Synthesis and evaluation of radiolabeled piperazine derivatives of vesamicol as SPECT agents for cholinergic neurons.

To diagnose and investigate neurodegenerative diseases affecting cholinergic neuron density, piperazine derivatives of vesamicol were synthesized and evaluated. Previously, we reported that trans-5-iodo-2-hydroxy-3-[4-phenylpiperazinyl] tetralin (DRC140, 1) possessed high selectivity for vesicular acetylcholine transporter (VAChT). In present study of the effect of alkyl substituents, we observed that the introduction of a methyl group into the ortho or meta positions of the phenyl group of 1 increased affinity for VAChT. trans-5-Iodo-2-hydroxy-3-[4-[2-methylphenyl] piperazinyl]tetralin (2) displayed high affinity and specificity for VAChT. The regional distributions of radioactivity in the rat brain correlated well with known patterns of central cholinergic innervation. [(123)I]2 is a potentially useful compound for SPECT imaging.

Piperazine analog of vesamicol: in vitro and in vivo characterization for vesicular acetylcholine transporter.

The probes to detect vesicular acetylcholine transporter (VAChT) in vivo are important to evaluate the mapping and function in cholinergic system. To develop high-specific and high-affinity radiotracer for single photon emission computed tomography, we investigated piperazine analogs which replaced the piperidine ring of (-)-vesamicol with a piperazine ring. We found that the piperazine analog of iodobenzovesamicol, trans-5-iodo-2-hydroxy-3-[4-phenylpiperazinyl] tetralin (DRC140), had high affinity for VAChT in rat brain. We carried out binding assay in subcellular fraction of the rat brain. The highest B(max) for [(125)I]-DRC140 binding was observed in the synaptic vesicle fraction (1,751 fmol/mg protein), followed by the crude vesicle (821 fmol/mg protein) and the P2 fraction (187 fmol/mg protein). These K(d) values were similar to the affinity of highly purified synaptic vesicular fraction (K(d) = 0.3 nM) with a one-site model. The possibility that [(125)I]-DRC140 recognizes sigma receptor was excluded by our finding large inhibition constants (K(i) = 849 nM for haloperidol, K(i) = 3,052 nM for 1,3-di(2-tolyl)guanidine). In vivo distribution studies with the [(123)I]-DRC140 in rats showed a rapid brain uptake. The highest brain area was in striatum, followed by frontal cortex, occipital cortex, and hippocampus. The lowest brain area was cerebellum. The radioactivity of high-accumulated areas in ex vivo autoradiography was reduced by a preinjection of (-)-vesamicol and these levels were reduced to the radioactivity in cerebellum. These results show that [(125)I]-DRC140 can provide extremely high specific tracer with excellent brain permeability as a ligand for single photon emission computed tomography.

Direct 99mTc labeling of human immunoglobulin with an insoluble macromolecular Sn(II) complex.

An insoluble macromolecular Sn(II)(R-Sn) complex which strongly binds Sn(II) by chelation was applied to the direct 99mTc labeling of human immunoglobulin(IgG) to minimize the influence of Sn(II). 99mTc labeling was achieved at greater than 90% yield simply by the short-term mixing of IgGa containing > 2-SH groups per IgG molecule, 99mTc pertechnetate, and the R-Sn complex in pH 7 solution. The 99mTc-IgGa obtained by this labeling method has high stability based on the thiol-specific binding of 99mTc without transchelation from another weakly bound 99mTc-complex.