Molecular neuroimaging is today considered essential for evaluation of novel CNS drugs; it is used to quantify blood-brain barrier permeability, verify interaction with key target and determine the drug dose resulting in 50% occupancy, IC₅₀. In spite of this, there has been limited data available to inform on how to optimize study designs. Through simulations, we here evaluate how IC₅₀ estimation is affected by the (i) range of drug doses administered, (ii) number of subjects included, and (iii) level of noise in the plasma drug concentration measurements. Receptor occupancy is determined from PET distribution volumes using two different methods: the Lassen plot and Likelihood estimation of occupancy (LEO). We also introduce and evaluate a new likelihood-based estimator for direct estimation of IC₅₀ from PET distribution volumes. For estimation of IC₅₀, we find very limited added benefit in scanning individuals who are given drug doses corresponding to less than 40% receptor occupancy. In the range of typical PET sample sizes (5–20 subjects) each extra individual clearly reduces the error of the IC₅₀ estimate. In all simulations, likelihood-based methods gave more precise IC₅₀ estimates than the Lassen plot; four times the number of subjects were required for the Lassen plot to reach the same IC₅₀ precision as LEO.