*2.6. Molecular Docking and Molecular Dynamic Simulation (MDS)*

The molecular docking was performed with bioinformatics tools MGL Tools [36], Auto Dock Vina [37], Molecular Operating Environment software (MOE), and Discovery Studio visualization tool. The PDB structure for the BSA (PDB ID: 6qs9) obtained from Protein Data Bank was used for the molecular docking [38]. The molecular docking was carried out for the three binding subdomains of BSA, and three runs were carried out for each binding Site. The top-scoring conformations were selected and analyzed for the interaction. MOE default parameters were used for docking analysis. The molecular dynamic simulation was carried out with NAMD 2.13 Suite (http://www.ks.uiuc.edu/Research/namd (accessed on 20 September 2021)) [39]. The ligand structures were immersed in the TIP3P water box. The charges neutralization was carried out by adding Na<sup>+</sup> or Cl<sup>−</sup> with their maximum concentration of 0.15 M. The minimization of the system was carried out in MOE. The default MOE parameters were used for the minimization, which included Amber10: EHT force field. Partial Charge on LL toms was calculated using Amber10: EHT force field. The system's energy was minimized to an RMS gradient of 0.1 kcal/mol/A2. As per the MOE manual, an RMS of 0.1 is sufficient for structure minimization for molecular dynamic simulation. The minimized system was gradually heated. The temperature raised from 0 K to 310 K at an incremental level of 50 K for 100 ps and equilibrated at each temperature step followed by an unrestrained run of 20 ns. Constrained light bond with 2 fs time step. Long-range electrostatics were treated via the particle mesh Ewald method. The grid box formed had following parameters and dimensions: solvent molecules—26,824, 1.009 g/cm<sup>3</sup> ; Space group—P1, Triclinic, 1, 1×; size—(105.8, 83.9, 102.2); cell shape—(90.0, 90.0, 90.0). The parameters RMSD and RMSF were obtained for the Protein BSA and the protein in the

presence of ligand (BSA-ERL) systems. The trajectory was visualized and analyzed with VMD 1.9.3 tool.
