Pre-clinical validation of a novel alpha-7 nicotinic receptor radiotracer, [3H]AZ11637326: Target localization, biodistribution and ligand occupancy in the rat brain

The alpha-7 neuronal nicotinic receptor is a novel pharmacological target for psychiatric and cognitive disorders. Selective radiotracer tools for pre-clinical receptor occupancy can facilitate the interpretation of the biological actions of small molecules at a target receptor. We discovered a high affinity nicotinic alpha-7 subtype-selective ligand, AZ11637326, with physical–chemical and pharmacokinetic properties suitable for an in vivo radioligand tool. [3H]AZ11637326 synthesis by tritiodehalogenation of the corresponding tribromide precursor yielded a high specific activity radiotracer with high affinity alpha-7 receptor binding in the rat hippocampus determined by autoradiography (Kd = 0.2 nM). When [3H]AZ11637326 was administered to rats by intravenous bolus, rapid uptake was measured in the brain followed by a 3–4 fold greater specific binding in regions containing the alpha-7 receptor (frontal cortex, hippocampus, hypothalamus and midbrain) when compared to non-target regions (striatum and cerebellum). Systemic administration of the high affinity alpha-7 receptor antagonist, methyllycaconitine (MLA), or pretreatment with alpha-7 selective agonists (AR-R17779, PyrQTC, DBCO-4-POM, and DBCO-3-POM) significantly blocked the alpha-7 specific binding of [3H]AZ11637326 in the rat brain. The rank order of ligand ED50 values for in vivo alpha-7 receptor occupancy in rat hippocampus was: DBCO-4-POM > DBCO-3-POM ∼ MLA > PyrQTC > AR-R17779. The occupancy affinity shift was consistent with in vitro binding affinity in autoradiography. Our studies established the optimal conditions for [3H]AZ11637326 in vivo specific binding in the rat brain and support the use of [3H]AZ11637326 as a pre-clinical tool for assessment of novel alpha-7 compounds in drug discovery.

► The alpha-7 neuronal nicotinic receptor is a drug target for CNS disorders.
► Pre-clinical receptor occupancy at a target receptor will aid drug discovery.
► We report a high affinity alpha-7 subtype-selective radiotracer, [3H]AZ11637326.
► CNS biodistribution and pharmacological selectivity in vivo is optimal in rat.
► Selective alpha-7 agonist occupancy in vivo matches functional data trends.