CORE--IN VITRO TESTING OF AGT INHIBTORS

核心B将测试化合物灭活AGT和渲染的能力 体外培养的肿瘤细胞对BCNU敏感。三种类型的 将进行的化验如下：(A)新的AGT潜在抑制剂 将使用纯化的重组人进行AGT灭活潜力测试 人类AGT。被设计为亲药物的化合物也将在 激活酶的存在/(B)潜在AGT的能力 体外抑制HT29结肠癌细胞AGT活性的抑制剂 将被确定；以及(C)AGT抑制剂的敏化能力 BCNU对肺、结肠、乳腺和前列腺肿瘤细胞的杀伤作用 量过了。 被检测的化合物包括第一阶段产生的化合物。 支持，根据第一阶段产生的数据设计的化合物 根据方案1中的研究设计的载体和化合物。 在这个核心中确定的有用的抑制剂将被提供 到建议书中的所有计划，无论是直接还是通过核心C。 这些研究将表明化合物是否具有固有的能力 使AGT失活以及它们在细胞中是否活跃，从而提供 摄取和代谢转化为更活跃的初步信息 或不太活跃的物种。被测试的化合物将被设计成 提供比BG更有效或更耐新陈代谢的化合物， 在肿瘤中作为前药产生AGT抑制剂的化合物， 可能使抗BG突变体失活的化合物和 将适合于区域治疗，凭借其高效力和 新陈代谢速度快。

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.