The present research explores the urease inhibitory potential of a designed set of quinoline-2-one/chalcone hybrid derivatives (4a-e). The biological assessment demonstrated that these molecules exhibit strong urease inhibition, with IC50 values ranging from 0.58-1.90 µM. Their inhibitory potency notably surpasses that of the reference compound, thiourea (IC50 = 21.50 µM). Within this series, compounds 4a, 4c, and 4e emerged as the most active inhibitors, displaying IC50 values of 0.93 ± 0.08 µM, 0.75 ± 0.06 µM, and 0.58 ± 0.04 µM, respectively. In addition, the anti-Helicobacter pylori potential of these hybrids was assessed against three metronidazole-resistant H. pylori strains using the disk diffusion technique. The outcomes revealed a strong association, indicating that methoxy-bearing analogs 4c and 4e identified as the most effective urease inhibitors also exhibited pronounced antibacterial action against H. pylori. Collectively, these observations underscore the promising dual therapeutic profile of this hybrid framework.