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: 8692642
• 负责人: Yu Chen
• 金额: $ 37.3万
• 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/8692642
• 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.

说明(申请人提供)：丝氨酸内酰胺酶的产生是革兰氏阴性细菌对内酰胺类抗生素的主要耐药机制之一，这些抗生素包括广泛使用的青霉素和头孢菌素，以及碳青霉烯类等最后使用的抗生素。内酰胺酶的持续突变凸显了开发新型内酰胺酶抑制剂的迫切需要。我们建议通过靶向保守的结构基序，特别是那些对广谱内酰胺酶活性至关重要的结构基序，来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被确定。具体地说，以CTX-M A类内酰胺酶为模型系统，我们的目标是：1)应用基于片段和结构指导的方法开发新型内酰胺酶抑制剂化学类型；2)用超高分辨率和室温X射线结晶学研究耐药性和配体结合；以及3)研究非共价抑制剂耐药性突变的进化。这些实验将导致新的具有临床潜力的内酰胺酶抑制剂，同时提供对与耐药进化相关的内酰胺酶突变的更深入的了解。