检测β-内酰胺酶的荧光小分子探针的设计、合成及其在结核杆菌检测中的应用

Tuberculosis is one of the most deadly diseases, infecting one-third of the world’s population. Early detection is critical to the prevention and control of tuberculosis. Fast, high sensitivity, high specificity and low-cost method for detecting Mycobacterium tuberculosis is one of the major difficulties of current anti-tuberculosis research. Currently, it is still challenging for the diagnosis of Mycobacterium tuberculosis via fluorescence molecular probes in our country. This research project mainly aims at developing new near-infrared fluorescent molecular probes for specific detection of Blac type β-lactamase secreted by Mycobacterium tuberculosis, which is based on the mechanism of cephalosporin hydrolysis by β-lactamases yielding hydrolysed β-lactam and being concomitant with the loss of a 3’-leaving group. Substituents in the β-lactam ring will be screened to solve the major problem for the selective reaction of fluorescent probe with Blac type β-lactamase among other β-lactamases. Further research on the current serious drug resistance of Mycobacterium tuberculosis with β-lactam antibiotics will be carried out. All of the results will help the research and development of the rapid and accurate diagnosis of Mycobacterium tuberculosis in future.

结核病是最致命的疾病之一，影响着全球三分之一的人口。早期的检测对结核病的预防和控制非常重要。快速、灵敏度高、专一性强、低成本的结核杆菌的检测方法是目前抗结核病研究的难点之一。目前应用荧光分子探针来早期检测结核杆菌的研究在国内亟待开发。本项目课题的研究着眼于发展一种近红外荧光分子探针来特异性检测结核杆菌分泌的Blac型β-内酰胺酶；课题研究基于β-内酰胺环被β-内酰胺酶水解开环并脱去3-位取代基这一原理来设计探针；这一课题的研究将通过修饰β-内酰胺环的取代基来解决荧光探针对Blac型β-内酰胺酶与其他β-内酰胺酶选择性的难题，而且还可以针对目前结核杆菌对β-内酰胺类抗生素存在严重耐药性问题开展进一步深入研究，为快速准确检测结核杆菌的研究与开发奠定一定的基础。

选择性检测Blac型β-内酰胺酶在结核病的预防和治疗具有重要的科学意义。本项目利用荧光探针的优良性质，设计并合成了两个光稳定性好，在水溶液中量子产率高的近红外小分子荧光探针，具有高选择性和挺好的稳定性，并且特异性检测结核分枝杆菌分泌的Blac型β-内酰胺酶，同时深入地研究了荧光分子探针选择性结合Blac型β-内酰胺酶的机理，为荧光分子探针技术应用于结核分枝杆菌检测领域方面提供坚实的基础，具有挺好的实用价值。同时我们设计并合成了一系列选择性检测二氧化硫、硫化氢、钯、苯硫酚、半胱氨酸、高半胱氨酸和谷胱甘肽等方面的荧光探针，并应用到实际样品的检测中。这些研究成果将扩展、推动小分子探针在生命科学和环境领域的应用，促进生命科学的发展。