Rapid and Generic Discovery of Small Molecule Ligands Targeting RNA

描述(由申请人提供)：我们寻求创建一种快速和通用的方法来发现小分子配体，这些小分子配体可以与RNA特异结合，并产生精确和有用的生物效应。这一挑战的困难体现在观察到，在过去30年里，通过人类的聪明才智从头开始开发的与RNA结合的高效小分子配体的数量很少，可能最多只有两到三个分子。这项提议的总体愿景是使配体发现过程既简单又普遍适用于任意的RNA靶标。为了实现这一目标，将利用SHAPE(通过引物延伸分析的选择性2‘-羟基酰化)化学探测方法-发明、机械验证并被项目实验室广泛应用-转化为高通量和高含量的RNA配体发现技术。大型RNA靶标的核苷酸分辨筛选信息将使用创新的、完全自动化的分析算法进行解释，这些算法利用SHAPE探测异常丰富的信息输出，使非专家用户能够快速识别靶标。到资助期结束时，我们预计使用基于片段的文库来靶向RNA的小分子将不会比传统的蛋白质靶向更困难，甚至可能被证明更直接。具体地说，我们希望建立对RNA的广泛药效性的估计，表征用于靶向RNA的选择性和有用的小分子片段的性质，并鉴定多种小分子化合物，作为结构探测工具和开发高效抗病毒疗法的先导。

DESCRIPTION (provided by applicant): We seek to create a rapid and generic approach for discovery of small molecule ligands that bind RNA specifically and elicit precise and useful biological effects. The difficulty of this challenge is exemplified by the observation that the number of highly effective small molecule ligands that bind RNA that were developed de novo by human ingenuity over the past 30 years is miniscule, perhaps two or three molecules at most. The overarching vision of this proposal is to make the process of ligand discovery both straightforward and generically applicable to arbitrary RNA targets. This goal will be pursued by leveraging the SHAPE (selective 2'-hydroxyl acylation analyzed by primer extension) chemical probing approach - invented, mechanistically validated, and broadly applied by the project laboratory - into a high-throughput and high- content ligand discovery technology for RNA. Nucleotide-resolution screening information for large RNA targets will be interpreted using innovative, and fully automated, analysis algorithms that leverage the exceptionally information-rich output of SHAPE probing to make possible rapid target identification by non-expert users. By the end of the funding period, we envision that small molecule targeting of RNA using fragment-based libraries will be no more difficult, and may even prove more straightforward than, conventional targeting of proteins. Specifically, we expect to establish estimates for the broad-based drugability of RNA, to characterize the properties of selective and useful small molecule fragments for targeting RNA, and to identify multiple small molecule compounds useful as structure-probing tools and as leads for developing highly potent antiviral therapeutics.