Heteroaryl derivatives of suvorexant as OX1R selective PET ligand candidates: Cu-mediated 18F-fluorination of boroxines, in vitro and initial in vivo evaluation

Abstract Background The orexin receptor (OXR) plays a role in drug addiction and is aberrantly expressed in colorectal tumors. Subtype-selective OXR PET ligands suitable for in vivo use have not yet been reported. This work reports the development of 18 F-labeled OXR PET ligand candidates derived from the OXR antagonist suvorexant and the OX1R-selective antagonist JH112. Results Computational analysis predicted that fluorine substitution (1e) and introduction of the fluorobenzothiazole scaffold (1f) would be suitable for maintaining high OX1R affinity. After multi-step synthesis of 1a–1f, in vitro OXR binding studies confirmed the molecular dynamics calculations and revealed single-digit nanomolar OX1R affinities for 1a–f, ranging from 0.69 to 2.5 nM. The benzothiazole 1f showed high OX1R affinity (K i = 0.69 nM), along with 77-fold subtype selectivity over OX2R. Cu-mediated 18 F-fluorination of boroxine precursors allowed for a shortened reaction time of 5 min to provide the non-selective OXR ligand 18 F1c and its selective OX1R congener 18 F1f in activity yields of 14% and 22%, respectively, within a total synthesis time of 52–76 min. 18 F1c and 18 F1f were stable in plasma and serum in vitro, with logD 7.4 of 2.28 ( 18 F1c) and 2.37 ( 18 F1f), and high plasma protein binding of 66% and 77%, respectively. Dynamic PET imaging in rats showed similar brain uptake of 18 F1c (0.17%ID/g) and 18 F1f (0.15%ID/g). However, preinjection of suvorexant did not significantly block 18 F1c or 18 F1f uptake in the rat brain. Pretreatment with cyclosporine A to study the role of P-glycoprotein (P-gp) in limiting brain accumulation moderately increased brain uptake of 18 F1c and 18 F1f. Accordingly, in vitro experiments demonstrated that the P-gp inhibitor zosuquidar only moderately inhibited polarized, basal to apical transport of 1c (p < 0.05) and had no effect on the transport of 1f, indicating that P-gp does not play a relevant role in brain accumulation of 18 F1c and 18 F1f in vivo. Conclusions The in vitro and in vivo results of 18 F1c and 18 F1f provide a solid basis for further development of suitable OXR PET ligands for brain imaging.