Based on the relationship DG = -RTlnKEq, this strain will decrease the affinity of binding by approximately 5-fold for each kcal/mol of strain energy as compared to a hypothetical equivalent ligand that binds similarly but without strain. To the extent that the bound conformation is retained among those available, macrocyclization will conformationally pre-organize the compound and thereby reduce the entropic penalty associated with binding to the target. Adding rigidifying features to the macrocycle, thereby further decreasing the conformations available, can further reduce the entropic penalty for binding and thus increase affinity. However, if this rigidification favors a conformation that is dissimilar to the bound conformation, a component of strain will be introduced upon binding that will reduce affinity. Maximizing the affinity increase achieved through macrocyclization thus requires maximizing conformational preorganization in the unbound state while minimizing strain in the bound state.