Hydrazonophenol, a Food Vacuole-Targeted and Ferriprotoporphyrin IX-Interacting Chemotype Prevents Drug-Resistant Malaria

The rapid emergence of resistance against frontline antimalarial drugs essentially warrants the identification of new-generation antimalarials. Here, we describe the synthesis of (<i>E</i>)-2-isopropyl-5-methyl-4-((2-(pyridin-4-yl)­hydrazono)­methyl)­phenol (<b>18</b>), which binds ferriprotoporphyrin-IX (Fe^III-PPIX) (<i>K</i>_d = 33 nM) and offers antimalarial activity against chloroquine-resistant and sensitive strains of <i>Plasmodium falciparum</i> in vitro. Structure–function analysis reveals that compound <b>18</b> binds Fe^III-PPIX through the −CN–NH– moiety and 2-pyridyl substitution at the hydrazine counterpart plays a critical role in antimalarial efficacy. Live cell confocal imaging using a fluorophore-tagged compound confirms its accumulation inside the acidic food vacuole (FV) of <i>P. falciparum</i>. Furthermore, this compound concentration-dependently elevates the pH in FV, implicating a plausible interference with Fe^III-PPIX crystallization (hemozoin formation) by a dual function: increasing the pH and binding free Fe^III-PPIX. Different off-target bioassays reduce the possibility of the promiscuous nature of compound <b>18</b>. Compound <b>18</b> also exhibits potent in vivo antimalarial activity against chloroquine-resistant <i>P. yoelii</i> and <i>P. berghei</i> ANKA (causing cerebral malaria) in mice with negligible toxicity.