We perform the annihilation of the electrostatic interactions of the guest in the bulk and binding site in separate systems over 11 equally-spaced λ values. The vdW interactions between the ligand and the protein are decoupled with the soft-core potential from Pham and Shirts 46 across 21 equally-spaced λ values. We applied Boresch-style restraints on the ligand in reference to the protein to restrict its translational and orientational motion during the alchemical transformation. To simplify the procedure, we used the MDRestraintsGenerator correction at 1 M is done analytically. We use a force constant of 5 kcal/mol/ Å2 and 100 kcal/mol/rad 2 for the distance and angle restraints, respectively. For each ABFE window, we performed an energy minimization followed by 2 ns and 20 ns of equilibration and production run, respectively. The free energy is recovered using the TI method, and the error in ∆G b was estimated with bootstrap resampling similar to the host-guest calculations.