Evolution and inhibition of beta-lactamase activity in antibiotic resistance

抗生素耐药性中β-内酰胺酶活性的进化和抑制

• 批准号: 8579069
• 负责人: Yu Chen
• 金额: $ 36.24万
• 依托单位: UNIVERSITY OF SOUTH FLORIDA
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-07-01 至 2018-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8579069
• 关键词: Active Sites; Affect; Affinity; Anabolism; Anti-Bacterial Agents; Antibiotic Resistance; Antibiotics; Bacteria; Bacterial Infections; Binding; Biochemical; Biological Models; Carbapenems; Catalysis; Cefotaxime; Ceftazidime; Cell Wall; Cephalosporins; Chemicals; Clinical; Collaborations; Cyclophosphamide; Development; Engineering; Enzymes; Escherichia coli; Evolution; Goals; Gram-Negative Bacteria; Health; Hydrolysis; In Vitro; Investments; Lactamase; Lactams; Lead; Ligand Binding; Ligands; Monobactams; Movement; Multi-Drug Resistance; Mutagenesis; Mutation; Penicillin-Binding Proteins; Penicillins; Pharmaceutical Chemistry; Pharmaceutical Preparations; Positioning Attribute; Predisposition; Production; Property; Proteins; Protons; Research Personnel; Resistance; Resolution; Resort; Resources; Sampling; Serine; Site-Directed Mutagenesis; Staging; Structure; Synthesis Chemistry; Temperature; Testing; University Hospitals; X ray diffraction analysis; X-Ray Crystallography; X-Ray Diffraction; analog; bacterial resistance; base; beta-Lactamase; chemotherapeutic agent; clinically relevant; design; drug discovery; enzyme mechanism; inhibitor/antagonist; insight; lead series; mutant; novel; pathogen; pressure; protonation; prototype; public health relevance; research study; resistance mechanism; resistance mutation; suicide substrates

DESCRIPTION (provided by applicant): The production of serine ¿-lactamase is one of the primary resistance mechanisms used by Gram-negative bacterial pathogens against ¿-lactam antibiotics, which include the widely used penicillins and cephalosporins, as well as last resort antibiotics such as the carbapenems. The development of novel ¿-lactamase inhibitors is a pressing need underscored by the continuing mutation of ¿-lactamases. We propose the development of high affinity non-covalent ¿-lactamase inhibitors by targeting conserved structural motifs, particularly those essential for extended spectrum ¿-lactamase activity. Prototypes of these inhibitors have already been identified. Specifically, using the CTX-M Class A ¿-lactamases as a model system, we aim to: 1) apply a fragment-based and structure-guided approach to develop novel ¿-lactamase inhibitor chemotypes; 2) study resistance and ligand binding by ultrahigh-resolution and room-temperature X-ray crystallography; and 3) investigate the evolution of resistance mutations against non-covalent inhibitors. These experiments will lead to new ¿- lactamase inhibitors with clinical potential, while providing a deeper understanding of ¿-lactamase mutations relevant to resistance evolution.

描述（申请人提供）：产生丝氨酸-内酰胺酶是革兰氏阴性细菌病原体对-内酰胺抗生素的主要耐药机制之一，其中包括广泛使用的青霉素和头孢菌素，以及最后的抗生素，如碳青霉烯类。新型<$-内酰胺酶抑制剂的开发是<$-内酰胺酶持续突变所强调的迫切需要。我们建议通过靶向保守的结构基序，特别是那些对超广谱<$-内酰胺酶活性至关重要的结构基序，开发高亲和力的非共价<$-内酰胺酶抑制剂。这些抑制剂的原型已经确定。具体而言，使用CTX-M A类<$-内酰胺酶作为模型系统，我们的目标是：1）应用基于片段和结构指导的方法开发新的<$-内酰胺酶抑制剂化学型; 2）通过超高分辨率和室温X射线晶体学研究耐药性和配体结合; 3）研究非共价抑制剂的耐药性突变的演变。这些实验将导致新的具有临床潜力的<$-内酰胺酶抑制剂，同时提供与耐药性演变相关的<$-内酰胺酶突变的更深入了解。