Bioassay Guided Isolation of Hypoglycemic Compounds from Philippine Medicinal Pla

生物测定引导下从菲律宾药用植物中分离降血糖化合物

• 批准号: 8034313
• 负责人: LEAH TOLOSA
• 金额: $ 4.64万
• 依托单位: UNIVERSITY OF MARYLAND BALTIMORE COUNTY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-02-22 至 2012-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8034313
• 关键词: Affect; Ally; Analytical Chemistry; Animals; Binding Proteins; Biochemistry; Biodiversity; Biological; Biological Assay; Biological Factors; Biological Markers; Biosensing Techniques; Biosensor; Blood; Blood Glucose; Blood specimen; Buffers; Caring; Cell Culture Techniques; Chemical Structure; Chronic Disease; Collaborations; Complex; Crude Extracts; Data; Data Storage and Retrieval; Developed Countries; Developing Countries; Development; Devices; Diabetes Mellitus; Dyes; Electrical Engineering; Electronics; Elements; Ensure; Enzymes; Event; Fatty Acids; Fermentation; Filipino; Fluorescence; Fluorescence Spectroscopy; Future; Geography; Glean; Glucose; Glutamates; Glutamine; Glycerol; Grant; Health; Heart Diseases; Herbal Medicine; High Pressure Liquid Chromatography; Hot Spot; Housing; Human; Hypoglycemia; Imagery; Implant; In Situ; Indigenous; Individual; Kidney Diseases; Lead; Length; Ligand Binding; Maintenance; Marines; Measures; Medicinal Plants; Medicine; Membrane; Metabolic; Methods; Microdialysis; Microfluidics; Monitor; Mus; Non-Insulin-Dependent Diabetes Mellitus; Nonesterified Fatty Acids; Pattern; Pharmaceutical Preparations; Pharmacologic Substance; Philippines; Planets; Plant Components; Plants; Principal Investigator; Property; Proteins; Protocols documentation; Pump; Pyruvate; Quality Control; Reading; Reagent; Recording of previous events; Reporting; Retrieval; Sampling; Scheme; Screening procedure; Series; Signal Transduction; Site; Skin; Source; Surface; Symptoms; System; Tail; Techniques; Testing; Time; Tissue Sample; Traditional Medicine; Universities; Venipunctures; Work; Writing; base; cost; design; diabetic; drug development; experience; human subject; instrument; interest; male; monitoring device; multidisciplinary; novel; parent grant; parent project; prevent; programs; response; sensor; transmission process; trend; user-friendly; wasting

DESCRIPTION (provided by applicant): The Philippines is one of the planet's biodiversity hot spots. Of 6,000 endemic plants, it is believed that traditional doctors and herbalists use about 1,500 as medicinal plants of which only 1% has been characterized for their active components. Despite this paucity of information, in developing countries such as the Philippines where drugs imported from the developing world are beyond the purchasing power of the majority, herbal medicines remain as the main source of remedies for many ailments. Thus, the long-term objectives of this study is to definitively identify plants that are anti-diabetic based on bioassays for metabolite levels, as well as provide a novel method of quantifying efficacy of herbal medicines based on bioassay guided responses of experimental animals. The former will have an immediate impact in the availability of effective and affordable remedies for diabetes in the Philippines. The latter will provide a means of fact-based quality control of herbal medicines that at present is not always available even in the US. In order to arrive at these objectives, we propose to screen 10+ indigenous plants that have been reported to possess anti-diabetic properties. Screening will be executed at the University of the Philippines Diliman. The screening of crude extracts, fractions and isolates will be bioassay directed using a set of biosensors for glucose, glutamine and fatty acids. These biosensors are currently being developed in the Tolosa lab under project grant R01DK072465 for a device for continuous metabolite monitoring. Although the final device is not yet ready, the individual protein- based sensors can be used in an assay format to test for metabolites in blood. Thus, we propose to develop the protocols for measuring metabolite levels in Male Swiss Webster mice in response to a high glucose load. Using this bioassay protocol, we will screen and identify two plants with the most potent hypoglycemic activity from identified Philippine medicinal plants that have been folkloric ally reported for the treatment of diabetes. Crude extracts will be successively fractionated until the hypoglycemic constituents(s) are isolated from the two most potent Philippine medicinal plants. The chemical structures of these pure compounds will be determined by standard spectroscopic techniques. The novelty of this project is that rather than measuring a single metabolite (glucose), we will multiplex glucose, glutamine and fatty acids to see ratios and trends of these metabolites in response to a high glucose load, and then to the anti-diabetic principle(s). It should be interesting to see if patterns can be gleaned from these trends to further classify anti-diabetic compounds into groups based on their activity. PHS 398/2590 (Rev. 11/07) Page Continuation Format Page PUBLIC HEALTH RELEVANCE: The geography of the Philippine archipelago makes it one of few places on earth with a rich diversity of terrestrial and marine species, many of which have yet to be discovered or characterized. Because of this, Filipinos have a very long history of employing many natural products for various uses including for medicinal purposes. As pharmaceuticals imported from developed countries become more expensive and beyond the purchasing capacity of most Filipinos, fact-based determination of efficacy of readily available herbal medicines is an imperative. Here we propose to screen several Philippine indigenous plants used in traditional Filipino medicine for the treatment of diabetes and its symptoms. These plants will be screened based on their capacity to lower blood glucose levels and affect two other metabolites, glutamine and fatty acids. At the end of the project, we hope to isolate, purify and identify the active principle(s) in the two most active plant species. An immediate impact of this study is the definitive identification of plants that can be readily availed by Filipinos in preventative or maintenance care of diabetes. On the long term, identification of the chemical structures of the active components of these plants will provide lead compounds for further drug development. PHS 398/2590 (Rev. 11/07) Page Continuation Format Page

