Exploring novel inhibitors for Babesia bigemina lactate dehydrogenase: a computational structural biology perspective.

Babesia bigemina is an apicomplexan parasite responsible for causing "Texas fever" in bovines. Current treatments for bovine babesiosis are hindered by several limitations, including toxicity, insufficient efficacy in eliminating the parasite, and the potential for resistance development. A promising approach to overcome these challenges is the identification of compounds that specifically target essential metabolic pathways unique to the parasite. One such target is lactate dehydrogenase (LDH), a critical enzyme involved in the regulation of anaerobic glycolysis. Notably, Babesia bigemina LDH (BbigLDH) exhibits a five-amino acid insertion in the active site, a feature that differentiates it from the host's LDH. This structural divergence makes apicomplexan LDH an attractive and potentially selective drug target for therapeutic intervention. In this study, a structure-based drug discovery approach was implemented to find novel inhibitor candidates. Potential candidates were identified using a virtual screening workflow. The compounds with favorable docking scores were filtered using the QM-polarized ligand docking and induced fit docking methods. As a result, 20 novel compounds were identified that bind to the active site of BbigLDH but show low affinity to the host LDHs. Molecular dynamics simulations of the complexes (8.8 µs in total) were performed, and binding free energies were calculated. As a result, protein structures containing compounds C9, C16 and C18 maintained their stability throughout 1 µs simulations with low binding free energies and conserved interactions with known catalytic residues. Therefore, these three compounds deserve further investigation to better understand their mode of action and therapeutic potential for babesiosis. The results of this study elucidate the structural features of the BbigLDH enzyme and provide novel LDH binders that may pave the way for further research into the development of parasite-specific LDH inhibitors.