title: Radiosynthesis of [123I]N-methyl-4-iododexetimide and [123I]N-methyl-4-iodolevetimide: in vitro and in vivo characterisation of binding to muscarinic receptors in the rat heart.
abstract: [123I]N-methyl-4-iododexetimide, [123I]MIDEX, and its pharmacologically inactive enantiomer [123I]N-methyl-4-iodolevetimide, [123I]MILEV were prepared via electrophilic iododesilylation using Chloramine-T as oxidising agent followed by N-methylation using methyl iodide. The radiotracers were purified with semi-preparative HPLC with radiochemical yields of 80 +/- 11% (n = 6). The average time of synthesis was 100 min with specific activity > 2000 Ci/mmol. In vitro, the binding of [123I]MIDEX, after addition of carrier, measured on homogenates of rat atrium was Bmax = 4.5 +/- 0.4 pmol/mg protein, Kd = 3.3 +/- 0.2 nM while in the ventricle Bmax = 2.3 +/- 0.2 pmol/mg protein, Kd = 4.0 +/- 1.4 nM. In vitro, the binding of [123I]MILEV was non-specific. The in vivo biodistribution of [123I]MIDEX showed high uptake in the atrium (3.2% ID/g) and left and right ventricles (2.2, 2.5% ID/g respectively) at 5 min followed by clearance. High heart-to-lung and moderate liver-to-lung ratios were obtained during 60 min. Radioactivity in the atrium and ventricles was reduced by pre-administration of the m-AChR antagonist MQNB (1 mg/kg). Pretreatment of rats with other m-AChR ligands, pirenzapine (M1), methoctramine (M2) and 4-DAMP (M3) also resulted in reduction of [123I]MIDEX uptake with methoctramine being the most potent. [123I]MIDEX distribution in the rat heart was not significantly inhibited by pre-administration of selective adrenergic drugs. The uptake was highly stereoselective since the inactive enantiomer, [123I]MILEV, demonstrated very low myocardial retention. The stability of [123I]MIDEX was evaluated by performing a metabolite study on atrium samples which revealed unchanged radiotracer 60 min postinjection. These results suggest that [123I]MIDEX may be a useful single photon agent for in vivo imaging of myocardial m-AChR in humans with [123I]MILEV offering the potential of assessing non-specific binding of the active tracer.
authors.affiliation: Biomedicine and Health Program, Australia Nuclear Science and Technology Organisation, Sydney.
authors.forename: M
authors.initials: M
authors.lastname: Kassiou
authors.forename: K
authors.initials: K
authors.lastname: Mardon
authors.forename: A G
authors.initials: AG
authors.lastname: Katsifis
authors.forename: L
authors.initials: L
authors.lastname: Najdovski
authors.forename: B
authors.initials: B
authors.lastname: Dikic
authors.forename: F
authors.initials: F
authors.lastname: Mattner
authors.forename: R M
authors.initials: RM
authors.lastname: Lambrecht
authors.forename: R J
authors.initials: RJ
authors.lastname: Hicks
authors.forename: P
authors.initials: P
authors.lastname: Eu
authors.forename: C
authors.initials: C
authors.lastname: Loc'h
mesh_terms: D000818:Animals; D001667:Binding, Competitive / Q000187:drug effects; D018678:Cholinergic Agents / Q000138:chemical synthesis* / Q000494:pharmacology; D002851:Chromatography, High Pressure Liquid; D003909:Dexetimide / Q000031:analogs & derivatives* / Q000138:chemical synthesis / Q000494:pharmacology; D006325:Heart Atria / Q000378:metabolism; D006352:Heart Ventricles / Q000378:metabolism; D007457:Iodine Radioisotopes; D007553:Isotope Labeling; D008297:Male; D009206:Myocardium / Q000378:metabolism*; D051381:Rats; D017208:Rats, Wistar; D011976:Receptors, Muscarinic / Q000187:drug effects / Q000378:metabolism*; D014018:Tissue Distribution
chemical_list: D018678:Cholinergic Agents; D007457:Iodine Radioisotopes; C094599:N-methyl-4-iododexetimide; D011976:Receptors, Muscarinic; D003909:Dexetimide