Isoniazid: Radical-induced oxidation and reduction chemistry

Abstract

Isoniazid is a potent and selective therapeutic prodrug agent used to treat infections by Mycobacterium tuberculosis. Although it has been used clinically for over five decades its full mechanism of action is still being elucidated. Essential to its mechanism of action is the activation of isoniazid to a reactive intermediate, the isonicotinyl acyl radical, by the catalase-peroxidase KatG. The isonicotinyl acyl radical then reacts with NAD producing an inhibitor of the NADH-dependent enoyl ACP reductase responsible for mycolic acid synthesis as its primary target. However, the initial oxidation of isoniazid by KatG has also revealed alternative reaction pathways leading to an array of carbon-, oxygen-, and nitrogen-centered radical intermediates. It has also been reported that isoniazid produces nitric oxide in the presence of KatG and hydrogen peroxide. In this study, the temperature-dependent rate constants for the hydroxyl radical oxidation and solvated electron reduction of isoniazid and two model compounds have been studied. Based on these data the initial oxidation of isoniazid by the hydroxyl radical has been shown to predominantly occur at the primary nitrogen of the hydrazyl moiety, consistent with the postulated mechanism for the formation of the isonicotinyl radical. The hydrated electron reduction occurred mostly at the pyridine ring. Concomitant EPR spin-trap measurements under a variety of oxidizing and reducing conditions did not show any evidence of nitric oxide production as had been previously reported. Finally, examination of the transient absorption spectra obtained for hydrated electron reaction with isoniazid demonstrated for the first time an initial reduced transient identified as the isonicotinyl acyl radical produced from isoniazid.