Dynamic stability-driven machine learning improves binding pose identification on human serum albumin.

Accurate identification of ligand binding modes on human serum albumin (HSA) is important for understanding drug-protein interactions that influence pharmacokinetics, including site-specific competition and protein-binding displacement. However, because HSA is a large, multisite binding protein that permits diverse binding configurations, conventional molecular docking often struggles to distinguish crystal-like poses from nonspecific surface-binding poses using static scoring functions alone. Here, we developed a machine-learning framework for HSA binding-pose identification that incorporates dynamic stability information derived from short molecular dynamics (MD) simulations. Time-dependent interaction patterns observed during MD trajectories, including contact persistence, distance fluctuations, and interaction energy-related descriptors, were quantified as interpretable features and integrated with docking scores. Using Leave-One-PDB-Out cross-validation on 31 HSA-ligand crystal structures, models incorporating MD-derived features significantly outperformed the docking score-only model under both pose-level (root-mean-square deviation [RMSD] ≤ 2.5 Å) and site-level (RMSD ≤ 5.0 Å) criteria (Holm-corrected p < 0.001). External validation using a temporally independent nateglinide-HSA complex further supported the utility of the framework for discriminating plausible binding modes within the HSA setting. Notably, informative dynamic features were obtained from 5 ns MD simulations, indicating that early-stage dynamic behavior is sufficient to improve pose discrimination. These findings show that short MD simulations can provide practically useful and interpretable features for machine-learning-based binding-pose identification on HSA. While broader validation will be required to establish applicability beyond HSA, the proposed framework offers a practical strategy for improving binding-mode analysis in pharmacokinetically relevant HSA studies.