Determining the mechanism of inhibition of metallo-b-lactamase inhibitors

确定金属-b-内酰胺酶抑制剂的抑制机制

• 批准号: 9812399
• 负责人: MICHAEL W CROWDER
• 金额: $ 43.35万
• 依托单位: MIAMI UNIVERSITY OXFORD
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-08-01 至 2022-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9812399
• 关键词: Active Sites; Address; Anti-Bacterial Agents; Antibiotic Resistance; Antibiotics; Antimicrobial Resistance; Bacterial Antibiotic Resistance; Bacterial Infections; Binding; Binding Sites; Biochemical; Calorimetry; Carbapenems; Clinical; Clinical Trials; Cobalt; Complex; Data; Development; Dialysis procedure; Electron Spin Resonance Spectroscopy; Electrons; Enterobacteriaceae; Enzymatic Biochemistry; Enzyme Inhibitor Drugs; Enzyme Kinetics; Enzymes; Equilibrium; Evolution; Future; Genes; Glean; Goals; Ions; Kinetics; Knowledge; Mass Spectrum Analysis; Measures; Mediating; Metal Ion Binding; Metals; Molecular Conformation; NMR Spectroscopy; Online Systems; Organism; Pathway interactions; Peer Review; Plasmids; Proteins; Publications; Publishing; Recombinants; Reporting; Research; Research Personnel; Resistance; Resort; Spectrum Analysis; Structure; Structure-Activity Relationship; Students; System; Techniques; Therapeutic; Time; Titrations; Toxic effect; Training; United States National Institutes of Health; Work; Zinc; analog; bacterial resistance; base; beta-Lactamase; beta-Lactams; candidate selection; clinical application; clinical candidate; clinical development; clinically relevant; experimental study; functional group; global health; inhibitor/antagonist; interdisciplinary approach; meetings; metalloenzyme; novel; overexpression; pathogenic bacteria; programs; scaffold; spectroscopic survey; stoichiometry; undergraduate student; web site

PROJECT SUMMARY/ABSTRACT Metallo-b-lactamases (MBLs) are bacterial enzymes that render pathogenic bacteria resistant to the largest class of antibiotics. Although a number of MBL inhibitors have been published, clear structure-activity relationships (SARs) are often difficult to discern, and none of these inhibitors has advanced to clinical use. We propose that the disconnect between the discovery of inhibitors and the development of clinically-useful compounds is due, in part, to a lack of understanding the mechanisms whereby these compounds work, and by the predominance of metal-stripping mechanisms, which are inherently non-specific and have associated toxicity issues. To directly address these issues, we propose to discern the mechanism of inhibition of several under-defined MBL inhibitors. Specific aim #1 will characterize inhibition mechanisms of several MBL inhibitor classes. Many reported MBL inhibitors contain functional groups with potential to bind metal ions, but their mechanisms remain undefined. Here, we use a multidisciplinary approach to determine if selected classes of MBL inhibitors work by metal- stripping, by ternary complex formation (inhibitor:metal ion:protein) or by other mechanisms. This information will enable more effective SAR studies and allow better selection of candidates more suitable for clinical applications. Specific aim #2 will develop two novel techniques to probe inhibitor binding to MBLs. One technique is EPR- based and is predicted to yield information about an invariant b-hairpin loop that resides over the MBL active sites. The second technique is native MS-based and is predicted to yield direct information about the mechanism of inhibition using lower concentrations of enzyme/inhibitor. The long-term goal of this research program is to identify the most promising scaffolds for MBL inhibitors, which can be given in combination with existing b-lactam containing antibiotics to treat antibiotic resistant bacterial infections. Through clearly determining the mechanism of action of promising MBL inhibitor scaffolds, we will provide a clearer pathway for these inhibitors to proceed to clinical trials.

项目摘要/摘要 金属-B-乳糖酶（MBL）是细菌酶，使致病性细菌具有抗性最大的类别 抗生素。尽管已经发布了许多MBL抑制剂，但明确的结构活动关系 （SARS）通常很难辨别出，这些抑制剂都没有发展为临床用途。我们提出了这一点 发现抑制剂的发现与临床使用化合物的开发之间的断开是， 在某种程度上，由于缺乏理解这些化合物起作用的机制，并以优势为主 金属剥离机制的固有非特异性并且存在相关的毒性问题。直接 解决这些问题，我们建议辨别抑制几种不明显MBL抑制剂的机制。 具体目标＃1将表征几种MBL抑制剂类别的抑制机制。许多人报告了MBL 抑制剂包含具有结合金属离子的潜力的官能团，但其机制仍然不确定。 在这里，我们使用多学科方法来确定金属的选定类MBL抑制剂是否起作用 剥离，由三元复合物形成（抑制剂：金属离子：蛋白质）或其他机制。此信息 将实现更有效的SAR研究，并允许更好地选择更适合临床的候选者 申请。 特定的目标＃2将开发两种新技术来探测抑制剂与MBL的结合。一种技术是EPR- 基于MBL Active在MBL上 站点。第二种技术是基于天然MS的技术，预计将产生有关机制的直接信息 使用较低浓度的酶/抑制剂抑制作用。 该研究计划的长期目标是确定MBL抑制剂最有希望的脚手架，这 可以与现有的B-内酰胺（含有抗生素治疗抗生素耐药细菌）结合给出 感染。通过明确确定有希望的MBL抑制剂支架的作用机理，我们将 为这些抑制剂提供了更清晰的途径，以进行临床试验。