To assess the four compounds' binding affinity, we calculated free energy for each system. GMX_MMPBSA (v1.5.2) tool was used in conjunction with GROMACS to perform such calculations. GMX_MMPBSA is based on AMBER's robust MMPBSA.py tool . The first 50 ns were assumed to be an equilibration phase and were excluded from the analyses. Therefore, the free energy calculations were carried out for the last 50 ns of the simulations divided into 5000 frames. GMX_MMPBSA removed the PBC prior to the calculations. The analyses were performed on an interval of 2 frames, ending with a total of 2501 frames included in the calculations for each system. The free energy calculations were carried out using the Poisson-Boltzmann (PB) approach. AMBER's topologies were generated using the new ff19SB for the proteins and the gaff forcefield for the ligands. Sodium and Chloride ions were represented by the frcmod.ions1lm_126_tip3p (Li/Merz ion parameters for +1 and -1 ions in TIP3P water).