Discovery of novel repurposed anthelminthics against Trichinella spiralis and albendazole-resistant nematodes through metabolomics-guided virtual screening

Trichinella spiralis is a parasitic nematode that causes trichinellosis, leading to weakness, muscle pain, facial edema, and potentially death . Albendazole (ABZ) is the current drug of choice; however, resistance has been increasingly reported, highlighting the urgent need for novel therapeutics. Developing new drugs is costly and time-consuming, making computational approaches and drug repurposing attractive alternatives. In this study, we combined metabolomics and virtual screening to identify potential anthelmintic candidates for trichinellosis and ABZ-resistant nematodes. Both adult and larval T. spiralis were treated with ABZ, then untargeted metabolomics were performed. Over 11,000 features were detected using the XCMS platform, with 122 and 133 metabolites significantly altered in adults and larvae, respectively. Pathway analysis with MetaboAnalyst identified fatty acid degradation as a key pathway affected by ABZ. Since this pathway is essential for worm lipid metabolism, we targeted the enzymes carnitine palmitoyltransferase (CPT) 1 and 2, which transport fatty acids into mitochondria. Virtual screening against parasite and human CPTs identified 87 compounds that selectively bind the parasite proteins. Lumacaftor, entrectinib, and fluspirilene showed the lowest binding energies and were tested in vitro . In larval T. spiralis , fluspirilene and entrectinib killed the parasites at concentrations of 266.8 and 442.6 μg/ml, respectively, while lumacaftor showed no activity. Importantly, entrectinib also killed both ABZ-sensitive and ABZ-resistant Caenorhabditis elegans at lower concentrations than ABZ, demonstrating potent activity against resistant nematodes. Our findings suggest that entrectinib is a promising candidate for a novel anthelmintic, with potential to overcome ABZ resistance in trichinellosis and other parasitic infections. • Metabolomics of Trichinella spiralis revealed fatty acid degradation as a major pathway disrupted by albendazole (ABZ). • Carnitine palmitoyltransferase (CPT) 1 and 2 were identified as key targets for in silico screening of repurposed drugs. • Computational docking yielded 87 compounds with selective affinity for parasite CPTs over human homologs. • Entrectinib and fluspirilene exhibited strong in vitro activity against T. spiralis muscle larvae. • Entrectinib killed both ABZ-sensitive and -resistant nematodes, showing its potent activity against resistant worms.