新型抗结核药物BBD的筛选及作用机制初探  

Selection of a novel anti-tuberculosis drug,bis-biguanide dihydrochloride,and its mechanism to inhibit intracellular replication of Mycobacterium tuberculosis

作　　者：金文龙[1,2] 黄慧嫦[1,2] 王菲菲[3] 沈洪波[2] 

机构地区：[1]上海大学生命科学院,上海200444 [2]中国科学院上海巴斯德研究所 [3]复旦大学上海医学院基础医学院病原生物学系

出　　处：《中国病原生物学杂志》2017年第7期595-600,共6页Journal of Pathogen Biology

基　　金：国家自然科学基金项目(No.KRF101397);上海市自然科学基金项目(No.14ZR1444300)

摘　　要：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。Objectives To screen novel anti-mycobacterial agents and to study the mechanism of action of the potent anti-mycobacterial agent bis-biguanide dihydrochloride （BBD）. Methods A host cell-based based high-throughput screening （HTS） assay was created. High-performance liquid chromatography （HPLC） as well as pharmacokinetic exper- iments with SD rats were performed to analyze the characteristics of BBD. Results BBD was identified as a potent an- ti-mycobacterial agent. Further characterization indicated that BBD inhibited the intracellular and extracellular growth of Mycobacteriurn srnegrnatis and slow-growing M. boris BCG. BBD also potently inhibited replication of clinically isolates of M. tuberculosis and multidrug-resistant strains of M. tuberculosis. BBD can pass through the cell membrane to inhibit the intracellular growth of M. tuberculosis. Label-free quantification revealed that BBD mainly inhibits the expression of pro- teins related to the cell membrane and ribosomes of M. tuberculosis, and especially the functioning of ribosomes and meta- bolic pathways such as oxidative phosphorylation. Pharmacokinetic experiments in SD rat indicated that the concentration of BBD in plasma was more than 3.5 times the MIC. This means that the concentration of BBD in vivo is high enough to inhibit the growth of M. tuberculosis. Conclusion The current findings warrant further study to develop BBD as a new and effective drug against tuberculosis and multidrug-resistant tuberculosis.

关 键 词：抗结核药物 BBD 结核分枝杆菌 抑制 

分 类 号：R378.911[医药卫生—病原生物学]