DEVELOPMENTAL PHARMACOLOGY OF NEW ANTICANCER AGENTS

该计划涵盖多种药理学、细胞、 生化和分子程序的基础不仅仅是理解 不仅可以了解新型抗癌药物的作用机制， 为临床方案的设计提供重要的输入。 药理学研究将定义小鼠的药代动力学以及 正常人的血清结合特征、药物分布和代谢 和肿瘤组织。 我们拥有最先进的仪器实验室 可用于代谢物鉴定。 细胞药代动力学将 定义药物的摄取、流出和细胞内分布 研究。 细胞和生化研究主要针对 确定化合物是否按照 6 个标准中的任何一个起作用 机制：烷基化、DNA 结合、拓扑异构酶抑制、蛋白质 合成抑制、抗代谢作用或抗有丝分裂作用。 细胞研究也将定义体外剂量反应关系 作为细胞毒性和进展的年龄反应特征 延迟。 我们的癌症中心拥有先进的流式细胞术核心 中心完全有能力进行这些和其他研究，包括细胞周期蛋白水平 有丝分裂蛋白表达的测定和定量。 一个 一系列生化检测将检查药物对大分子的影响 合成抑制位点。 分子研究针对 DNA 效应 检查结合以及损伤和修复动力学；后者 通过碱性洗脱进行评估。 另外，直接影响 将评估拓扑异构酶。 标准和较新的脉冲 场电泳技术使我们能够评估 通过细胞凋亡机制介导的药物细胞毒性。 较新的检测方法 针对药物对膜结构的影响将由 多种荧光探针。 目前可用的抗性系为 以及专门开发的对新试剂有抵抗力的品系 用于确定药物作用机制和交叉模式 阻力。

This Program encompasses a wide variety of pharmacologic, cellular, biochemical and molecular procedures basic not only to an understanding of the mechanism of action of the new anticancer agents, but also to provide important input into the design of clinical protocols. Pharmacology studies will define pharmacokinetics in mice as well as serum binding characteristics, drug distribution and metabolism in normal and tumor tissues. We have a state-of-the art instrumentation laboratory available for metabolite identification. Cellular pharmacokinetics will define the uptake, efflux and intracellular distribution of agents under study. The cellular and biochemical studies are primarily directed at determining whether the compound acts by any of the 6 standard mechanisms: alkylation, DNA binding, topoisomerase inhibition, protein synthesis inhibition, antimetabolite action or antimitotic effect. Cellular studies will define dose-response relationships in vitro as well as age-response characteristics for both cytotoxicity and progression delay. We have a sophisticated flow cytometry Core within our Cancer Center fully capable of these and other studies including cyclin level determination and quantitation of expression of mitotic proteins. A series of biochemical assays will examine drug effects on macromolecular synthesis inhibition loci. Molecular studies are directed at DNA effects examining binding as well as damage and repair kinetics; the latter assessed by alkaline elution. In addition, direct effect on topoisomerase enzymes will be assessed. Both standard and newer pulse- field electrophoresis techniques provides us with the ability to assess drug cytotoxicity mediated through an apoptotic mechanism. Newer assays directed at drug effects on membrane structures will be assessed by a variety of fluorescent probes. Presently available resistant lines as well as specifically developed lines resistant to the new agent will be used to determine mechanism of drug action and patterns of cross resistance.