Natural Product HCV Drugs from Rare Plant-Microbe Interactions

来自稀有植物-微生物相互作用的天然产物 HCV 药物

• 批准号: 9230277
• 负责人: Mark T Hamann
• 金额: $ 29.81万
• 依托单位: MEDICAL UNIVERSITY OF SOUTH CAROLINA
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-09-30 至 2017-09-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9230277
• 关键词: Natural; Product; HCV; Drugs; Rare

The key objective of this project involves the exploration of US rare and endangered plants and their unique host-microbiome community. These communities will be studied for their biosynthesis and production of new classes of natural products (NPs) HCV drug leads with unique MOAs. The project involves a well established collaborative relationship with the laboratories of Prof Mark Hamann and at the University of Mississippi and Tony Whitaker with RFS Pharma and Raymond Schinazi at Emory University/Atlanta VA. The collective expertise provides a unique opportunity to explore new sources for treatments for HCV which are resistant to current forms of treatment. The specific aims of this program include: 1. A collection and phylogenetic analysis of 100 endangered plant associated bacteria and fungi strains each year. Phylogenetically diverse and unique US endangered plant species (20) will be evaluated for bacteria and fungi which inhibit HCV. This will provide 500 unique bacteria and fungi over the course of the five year project period. 2. Innovative epigenetic tools will be applied to stimulate production of secondary metabolites from silent genes. These will include regulatory tools available commercially as well as those derived from the host endangered plant itself. Each new strain will be subjected to 10 or more epigenetic tools. Highly sensitive HPLC with UV-TOF-ELS detection and small volume NMR will be utilized to evaluate NP production from genes silent under standard culture conditions. 3. A unique HCV real time PCR assay is featured in this grant application as a primary screen to identify extracts obtained from Aim 2 with HCV selectivity for fractionation in aim 4. 4. Active leads from Aim 3 will be purified using HPLC with MS, UV and ELSD detection. Repeated bioassays using real time PCR will be completed until HCV selective metabolites are generated. Natural product structures will be assigned using integrated NMR, HPLC-MS, UV, IR and X-ray crystallographic analysis.

该项目的主要目标包括探索美国珍稀濒危植物及其独特的宿主微生物群落。将研究这些群落的生物合成和新型天然产物 (NP) 的生产，以及具有独特 MOA 的 HCV 药物先导物。该项目与密西西比大学的 Mark Hamann 教授的实验室、RFS Pharma 的 Tony Whitaker 实验室以及埃默里大学/弗吉尼亚州亚特兰大的 Raymond Schinazi 实验室建立了良好的合作关系。 集体的专业知识提供了一个独特的机会来探索对当前治疗形式有抵抗力的丙型肝炎治疗的新来源。 该计划的具体目标包括： 1. 每年收集100个濒危植物相关细菌和真菌菌株并进行系统发育分析。 将评估系统发育多样化且独特的美国濒危植物物种 (20) 中抑制 HCV 的细菌和真菌。 这将在五年的项目期间提供 500 种独特的细菌和真菌。 2. 将应用创新的表观遗传学工具来刺激沉默基因产生次级代谢产物。 这些将包括商业上可用的监管工具以及来自濒危宿主植物本身的监管工具。 每个新菌株都将接受 10 种或更多的表观遗传工具。 具有 UV-TOF-ELS 检测和小体积 NMR 的高灵敏度 HPLC 将用于评估标准培养条件下沉默基因的 NP 生产。 3. 本拨款申请中采用了独特的 HCV 实时 PCR 测定作为主要筛选，以鉴定从 Aim 2 中获得的提取物，并具有 HCV 选择性，用于目标 4 中的分级分离。 4. 来自 Aim 3 的活性先导化合物将使用 HPLC 和 MS 进行纯化， UV 和 ELSD 检测。 使用实时 PCR 的重复生物测定将完成，直到产生 HCV 选择性代谢物。 将使用集成 NMR、HPLC-MS、UV、IR 和 X 射线晶体分析来确定天然产物结构。