CORE--IN VITRO TESTING OF AGT INHIBTORS

核心B将测试化合物的抗AGT能力， 培养的肿瘤细胞对BCNU的杀伤敏感。 的三类 将进行的测定是：（a）AGT的新的潜在抑制剂 将使用纯化的重组体检测AGT灭活潜力 人AGT。 设计为前药的化合物也将在 活化酶的存在/（B）潜在AGT的能力 抑制剂在培养的HT 29结肠癌细胞中使AGT失活 （c）AGT抑制剂致敏的能力， 肺癌、结肠癌、乳腺癌和前列腺癌细胞将被BCNU杀死， 测定了 待分析的化合物包括第一阶段产生的化合物 根据第一阶段产生的数据设计的化合物 的支持和化合物设计的基础上，在程序1的研究。 将提供在该核心中确定的有用抑制剂 直接或通过核心C提交给提案中的所有程序。 这些研究将表明化合物是否具有内在的能力， 研究AGT以及它们是否在细胞中活跃，从而提供 关于摄取和代谢转化为更活跃的 或不太活跃的物种。 待测试的化合物将被设计成 提供比BG更有效或更耐代谢的化合物， 在肿瘤中作为产生AGT抑制剂的前药的化合物， 可能抑制BG抗性突变体的化合物和 由于其高效力和 快速的新陈代谢。

Core B will test compounds for the ability to inactivate AGT and to render tumor cells in culture sensitive to killing by BCNU. The three types of assay that will be carried out are: (a) new potential inhibitors of AGT will be tested for AGT inactivating potential using purified recombinant human AGT. Compounds designed as pro-drugs will also be tested in the presence of activating enzymes/ (b) the ability of the potential AGT inhibitors to inactive the AGT in HT29 colon carcinoma cells in culture will be determined; and (c) The ability of the AGT inhibitors to sensitize lung, colon, breast and prostate tumor cells to killing by BCNU will be measured. The compounds to be assayed include those produced during the first period of support, compounds designed based on data generated in the first period of support and compounds designed based on the studies in Program 1. Useful inhibitors which are identified in this core will be made available to all programs in the proposal either directly or via core C. These studies will indicate whether compounds have inherent ability to inactivate the AGT and whether they are active in cells thus providing preliminary information on uptake and metabolic conversion to more active or less active species. The compounds to be tested will be designed to provide compounds that are more potent or resistant to metabolism than BG, compounds that act as prodrugs generating AGT inhibitors in tumors, compounds likely to inactivate the BG-resistant mutants and compounds that would be suitable for regional therapy by virtue of a high potency and a rapid rate of metabolism.