ABSTRACT
The α3ß4α5 nAChR has been recently shown to be a useful target for smoking cessation pharmacotherapies. Herein, we report on the development and characterization of the α3ß4α5 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 α3ß4 and α3ß4α5 nicotinic receptors. It was demonstrated that while no subtype selectivity was observed for nicotine and nornicotine, anabasine was selective for the α3ß4α5 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 α3ß4 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 α3ß4 and α3ß4α5 nAChRs.
Subject(s)
Chemistry Techniques, Analytical/methods , Chromatography, Affinity , Plant Extracts/chemistry , Receptors, Nicotinic/chemistry , Alkaloids/chemistry , Anabasine/chemistry , Binding Sites , Fabaceae/chemistry , Lycopodiaceae/chemistry , Nicotine/analogs & derivatives , Nicotine/chemistry , Smoke/analysisSubject(s)
Adenosine Triphosphate , Europium , Luminescence , Models, Biological , Spectrometry, FluorescenceABSTRACT
Model melanins synthesized from adrenochrome, DOPA and pyrocatechol quench the peroxyradical-coupled chemiluminescence initiated by electrolysis, photolysis and redox reactions in several selected systems. Calculations of the selfadsorption (inner filter effect) of melanins that decreases chemiluminescence intensity as well as changes in absorption spectra of photolabile substrate-probes such as cytochrome c and rhodamine, lead to the conclusion that melanins scavenge peroxyradicals and inhibit chemiluminescence. The inhibition effect (beta-value) is much higher than that of most powerful antioxidants in all chemiluminescence systems excluding the luminol-hydrogen peroxide one. Donor-acceptor properties of melanins, pertinent to their antioxidizing activity, are discussed.
