In Silico Targeting of Trypanothione Reductase and Glycerol-3-Phosphate Dehydrogenase in Leishmania

Leishmaniasis remains a neglected tropical disease with treatment limitations driven by toxicity, cost, and emerging resistance. Trypanothione reductase (TryR) and glycerol-3-phosphate dehydrogenase (GPDH) are essential Leishmania enzymes supporting redox homeostasis and energy/redox-linked metabolism, providing a biologically grounded rationale for dual-target inhibition. We applied an integrated in silico workflow to prioritize candidate inhibitors using ADMET prediction (SwissADME/pkCSM), molecular docking (AutoDock Vina), and 100 ns molecular dynamics (MD) simulations; human GPDH was included as a negative control to probe potential off-target liability. ADMET screening identified 41 drug-like candidates, with most predicted to have high GI absorption and low toxicity flags across assessed endpoints (computational predictions interpreted cautiously). Docking highlighted two leading compounds. CID 6529858 showed the most favorable predicted binding to Leishmania GPDH (−8.9 kcal/mol) with a modest parasite-favored score difference versus human GPDH (−7.2 kcal/mol; Δ = −1.7 kcal/mol), while eupatorin (CID: 97214) displayed dual-target potential (TryR −7.5 kcal/mol; Leishmania GPDH −8.2 kcal/mol; human GPDH −7.2 kcal/mol; Δ = −1.0 kcal/mol). In MD, key complexes remained stable: CID 6529858 exhibited low GPDH backbone deviation (~0.25–0.40 nm), and eupatorin showed the most stable TryR trajectory (average RMSD ~0.45 nm), supported by generally low residue fluctuations across complexes. PCA further suggested ligand-associated restriction of large-scale motions (e.g., GPDH PC1 = 27.38%; TryR PC1 = 18.1%). Overall, these results nominate eupatorin as a promising dual-target lead and CID 6529858 as a strong GPDH-focused scaffold, warranting experimental enzyme inhibition, antiparasitic efficacy, and host–cell cytotoxicity testing to confirm potency and selectivity.