ISOLATION, STRUCTURE, SYNTHETIC MODIFICATION, AND BIOLOGICAL EVALUATION

分离、结构、合成修饰和生物学评价

• 批准号: 6396805
• 负责人: MANSUKH C WANI
• 金额: --
• 依托单位: UNIVERSITY OF ILLINOIS AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-09-01 至 2000-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6396805
• 关键词: DNA damage; DNA gyrase; DNA topoisomerases; X ray spectrometry; antineoplastics; apoptosis; bioassay; biological products; chemical structure function; chemical synthesis; cooperative study; drug design /synthesis /production; drug screening /evaluation; enzyme inhibitors; infrared spectrometry; mass spectrometry; medicinal plants; nuclear magnetic resonance spectroscopy; plant extracts; plants; solvent extraction; tissue /cell culture; ultraviolet spectrometry

The RTI-NPP group has been involved for 35 years in a search for natural products with anticancer activity based on the philosophy that secondary metabolites of plant, marine, or fungal origin may have valuable anticancer activity. Early studies resulted in isolation of camptothecin and taxol now in extensive clinical use, directly or as analogs, in anticancer chemotherapy. RTI wishes to continue this long-term effort as part of a consortium response to an NCI National Cooperative Natural Product Drug Discovery Groups (NCNPDDG) Grant Application involving the University of Illinois at Chicago (UIC), Bristol-Myers Squibb (BMS), and the Research Triangle Institute (RTI). RTI will test the extracts prepared at RTI or received from other partners of the consortium in the bioassay systems described below. RTI will utilize its extensive experience on isolation of active compounds from plant samples guided by bioassay to fractionate extracts of 0.5-1.0 kg of plant material employing the "in house" assays. In addition, RTI will fractionate samples active in other assays carried out at BMS. The structure of pure compounds will be obtained utilizing all modern methods including UV, IR, MS, NMR, and X-Ray. when larger quantities of active plants are collected and assigned to RTI, quantities of plant sample up to 10-20 kg will be extracted and fractionated. Pure samples thus obtained will be supplied as requested by the Principal Investigator for "in vivo" testing by BMS or for "in vitro" testing by RTI, UIC, and BMS. RTI also proposes to carry out modifications of lead structures by partial synthesis and "in vitro" evaluation of analogs in order to generate SAR data. In this manner RTI and its consortium partners, UIC and BMS, hope to make a major contribution to cancer chemotherapy. The following assays will be employed at RTI to support discovery and purification of plant products with potential anticancer activity: (i) DNA nicking (Detection of compounds that cause DNA strand scissions in the presence of Cu++; (ii) Inhibition of topoisomerase I (inhibition of catalytic relaxation and stabilization of cleavable complex assays); (iii) Inhibition of topoisomerase II (inhibition of catalytic relaxation and stabilization of cleavable complex assays); (iv) Apoptosis induction in cultured human cell lines (detection and quantitation of high- molecular-weight fragments that are internally generated early in the presence of apoptosis induction. The first three assays listed will utilize the organic phase of a chloroform:water partition of the methanolic plant extract. This treatment effectively removes tannins, which interfere with these assays, from the organic partition. Assay number four will utilize the aqueous phase of that partition.

RTI-NPP组已参与了35年的搜索自然 具有抗癌活性的产品基于次要的理念 植物，海洋或真菌起源的代谢物可能具有宝贵 抗癌活性。早期研究导致甲状腺素分离 现在直接或作为类似物的紫杉醇在广泛的临床使用中 抗癌化疗。 RTI希望继续这项长期努力 财团对NCI国家合作天然的响应的一部分 产品药物发现小组（NCNPDG）授予涉及 伊利诺伊大学芝加哥大学（UIC），布里斯托尔美犬（BMS）和 研究三角研究所（RTI）。 RTI将测试提取物 在RTI准备或从财团的其他合作伙伴中收到 下面描述的生物测定系统。 RTI将利用其广泛的 从植物样品中隔离活性化合物的经验 生物测定到0.5-1.0 kg植物材料的分级提取物 使用“内部”测定法。此外，RTI将分数 在BMS进行的其他测定中活跃的样品。纯的结构 使用所有现代方法，包括紫外线，IR， MS，NMR和X射线。当收集大量活性植物时 并分配给RTI，最多10-20公斤的植物样品将是 提取和分级。因此，将提供纯样品 按照BMS的“体内”测试的首席研究员的要求 或用于RTI，UIC和BMS的“体外”测试。 RTI也建议 通过部分合成对铅结构进行修改，并在 体外“对类似物的评估以生成SAR数据。 RTI及其财团合作伙伴UIC和BMS希望成为专业 对癌症化疗的贡献。 以下测定将在RTI上用于支持发现和 具有潜在抗癌活性的植物产品的纯化：（i） DNA入口（检测引起DNA链剪s的化合物 Cu ++的存在； （ii）抑制拓扑异构酶I（抑制 催化松弛和​​可切合复杂测定法的稳定）； （iii）抑制拓扑异构酶II（抑制催化弛豫 并稳定可裂解的复杂测定法）； （iv）凋亡诱导 在培养的人类细胞系中（检测和定量高 分子重量片段是在内部生成的早期生成的 凋亡诱导的存在。列出的前三个测定法 利用氯仿的有机相： 甲醇植物提取物。这种治疗有效地去除了单宁， 从有机分区中干扰这些测定法。测定 排名第四将利用该分区的水相。