TARGET PROMOTED ALKYLATION OF NUCLEIC ACIDS

对DNA烷基化的研究兴趣继续保持， 关于核酸结构、毒理学和 药理学烷化剂在化疗中的重要性 被低估，尽管与此相关的严重并发症 治疗DNA的选择性修饰通常归因于 改性剂的固有化学性质和结合性质， DNA通常充当被动受体和反应靶。的 DNA作为催化剂并控制自身修饰的潜力 尽管在这方面取得了重大进展， RNA催化的相关领域。该提案旨在揭示一个 新的、也许是通用的试剂激活机制，应该 拓宽了我们对潜在的诱变、化疗和 诊断反应物 以往的研究表明，双链DNA可以激活甲硅烷基 用于烷基化和交联的苯酚衍生物。这些衍生物 最初被构建用于氟化物依赖性反应，但 发现与靶核苷酸结合后自发反应 序列的这个过程的机制还有待发现。的 拟议的调查将开始定位的地区， 负责激活过程的DNA。余留机制将 然后由其官能团和构象部分定义 要求.这些反过来将通过检查影响来确定 的核苷酸类似物纳入区域的激活。 最后，基于硅烷基的化学将被扩展到新试剂 设计和应用，以提高其在体外和体内的未来效用。 vivo.因此，靶修饰的效率和特异性 将在DNA，RNA和蛋白质复合物中进行检查， 定义生物结构和反应性。我们线索的效率 还将用各种细胞系测试化合物， 涉及临床上使用的烷化剂。

Research interest in DNA alkylation continues to be sustained by numerous issues concerning nucleic acid structure, toxicology, and pharmacology. The importance of alkylating agents in chemotherapy cannot be underestimated despite the serious complications associated with such treatment. Selective modification of DNA is generally ascribed to the intrinsic chemistry and binding properties of the modifying agent since DNA typically acts as a passive receptor and target of reaction. The potential for DNA to act as a catalyst and control its own modification has received limited attention despite substantial advances in the related field of RNA catalysis. This proposal is designed to reveal a new and perhaps general mechanism of reagent activation that should broaden our understanding of potential mutagenic, chemotherapeutic and diagnostic reactants. Previous studies have demonstrated that duplex DNA can activate silyl phenol derivatives for alkylation and cross-linking. These derivatives had originally been constructed for fluoride-dependent reaction but were found to react spontaneously after binding to target nucleotide sequences. The mechanism of this process remains to be discovered. The proposed investigations will begin by localizing the region of duplex DNA that is responsible for the activation process. The mechanism will then be defined in part by its functional group and conformational requirements. These in turn will be identified by examining the effect of nucleotide analogues incorporated into the region of activation. Finally, the silyl-based chemistry will be extended to new reagent design and application to enhance its future utility in vitro and in vivo. Accordingly, the efficiency and specificity of target modification will be examined in DNA, RNA and protein complexes as a method to help define biological structure and reactivity. The efficiency of our lead compounds will also be tested with a variety of cell lines and compared to alkylating agents that are used clinically.