AMINOGLYCOSIDE/RNA INTERACTIONS AND CORNEAL INFECTIONS

氨基糖苷/RNA 相互作用和角膜感染

• 批准号: 6329574
• 负责人: ROBERT R RANDO
• 金额: $ 27.23万
• 依托单位: HARVARD MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-12-01 至 2003-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6329574
• 关键词: RNA; RNA binding protein; aminoglycoside antibiotics; binding sites; chemical binding; cornea disorder; drug adverse effect; drug design /synthesis /production; drug interactions; eye disorder chemotherapy; eye infections; gentamicins; molecular shape; neomycin; nuclear magnetic resonance spectroscopy; nucleic acid inhibitor; nucleic acid structure

RNA molecules are able to form precise three-dimensional structures which can be binding-sites for small organic molecules. An understanding of the rules that govern RNA/small molecule recognition processes rules would allow for an approach to our long-term goal of the de novo design of antagonists directed at particular RNA structures, in much the same way that inhibitors are designed for protein-based enzymes and receptors. Specific inhibitors designed to inhibit RNA molecules could be of enormous interest in ophthalmology and generally in medicine, in the design, for example, of small-molecules that can specifically interfere with the expression of mutant proteins that can lead to retinal degeneration, and in the design of small molecules that can antagonize RNA molecules from infectious disease producing organisms. This proposal describes approaches to gaining an understanding of the rules by which certain classes of naturally occurring RNA antagonists, the aminoglycosides, are recognized by specific RNA molecules. Random RNA molecules are selected by column methods to bind to defined aminoglycoside containing antibiotics with high affinity and specificity. New quantitative fluorescence methods are developed to determine the affinities and stoichiometries of antibiotic binding to the selected RNA aptamers. Novel chemical approaches are developed to reveal the regions of the aptamers which define the binding-sites for the specific antibiotic. High field NMR structural studies on the high affinity binding aptamers are planned. Those motifs in the specific RNA aptamers which recognize particular aminoglycosides will be determined and used as a guide for the future design of specific antagonists directed against naturally occurring RNA molecules. Two biologically occurring RNA molecules of particular interest in this context are the procaryotic 16S rRNA decoding region and the HIV-RRE transcriptional activator region. Quantitative structure-activity studies on aminoglycoside binding to the decoding and RRE regions leads to the design of novel aminoglycosidemimetic diversity libraries containing l,3(2)-hydroxylamine moieties. Specific antagonists directed against the decoding and RRE regions of RNA will be prepared and studies quantitatively. These antagonists are expected to be starting points for the design of drugs useful in the treatment of corneal infections. Overall, the studies described herein will serve as the basis for a general program on the design of specific RNA antagonists.

RNA分子能够形成精确的三维结构，这些结构可以是小有机分子的结合位点。 对控制RNA/小分子识别过程规则规则的规则的理解将允许我们对针对特定RNA结构的拮抗剂从头设计的长期目标采取一种方法，其方式与抑制剂的设计方式几乎相同。 Specific inhibitors designed to inhibit RNA molecules could be of enormous interest in ophthalmology and generally in medicine, in the design, for example, of small-molecules that can specifically interfere with the expression of mutant proteins that can lead to retinal degeneration, and in the design of small molecules that can antagonize RNA molecules from infectious disease producing organisms.该建议描述了了解某些天然发生的RNA拮抗剂（氨基糖苷）被特定的RNA分子识别的方法的方法。 通过列方法选择随机RNA分子，以与含有高亲和力和特异性的含有抗生素的定义的氨基糖苷结合。 开发了新的定量荧光方法，以确定抗生素结合与所选RNA适体的亲和力和石化。 开发了新型的化学方法来揭示适体的区域，这些区域定义了特定抗生素的结合位点。 计划对高亲和力结合适体的高场NMR结构研究。 特定RNA适体中识别特定氨基糖苷的特定基序将被确定并用作针对天然存在的RNA分子的特定拮抗剂的未来设计。在这种情况下，有两个具有生物学的RNA分子是procaryotic 16S rRNA解码区域和HIV-RRE转录激活剂区域。 对氨基糖苷与解码和RRE区域结合的定量结构活性研究导致设计新型的含有L，3（2）羟胺基团的新型氨基糖酶异常多样性库。 针对RNA的解码区域和RRE区域的特定拮抗剂将准备并进行定量研究。预计这些拮抗剂将是设计用于治疗角膜感染的药物的起点。 总体而言，本文描述的研究将作为特定RNA拮抗剂设计一般计划的基础。