Evaluating aziridinyl nitrobenzamide compounds as leishmanicidal prodrugs
370 - 377 (7)
Antimicrobial Agents and Chemotherapy
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Many of the nitroaromatic agents used in medicine function as prodrugs and must undergo activation before exerting their toxic effects. In most cases this is catalysed by FMN-dependent type I nitroreductases (NTRs), a class of enzyme absent from higher eukaryotes but expressed by bacteria and several eukaryotic microbes including trypanosomes and Leishmania. Here, we utilize this difference to evaluate whether a library of aziridinyl nitrobenzamides have activity against Leishmania major. Biochemical screens using purified L. major NTR (LmNTR) revealed that compounds containing an aziridinyl-2,4-dinitrobenzyl core were effective substrates for the enzyme and showed that the 4-nitro group was important for this activity. To facilitate drug screening against intracellular amastigote parasites, we generated leishmanial cells that expressed the luciferase reporter gene and optimized a mammalian infection model in a 96-well plate format. A subset of aziridinyl-2,4-dinitrobenzyl compounds possessing a 5-amide substituent displayed significant growth inhibitory properties against the parasite, with the most potent agents generating 50 % inhibitory concentrations of <100 nM towards the intracellular form. This antimicrobial activity was shown to be LmNTR specific since L. major NTR+/- heterozygote parasites were slightly resistance to the most aziridinyl dinitrobenzyl agents tested. When the most potent leishmanicidal agents were screened against the mammalian cells in which the amastigote parasites were propagated, no growth inhibitory effect was observed at concentration up to 100 M. We conclude that the aziridinyl nitrobenzamides represent a new lead structure that may have the potential to treat leishmanial infections.