中文摘要：

目的以L12-L10相互作用为靶点筛选具有抗结核活性的先导化合物。方法应用酵母双杂交模型AH109（pAD—L12＋pBD—L10）通过生长抑制方法筛选阳性化合物，以AH109（pAD—T＋pBD-53）作为对照；通过96孔板法检测阳性化合物对耻垢分枝杆菌的抑制活性；应用定量微孔板快速显色法（MABA）检测抗结核杆菌活性；通过β-半乳糖苷酶活性定量检测判断阳性化合物在模型上对L12-L10相互作用的阻断活性；应用体外蛋白表达系统检测阳性化合物对蛋白表达的抑制作用；应用平皿二倍稀释法检测阳性化合物的药敏作用。结果筛选到4个在模型上具有活性的阳性化合物，其对耻垢分枝杆菌具有比较好的抑制活性，其最小抑制浓度（MIC）在3．125～12．5gg／ml之间；其中IBM—T275对结核分枝杆菌标准株和临床分离株均具有比较好的抑制活性，MIC在5～10gg／ml之间，而对细菌抑制活性较低，其MIC均在64μg／ml以上；IBM．T275能够抑制酵母模型内β-半乳糖苷酶的表达，并且能够体外抑制蛋白表达，其IC如为12．57μg／ml。结论筛选到1个具有抗结核杆菌活性的阳性化合物，其抗结核活性可能与阻断L12-L10蛋白相互作用相关。

英文摘要：

Objeetive To screen potential small-molecular inhibitors of L 12-L 10 interaction with anti-tuberculosis activity. Methods A yeast two-hybrid system was used to identify small molecules that block the interaction between L 12 and L 10 proteins. The minimum inhibitory concentration （MIC） was measured in sterile 96-well microplates. Anti-tuberculosis activity of compound IBM-T275 was determined by microplate alamar blue assay （MABA）. The t3-gal activity was measured to detect the disruption of L12-L10 interaction by compound IBM-T275. Translation inhibition by compound IBM-T275 was assessed using an in vitro ceil-free translation system. The MICs of compound IBM-T275 against ATCC strains and clinical isolates were determined using the agar dilution method. Results Four compounds were found to inhibit the growth of AH109 （pAD-L12 ＋ pBD-L10） more dramatically than that of AH109 （pAD-T ＋ pBD-53）, and the MICs range was 3.125-12.5 gg/ml on mc2155. Among the hit compounds, IBM-T275 showed anti-tuberculosis activity on standard strain and clinical strains with MICs range at 5 - 10 μg/ml, but showed high MICs on other bacterial strains （MICs 〉 64 gg/ml）. Compound IBM-T275 could inhibit 13-gal activity in the model as well as the in vitro protein translation with IC50 of 12.57μg/ml. Conclusion A new compound was found to be an inhibitor of L12-L10 interaction with anti-tuberculosis activity through yeast two-hybrid screening system.