Identification and regional distribution in rat brain of radiometabolites of the dopamine transporter PET radioligand [11C]PE2I

Purpose We aimed to determine the composition of radioactivity in rat brain after intravenous administration of the dopamine transporter radioligand, [C-11]PE2I. Methods PET time-activity curves (TACs) and regional brain distribution ex vivo were measured using no-carrier-added [C-11]PE2I. Carrier-added [C-11]PE2I was administered to identify metabolites with high-performance liquid radiochromatography (RC) or RC with mass spectrometry (LC-MS and MS-MS). The stability of [C-11]PE2I was assessed in rat brain homogenates. Results After peak brain uptake of no-carrier-added [C-11]PE2I, there was differential washout rate from striata and cerebellum. Thirty minutes after injection, [C-11]PE2I represented 10.9 +/- 2.9% of the radioactivity in plasma, 67.1 +/- 11.0% in cerebellum, and 92.5 +/- 3.2% in striata, and was accompanied by two less lipophilic radiometabolites. [C-11]PE2I was stable in rat brain homogenate for at least 1 h at 37 degrees C. LC-MS identified hydroxylated PE2I (1) (m/z 442) and carboxyl-desmethyl-PE2I (2) (m/z 456) in brain. MS-MS of 1 gave an m/z 442 -> 424 transition due to H2O elimination, so verifying the presence of a benzyl alcohol group. Metabolite 2 was the benzoic acid derivative. Ratios of ex vivo measurements of [C-11]PE2I, [C-11]1, and [C-11]2 in striata to their cognates in cerebellum were 6.1 +/- 3.4, 3.7 +/- 2.2 and 1.33 +/- 0.38, respectively, showing binding selectivity of metabolite [C-11]1 to striata. Conclusion Radiometabolites [C-11]1 and [C-11]2 were characterized as the 4-hydroxymethyl and 4-carboxyl analogs of [C-11]PE2I, respectively. The presence of the pharmacologically active [C-11]1 and the inactive [C-11]2 is a serious impediment to successful biomathematical analysis.