Synthesis and evaluation of two new candidate high-affinity full agonist PET radioligands for imaging 5-HT1B receptors

Abstract

Introduction: The serotonin 1B receptor subtype is of interest in the pathophysiology and treatment of depression, anxiety, and migraine. Over recent years 5-HT_1B receptor binding in human brain has been examined with PET using radioligands that are partial but not full agonists. To explore how the intrinsic activity of a PET radioligand may affect imaging performance, two high-affinity full 5-HT_1B receptor agonists (AZ11136118, 4; and AZ11895987, 5) were selected from a large compound library and radiolabeled for PET examination in non-human primates.

Methods: [¹¹C]4 was obtained through Pd(0)-mediated insertion of [¹¹C]carbon monoxide between prepared iodoarene and homochiral amine precursors. [¹¹C]5 was obtained through N-¹¹C-methylation of N-desmethyl precursor 6 with [¹¹C]methyl triflate. [¹¹C]4 and [¹¹C]5 were studied with PET in rhesus or cynomolgus monkey. [¹¹C]4 was studied with PET in mice and rats to measure brain uptake and specific binding. Ex-vivo experiments in rats were performed to identify whether there were radiometabolites in brain. Physiochemical parameters for [¹¹C]4 (pKa, logD and conformational energetics) were evaluated.

Results: Both [¹¹C]4 and [¹¹C]5 were successfully produced in high radiochemical purity and in adequate amounts for PET experiments. After intravenous injection of [¹¹C]4, brain radioactivity peaked at a low level (0.2 SUV). Pretreatment with tariquidar, an inhibitor of the brain P-gp efflux transporter, increased brain exposure four-fold whereas pretreatment with a high pharmacological dose of the 5-HT_1B antagonist, AR-A000002, had no effect on the binding. Ex-vivo experiments in rats showed no radiometabolites entering brain. [¹¹C]5 also failed to enter monkey brain under baseline conditions.

Conclusions: [¹¹C]4 and [¹¹C]5 show too low brain uptake and specific binding to be useful PET radioligands. Low brain uptake is partly ascribed to efflux transporter action as well as unfavorable conformations.

Keywords: Agonist; Carbon-11; Carbonylation; Radioligand; Serotonin subtype 1B receptor.