To summarize, this synthetic strategy resulted in the building of a chemical library of various substituted aminated 3-benzylmenadiones. Besides the early hit 4a, which displayed a potent activity against intracellular T. cruzi amastigotes (mean IC50 = 1.29 µM, 6 replicates), four compounds (9a, 11b, 15f and 19c) displayed IC50 values ≤ 5 µM, in the same range as Bz (2.25 µM) (Table ). Most of the tested compounds showed quite significant toxicity towards the possible replacement of the benzylic function Bn1 with its pyridine analog (Figure ). A slight decrease in the molecule's inhibition could be observed, yet it allowed to reduce its lipophilicity. It can also be interesting to consider the importance of the methylene moiety on the substituted piperazine. As matter of fact, we were able to demonstrate that the complete deletion of this carbon atom rendered the compound as inactive. Finally, the introduction of other piperazine bioisosteres to improve metabolic stability and potentially increase the naphthoquinone's solubility could also direct towards a possible lead compound. In summary, the discovery of this new hit and the elucidation of the molecule's key positions will serve for further structural refinement with improved solubility and activity.