Interaction of the NSAID cyclooxygenase inhibitor, acemetacin, with human serum albumin: an experimental and computational modelling investigation

) showed an enthalpy-driven process involving hydrogen bonds, van der Waals, and hydrophobic interactions. A 7.1% increase in the α-helical content, local perturbations around protein fluorophores and morphological alterations were noticed in the presence of ACM, as affirmed by circular dichroism, 3D fluorescence and atomic force microscopy. Molecular docking predicted the binding preference of ACM towards Site I over Site III, while the molecular dynamics simulation over 100 ns supported a stable ACM-HSA complex with an average RMSD of 0.32 nm and low RMSF fluctuations. This was well supported by competitive displacement results using site-specific probes, confirming Sudlow's Site I as the primary binding locus. The observed moderate binding affinity, site-selective binding behavior, and structural stability suggest effective serum transport and influence on tissue distribution, with potential implications for drug-drug interactions and therapeutic efficacy.