛을 내는 Adipored Reagent(PT-7009, Takara, Japan)와 PBS의 혼합물을 1㎖/well씩 분주하여 20분간 염색하였다. 염색여부를 현미경으로 확인한 후 Microplate Fluorescence Reader( BIO-TEK, FLX-800, USA) 에서 485nm의 파장을 주어 535nm 파장의 양을 감지하였다. 이때 535nm 파장의 에너지가 높을수록 세포 내에 많은 지질이 축적된 것으로 볼 수 있으며, 그 결과를 도 3에 나타내었다.To determine the effect of AQE on lipid accumulation in differentiated 3T3-L1 adipocytes, microscopic examination was performed to confirm the accumulation of sufficient lipids, and after removing the medium, only intracellular lipids were stained to light specific wavelengths. A mixture of Adipored Reagent (PT-7009, Takara, Japan) and PBS to give was stained for 20 minutes by dispensing 1mL / well. After staining with a microscope, the microplate fluorescence reader (BIO-TEK, FLX-800, USA) gave a wavelength of 485nm to detect the amount of 535nm wavelength. At this time, the higher the energy of 535nm wavelength can be seen that the accumulation of a lot of lipids in the cell, the results are shown in FIG.
본 실험 결과, 3T3-L1 세포 분화 에서, AQE의 지방세포 내 트리아실 글리세롤생성 억제 활성을 평가한 결과는 도 3에 제시하였다. AQE에서 지방세포내 트리아실 글리세롤(Triacylglycerol) 생성 억제된 것이 확인되었다.
As a result of this experiment, the results of evaluating triacyl glycerol production inhibitory activity of adipocytes of AQE in 3T3-L1 cell differentiation are shown in FIG. 3. It was confirmed that AQE inhibited triacylglycerol production in adipocytes.
실험예Experimental Example 4. 배지로 용출된 글리세롤 양 측정 실험 4. Experiment to measure the amount of glycerol eluted into the medium
상기 실시예에서 얻은 AQE에 대한 지방 분해에 미치는 영향을 확인하기 위하여 문헌에 기재된 방법을 응용하여 하기와 같이 실험을 수행하였다 (Renu G.J, Lucy G.A. Ectopic Expression of Hel-N1, an RNA-Binding Protein, increases Glucose Transporter (GLUT1) Expression in 3T3-L1 Adipocytes. Molecular and cellular biololgy, p.95462, 1997).In order to confirm the effect on lipolysis on AQE obtained in the above example, the experiment was performed by applying the method described in the literature (Renu GJ, Lucy GA Ectopic Expression of Hel-N1, an RNA-Binding Protein, increases Glucose Transporter (GLUT1) Expression in 3T3-L1 Adipocytes.Molecular and cellular biololgy, p. 95462, 1997).
처리시약이 분화된 지방세포의 지방 분해에 미치는 영향을 배지내 글리세롤 함량을 측정함으로서 조사하였다. 방출된 glycerol은 ATP의 존재 하에서 glycerol kinase에 의해 glycerol-1-phosphate로 전환되며 이는 glycerol phosphate oxidase에 의해 dihydroxyacetone phosphate와 hydrogen peroxide(H2O2)로 산화된다. 생성된 H2O2는 4-aminoantipyrine과 sodium N-ethyl-N-(3-sulfopropyl) m-anisidine 존재하에서 peroxidase에 의해 자주색을 띠는 quinoneimine로 전환된다. 따라서 생성된 quinoneimine에 의해 540nm에서 흡광도의 증가는 배지 내에서 글리세롤 농도 증가를 나타낸다. Glycerol 함량은 유리 글리세롤을 표준시약으로 사용한 표준곡선을 작성함으로써 측정하였다.The effect of treatment reagents on lipolysis of differentiated adipocytes was investigated by measuring the content of glycerol in the medium. The released glycerol is converted to glycerol-1-phosphate by glycerol kinase in the presence of ATP, which is oxidized to dihydroxyacetone phosphate and hydrogen peroxide (H 2 O 2 ) by glycerol phosphate oxidase. The resulting H 2 O 2 is converted to purple quinoneimine by peroxidase in the presence of 4-aminoantipyrine and sodium N-ethyl-N- (3-sulfopropyl) m-anisidine. Thus, the increase in absorbance at 540 nm by the produced quinoneimine indicates an increase in glycerol concentration in the medium. Glycerol content was determined by preparing a standard curve using free glycerol as a standard reagent.
