Department of Chemistry & Biochemistry, Ohio University, Athens, OH, 45701, USA.
Department of Chemistry & Biochemistry, Ohio University, Athens, OH, 45701, USA; Honors Tutorial College, Ohio University, Athens, OH, 45701, USA.
Biochem Biophys Res Commun. 2023 Feb 12;644:55-61. doi: 10.1016/j.bbrc.2022.12.080. Epub 2022 Dec 29.
RNA structure plays an important role in regulating cellular function and there is a significant emerging interest in targeting RNA for drug discovery. Here we report the identification of 4-aminoquinolines as modulators of RNA structure and function. Aminoquinolines have a broad range of pharmacological activities, but their specific mechanism of action is often not fully understood. Using electrophoretic mobility shift assays and enzymatic probing we identified 4-aminoquinolines that bind the stem-loop II motif (s2m) of SARS-CoV-2 RNA site-specifically and induce dimerization. Using fluorescence-based RNA binding and T-box riboswitch functional assays we identified that hydroxychloroquine binds the T-box riboswitch antiterminator RNA element and inhibits riboswitch function. Based on its structure and riboswitch dose-response activity we identified that the antagonist activity of hydroxychloroquine is consistent with it being a conformationally restricted analog of the polyamine spermidine. Given the known role that polyamines play in RNA function, the identification of an RNA binding ligand with the pharmacophore of a conformationally restricted polyamine has significant implications for further elucidation of RNA structure-function relationships and RNA-targeted drug discovery.
RNA 结构在调节细胞功能方面起着重要作用，因此针对 RNA 进行药物研发的兴趣日益浓厚。在这里，我们报告了鉴定 4-氨基喹啉类化合物作为 RNA 结构和功能调节剂的研究。氨基喹啉类化合物具有广泛的药理活性，但它们的具体作用机制通常并不完全清楚。我们使用电泳迁移率变动分析和酶探测实验鉴定了 4-氨基喹啉类化合物，这些化合物能够特异性地结合 SARS-CoV-2 RNA 的茎环 II 模体 (s2m)，并诱导二聚化。通过荧光 RNA 结合和 T 框核糖体开关功能测定，我们鉴定出羟氯喹结合 T 框核糖体开关抗终止 RNA 元件并抑制核糖体开关功能。根据其结构和核糖体开关剂量反应活性，我们鉴定出羟氯喹的拮抗剂活性与其作为多胺亚精胺的构象受限类似物一致。鉴于多胺在 RNA 功能中的已知作用，具有构象受限多胺药效团的 RNA 结合配体的鉴定对进一步阐明 RNA 结构-功能关系和 RNA 靶向药物发现具有重要意义。
