Unveiling the Antibacterial Efficacy of a Benzonitrile Small Molecule, IITR00210, in <i>Shigella</i> Infection

The escalating prevalence of bacterial infections and the rapid emergence of multidrug-resistant Gram-negative bacterial pathogens highlight an urgent demand for effective antibacterial agents. In this study, we report our findings on IITR00210, a small molecule belonging to the nitrile class. The small molecule demonstrates broad-spectrum activity against bacterial pathogens, specifically against enteric pathogens, and exhibits antibiofilm activity. IITR00210 displays potent bactericidal activity against enteropathogens, resulting in a reduction of bacterial counts greater than 3 Log₁₀ CFU in time-kill kinetic assays. Mechanistic investigations revealed that IITR00210 induces bacterial cell envelope stress, leading to the alteration of the overall proton motive force (PMF). The disruption of PMF causes intracellular ATP dissipation and ultimately promotes cell death. The cell envelope stress generated in the presence of IITR00210 leads to a translational aberration. Importantly, IITR00210 exhibits a safe profile in <i>in vitro</i> and <i>in vivo</i> settings. The small molecule further showed potent intracellular antibacterial activity in polymorphonuclear cells infected with enteric pathogens and antiadhesion activity in mammalian cell lines. IITR00210 proves to be a promising therapeutic candidate, displaying a lack of stable resistance development, and it exhibited efficacy in the treatment of bacterial infections in a shigellosis murine model.