Identification of benzamides derivatives of norfloxacin as promising microRNA-21 inhibitors via repressing its transcription

MicroRNA-21 is a carcinogenic microRNA, whose overexpression arises in a variety of tumor tissues. Hence, microRNA-21 a prospective target for cancer treatment, and regulation of microRNA-21 by small molecule inhibitors is deemed as a promising approach for tumor therapy. In this work, to discover potent microRNA-21 inhibitor, series of 4-(N-norfloxacin-acyl)aminobenzamides were designed and synthesized, and their inhibitory effects were appraised by utilizing dual luciferase reporter assays. The results indicated that compound A7 was the most efficient microRNA-21 small molecule inhibitor. What's more, A7 suppressed the migration of Hela cells and the colony formation of Hela and HCT-116 cells as well as promoted apoptosis of Hela cells. In the mechanism study, results of RT-qPCR certified that A7 could reduce the level of mature microRNA-21 via disrupting its expression at the transcriptional level of its primary form pri-miR-21, which was distinct from most previous inhibitors directly binding with pre-miR-21. Noticeably, Western blotting and RT-qPCR uncovered A7 could upregulate the expression PTEN, EGR1 and SLIT2, which are the downstream functional targets of microRNA-21. These findings demonstrated that A7 was a promising microRNA-21 small molecule inhibitor and 4-(N-norfloxacin-acyl) aminobenzamide can serve as a new scaffold for discovery of potent microRNA-21 inhibitor.

Automated summary

In this study, a series of 4-(N-norfloxacin-acyl)aminobenzamides were designed and synthesized, and compound A7 was identified as the most efficient microRNA-21 small molecule inhibitor. A7 demonstrated inhibitory effects on migration, colony formation, and apoptosis in Hela cells. Mechanistically, A7 reduced the levels of mature microRNA-21 by disrupting its transcription at the level of its primary form pri-miR-21. Furthermore, A7 upregulated the expression of downstream functional targets PTEN, EGR1 and SLIT2 of microRNA-21.