Synthesis and in vitro evaluation of new benzovesamicol analogues as potential imaging probes for the vesicular acetylcholine transporter

Abstract

Our goal was to synthesize new stereospecific benzovesamicol analogues, which could potentially be used as SPECT or PET radioligands for the vesicular acetylcholine transporter (VAChT). This paper describes the chemical synthesis, resolution and determination of binding affinity for four enantiomeric pairs of derivatives. Their intrinsic affinities were determined by competition against binding of [ 3H]vesamicol to human VAChT. Of the eight enantiomers, ( E)-( R, R)-5-AOIBV [( R, R)- 3], and ( R, R)-5-FPOBV [( R, R)- 4] displayed the highest binding affinities for VAChT ( K d = 0.45 and 0.77 nM, respectively), which indicated that an elongation of the chain from 5-idodo as in the case of 5-iodobenzovesamicol (5-IBVM), to a 5-( E)-3-iodoallyloxy or 5-fluoropropoxy substituent, as in 5-AOIBV and 5-FPOBV, respectively, was very well tolerated at the vesamicol binding site. The enantiomer ( R, R)-4-MAIBV [( R, R)- 16], which retains the basic structure of (−)-5-IBVM but possess an additional aminomethyl substituent in the 4-position of the piperidine ring, displayed lower binding affinity ( K d = 8.8 nM). Nevertheless, the result suggests that substitution at this position may be an interesting alternative to investigate for development of new benzovesamicol analogues. As expected, the corresponding ( S, S) enantiomers displayed lower K d values, they were approximately 10-fold lower in the case of ( S, S)-5-FPOBV ( K d = 8.4 nM) and ( E)-( S, S)-5-AOIBV ( K d = 4.3 nM). ( R, R)- 3, and ( R, R)- 4 showed the same high affinity for VAChT as (−)-5-IBVM and may be suitable as imaging agents of cholinergic nerve terminals.