In the present implementation, Δd and Δt are provided by the user, with defaults set to 0.05 Å and, 0.24 ps, respectively. The other critical parameter is the strength of the RAMD force (K) in kcal/mol/Å. This parameter is the only one that we have found to require adjustment for each system series because it is intuitively connected to the steepness of the barrier for the unbinding process. To avoid time-consuming tuning of this parameter, we slowly increase the force constant throughout the RAMD simulations to ensure that we achieve unbinding even in cases where the initial constant is too low. In Desmond, the RAMD force is approximated with a moving harmonic potential which is dynamically positioned to produce an equivalent force. In order to reduce correlation of RAMD simulations starting from the same initial configuration, we sample 50 ps of unbiased MD for each of our 30 RAMD replicas, starting each with different initial velocities. Next, RAMD sampling is initiated starting with force constant of 15 kcal/mol/Å 2 which increases linearly by 5 kcal/mol/Å 2 /ns until the ligand unbinds.