Metal Probes and Drug Binding to DNA

该提案构成了MERIT奖研究计划的竞争性更新， 将金属嵌入剂应用于靶向核酸。已经设计了金属络合物， 通过形状选择或通过辅助的功能化以高特异性在DMA位点反应， 配体以获得与DMA的非共价接触的整体。我们还设计了笨重的铑 嵌入剂，其通过光活化而特异性靶向DMA双链体中的单碱基错配， 链断裂我们现在提出这些金属配合物作为新的诊断剂的应用， 可能作为化疗剂。具有大体积嵌入配体的金属络合物靶向 将合成含有荧光团、肽、切割官能团和金属化的错配 地点，特别是Pt。将对与寡核苷酸结合的复合物进行晶体学和NMR研究。 用于表征金属结合结构。值得注意的是，DMA错配特异性靶向剂将是 应用于探针错配修复（MMR）缺陷细胞系与表达细胞系中的错配频率， 检查错配是否优先定位在不同的基因中。由于MMR缺陷与 这些实验为开发灵敏、直接的诊断方法奠定了基础。 用于组织中的癌性转化的测定。Rh和Ru配合物发光的系统研究 通过流式细胞术和共聚焦显微镜将进行，以确定控制因素 功能化的或作为更复杂的缀合物的三（金属）螯合物的细胞摄取。与 靶向错配的发光金属络合物，MMR缺陷细胞的直接诊断， 提出了鉴于核摄取结果，还将在细胞毒性和细胞毒性中检查金属络合物。 在MMR缺陷型细胞系与成熟型细胞系中的增殖测定。通过结合DMA失配定位 本发明涉及一种具有强效DMAAdamaging剂的药物，其优先损伤MMR缺陷细胞，那些最 易受癌变的影响，我们计划研究一种全新的途径， 抗癌剂。

This proposal constitutes a competitive renewal of a MERIT award research program to design and apply metallointercalators in targeting nucleic acids. Metal complexes have been designed that bind and react at DMA sites with high specificity either through shape-selection or through functionalization of ancillary ligands to obtain an ensemble of non-covalent contacts with DMA. We have also designed bulky rhodium intercalators that specifically target single base mismatches in the DMAduplex through photoactivated strand cleavage. We now propose the application of these metal complexes as new diagnostic agents and potentially as chemotherapeutic agents. Metal complexes with bulky intercalating ligands targeted to mismatches will be synthesized containing fluorophores, peptides, cleaving functionalities and metalation sites, notably Pt. Crystallographic and NMR studies of the complexes bound to oligonucleotides will be utilized to characterize metal-bound structures. Significantly, DMA mismatch-specific targeting agents will be applied to probe mismatch frequencies in mismatch repair (MMR) deficient versus proficent cell lines and to examine if mismatches are preferentially localized in different genes. Since MMR-deficiency is associated with cancerous transformation, these experiments form the basis for developing sensitive, direct diagnostic assays for cancerous transformations in tissues. Systematic studies with luminescent Rh and Ru complexes through both flow cytometry and confocal microscopy will be conducted to determine factors governing cellular uptake of the tris(metal) chelates, either functionalized or as more complex conjugates. With luminescent metal complexes targeted to mismatches, direct diagnostics of MMR-deficient cells are proposed. Given nuclear uptake results, metal complexes will also be examined in cytotoxicity and cell proliferation assays in MMR deficient versus proficient cell lines. By combining a DMA mismatch-targeting agent with a potent DMAdamaging agent to preferentially damage MMR-deficient cells, those most susceptible to cancerous transformation, we plan to examine a completely new route to rationally designed anticancer agents.