Novel Thienopyrimidine-Based PET Tracers for P2Y(12) Receptor Imaging in the Brain

[Image: see text] The P2Y(12) receptor (P2Y(12)R) is uniquely expressed on microglia in the brain, and its expression level directly depends on the microglial activation state. Therefore, P2Y(12)R provides a promising imaging marker for distinguishing the pro- and anti-inflammatory microglial phenotypes, both of which play crucial roles in neuroinflammatory diseases. In this study, three P2Y(12)R antagonists were selected from the literature, radiolabeled with carbon-11 or fluorine-18, and evaluated in healthy Wistar rats. Brain imaging was performed with and without blocking of efflux transporters P-glycoprotein and breast cancer resistance protein using tariquidar. Low brain uptake in healthy rats was observed for all tracers at baseline conditions, whereas blocking of efflux transporters resulted in a strong (6–7 fold) increase in brain uptake for both of them. Binding of the most promising tracer, [(18)F]3, was further evaluated by in vitro autoradiography on rat brain sections, ex vivo metabolite studies, and in vivo P2Y(12)R blocking studies. In vitro binding of [(18)F]3 on rat brain sections indicated high P2Y(12)R targeting with approximately 70% selective and specific binding. At 60 min post-injection, over 95% of radioactivity in the brain accounted for an intact tracer. In blood plasma, still 40% intact tracer was found, and formed metabolites did not enter the brain. A moderate P2Y(12)R blocking effect was observed in vivo by positron emission tomography (PET) imaging with [(18)F]3 (p = 0.04). To conclude, three potential P2Y(12)R PET tracers were obtained and analyzed for P2Y(12)R targeting in the brain. Unfortunately, the brain uptake appeared low. Future work will focus on the design of P2Y(12)R inhibitors with improved physicochemical characteristics to reduce efflux transport and increase brain penetration.