HUMAN TOPO I--AN EXPLOITABLE ANTICANCER DRUG TARGET

这项建议的目的是加强临床开发 人DNA拓扑异构酶-I的羟乙胺和其他抑制剂 阐述了该制剂的细胞生物化学方面。这 研究将与NCI赞助的第一阶段一起进行 Hycamptamin的临床和药理试验将在 M.D.安德森癌症中心。 这项提议的具体目标包括DNA的量化 人肿瘤细胞中拓扑异构酶-I的研究 容易接近的肿瘤带瘤部位。这些分析将使用 单特异性抗人拓扑异构酶-I抗体及免疫印迹分析。 其次，拓扑异构酶-I对羟基甲胺的敏感性 这些活组织检查将在体外暴露于药物后进行定量。 使用条带耗尽免疫印迹。第三，乙基苯丙胺 拓扑异构酶-I对患者体内肿瘤易感性的研究 将通过重复免疫印迹程序使用 喜树碱后立即进行第二次肿瘤活组织检查 行政管理。最后，我们将比较体外和体内的 拓扑异构酶-I药物敏感性的血浆测定法 药理和临床反应。基于一般生物化学 要采用的原则，在这次试验中制定的方法将是 适用于随后的乙醇胺第二阶段试验和 人DNA拓扑异构酶I和II的其他抑制剂的研究进展。

The aim of this proposal is to augment the clinical development of hycamptamine and other inhibitors of human DNA topoisomerase-I by addressing aspects of the cellular biochemistry of this agent. This study will be performed in conjunction with the NCI-sponsored phase I clinical and pharmacologic trial of hycamptamine to be conducted at the M.D. Anderson Cancer Center. The specific aims of this proposal include the quantification of DNA topoisomerase-I in human tumor cells obtained from needle aspirates of accessible tumor-bearing sites. These assays will be performed using a monospecific antihuman topoisomerase-I antibody and immunoblot analysis. Secondly, the hycamptamine susceptibility of the topoisomerase-I from these biopsies will be quantitated following exposure to drug in vitro using band-depletion immunoblotting. Thirdly, the hycamptamine susceptibility of the topoisomerase-I in the patient's tumor in vivo will be quantitated by repeating the immunoblotting procedure using a second tumor biopsy obtained immediately after hycamptamine administration. Finally, we will compare the in vitro and in vivo estimates of topoisomerase-I drug susceptibility with the plasma pharmacology and clinical response. Based on the general biochemical principles to be employed, the approach developed in this trial will be applicable to subsequent Phase II trials of hycamptamine and to the development of other inhibitors of human DNA topoisomerase I and II.