(3-iodobenzyl)trozamicol and vesamicol

Vesamicol derivatives are promising candidates as ligands for the vesicular acetylcholine transporter (VAChT) to enable in vivo imaging of cholinergic deficiencies if applied as positron emission tomography radiotracers. So far, optimization of the binding affinity of vesamicol-type ligands was hampered by the lack of respective quantitative structure-activity relationships. We developed the first quantitative model to predict, from molecular structure, the binding affinity of vesamicol-type ligands toward VAChT employing comparative molecular field analysis (CoMFA) for a set of 37 ligands, covering three different structural types (4-phenylpiperidine, spiro, and tropan derivatives of vesamicol). The prediction capability was assessed by leave-one-out cross-validation (LOO) and through leaving out and predicting 50% of the compounds selected such that both the training and the prediction sets cover almost the whole range of experimental data. The statistics indicate a significant prediction power of the models ( q (2) (LOO) = 0.66, q (2) (50% out) = 0.59-0.74). The discussion includes detailed analyses of CoMFA regions critical for ligand-VAChT binding, identifying structural implications for high binding affinity.

Topics: Binding Sites; Fluorobenzenes; Iodobenzenes; Ligands; Models, Molecular; Molecular Structure; Piperidines; Quantitative Structure-Activity Relationship; Reproducibility of Results; Static Electricity; Stereoisomerism; Vesicular Acetylcholine Transport Proteins

m-(Iodobenzyl)trozamicol (MIBT) is a recently discovered vesamicol analogue. It has been shown that radiolabelled [125I]MIBT can be used as a marker of cholinergic innervation in the heart as well as in the brain. The purpose of this study was to analyze the direct effects of MIBT on the atrioventricular and intraventricular conduction in addition to the coronary blood flow using the canine isolated, blood-perfused atrioventricular node preparation. Intracoronary administration of MIBT suppressed the atrioventricular and intraventricular conduction, while it increased the coronary blood flow. The effect and duration of action on the intraventricular conduction was less pronounced compared with other effects. Moreover, the doses of MIBT needed to cause negative dromotropic and coronary vasodilator effects in this study was much greater than those needed for imaging the cardiac cholinergic innervation. Pretreatment of the preparations with a muscarinic receptor antagonist, atropine, did not block these effects of MIBT, suggesting that MIBT may possess muscarinic receptor-independent ion channel activity in the cardiac conduction system and coronary arteries.

Topics: Animals; Atrioventricular Node; Atropine; Cholinergic Antagonists; Coronary Circulation; Dogs; Dose-Response Relationship, Drug; Iodobenzenes; Perfusion; Piperidines; Receptors, Cholinergic

The vesamicol analogue, meta-[(125)I]iodobenzyltrozamicol [(+)-[(125)I]MIBT] was evaluated as a probe for the in vitro labeling of the vesicular acetylcholine transporter in primate brain. In the striatum, (+)-[(125)I]MIBT bound a single high-affinity site with a Kd value of 4.4 +/- 0.7 nM. Competition for (+)-[(125)I]MIBT binding to the striatum by a group of vesamicol analogues displayed a pharmacological profile similar to the rank order of potency previously observed for the vesicular acetylcholine transporter on Torpedo synaptic vesicles. High-affinity binding of (+)-[(125)I]MIBT in the occipital cortex was characterized by a Kd value of 4.6 +/- 1.1 nM. However, the rank order of potency for inhibition of (+)-[(125)I]MIBT binding to the occipital cortex by the same test compounds differed from that observed in the striatum. The results suggest that (+)-[(125)I]MIBT is a reliable probe of the vesicular acetylcholine transporter in primate striatum, but its binding in primate occipital cortex is more complex.

Topics: Acetylcholine; Animals; Brain Chemistry; Carrier Proteins; Iodobenzenes; Ligands; Macaca mulatta; Male; Membrane Transport Proteins; Neostriatum; Neuromuscular Depolarizing Agents; Piperidines; Radioligand Assay; Synaptic Vesicles; Vesicular Acetylcholine Transport Proteins; Vesicular Transport Proteins

Racemic (m-iodobenzyl)trozamicol (6, MIBT), a high-affinity vesamicol receptor ligand, was radiolabeled, resolved, and evaluated in rats. Following iv injection, (+)- and (-)-[125I]MIBT achieved initial brain levels of 0.57 and 0.92% dose/g of tissue, respectively. The level of (+)-[125I]MIBT subsequently declined by 74% within 3 h, while that of (-)-[125I]MIBT remained stable for the duration. Ex vivo autoradiographic mapping of (-)-[125I]MIBT distribution in rat brain revealed a pattern which was inconsistent with central cholinergic innervation. However, high levels of (+)-[125I]MIBT were observed over the amygdala, striatum, nucleus accumbens, olfactory tubercle, and nuclei of the fifth and seventh cranial nerves, while moderate to low levels were detected within the cortex, hippocampus, and cerebellum. Thus, the distribution of (+)-[125I]MIBT parallels that of other presynaptic cholinergic markers. Co-injection of (+)-[125I]MIBT with 4-aminobenzovesamicol (2b), a potent vesamicol receptor ligand, reduced the levels of radiotracer in the striatum, cortex, and cerebellum by 58, 35, and 9%, respectively. Thus, (+)-[125I]MIBT binds to vesamicol receptors in vivo. In contrast, coadministration of (+)-[125I]MIBT with haloperidol (0.5 mumol/kg), reduced radiotracer levels in the cortex and cerebellum by 34 and 59%, respectively, while increasing the levels in the striatum by 32%. We conclude that although the distribution of (+)-[125I]MIBT qualitatively reflects cholinergic innervation, a fraction of radiotracer in the cortex and cerebellum is bound to sigma receptors.

Topics: Animals; Autoradiography; Brain; Cerebellum; Cerebral Cortex; Choline; Corpus Striatum; Iodine Radioisotopes; Iodobenzenes; Isotope Labeling; Kinetics; Male; Molecular Structure; Piperidines; Rats; Rats, Wistar; Receptors, sigma; Tissue Distribution