In an effort for fighting with dreadful drug resistance, iminotetraberberine was hybridized with metronidazole to construct a unique type of potential broad-spectrum antibacterial iminotetrahydroberberine-corbelled metronidazoles. Some prepared hybrids exerted promising inhibitory effects against the tested microorganisms in comparison to the natural berberine, clinical metronidazole and norfloxacin. Noticeably, phenyl oxime derivative 8e displayed a broad antibacterial spectrum with a quite low MIC value of 0.024 mM against P. aeruginosa, being 63-, 62- and 2-fold to berberine, metronidazole and norfloxacin, respectively. The active compound 8e with low cytotoxicity under effective bacteriostatic concentration could decrease biofilm viability and show much lower trend to induce the resistant development than norfloxacin in the tested period. Mechanism investigation showed that compound 8e could disturb the bacterial membrane to lead to the leakage of cellular contents, thus exerting potent antibacterial potency. It was also revealed that compound 8e could interact with penicillin binding protein via multi-site non-covalent binding in docking simulation. The above results manifested that iminotetrahydroberberine-corbelled metronidazoles might bring hope for the exploitation of new broad-spectrum antibacterial agents with a membrane-destruction mechanism. CI - Copyright (c) 2022 Elsevier Ltd. All rights reserved.