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特异性结合RNA的小分子配体，并引发精确和有用的生物效应。我们观察到，在过去的30年里，人类聪明才智重新开发的结合RNA的高效小分子配体数量微乎其微，最多可能只有两三个分子。这一观察结果说明了这一挑战的难度。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RNA靶标。这一目标将通过利用项目实验室发明、机械验证和广泛应用的SHAPE（通过引物延伸分析的选择性2'-羟基酰化）化学探测方法，成为RNA的高通量和高含量配体发现技术来实现。大RNA靶标的核苷酸分辨率筛选信息将使用创新的全自动分析算法进行解释，该算法利用SHAPE探测异常丰富的信息输出，使非专业用户能够快速识别靶标。在资助期结束时，我们设想使用基于片段的文库来靶向RNA的小分子将不会更困难，甚至可能比传统的靶向蛋白质更直接。具体来说，我们期望对RNA的广泛药物性进行估计，表征靶向RNA的选择性和有用的小分子片段的特性，并确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治疗的先导。

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. PUBLIC HEALTH RELEVANCE: This unconventional and innovative proposal is focused on inventing new ways to create drugs that work by binding to RNA. Because many serious human illnesses involve RNA components - including the RNA genomes of viruses like HIV and HCV, the specialized RNA recognition elements in cancer-causing genes, and the pathogenic RNA structures that cause currently incurable hereditary tri-nucleotide expansion diseases - the potential impact of this project is exceptionally wide-ranging.