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Regarding the affinity of the fluoro compound 27 , it was anticipated that the hydroxyl group of the tyrosine side chain of 25 makes a hydrogen-bond with Glu494 of IRAP, an interaction that is lost in the case of 27 according to the free energy perturbation (FEP) analysis (Jespers et al., 2019), explaining the reduction in binding affinity of compound 27 as compared to the analogous compound 25