HUMAN TOPO I--AN EXPLOITABLE ANTICANCER DRUG TARGET

该提案的目的是加强临床开发， 盐酸喜树碱和其它人DNA拓扑异构酶-I抑制剂， 涉及该试剂的细胞生物化学方面。 这 研究将与NCI申办的I期研究联合进行 将于2005年12月在纽约进行的海坎普他明临床和药理试验 M.D.安德森癌症中心。 该提案的具体目标包括DNA的定量 拓扑异构酶-I在人肿瘤细胞中的表达 可触及的肿瘤部位。 这些试验将使用 单特异性抗人拓扑异构酶-I抗体和免疫印迹分析。 第二，拓扑异构酶-I的盐酸喜树碱敏感性， 这些活组织检查将在体外暴露于药物后定量 使用条带耗尽免疫印迹法。 第三，海坎普他明 患者体内肿瘤中拓扑异构酶-I的易感性 将通过重复免疫印迹程序进行定量， 在盐酸喜树碱给药后立即进行第二次肿瘤活检 局 最后，我们将比较体外和体内 血浆拓扑异构酶-I药物敏感性的估计 药理学和临床反应。 基于一般的生化 原则，在这项试验中开发的方法将是 适用于随后的海坎他明II期试验和 开发人DNA拓扑异构酶I和II的其他抑制剂。

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.