金属β-内酰胺酶(MβLs)包括NDM-1引起的细菌耐药导致临床医学危机，以MβLs及其耐药菌为靶标设计合成广谱抑制剂和光敏化合物是目前人们密切关注的热点。本项目拟表达纯化每个组别的MβLs，根据其催化底物水解形成相同过渡态的机理特征，设计合成和表征一系列具有识别MβLs及其耐药菌的过渡态类似物型β-内膦酰胺，构建一系列对MβLs耐药菌具有光动力学活性的荧光β-内酰胺，动力学评价其对MβLs的抑制能力和抑制类型，MIC测试其与抗生素联合抑制MβLs耐药菌（如铜绿假单胞菌）的协同效应，微量热法测定其抑制MβLs的热力学参数，光动力学法测定其对MβLs耐药菌的灭活率等。用单晶X-衍射等现代光谱技术,在分子层次上系统地研究抑制剂与MβLs的作用及其构效关系。通过本项目研究，创造和发现对MβLs及其耐药菌有广谱抑制作用的β-内磷酰胺和荧光β-内酰胺类化合物,为发展新型抗菌药物提供重要信息。

The antibiotic resistance in bacteria induced by metallo-β-lactamases (MβLs) including the NDM-1 has resulted in the clinical crisis, people pay close attention to the design and synthesis of broad-spectrum inhibitors and photosensitive compounds with MβLs and the drug-resistant bacteria as targets. The project intends to do expression and purification of the MβLs from each subclass of them, based on the same transition state mechanism of MβLs catalyzing substrate hydrolysis, design, synthesis and characterization of a series of β-phospholactams as the transition state analogs which can identify MβLs and antibiotic resistant bacteria, construction of a series of the fluorescence β-lactams with the photodynamic inactivation of the antibiotic resistant bacteria with MβLs, evaluation of the inhibition activity and inhibition type of the β-phospholactams with steady-state kinetics, testing their synergies in combination with antibiotics to inhibit MβLs and the antibiotic resistant bacteria (eg Pseudomonas aeruginosa) with MIC method, determination of the thermodynamic parameters of them inhibiting MβLs with the microcalorimetric titration, and the photodynamic method for determination of the inactivation rate of the fluorescence β-lactams against the antibiotic resistant bacteria. With modern spectroscopic techniques including single crystal x-ray spectroscopy, at the molecular level, systematic studies on the interaction of the inhibitors and MβLs and the structure-activity relationship. Through this project, creating and finding the β-phospholactam and fluorescence β-lactam compounds as broad-spectrum inhibitors of MβLs and drug-resistant bacteria for the development of new antimicrobial agents to provide information of significant value.