DEVELOPMENTAL PHARMACOLOGY OF NEW ANTICANCER AGENTS

该计划涵盖了广泛的药理学、细胞学、 生物化学和分子程序不仅是理解 研究新型抗癌剂的作用机制，也是为了 为临床方案的设计提供重要的意见。 药理学研究将定义小鼠的药代动力学以及 正常人血清结合特性、药物分布及代谢 和肿瘤组织。我们有一个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 确定药物在以下条件下的摄取、外流和细胞内分布 学习。细胞和生化研究主要针对的是 确定该化合物是否按照6个标准中的任何一个标准起作用 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。 细胞研究也将在体外确定剂量-反应关系 作为细胞毒性和进展的年龄反应特征 延迟。我们的癌症体内有一个复杂的流式细胞术核心 中心完全有能力进行这些研究和其他研究，包括周期素水平 有丝分裂蛋白表达的测定和定量。一个 一系列生化分析将检测药物对大分子的影响 合成抑制基因。分子研究针对的是DNA效应 检查绑定以及损伤和修复动力学；后者 用碱洗脱法进行评估。此外，对 将对拓扑异构酶进行评估。无论是标准脉搏还是新脉搏- 现场电泳技术为我们提供了评估 药物的细胞毒作用是通过细胞凋亡机制来实现的。较新的化验方法 针对膜结构上的药物效应，将通过 各种荧光探头。目前可用的抗性品系为 以及专门培育的抗新剂品系将 用于确定药物的作用机制和交叉模式 抵抗。

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.