描述(申请人提供)：菲律宾是地球上生物多样性的热点之一。在6000种特有植物中，据信传统医生和草药医生使用约1500种作为药用植物，其中只有1%的植物具有活性成分。尽管信息如此匮乏，但在菲律宾等发展中国家，从发展中国家进口的药品超出了大多数人的购买力，草药仍然是许多疾病的主要治疗来源。因此，本研究的长期目标是基于代谢产物水平的生物测定确定抗糖尿病的植物，并提供一种基于实验动物的生物测定引导反应来量化草药疗效的新方法。前者将对菲律宾获得有效和负担得起的糖尿病药物产生立竿见影的影响。后者将提供一种基于事实的草药质量控制手段，目前即使在美国也不总是可以获得这种手段。为了达到这些目标，我们建议筛选10+种已被报道具有抗糖尿病特性的本土植物。筛选工作将在菲律宾迪里曼大学进行。粗提物、组分和分离物的筛选将使用一套葡萄糖、谷氨酰胺和脂肪酸的生物传感器进行生物检测。这些生物传感器目前正在Tolosa实验室的R01DK072465项目资助下开发，用于持续监测代谢物的设备。虽然最终的设备还没有准备好，但单个基于蛋白质的传感器可以用于检测血液中的代谢物。因此，我们建议开发测量雄性瑞士韦氏小鼠对高糖负荷反应的代谢物水平的方案。使用这一生物测定方案，我们将从菲律宾已确认的药用植物中筛选并鉴定出两种具有最强降血糖活性的植物，这些植物已被民间报道用于治疗糖尿病。粗提物将被连续分级，直到从两种最有效的菲律宾药用植物中分离出降血糖成分(S)。这些纯化合物的化学结构将由标准的光谱技术确定。这个项目的新奇之处在于，我们不是测量单一的代谢物(葡萄糖)，而是将葡萄糖、谷氨酰胺和脂肪酸复合，以了解这些代谢物在高糖负荷下的比率和趋势，然后是抗糖尿病原则(S)。看看能否从这些趋势中收集模式，根据抗糖尿病化合物的活性进一步将其归类，应该是一件有趣的事情。PHS 398/2590(11/07版)页面续格式页面 与公共卫生相关：菲律宾群岛的地理位置使其成为地球上少数几个拥有丰富的陆地和海洋物种多样性的地方之一，其中许多物种尚未被发现或表征。正因为如此，菲律宾人有着非常悠久的历史，将许多天然产品用于各种用途，包括医疗目的。由于从发达国家进口的药品变得更加昂贵，超出了大多数菲律宾人的购买能力，因此必须根据事实确定现成草药的疗效。在这里，我们建议筛选几种菲律宾本土植物，用于治疗糖尿病及其症状的菲律宾传统药物。这些植物将根据它们降低血糖水平和影响另外两种代谢物谷氨酰胺和脂肪酸的能力进行筛选。在项目结束时，我们希望分离、纯化和鉴定这两个最活跃的植物物种中的活性成分(S)。这项研究的立竿见影的效果是确定了菲律宾人在糖尿病预防或维持护理中可以轻易利用的植物。从长远来看，鉴定这些植物活性成分的化学结构将为进一步的药物开发提供先导化合物。PHS 398/2590(11/07版)页面续格式页面