ISOLATION, STRUCTURE, SYNTHETIC MODIFICATION, AND BIOLOGICAL EVALUATION

分离、结构、合成修饰和生物学评价

• 批准号: 6237151
• 负责人: MANSUKH C WANI
• 金额: $ 15.76万
• 依托单位: UNIVERSITY OF ILLINOIS AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-09-01 至 1998-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6237151
• 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 小组 35 年来一直致力于寻找天然物质 具有抗癌活性的产品基于以下理念：次要 植物、海洋或真菌来源的代谢物可能具有有价值的 抗癌活性。早期研究导致喜树碱的分离 和紫杉醇现在在临床上广泛使用，直接或作为类似物， 抗癌化疗。 RTI 希望继续这一长期努力 这是财团响应 NCI 国家自然合作社的一部分 产品药物发现小组 (NCNPDDG) 拨款申请涉及 伊利诺伊大学芝加哥分校 (UIC)、百时美施贵宝 (BMS) 和 三角研究所 (RTI)。 RTI 将测试提取物 在 RTI 准备或从联盟的其他合作伙伴处收到 生物测定系统如下所述。 RTI 将利用其广泛的 从植物样品中分离活性化合物的经验 对 0.5-1.0 kg 植物材料的分级提取物进行生物测定 采用“内部”测定。此外，RTI 将分馏 在 BMS 进行的其他测定中活跃的样品。纯的结构 化合物将利用所有现代方法获得，包括紫外线、红外线、 MS、NMR 和 X 射线。当收集大量活性植物时 并分配给 RTI，植物样品数量最多为 10-20 公斤 提取并分馏。将提供由此获得的纯样品 应首席研究员的要求，进行 BMS 的“体内”测试 或通过 RTI、UIC 和 BMS 进行“体外”测试。 RTI 还建议 通过部分合成和“在 体外”评估类似物以生成 SAR 数据。在此 RTI 及其财团合作伙伴 UIC 和 BMS 希望通过这种方式做出重大贡献 对癌症化疗的贡献。 RTI 将采用以下测定来支持发现和 具有潜在抗癌活性的植物产品的纯化：(i) DNA 切口（检测导致 DNA 链断裂的化合物） Cu++ 的存在； (ii) 拓扑异构酶 I 的抑制（抑制 可裂解复合物检测的催化松弛和​​稳定）； (iii) 拓扑异构酶 II 的抑制（催化弛豫的抑制 和可裂解复合物检测的稳定性）； (iv) 细胞凋亡诱导 在培养的人类细胞系中（高- 早期内部产生的分子量片段 存在细胞凋亡诱导。列出的前三个测定将 利用氯仿：水分配的有机相 甲醇植物提取物。这种处理可有效去除单宁， 来自有机分区的干扰这些测定的物质。测定 第四号将利用该分区的水相。