Before revision:  We find that tauRAMD yields the ranking of dissociation rates for the simulated ligands, protein mutants and temperature conditions in good accordance with experiments . Comparison of the unbinding paths and metastable states of benzene from the L99A mutant reveals a good agreement of tauRAMD with much more computationally demanding conventional MD simulations. Moreover, mapping of the egress routes for all the systems studied shows that visiting more metastable states slows dissociation and indicates that accurate dissociation rates can be computed without exhaustive sampling, as long as the main egress route is sampled. Overall, this study provides physical insights that could be exploited in the rational optimization of the kinetic properties of drug candidates.
After revision:  We find that tauRAMD yields the ranking of dissociation rates for the simulated ligands, protein mutants and temperature conditions in good accordance with experiments and thereby allows a comprehensive characterization of the ligand egress routes and determinants of tau. Although ligand dissociation by multiple egress routes is observed, we find that egress via the predominant route determines the value of tau. We also find that the presence of metastable states along egress pathways slows down protein-ligand dissociation. These physical insights could be exploited in the rational optimization of the kinetic properties of drug candidates.