ISOLATION, STRUCTURE, SYNTHETIC MODIFICATION, AND BIOLOGICAL EVALUATION

RTI-NPP小组35年来一直致力于寻找天然的 具有抗癌活性的产品基于二次 植物、海洋或真菌来源的代谢物可能有价值 抗癌活性。早期的研究导致了喜树碱的分离 和紫杉醇现在广泛的临床使用，直接或作为类似物，在 抗癌化疗。RTI希望继续这一长期努力，因为 财团对NCI国家自然合作项目的回应 产品药物发现小组(NCNPDDG)批准的申请涉及 伊利诺伊大学芝加哥分校(UIC)、百时美施贵宝(BMS)和 三角研究院(RTI)。RTI将对提取物进行测试 在RTI准备的或从财团的其他合作伙伴在 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物质的体会 0.5-1.0公斤植物原料分级提取物的生物测定 采用“内部”分析方法。此外，RTI将分拆 在BMS进行的其他化验中活跃的样品。纯净的结构 化合物将利用所有现代方法获得，包括UV，IR， MS、核磁共振和X-射线衍射仪。当收集到大量活跃的植物时 并分配给RTI，数量不超过10-20公斤的植物样本将 提取并分馏。这样获得的纯样品将被提供 应首席调查员的要求，进行BMS的活体测试 或用于RTI、UIC和BMS的“体外”检测。RTI还建议 通过部分合成和“In”对铅结构进行修饰 为了产生合成孔径雷达数据，对类似物进行“体外”评估。 方式RTI及其财团合作伙伴，UIC和BMS，希望取得重大进展 对癌症化疗的贡献。 RTI将采用以下分析方法来支持发现和 具有潜在抗癌活性的植物产物的提纯：(一) DNA刻痕(检测导致DNA链断裂的化合物 铜的存在；(Ii)抑制拓扑异构酶I(抑制 可裂解络合物分析的催化松弛和稳定)； (3)抑制拓扑异构酶II(抑制催化松弛 和可裂解复合体的稳定性)；(Iv)诱导细胞凋亡 在培养的人类细胞系中(检测和定量高- 在早期由内部产生的分子量片段 存在诱导细胞凋亡的现象。列出的前三种化验结果将 利用氯仿的有机相：水的分配 植物甲醇萃取物。这种处理有效地去除了单宁， 它们干扰了这些分析，从有机分配。化验 四号将利用该隔板的水相。

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.