Prodrug activation in malaria parasites mediated by an imported erythrocyte esterase, acylpeptide hydrolase (APEH)

• DOI: 10.1073/pnas.2417682122
• Publication Date: 2025-03-04T15:51:56Z
• Authors: Sesh A. Sundararaman; Justin J. Miller; Ellora C. Daley; Kelsey A. O’Brien; Paulina Kasak; Abigail M. Daniels; Rachel L. Edwards; Kenneth M. Heidel; Darean A. Bague; Madeleine A. Wilson; Andrew J. Koelper; Elexi C. Kourtoglou; Alex D. White; Sloan A. August; Georgia A. Apple; Regis W. Rouamba; Anthony J. Durand; John J. Esteb; Florian L. Muller; R. Jeremy Johnson; Geoffrey C. Hoops; Cynthia S. Dowd; Audrey R. Odom John
• Keywords: Esterase; Carboxylesterase; Hydrolase

The continued emergence of antimalarial drug resistance highlights the need to develop new therapies. Unfortunately, development is often hampered by undesirable drug-like properties lead compounds. Prodrug approaches temporarily mask compound features, improving bioavailability and target penetration. We have found that lipophilic diester prodrugs phosphonic acid antibiotics, such as fosmidomycin (Fsm), exhibit significantly higher potency than their parent compounds [R.L. Edwards et al. , Sci. Rep. 7 8400 (2017)]. However, activating enzymes for these were unknown. Here, we show an erythrocyte enzyme, acylpeptide hydrolase (APEH), major enzyme multiple ester prodrugs. Surprisingly, this taken up malaria parasite, Plasmodium falciparum where it localizes parasite cytoplasm retains enzymatic activity. Using a fluorogenic library, characterize structure–activity relationship APEH compare P. esterases. parasite-internalized plays important role in activation substrates with branching at alpha carbon, keeping its exopeptidase Our findings highlight mechanism antimicrobial prodrug activation, relying on host-derived yield microbial target. Mutations prodrug-activating are common [E. S. Istvan Nat. Commun. 8 14240 (2017); K. M. V. Sindhe mBio 11 e02640-19 (2020); J. H. Butler Acs Infect Dis. 6 2994–3003 (2020)]. Leveraging internalized host would circumvent this, enabling design barriers resistance.