Targeting Nucleic Acids with an Integrated Virtual and Actual Screen

描述(由申请人提供)：DNA仍然是小分子治疗剂的代表不足的靶标。越来越多的证据表明，非B-DNA结构在基因表达和端粒功能中发挥着重要作用。针对这些结构元素是开发新的治疗剂的一种有吸引力的创新策略。在这样的抗基因策略中使用小分子是有前景的，原因有很多。在基因扩增之前对其进行靶向以产生多个mRNA和蛋白质分子是有利的，因为要打击的靶点更少。此外，更容易从化学上操纵小分子(而不是治疗性DNA或RNA分子)，以优化结合和药理特性。我们寻求更新我们的创新和高效计划，以开发一个集成的虚拟和实际筛选平台，用于发现选择性地与具有生物学意义的独特DNA结构结合的新先导化合物。在最初的筹资期间取得了显著进展，我们实现了最初提案中提出的大部分具体目标。我们已经发现了几种化合物，它们选择性地结合到特定的四链结构上，这是目前人们非常感兴趣的目标。我们建议进行研究，继续开发和优化综合筛查平台。在下一个资助期，我们将专注于发现选择性地与具有重要生物学意义的四链和DNA-RNA杂交结构结合的先导化合物。为了做到这一点，我们将使用一种将分子动力学模拟与严格的实验验证相结合的新方法来开发复杂四链结构的结构模型。我们将表征我们在最初的资助期发现的新型四链粘合剂的生物物理和功能特性。具体目标包括：1.虚拟筛选平台的开发和完善。2.高通量配基结合分析方法的发展。3.发现与具有重要功能的核酸结构结合的先导化合物。4.在最初筹资期间发现的新型G-四链结合剂的生物物理和生物学特性。拟议的研究将为发现新的先导化合物提供一个改进的综合平台，并彻底表征几个新发现的具有独特化学支架的四链粘结剂。

DESCRIPTION (provided by applicant): DNA remains an underrepresented target for small molecule therapeutic agents. There is mounting evidence to indicate that non-B DNA structures play prominent roles in gene expression and in the function of telomeres. Targeting these structural elements is an attractive and innovative strategy for the development of new therapeutic agents. The use of small molecules in such an antigene strategy is promising for many reasons. Targeting the gene before its amplification to produce multiple mRNA and protein molecules is advantageous because there are fewer targets to hit. In addition, is easier to manipulate small molecules (rather than therapeutic DNA or RNA molecules) chemically to optimize binding and pharmacological properties. We seek renewal for our innovative and highly productive program to develop an integrated virtual and actual screening platform for the discovery of new lead compounds that bind selectively to unique DNA structures of biological significance. Progress during the initial funding period was outstanding, and we achieved most of the specific aims proposed in our initial proposal. We have discovered several compounds that bind selectively to particular quadruplex structures, a target of intense current interest. We propose studies that will continue to develop and optimize the integrated screening platform. During the next funding period, we will focus on the discovery of lead compounds that bind selectively to biologically important quadruplex and DNA-RNA hybrid structures. In order to do this, we will develop structural models for complex quadruplex structures using a novel approach that integrates molecular dynamics simulations with rigorous experimental validation. We will characterize the biophysical and functional properties of the novel quadruplex binders we have discovered during the initial funding period. Specific aims include: 1. Development and refinement of the virtual screening platform. 2. Development of higher-throughput assays for ligand binding. 3. Discovery of lead compounds that bind to functionally important nucleic acid structures. 4. Biophysical and biological characterization of novel G-quadruplex binding agents discovered during the initial funding period. The proposed studies will deliver an improved integrated platform for the discovery of novel lead compounds and a thorough characterization of several newly-discovered quadruplex binders with unique chemical scaffolds.