TARGET PROMOTED ALKYLATION OF NUCLEIC ACIDS

目标促进核酸烷基化

• 批准号: 6164315
• 负责人: STEVEN E ROKITA
• 金额: $ 23.96万
• 依托单位: UNIV OF MARYLAND, COLLEGE PARK
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-05-01 至 2003-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6164315
• 关键词: alkylation; binding sites; cell line; chemical binding; chemical group; chemical structure function; conformation; crosslink; nucleic acid structure; nucleotide analog; phenols

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.

对 DNA 烷基化的研究兴趣继续维持 有关核酸结构、毒理学和 药理。烷化剂在化疗中的重要性不可忽视 尽管与此类相关的严重并发症，但仍被低估 治疗。 DNA 的选择性修饰通常归因于 改性剂的固有化学和结合特性 DNA 通常充当被动受体和反应目标。这 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，但受到的关注仍然有限 RNA催化相关领域。该提案旨在揭示 新的也许是通用的试剂激活机制应该 拓宽我们对潜在诱变、化疗和 诊断反应物。 先前的研究表明，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。这些衍生品 最初是为氟化物依赖性反应而构建的，但后来被 发现与目标核苷酸结合后自发反应 序列。这一过程的机制仍有待探索。这 拟议的调查将从定位复式区域开始 负责激活过程的 DNA。该机制将 然后部分由其官能团和构象定义 要求。这些将通过检查效果来确定 核苷酸类似物并入激活区域。 最后，基于甲硅烷基的化学将扩展到新的试剂 设计和应用以增强其未来在体外和体内的实用性 体内。因此，目标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，作为帮助 定义生物结构和反应性。我们领导的效率 化合物还将用多种细胞系进行测试并进行比较 临床上使用的烷化剂。