Abstract
The two major brain nicotinic acetylcholine receptors (nAChRs) involved in cognitive function are the α4β2 and α7 nAChRs. A "methyl scan" of the pyrrolidinyl ring was used to detect differences in the interactions of nicotine with these two receptors. Each methylnicotine was investigated using voltage-clamp and radioligand binding techniques. Methylation at each ring carbon elicited unique changes in nicotine’s receptor interactions. Replacing the 1'-N methyl with an ethyl group or adding a second 1'-N methyl group significantly reduced interaction with α4β2 but not α7 receptors. 2'-methylation uniquely enhanced binding and agonist potency at α7 receptors. Although 3'and 5'trans-methylations were much better tolerated by α7 receptors than α4β2 receptors, both 4’ methylations decreased potency and efficacy at α7 receptors much more than at α4β2 receptors. Cis-5'-methylnicotine lacked agonist activity and displayed a low affinity at both receptors, but trans-5'methylnicotine retained considerable α7 receptor activity. Differences between the two 5’-methylated analogs of the potent pyridyl oxymethylene-bridged nicotine analog A84543 were consistent with what was found for the 5'-methylnicotines. Computer docking of the methylnicotines to the Lymnaea AChBP crystal structure containing two "persistent" waters predicted most of the changes in receptor affinity that were observed with methylation, particularly the lower affinities of the cis-methylnicotines. The much smaller effects of 1', 3'- and 5'-methylations and the greater effects of 2'- and 4'-methylations on nicotine α7 nAChR interaction might be exploited for the design of new drugs based on the nicotine scaffold.
SIGNIFICANCE STATEMENT Using a comprehensive methyl scan approach we show that the orthosteric binding sites for acetylcholine and nicotine in the two major brain nicotinic acetylcholine receptors interact differently with the pyrrolidinium ring of nicotine, and suggested reasons for the higher affinity of nicotine for the heteromeric receptor. Potential sites for nicotine structure modification have been identified that may be useful in the design of new drugs targeting these receptors.
- Acetylcholine receptors
- Cholinergic pharmacology
- Ligand-gated ion channels
- Molecular modeling
- Nicotinic cholinergic receptors
- Structure-activity relationships
- The American Society for Pharmacology and Experimental Therapeutics