In vivo binding, pharmacokinetics and metabolism of the selective M(2) muscarinic antagonists [H-3]AF-DX 116 and [H-3]AF-DX 184 in the anesthetized rat

The pharmacokinetics, in vivo binding and metabolism of two M2 muscarinic receptor antagonists, [3H]AF-DX 116 and [3H]AF-DX 384, were studied in anesthetized rats, which received either the tracer alone or following a saturating injection of atropine. Both radioligands were cleared from the circulation with distribution half-lives of 17 and 14 sec and elimination half-lives of 17 and 40 min for [3H]AF-DX 116 and [3H]AF-DX 384, respectively. A radioactive distribution, predominant in peripheral organs when compared to brain, was found at each time studied after tracer injection. Atropine-displaceable tracer uptake was evidenced at 20–40 min in brain (31%), submandibular glands (26%), spleen (37%) and notably heart (55%) for [3H]AF-DX 116 but only in heart (50%) for [3H]AF-DX 384 at 10–20 min. Regional brain sampling revealed a relatively uniform distribution of [3H]AF-DX 384 and a −45% atropine saturation effect (i.e., specific binding) in the thalamus 20 min after injection. Sequential thin-layer chromatographic studies performed on tissue extracts demonstrated the rapid appearance of labeled metabolites of both radiotracers in brain (but less so in liver) and especially in cardiac tissues, where almost 70% of total radioactivity still corresponded to authentic tracer 40 min after injection. Thus, based on their low blood-brain barrier permeability and the high presence of labeled metabolites in the central nervous system, AF-DX 116 and AF-DX 384 might be more helpful in the study of M2 muscarinic receptors present in heart rather than brain. Labeled with positron emittors, these M2 antagonists might be applicable to the pathophysiological study of disease states, such as cardiomyopathies.