분화된 3T3-L1 세포에 시료를 처리한 후 48시간 배양한 배지를 에펜도르프 튜브(eppendorf tube)에 넣고 70℃에서 10분간 가열하여 세포로부터 유리된 효소들을 불활성화 시켰다. 50μl의 배지를 글리세롤 시약(glycerol reagent, Sigma F6428)에 첨가하여 1분간 반응시킨 후 1ml 큐벳(cuvette)에 옮겨 넣고 흡광도를 540nm에서 측정하였다.After treating the sample to differentiated 3T3-L1 cells, the culture medium was incubated for 48 hours in an Eppendorf tube and heated at 70 ° C. for 10 minutes to inactivate enzymes released from the cells. 50 μl of the medium was added to a glycerol reagent (Sigma F6428), reacted for 1 minute, and then transferred to a 1 ml cuvette, and the absorbance was measured at 540 nm.
3T3-L1 세포 분화에서, AQE의 처리에 따른 세포 배양 배지에 용출된 유리 글리세롤 함량을 평가한 결과는 도 4에 제시하였다. In 3T3-L1 cell differentiation, the results of evaluating the free glycerol content eluted in the cell culture medium following the treatment of AQE are shown in FIG. 4.
본 실험 결과, AQE가 배지로 용출된 유리 글리세롤(free glycerol) 함량을 억제되었다( 도 4 참조).
As a result of this experiment, AQE inhibited the content of free glycerol eluted into the medium (see FIG. 4).
실험예Experimental Example 5. 지방세포로 유입된 5. Entered into fat cells 포도당 양Glucose amount 측정 실험 Measurement experiment
상기 실시예에서 얻은 AQE에 대한 지방세포로의 포도당의 유입 양을 측정하기 위하여 문헌에 기재된 방법을 응용하여 하기와 같이 실험을 수행하였다(Nugent C, Prins JB, Whitehead JP. Potentiation of glucose uptake in 3T3-L1 adipocytes by PPAR gamma agonists is maintained in cells expressing a PPAR gamma dominant-negative mutant: evidence for selectivity in the downstream responses to PPAR gamma activation. Mol Endocrinol.15(10):1729-38, 2001).In order to measure the influx of glucose into adipocytes for the AQE obtained in the above example, the experiment was performed by applying the method described in the literature (Nugent C, Prins JB, Whitehead JP. Potentiation of glucose uptake in 3T3-). L1 adipocytes by PPAR gamma agonists is maintained in cells expressing a PPAR gamma dominant-negative mutant: evidence for selectivity in the downstream responses to PPAR gamma activation.Mol Endocrinol. 15 (10): 1729-38, 2001).
분화된 3T3-L1 세포로 유입되어 지방 합성에 이용될 수 있는 glucose의 량을 측정하기 위해 2-NBDG를 이용하였다. DMEM((Dulbeccos Modified Eagles Medium, Hyclone, SH30284)without phenol red)에서 2-NBDG((2-[N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl) amino]-2-deoxy-D-glucose, invitrogen, N13195)농도를 동등하게 유지하고 AQE 시료를 처리하였다. 6시간 배양 후 세포 내로 유입된 2-NBDG를 측정하기 위해 세포를 37℃에서 HEPES-buffered Kreb 용액(KR-H:NaCl 137mM, CaCl22H2O 1.85mM, MgSO47H2O 1.3mM KCl 4.8mM, HEPES 50mM)으로 세 번 세척하고 cell lysis buffer(E1941, Promega)를 이용하여 세포 용출액을 수집하였다. 1,600rpm에서 20분간 원심분리한 후 상층액을 취하여 Microplate Fluorescence Reader(BIO-TEK, FLX-800, USA)로 측정하였다.2-NBDG was used to measure the amount of glucose that could be introduced into differentiated 3T3-L1 cells and used for fat synthesis. 2-NBDG ((2- [N- (7-nitrobenz-2-oxa-1,3-diazol-4-yl) amino] -2 in DMEM ((Dulbeccos Modified Eagles Medium, Hyclone, SH30284) without phenol red) -deoxy-D-glucose, invitrogen, N13195) were maintained at the same concentration and treated with AQE samples After 6-hour incubation, cells were subjected to HEPES-buffered Kreb solution (KR) at 37 ° C. to measure 2-NBDG introduced into cells. -H: NaCl 137mM, CaCl 2 2H 2 O 1.85mM, MgSO 4 7H2O 1.3mM KCl 4.8mM, HEPES 50mM) and washed three times with cell lysis buffer (E1941, Promega) to collect the cell eluate. After centrifugation for 20 minutes at, the supernatant was taken and measured by Microplate Fluorescence Reader (BIO-TEK, FLX-800, USA).
