The present study explored anti-tubercular pyrrole derivatives against cancer targets using different in silico and in vitro approaches. Initially, nineteen anti-tubercular pyrrolyl benzohydrazide derivatives were screened against a potent cancer target PLK1 using an AutoDock Vina approach. Out of the nineteen derivatives, the two most potent derivatives C8 [N′-(4-(1H-pyrrol-1-yl) benzoyl)-3-chlorobenzohydrazide] and C18 [N′-(4-(1H-pyrrol-1-yl) benzoyl)-4-nitrobenzohydrazide], were subjected to molecular simulation analysis for a 100 ns trajectory. Further, these two derivatives were tested against A549, MCF-7, and HepG2 cell lines using an MTT proliferation assay. Apoptotic cell cycle and DAPI assays were also performed for C8 on A549 cell lines. Molecular dynamic analysis revealed that the stability of the C8–PLK1 protein complex during the 100 ns trajectory run was better than that of the C18–PLK1 protein complex. In addition, C8 showed lower IC50 values against the tested cell lines, in comparison to C18. Thus, C8 was selected for cell cycle, apoptosis, and DAPI analysis. Interestingly, C8 resulted in the significant cell cycle arrest of A549 cells at the G2/M phase, and annexin V-FITC/PI showed a significant increase (from 6.27% to 60.52%) in the percentage of apoptotic A549 cells. The present findings suggest that the anti-tubercular compound (C8) could be translated into a potent repurposed candidate against lung cancer. Nevertheless, in vivo assessment is necessary to further confirm the outcome and its clinical translation.