生物素合成途径介导的吡嗪酰胺抗结核治疗的靶标确证和机理研究

吡嗪酰胺（PZA）作为结核病的治疗药物已经被使用长达60余年，但是其分子作用机理并不完全清楚，尚存在一些争议。利用新近开发的药物靶标发现技术——蛋白质质谱串联的细胞热转变分析（MS-CETSA），项目申请人发现结核分枝杆菌（Mycobacterium tuberculosis）中的生物素（Biotin）合成途径中的一个关键酶BioX很可能是POA（PZA的代谢活化产物）的一个新的靶标分子。前期在细胞水平上的功能性实验也初步支持了这个假设。本项目拟从细胞、蛋白分子和动物实验等多个水平，对吡嗪酰胺通过生物素合成途径介导的抗结核作用展开详细的探索和研究。本项目旨在确证生物素合成途径与吡嗪酰胺治疗肺结核的相关性，明确吡嗪酰胺的直接作用靶标蛋白，进一步阐明其分子作用机制，从而为结核病治疗和新药开发提供新角度和新思路。

Pyrazinamide as one of the most important drugs to treat tuberculosis (TB) has been used for more than 60 years, however its mechanisms of action (MOAs) remain elusive and controversial. Utilizing the recently developed MS-CETSA methodology for drug-target deconvolution, we have discovered that one key enzyme in the biotin synthetic pathway called BioX could be a novel protein target for POA, which is the bioactive form of PZA. Some preliminary data performed on the liquid cell culture of mycobacteria has supported this hypothesis. In this proposal, we plan to further explore and investigate the idea that anti-mycobacterial effect of POA is through the biotin synthetic pathway, from the several different levels of cell culture, protein molecule and animal experiment. By conducting this research, we aim to confirm that the biotin synthetic pathway is indeed related to the anti-TB effect of POA, understand the direct protein target of POA, and further our understanding of its MOAs, thereby provide new perspectives for the treatment of TB and the development of new anti-TB drugs.