Vol. 299, Issue 2, 712-717, November 2001

Use of the -Imager for Rapid ex Vivo Autoradiography Exemplified with Central Nervous System Penetrating Neurokinin 3 Antagonists

Xavier Langlois, Paula te Riele, Cindy Wintmolders, Josée E. Leysen and Mirek Jurzak

Department of Receptor Pharmacology, Janssen Research Foundation, Beerse, Belgium

The neurokinin 3 (NK3) receptor antagonists represent a novel class of pharmacological agents, which are currently under evaluation for the treatment of psychiatric disorders. An efficient brain penetration is one of the main prerequisites to further evaluate compounds displaying high potency to bind the NK3 receptor. The present report describes a method for determining the in vivo occupancy of central NK3 receptors after peripheral administration of drugs. An ex vivo measurement of NK3 receptor occupancy by quantitative autoradiography employing [³H]senktide as the radioligand has been developed. The speed of the method, which is usually considered low due to the time dedicated to film exposure (from weeks to months), has been considerably increased by the use of the -imager. The high sensitivity of this new radioimager was used to visualize and quantitatively analyze the [³H]senktide binding sites in brain sections within hours. Using this method, we have demonstrated that the reference NK3 antagonist SR142801 dose dependently occupied the NK3 receptors in the gerbil brain after subcutaneous administration with an ED₅₀ of 0.85 mg/kg. The less active enantiomer SR142806 occupied the NK3 receptors only by 25% at the highest used dose of 10 mg/kg. These values are in accordance with the reported behavioral effects of the compounds. Our results indicate that ex vivo receptor occupancy measurements can be dependently used to predict the central activity of NK3 antagonists. More generally, the combination of ex vivo receptor autoradiography with the -imager detection constitutes a new and fast method to evaluate the brain penetration of drug candidates.