ISOLATION, STRUCTURE, SYNTHETIC MODIFICATION, AND BIOLOGICAL EVALUATION

RTI-NPP集团35年来一直致力于寻找天然 具有抗癌活性的产品，其理念是 植物、海洋或真菌来源的代谢物可能具有 抗癌活性。早期的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， NCI国家自然资源合作组织的一部分 产品药物发现组（NCNPDDG）资助申请，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将测试浸提液 在RTI准备或从其他合作伙伴的财团在 下面描述的生物测定系统。RTI将利用其广泛的 从植物样品中分离活性化合物的经验， 对0.5-1.0 kg植物材料的分馏提取物进行生物测定 使用“内部”测定法。此外，RTI将断裂 在BMS进行的其他试验中具有活性的样品。纯的结构 将利用所有现代方法包括UV，IR， MS、NMR和X射线。当收集到大量活跃的植物时， 并分配给RTI，植物样品的数量高达10-20公斤， 萃取和分馏。将提供由此获得的纯样品 根据主要研究者的要求，由BMS进行“体内”试验 或用于RTI、UIC和BMS的“体外”测试。RTI还建议， 通过部分合成和“在 类似物的“体外”评价以产生SAR数据。在这 RTI及其联盟合作伙伴UIC和BMS希望以一种主要的方式， 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物的纯化：（i） DNA切口（检测导致DNA链断裂的化合物， Cu++的存在;（ii）抑制拓扑异构酶I（抑制 可裂解复合物测定的催化弛豫和稳定化）; (iii)抑制拓扑异构酶II（抑制催化弛豫 和可裂解复合物测定的稳定化）;（iv）细胞凋亡诱导 在培养的人细胞系中（检测和定量高- 分子量的片段，是内部产生的早期， 存在凋亡诱导。列出的前三项检测将 利用氯仿的有机相：水分配的 甲醇植物提取物。这种处理有效地去除了单宁， 干扰这些分析的有机物。测定 4号将利用该分区的水相。

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.