Mycobacterium tuberculosis Raw sequence reads. Mycobacterium tuberculosis

1H-benzo[d]imidazole derivatives exhibit antitubercular activity in vitro as well as in vivo in human monocyte-derived macrophages in a nanomolar range of concentrations and are not toxic for human cells. The mode of action of 5-Bromo-2-(2-cyclohexylethyl)-1H-benzo[d]imidazole (EJMCh4) and 2-(2-cyclohexylethyl)-5,6-dimethyl-1H-benzo[d]imidazole (EJMCh6) compounds was identified by selection of resistant Mycobacterium tuberculosis mutants followed by whole genome sequencing as well as observed inhibition of mycolylation of arabinogalactan and synthesis of TDM indicated MmpL3 as a molecular target for their antitubercular activity.Bacterial strains:1. Mycobacterium tuberculosis H37Rv strain, IBMPAN stock2. 4/1 MmpL3 (V285A) mutant selected on EJMCh4 of Mycobacterium tuberculosis H37Rv strain, IBMPAN3. 4/3 MmpL3 (V195A) mutant selected on EJMCh4 of Mycobacterium tuberculosis H37Rv strain, IBMPAN4. 6A MmpL3 (V684A) mutant selected on EJMCh6 of Mycobacterium tuberculosis H37Rv strain, IBMPAN5. 6B MmpL3 (V240A) mutant selected on EJMCh6 of Mycobacterium tuberculosis H37Rv strain, IBMPAN6. 6C MmpL3 (G253E) mutant selected on EJMCh6 of Mycobacterium tuberculosis H37Rv strain, IBMPAN

1H-苯并[d]咪唑（1H-benzo[d]imidazole）衍生物在体外及人源单核细胞衍生巨噬细胞中均展现出纳摩尔级浓度的抗结核活性，且对人体细胞无毒性。5-溴-2-(2-环己基乙基)-1H-苯并[d]咪唑（EJMCh4）与2-(2-环己基乙基)-5,6-二甲基-1H-苯并[d]咪唑（EJMCh6）的作用机制通过以下方式得以阐明：首先筛选获得耐药结核分枝杆菌（Mycobacterium tuberculosis）突变株并开展全基因组测序，同时通过观察到其对阿拉伯半乳聚糖的分枝菌酰化作用以及TDM（海藻糖二霉菌酸酯）合成的抑制效果，证实MmpL3是这类化合物发挥抗结核活性的分子靶点。
细菌菌株：
1. 结核分枝杆菌H37Rv菌株，IBMPAN库存株
2. 以EJMCh4筛选得到的结核分枝杆菌H37Rv菌株4/1 MmpL3（V285A）突变株，IBMPAN
3. 以EJMCh4筛选得到的结核分枝杆菌H37Rv菌株4/3 MmpL3（V195A）突变株，IBMPAN
4. 以EJMCh6筛选得到的结核分枝杆菌H37Rv菌株6A MmpL3（V684A）突变株，IBMPAN
5. 以EJMCh6筛选得到的结核分枝杆菌H37Rv菌株6B MmpL3（V240A）突变株，IBMPAN
6. 以EJMCh6筛选得到的结核分枝杆菌H37Rv菌株6C MmpL3（G253E）突变株，IBMPAN