Abstract: Drug abuse represents a critical social issue that poses significant threats to public health and safety. As a highly favored model protein, bovine serum albumin (BSA) is frequently utilized to investigate the binding characteristics of drugs with proteins. Such investigations provide valuable insights into the transportation, distribution, and metabolism of drugs within biological systems. This study evaluated the binding properties of 5'-triphosphate disodium salt (Na₂ATP) and metal ions on loxoprofen sodium (LOXNa) with BSA by applying different spectroscopic and computational techniques. The multi-spectroscopic and molecular docking approaches reveal that novel Na₂ATP and LOXNa also interact in subdomain IIIA of of Sudlow site II by forming ground state complexes following a static quenching mechanism. The binding constant is approximately 103 M-1, and during the binding process, these two drugs exhibit positive synergistic drug effects. The complexation is spontaneous (ΔG ~ -29 kJ/mol), and enthalpy-driven (ΔH~ 82.16 kJ/mol and ΔS~ 365J/mol), and is primarily mediated by hydrogen bonding and van der Waal forces. Furthermore, the research revealed that the interaction between Na₂ATP-LOXNa and BSA could potentially alter the drug's transportation and release behavior within the body, ultimately influencing its bioavailability and therapeutic outcomes. Additionally, the existence of metal ions could have an impact on the stability and activity of the drug, offering crucial theoretical insights for enhancing our understanding of the interaction mechanisms between drugs and biological molecules, as well as for optimizing drug design strategies.

Key words: molecular docking, spectroscopy, bovine serum albumin, adenosine 5'-triphosphate disodium salt, loxoprofen sodium