본 실험결과, 3T3-L1 세포 분화에서 AQE의 처리에 따른 지방세포 내로 유입되는 포도당의 정도를 평가한 결과는 도 5에 제시하였다. AQE에서 포도당 유입이 감소됨을 확인 할 수 있었다.As a result of this experiment, the results of evaluating the degree of glucose introduced into the adipocytes following AQE treatment in 3T3-L1 cell differentiation are shown in FIG. 5. It was found that glucose influx was reduced in AQE.
하기에 본 발명의 조성물의 제제예를 설명하나, 본 발명은 이를 한정하고자 함이 아닌 단지 구체적으로 설명하고자 함이다.
Hereinafter, the formulation examples of the composition of the present invention will be described, but the present invention is not intended to limit the present invention but merely to explain in detail.
제제예Formulation example 1. One. 산제의Sanje 제조 Produce
AQE 20 mg
유당 100 mg
탈크 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
ACE 200 mgACE 200 mg
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
AQE 250 mgAQE 250 mg
락토오스 190 mgLactose 190 mg
마그네슘 스테아레이트 10 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
AQE 10 mg
만니톨 180 mg180 mg mannitol
주사용 멸균 증류수 2974 mgSterile sterilized water for injection 2974 mg
Na2HPO4,12H2O 26 mgNa2HPO4,12H2O 26 mg
통상의 주사제의 제조방법에 따라 1 앰플당(2㎖) 상기의 성분 함량으로 제조한다.
(2 ml) per 1 ampoule according to the usual injection preparation method.
제제예Formulation example 5. 5. 액제의Liquid 제조 Produce
AQE 10 gAQE 10 g
이성화당 10 g10 g of isomerized sugar
만니톨 5 g5 g of mannitol
정제수 적량Purified water
통상의 액제의 제조방법에 따라 정제수에 각각의 성분을 가하여 용해시키고 레몬향을 적량 가한 다음 상기의 성분을 혼합한 다음 정제수를 가하여 전체를 정제수를 가하여 전체 100㎖로 조절한 후 갈색병에 충진하여 멸균시켜 액제를 제조한다.
Each component was added to purified water in accordance with the usual liquid preparation method and dissolved, 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 ml with purified water, And sterilized to prepare a liquid preparation.
제제예Formulation example 6. 건강 식품의 제조 6. Manufacture of health food
AQE 5 mg
비타민 혼합물 적량Vitamin mixture quantity
비타민 A 아세테이트 70 ㎍70 [mu] g of vitamin A acetate
비타민 E 1.0 ㎎Vitamin E 1.0 mg
비타민 B1 0.13 ㎎0.13 mg vitamin B1
비타민 B2 0.15 ㎎0.15 mg of vitamin B2
비타민 B6 0.5 ㎎0.5 mg vitamin B6
비타민 B12 0.2 ㎍0.2 [mu] g vitamin B12
비타민 C 10 ㎎10 mg vitamin C
비오틴 10 ㎍Biotin 10 μg
니코틴산아미드 1.7 ㎎Nicotinic acid amide 1.7 mg
엽산 50 ㎍50 ㎍ of folic acid
판토텐산 칼슘 0.5 ㎎Calcium pantothenate 0.5 mg
무기질 혼합물 적량Mineral mixture quantity
황산제1철 1.75 ㎎1.75 mg of ferrous sulfate
산화아연 0.82 ㎎0.82 mg of zinc oxide
탄산마그네슘 25.3 ㎎Magnesium carbonate 25.3 mg
제1인산칼륨 15 ㎎15 mg of potassium phosphate monobasic
제2인산칼슘 55 ㎎Secondary calcium phosphate 55 mg
구연산칼륨 90 ㎎Potassium citrate 90 mg
탄산칼슘 100 ㎎100 mg of calcium carbonate
염화마그네슘 24.8 ㎎24.8 mg of magnesium chloride
상기의 비타민 및 미네랄 혼합물의 조성비는 비교적 건강식품에 적합한 성분을 바람직한 실시예로 혼합 조성하였지만, 그 배합비를 임의로 변형 실시하여도 무방하며, 통상의 건강식품 제조방법에 따라 상기의 성분을 혼합한 다음, 과립을 제조하고, 통상의 방법에 따라 건강식품 조성물 제조에 사용할 수 있다.
