Skelly_2025_Int.J.Mol.Sci_26_1975

Schistosomiasis, a parasitic disease caused by worms of the genus Schistosoma, affects >250 million people worldwide. With no available vaccine, treatment relies solely on one drug-praziquantel-underscoring the urgent need for new therapies. We identified a tegumental, non-neuronal acetylcholinesterase (AChE) from Schistosoma mansoni-SmTAChE-as a promising drug target. RNA interference confirmed its essential role in parasite survival, as gene suppression significantly reduced parasite recovery from infected animals. Here, we produced functionally active recombinant SmTAChE by using a mammalian expression system. Biochemical characterization confirmed its identity as a true acetylcholinesterase, with the highest turnover rate (K(cat) = 373 +/- 39 s(-1)) and catalytic efficiency (K(cat)/K(m) = 1.17 x 10(6) M(-1).S(-1)) for acetylthiocholine. Additionally, rSmTAChE was inhibited by classical AChE-specific inhibitors but not by a butyrylcholinesterase-specific inhibitor. To identify novel SmTAChE inhibitors, we developed a high-throughput chemical screening protocol (Z' factor > 0.9) and screened a 1894-compound validation library. Twelve compounds reproducibly inhibited rSmTAChE by >30% at 7.5 microM, including known AChE inhibitors like physostigmine and new selective inhibitors. Notably, compound #2 preferentially inhibited rSmTAChE (IC(50) = 0.74 microM) over human AChE (IC(50) = 151 microM), thus providing a foundation for developing parasite-specific therapies targeting SmTAChE and potentially leading to new treatments for schistosomiasis.