Development and characterization of the alpha 3 beta 4 alpha 5 nicotinic receptor cellular membrane affinity chromatography column and its application for on line screening of plant extracts

• Published in: Journal of Chromatography A Vol. 1431; pp. 138 - 144
• Main Authors: Ciesla, L, Okine, M, Rosenberg, A, Dossou, KSS, Toll, L, Wainer, I W, Moaddel, R
• Format: Journal Article
• Language: English
• Published: 29.01.2016
• Subjects: Affinity; Cellular; Chromatography; Extraction plants; Nicotine; Receptors; Screening; Selectivity
• ISSN: 0021-9673
• DOI: 10.1016/j.chroma.2015.12.065

The alpha 3 beta 4 alpha 5 nAChR has been recently shown to be a useful target for smoking cessation pharmacotherapies. Herein, we report on the development and characterization of the alpha 3 beta 4 alpha 5 nicotinic receptor column by frontal displacement chromatography. The binding affinity of the nicotine and minor alkaloids found in tobacco smoke condensates were determined for both the alpha 3 beta 4 and alpha 3 beta 4 alpha 5 nicotinic receptors. It was demonstrated that while no subtype selectivity was observed for nicotine and nornicotine, anabasine was selective for the alpha 3 beta 4 alpha 5 nicotinic receptor. The non-competitive inhibitor binding site was also studied and it was demonstrated while mecamylamine was not selective between subtypes, buproprion showed subtype selectivity for the alpha 3 beta 4 nicotinic receptor. The application of this methodology to complex mixtures was then carried out by screening aqueous-alcoholic solutions of targeted plant extracts, including Lycopodium clavatum L. (Lycopodiaceae) and Trigonella foenum graecum L. (Fabaceae) against both the alpha 3 beta 4 and alpha 3 beta 4 alpha 5 nAChRs.