In our calculations, we used a 'single trajectory protocol', where the unbound ligand and receptor structures are extracted from the same protein-ligand complex trajectory. Even though this is a computationally efficient way of performing the calculations, the simulations did not sample the complete unbound conformational space explored by the receptor and the ligand separately. This might be especially important for macrocycles, which have lots of flexible bonds and have a vast conformational space. A 'multiple trajectory protocol' where all the three end-states are simulated separately might have been a better approach.