RNA: New Antimicrobial Strategies

RNA：新的抗菌策略

• 批准号: 10253871
• 负责人: Adrian Ferre-D'Amare
• 金额: $ 91.85万
• 依托单位: NATIONAL HEART, LUNG, AND BLOOD INSTITUTE
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10253871
• 关键词: Adopted; Affinity; Alternative Splicing; Antibiotics; Bacillus; Binding; Biochemical; Catalytic RNA; Clostridium; Complex; Crystallization; Development; Dissociation; Equilibrium; Evolution; Folic Acid; Future; Genetic Transcription; Genus staphylococcus; In Vitro; Kinetics; Laboratories; Lead; Ligands; Listeria; Messenger RNA; Metabolic Pathway; Public Health; Purines; RNA; Regulator Genes; Reporting; Research; Signal Pathway; Specificity; Streptococcus; Stress; Structure; Study Subject; Time; Translations; Work; antimicrobial; base; design; emerging antibiotic resistance; functional group; guided inquiry; in vivo; insight; interest; mRNA Decay; novel; novel therapeutics; nucleobase; pathogenic bacteria; ribose phosphate; screening; small molecule; three dimensional structure; tool

In the past year, we reported on successful structure-guided discovery of small molecules targeting the ZTP riboswitch. This bacterial regulator is an mRNA domain that responds to ZTP, which accumulates during folate stress. Collaborating with the Schneekloth laboratory (NCI) we performed both, unbiased and structure-guided searches for new small molecules that bind the regulatory RNA, and trigger its function, in vitro and in vivo. We discovered that replacing the ribose-phosphate moiety of ZTP with simple heterocycles results in less polar compounds that nonetheless bind and activate the ZTP riboswitch comparably to the cognate ligand. The best of our current compounds binds to the riboswitch with approximately three times higher affinity than ZTP. Co-crystal structures show that these compounds take advantage of stacking of their heterocycle moiety on a conserved purine nucleobase of the riboswitch. Notably, the effective concentration of these compounds is considerably higher than their dissociation constant, supporting other observations indicating that this regulator is under kinetic control. In addition to producing lead compounds for the future development of anti-RNA antibiotics, this works underscores the importance of functional screening, as equilibrium binding studies may not yield the most active small molecules.

去年，我们报道了针对 ZTP 核糖开关的小分子的成功结构引导发现。 这种细菌调节剂是一个对 ZTP 做出反应的 mRNA 结构域，ZTP 在叶酸应激期间积累。 我们与 Schneekloth 实验室 (NCI) 合作，在体外和体内对能够结合调节 RNA 并触发其功能的新小分子进行了无偏见和结构引导的搜索。 我们发现，用简单的杂环取代 ZTP 的核糖磷酸部分会产生极性较小的化合物，但其与同源配体的结合和激活 ZTP 核糖开关的能力相同。 我们目前最好的化合物与核糖开关的结合亲和力比 ZTP 高大约三倍。 共晶结构表明这些化合物利用其杂环部分在核糖开关的保守嘌呤核碱基上的堆叠。 值得注意的是，这些化合物的有效浓度远高于其解离常数，这支持了表明该调节剂处于动力学控制之下的其他观察结果。 除了生产用于未来开发抗 RNA 抗生素的先导化合物外，这项工作还强调了功能筛选的重要性，因为平衡结合研究可能无法产生最活跃的小分子。