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目的筛选新型抗结核药物BBD并探讨其作用机制。方法建立基于分枝杆菌感染宿主细胞模型的高通量抗结核药物筛选体系进行药物筛选,通过高效液相HLPC以及SD大鼠的药物代谢动力学实验等研究筛选药物BBD的作用机制。结果筛选出一种新型抗结核药物BBD,该药能抑制细胞内、细胞外分枝杆菌的生长,甚至能抑制耐多药临床分离株结核菌的生长。BBD能够穿越A549细胞和THP-1细胞的细胞膜从而抑制胞内寄生菌的生长。Labelfree定量蛋白质组学分析BBD处理后牛型结核分支杆菌BCG的蛋白质组表达谱发生显著变化,变化主要集中在膜相关系统和核糖体,尤其对核糖体的功能和氧化磷酸化等代谢通路有明显的抑制作用。SD大鼠药物代谢动力学实验表明,用药期间血浆药物浓度最高为BBD体外MIC的3.5倍,半衰期约为4h。结论 BBD具有良好的穿膜效果,能快速被机体吸收。该药还能抑制胞内结核菌的生长,有望成为新型抗结核药物。
Objective To screen a new anti-tuberculosis drug BBD and explore its mechanism. Methods A high-throughput anti-TB drug screening system based on mycobacterial infection host cell model was established for drug screening. The mechanism of drug screening for BBD was studied by HLPC and pharmacokinetic experiments in SD rats. Results A novel anti-tuberculosis drug, BBD, was screened for its ability to inhibit the growth of intracellular and extracellular mycobacteria and even inhibit the growth of multidrug-resistant clinical isolates of M. tuberculosis. BBD can cross the cell membrane of A549 cells and THP-1 cells to inhibit the growth of intracellular parasites. Labelfree Quantitative Proteomics Analysis The proteome expression profile of the bovine Mycobacterium bovis BCG changed significantly after BBD treatment. The changes mainly focused on membrane-associated systems and ribosomes, especially for metabolic pathways such as ribosome function and oxidative phosphorylation Inhibition. Pharmacokinetic experiments in SD rats showed that the highest plasma drug concentration during treatment was 3.5-fold that of BBD in vitro with a half-life of about 4 hours. Conclusion BBD has a good transmembrane effect and can be quickly absorbed by the body. The drug can inhibit the growth of intracellular tuberculosis, is expected to become a new anti-TB drugs.