Radiosynthesis and Evaluation of 18 F-Labeled Deuterated Radioligand for Positron Emission Tomography Imaging of Cholesterol 24-Hydroxylase

High Resolution Image Download MS PowerPoint Slide Brain cholesterol homeostasis is critical for neuronal function and primarily regulated by cholesterol 24-hydroxylase (CYP46A1). Dysregulation of CYP46A1 has been implicated in Alzheimer’s disease (AD) and Huntington’s disease (HD). Building on the clinically validated positron emission tomography (PET) tracer [ 18 F]CHL-2205, we designed a deuterated isotopologue, CHL-2205 -d 3, targeting the amide N -methyl group to enhance stability and enable mechanistic studies. Compound 5 exhibited high CYP46A1 affinity (IC 50 = 0.38 nM; K i = 0.22 nM). Radiosynthesis via copper-mediated [ 18 F]fluorination afforded [ 18 F] 5 in 31.5 ± 1.5% non-decay-corrected radiochemical yield and high molar activity (>95 GBq/μmol). Autoradiography and PET imaging in mice demonstrated robust brain uptake, heterogeneous regional distribution, and specific target engagement. Radiometabolite analysis confirmed that brain radioactivity was mainly attributable to intact [ 18 F] 5, with a pharmacokinetics comparable to that of [ 18 F]CHL-2205. [ 18 F] 5 preserves [ 18 F]CHL-2205 imaging performance and provides a deuterated PET tool for quantitative bioanalysis and integrated PET–deuterium metabolic imaging (DMI) studies of brain cholesterol metabolism.