Although the composition ratio of the above-mentioned vitamin and mineral mixture is comparatively mixed with a composition suitable for health food as a preferred embodiment, the compounding ratio may be arbitrarily modified, and the above ingredients are mixed according to a conventional method for producing healthy foods , Granules can be prepared and used in the manufacture of health food compositions according to conventional methods.
제제예Formulation example 7. 건강 음료의 제조 7. Manufacture of health drinks
AQE 100 ㎖
구연산 1000 ㎎Citric acid 1000 mg
올리고당 100 g100 g of oligosaccharide
매실농축액 2 gPlum concentrate 2 g
타우린 1 gTaurine 1 g
정제수를 가하여 전체 900 ㎖Purified water was added to a total of 900 ml
통상의 건강음료 제조방법에 따라 상기의 성분을 혼합한 다음, 약 1시간동안 85℃에서 교반 가열한 후, 만들어진 용액을 여과하여 멸균된 2ℓ용기에 취득하여 밀봉 멸균한 뒤 냉장 보관한 다음 본 발명의 건강음료 조성물 제조에 사용한다. The above components were mixed according to a conventional health drink manufacturing method, and the mixture was heated for about 1 hour at 85 DEG C with stirring, and the solution thus prepared was filtered to obtain a sterilized 2-liter container, which was sealed and sterilized, ≪ / RTI >
상기 조성비는 비교적 기호음료에 적합한 성분을 바람직한 실시예로 혼합 조성하였지만, 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 blending ratio may be arbitrarily varied according to the regional and national preferences such as the demand level, the demanding country, and the intended use.
Claims (6)
A pharmaceutical composition for the prevention and treatment of obesity, containing 50-99% water and ethanol mixed solvent extracts of larvae as active ingredients.
Health functional food for the prevention and improvement of obesity, which contains 50-99% water and ethanol mixed solvent extracts of larvae as active ingredients.
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| KR101580018B1 (en) * | 2013-11-20 | 2015-12-23 | 임채규 | Composition of Functional Food for preventing and improving obesity |
| KR102153516B1 (en) * | 2018-01-11 | 2020-09-09 | 경북대학교 산학협력단 | Composition for preventing, treating or improving obesity comprising bioconverted fruit extract of Akebia quinata |
| KR102588562B1 (en) * | 2020-12-16 | 2023-10-12 | 주식회사 비비에프 | Health Functional beverage and manufacturing method of the same |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| KR100497945B1 (en) * | 2001-06-26 | 2005-06-29 | 한국 한의학 연구원 | Extract of herb for promoting release of growth hormone |
| KR100679290B1 (en) | 2005-06-30 | 2007-02-05 | 박정휘 | A composition comprising an extract of ?????201 crude drug complex as an effective ingredient treating or preventing obesity |
| KR20110128676A (en) * | 2010-05-24 | 2011-11-30 | (주)에스오씨 | Functional beverage for diet and its manufacturing mathod |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| KR100497945B1 (en) * | 2001-06-26 | 2005-06-29 | 한국 한의학 연구원 | Extract of herb for promoting release of growth hormone |
| KR100679290B1 (en) | 2005-06-30 | 2007-02-05 | 박정휘 | A composition comprising an extract of ?????201 crude drug complex as an effective ingredient treating or preventing obesity |
| KR20110128676A (en) * | 2010-05-24 | 2011-11-30 | (주)에스오씨 | Functional beverage for diet and its manufacturing mathod |
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| 주진만 등. 동의생리병리학회지. 2009년, 제23권, 제3호, 페이지 662-672 * |
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