The present investigation reports simple and eco-friendly biogenesis of silver nanoparticles (AgNPs) using leaf extracts of Asparagus racemosus and Sophora interrupta. UV-Vis spectra showed absorption peaks at 413 and 420 nm respectively corresponding to A. racemosus (AR-AgNPs) and S. interrupta (SI-AgNPs). TEM, DLS, XRD and TGA studies revealed that biogenic AgNPs are monodispersed, spherical shaped, thermally stable, FCC crystals and 4-15 nm in size. Zeta potential values were found to be − 28.9 and − 18.6 mV respectively corresponding to SI-AgNPs and AR-AgNPs. SI-AgNPs exhibited strong antibacterial activities against Escherichia coli, Klebsiella pneumoniae, Bacillus subtilis and Micrococcus luteus with inhibition zones of 11.8, 10.5, 9.8 and 9.1 mm respectively. Further SI-AgNPs exhibited significant cytotoxic properties against SKOV3, DU149, PC3 and A549 cells with IC 50 values of 120.87, 191.34, 235.06 and 314.27 µg/mL respectively. SI-AgNPs induce pronounced apoptotic effects on SKOV3 cells including loss of membrane integrity, chromatin condensation (pyknosis) and nuclear fragmentation (karyorrhexis). SI-AgNPs exhibited significant cell migration inhibition by reducing the wound healing of SKOV3 monolayers. Phytosynthesized AgNPs were found to be biocompatible towards normal (CHO) cells.Bethu Murali Sathyanarayana and Netala Vasudeva Reddy contributed equally to this work.