3H]linopirdine binding to rat brain membranes is not relevant for M-channel interaction

Abstract

Linopirdine was developed as a cognitive enhancing molecule and demonstrated to specifically block the potassium current generated by the brain specific KCNQ2–KCNQ3 proteins (M-channel). In this study we investigated the relevance of [ 3H]linopirdine binding in rat brain extracts to the interaction with the M-channel proteins. Our results confirm the presence of a high affinity site for [ 3H]linopirdine in rat brain tissues ( K D = 10 nM) but we also identified a high affinity binding site for [ 3H]linopirdine in rat liver tissues ( K D = 9 nM). Competition experiments showed that [ 3H]linopirdine is displaced by unlabelled linopirdine with comparable affinities from its binding sites on rat brain and rat liver membranes. [ 3H]linopirdine was completely displaced by a set of cytochrome P450 (CYP450) ligands suggesting that [ 3H]linopirdine binding to rat brain and liver membranes is linked to CYP450 interaction. The testing of CYP450 ligands on the M-channel activity, using a Rb + efflux assay on cells expressing the KCNQ2–KCNQ3 proteins, demonstrated that [ 3H]linopirdine binding results cannot be correlated to M-channel inhibition. The results obtained in this study demonstrate that [ 3H]linopirdine binding to rat brain and rat liver membranes is representative for CYP450 interaction and not relevant for the binding to the M-channel proteins.