Heterocycle Binding and Biology in the DNA Minor Groove

DNA 小沟中的杂环结合和生物学

• 批准号: 7008830
• 负责人: W David Wilson
• 金额: $ 31.97万
• 依托单位: GEORGIA STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-02-01 至 2010-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7008830
• 关键词: DNA; DNA topoisomerases; antiparasitic agents; cations; centromere; chemical binding; chemical structure function; combinatorial chemistry; divalent cations; drug screening /evaluation; enzyme activity; heterocyclic compounds; microorganism reproduction; nucleic acid sequence; nucleic acid structure; pharmacokinetics

DESCRIPTION (provided by applicant): Heterocyclic cations, developed by our collaborative research groups, bind specifically to extended AT sequences in the DNA minor groove. These compounds have shown clinical biological activity with low human toxicity against several parasitic microorganisms, and an orally available prodrug has progressed through Phase II clinical trials against several parasitic organisms. The compound is scheduled to begin Phase III trials against human trypanosomiasis in October 2004. Biophysical and in vivo biological studies clearly suggest that the target of action of the compounds is DNA and, in particular, the kinetoplast DNA (kDNA) of kinetoplastid microorganisms, such as trypanosomes and leishmania. The heterocyclic cations have shown in vivo ability to block transcription enhancer binding to AT rich sequences and to redistribute the topoisomerase II enzyme, which is critical for replication of the unique kDNA system into AT rich regions. Our hypothesis is that both of these effects require synergistic action of closely bound drug molecules in the AT rich kDNA. The research in this proposal is designed to develop new DNA-targeted antimicrobial drugs by providing a better understanding of the molecular basis of the biological action of the dications in parasites. We will use both parallel and combinatorial chemistry approaches to develop systematic and rational sets of new compounds to address specific questions about the heterocycle-DNA complexes. A battery of traditional, as well as novel biophysical methods, will be used to evaluate the interactions of the compounds with different DNA sequences, such as those that they target in the microorganisms. We are specifically focused on the effects on DNA structure and protein inhibition of cooperative binding of the compounds at closely spaced AT rich binding sites. Both long and short range effects of the compounds on DNA structure and properties will be evaluated. Detailed structural studies will be conducted with selected compounds that we feel can provide fundamental insight into the molecular recognition mechanisms. We will test the ability of the compounds to inhibit transcription enhancers that selectively target AT rich sequences by using model peptide systems, as well as in vivo analysis of protein inhibition. All new compounds will be tested against several microorganisms, and the results will be correlated with their ability to bind DNA and inhibit protein-DNA interactions and functions.

描述（由申请人提供）：由我们的合作研究小组开发的杂环阳离子，与 DNA 小沟中的延伸 AT 序列特异性结合。这些化合物已显示出临床生物活性，对多种寄生微生物具有低人体毒性，并且口服前药已通过针对多种寄生生物的 II 期临床试验。该化合物计划于 2004 年 10 月开始针对人类锥虫病的 III 期试验。生物物理和体内生物学研究清楚地表明，该化合物的作用目标是 DNA，特别是动质体微生物（如锥虫和利什曼原虫）的动质体 DNA (kDNA)。杂环阳离子在体内表现出阻断转录增强子与富含 AT 的序列结合并重新分配拓扑异构酶 II 的能力，这对于将独特的 kDNA 系统复制到富含 AT 的区域至关重要。我们的假设是，这两种效应都需要富含 AT 的 kDNA 中紧密结合的药物分子的协同作用。该提案中的研究旨在通过更好地了解寄生虫中的生物作用的分子基础来开发新的 DNA 靶向抗菌药物。我们将使用平行和组合化学方法来开发系统且合理的新化合物集，以解决有关杂环-DNA 复合物的具体问题。一系列传统和新颖的生物物理方法将用于评估化合物与不同 DNA 序列（例如它们在微生物中靶向的 DNA 序列）的相互作用。我们特别关注化合物在紧密排列的富含 AT 的结合位点上协同结合对 DNA 结构和蛋白质抑制的影响。将评估化合物对 DNA 结构和特性的长程和短程影响。我们将对选定的化合物进行详细的结构研究，我们认为这些化合物可以为分子识别机制提供基本的见解。我们将通过使用模型肽系统以及体内蛋白质抑制分析来测试这些化合物抑制选择性靶向富含 AT 序列的转录增强子的能力。所有新化合物都将针对多种微生物进行测试，结果将与它们结合 DNA 和抑制蛋白质-DNA 相互作用和功能的能力相关。