Dual Mechanism of Action of 5-Nitro-1,10-Phenanthroline against Mycobacterium tuberculosis

Abstract

New chemotherapeutic agents with novel mechanisms of action are urgently required to combat the challenge imposed by the emergence of drug-resistant mycobacteria. In this study, a phenotypic whole-cell screen identified 5-nitro-1,10-phenanthroline (5NP) as a lead compound. 5NP-resistant isolates harbored mutations that were mapped to <i>fbiB</i> and were also resistant to the bicyclic nitroimidazole PA-824. Mechanistic studies confirmed that 5NP is activated in an F₄₂₀-dependent manner, resulting in the formation of 1,10-phenanthroline and 1,10-phenanthrolin-5-amine as major metabolites in bacteria. Interestingly, 5NP also killed naturally resistant intracellular bacteria by inducing autophagy in macrophages. Structure-activity relationship studies revealed the essentiality of the nitro group for <i>in vitro</i> activity, and an analog, 3-methyl-6-nitro-1,10-phenanthroline, that had improved <i>in vitro</i> activity and <i>in vivo</i> efficacy in mice compared with that of 5NP was designed. These findings demonstrate that, in addition to a direct mechanism of action against <i>Mycobacterium tuberculosis</i>, 5NP also modulates the host machinery to kill intracellular pathogens.