Structure-Activity Relationships of Methoctramine-Related Polyamines as Muscular Nicotinic Receptor Noncompetitive Antagonists. 2. Role of Polymethylene Chain Lengths Separating Amine Functions and of Substituents on the Terminal Nitrogen Atoms.

Polymethylene tetraamine methoctramine (1) is a prototypical antimuscarinic ligand endowed with a significant affinity for muscular nAChRs. Thus, according to the universal template approach, structural modifications were performed on 1 in order to improve affinity and selectivity for the muscle-type nAChR. The polyamine derivatives synthesized were tested at both frog rectus and Torpedo nAChRs and at guinea pig left atria (M2) and ileum longitudinal muscle (M3) mAChRs. All of the compounds, like prototype 1, were noncompetitive antagonists of nicotinic receptors while being competitive antagonists at M2 and M3 mAChRs. The biological profile of polyamines 4-7 revealed that increasing the number of amine functions and the chain length separating these nitrogen atoms led to a significant improvement in potency at nAChRs. Moreover, the role of the number and type of amine functions in the interaction with nAChRs was further investigated through the synthesis of compounds 9 and 10. Tetraamines 8 and 11, bearing a rather rigid spacer between the nitrogen atoms instead of the very flexible polymethylene chain, displayed a profile similar to that of 1 at nAChRs, whereas a significant decrease in potency was observed at mAChRs. Tetraamine 12, bearing a 2-methoxyphenethyl group, was less potent than 1, whereas tetraamine 13, carrying a diphenylethyl moiety, was more potent than 1, confirming that an increase in size of the hydrophobic group on the terminal nitrogen atoms increases significantly the binding affinity for nAChRs. Tetraamines 14-17 were significantly more potent than prototype 2 at both frog rectus and Torpedo nAChRs, confirming that an increase in the distance between the amine functions results in a parallel increase in the affinity for nAChRs. To gain insight into the mode of interaction of polymethylene tetraamines with nAChRs, photolabile (19 and 20) and fluorescent (21 and 22) compounds were synthesized. A most intriguing finding was the observation that 19, which bears two identical azido groups on the terminal nitrogen atoms, was found to bind the Torpedo nAChR with a 1:1 stoichiometry, suggesting a U-shaped conformation in the receptor interaction. Moreover, the high potency shown by fluorescent compounds 21 and 22 appears promising for a further characterization of the polymethylene tetraamines binding site with the muscletype nAChR

Structure-activity relationships of methoctramine-related polyamines as muscular nicotinic receptor noncompetitive antagonists. 3. Effect of inserting the tetraamine backbone into a macrocyclic structure

The present article expands on the study of another aspect of structure-activity relationships of the polymethylene tetraamines, namely, the effect of inserting the tetraamine backbone into a macrocyclic structure. To this end, compounds 8-12 were designed by linking the two terminal nitrogen atoms of prototype methoctramine 2 to an aryl moiety. Alternatively, 2 was first modified to achieve compounds 6 and 7, which in turn were cyclized by linking the two terminal primary amine functions to a polyphenyl spacer, affording 13-20. All the compounds were tested on muscle-type nAChRs and most of them as well on AChE. Furthermore, selected compounds were tested also on peripheral M2 and M3 mAChRs. All these cyclic derivatives, like prototypes, were potent noncompetitive antagonists at both frog and Torpedo nAChRs, suggesting that polyamines do not need to be linear or in extended conformation to optimally interact with the nicotinic channel; rather, they may bind in a U-shaped conformation. Relative to muscarinic activity, macrocyclic compounds 10, 13, 14, and 20, in contrast with the profile displayed by 2, were almost devoid of affinity. It is derived that an aryl spacer is detrimental to the interaction of polyamines with mAChRs. Finally, all the diamine diamides investigated in this study were much less potent in inhibiting AChE activity than prototype 3, suggesting that a macrocyclic structure may not be suitable for AChE inhibition