title: Conditions for biphasic competition curves in radioligand binding for ligands subjected to metabolic transformation.
abstract: The conditions affording biphasic competition curves in radioligand binding for ligands subjected to metabolic transformation was analyzed theoretically. It was shown that when a competing ligand was subjected to transformation to a ligand that showed higher affinity than the parent compound, biphasic competition curves, which might wrongly be interpreted as indicating the presence of two receptor sites, could be observed in binding assays containing a homogenous receptor population. Biphasic competition curves were seen if the conversion of the competitor occurred according to zero and second order kinetics, as well as by enzymatic catalytic processes. However, when the conversion occurred according to a first order kinetics, the competition curves were uniphasic and resolved only into one-site fits, with the apparent affinity of the competitor reflecting the degree of conversion of the competitor to its metabolite. When the metabolic conversion resulted in a metabolite that showed lower affinity for the receptor than the parent compound, the competition curves became supersteep for conversions according to zero and second order kinetics, as well as for conversion by enzymatic catalytic processes.
authors.affiliation: Department of Pharmaceutical Biosciences, Uppsala University, Sweden. Jarl.Wikberg@farmbio.uu.se
authors.forename: J E
authors.initials: JE
authors.lastname: Wikberg
authors.forename: M
authors.initials: M
authors.lastname: Dambrova
authors.forename: S
authors.initials: S
authors.lastname: Uhlén
authors.forename: P
authors.initials: P
authors.lastname: Prusis
mesh_terms: D000818:Animals; D001667:Binding, Competitive; D001711:Biotransformation; D003198:Computer Simulation; D006801:Humans; D007700:Kinetics; D011869:Radioligand Assay