Garner Amanda L, Lorenz Daniel A, Gallagher Erin E
Department of Medicinal Chemistry, College of Pharmacy, University of Michigan, Ann Arbor, MI, United States; Program in Chemical Biology, University of Michigan, Ann Arbor, MI, United States.
Program in Chemical Biology, University of Michigan, Ann Arbor, MI, United States.
Methods Enzymol. 2019;623:85-99. doi: 10.1016/bs.mie.2019.04.020. Epub 2019 May 11.
Despite the great diversity of structure and function and relevance to human health, RNA remains an underexploited area of drug discovery. A major bottleneck toward this goal has been the identification of probes and drug leads that are specific for select RNAs and methods that will facilitate such discovery efforts. Our laboratory has recently developed an innovative approach for assaying RNA-small molecule interactions, catalytic enzyme-linked click chemistry assay or cat-ELCCA, which is a functional assay that takes advantage of the power of catalytic signal amplification combined with the selectivity and bioorthogonality of click chemistry. Importantly, through application of this platform assay technology to the challenging problem of identifying selective inhibitors of pre-microRNA maturation, we identified natural products as a potential source of such compounds. Herein we describe this methodology in addition to the downstream pipeline toward the discovery of natural product ligands for pre-microRNAs. Through cat-ELCCA, our goal is to discover novel ligands to facilitate our investigation of RNA recognition by small molecules.
尽管RNA在结构、功能以及与人类健康的相关性方面具有极大的多样性,但它仍是药物研发中一个未得到充分开发的领域。实现这一目标的一个主要瓶颈是识别对特定RNA具有特异性的探针和药物先导物,以及有助于此类研发工作的方法。我们实验室最近开发了一种用于检测RNA与小分子相互作用的创新方法,即催化酶联点击化学分析法(cat-ELCCA),这是一种功能分析方法,它利用了催化信号放大的能力,结合了点击化学的选择性和生物正交性。重要的是,通过将这种平台检测技术应用于识别前体微小RNA成熟的选择性抑制剂这一具有挑战性的问题,我们确定天然产物是此类化合物的潜在来源。在此,我们除了描述发现前体微小RNA天然产物配体的下游流程外,还描述了这种方法。通过cat-ELCCA,我们的目标是发现新型配体,以促进我们对小分子RNA识别的研究。