Development and Preclinical Characterization of [¹⁸F]H3-2406 and [¹⁸F]H3-2407 for Positron Emission Tomography...

The histamine subtype 3 receptor (H₃R) is a G protein-coupled receptor involved in various central nervous system (CNS) disorders. We herein describe the identification and preclinical evaluation of two H₃R antagonists: compounds 3 (H3-2406, K_i = 2.87 nM) and 4 (H3-2407, K_i = 3.15 nM) as potential PET radioligands. Both were radiolabeled with fluorine-18 using a copper-mediated method. Among them, [¹⁸F]3 showed high radiochemical yield (32.4%), molar activity (103 GBq/μmol), and moderate brain uptake (SUV = 2.4), with regional distribution matching known H₃R expression. [¹⁸F]3 also exhibited favorable pharmacokinetics, high metabolic stability and negligible efflux by transporter proteins. However, relatively high cerebellar uptake, along with dedicated blocking studies, suggested off-target binding to the sigma-1 receptor, likely due to limited selectivity over sigma-1 (69-fold). These findings highlight [¹⁸F]3 as a promising lead for H₃R imaging, with future work aimed at improving its selectivity and minimizing off-target binding.