Table 2.

GPR139 in silico mutant effects of 1a and 7c binding.

Mutation	Change of in vitro potency		In silico relative binding free energies (ΔΔG kcal/mol)
	1a	7c	1a			1a	7c
			Iteration 1	Iteration 2	Iteration 3	Final model	Final model
F109A3×33	Loss of function	>2000 fold decreased	−3.14 ± 0.84	0.66 ± 2.02	1.25 ± 1.03	6.52 ± 0.87	6.27 ± 0.66
H187A5×43	>34 fold decreased	138 fold decreased	−3.03 ± 1.04	−1.26 ± 1.60	0.56 ± 1.11	2.64 ± 0.93	1.72 ± 1.29
N271A7×38	Loss of function	Loss of function	0.25 ± 0.47	4.40 ± 0.54	3.73 ± 0.29	3.21 ± 0.45	2.71 ± 0.34

Iterative procedure of model optimization using molecular dynamics and the relative binding free energies obtained by free energy perturbation (FEP) calculations in comparison with in vitro potency as a scoring function. The FEP relative binding free energies that are in agreement with in vitro data are shown in bold.