Dual-targeting 5-nitroimidazole ethers eradicate H. pylori by induction of stress and disabling its adaptive response

Metronidazole represents a front line-drug against Helicobacter pylori . To unravel its largely unknown cellular targets, we performed activity-based protein profiling (ABPP). Surprisingly, an alkynylated ether probe (Metro-P3) exhibited a 60-fold enhanced potency. Chaperonin HpGroEL and thiol peroxidase HpTpx were identified as prominent Metro-P3 targets, the latter of which is essential for H. pylori survival under oxidative stress. Biological assays validated HpTpx inhibition as the origin of the activity boost, demonstrated by covalent modification of the catalytic cysteine and co-crystallization. Metro-P3 and refined ether analogs exhibited favorable pharmacological profiles with no cytotoxicity. The activity boost translated to an in vivo H. pylori mouse model demonstrating full eradication at 50-fold reduced dosing compared to metronidazole triple therapy. Our results highlight 5-nitroimidazole ethers as dual mode of action antibiotics which induce oxidative stress and inhibit the stress response leading to bacterial death.