Noncompetitive Inhibition of 5-HT3 Receptors by Citral, Linalool, and Eucalyptol Revealed by Nonlinear Mixed-Effects Modeling

• Published in: The Journal of pharmacology and experimental therapeutics Vol. 356; no. 3; p. 549
• Main Authors: Jarvis, Gavin E, Barbosa, Roseli, Thompson, Andrew J
• Format: Journal Article
• Language: English
• Published: United States 01-03-2016
• Subjects: Animals; Cyclohexanols - chemistry; Cyclohexanols - isolation & purification; Cyclohexanols - pharmacology; Female; Guinea Pigs; HEK293 Cells; Humans; Lippia; Male; Monoterpenes - chemistry; Monoterpenes - isolation & purification; Monoterpenes - pharmacology; Nonlinear Dynamics; Protein Structure, Secondary; Rats; Rats, Wistar; Receptors, Serotonin, 5-HT3 - chemistry; Receptors, Serotonin, 5-HT3 - metabolism; Serotonin 5-HT3 Receptor Antagonists - chemistry; Serotonin 5-HT3 Receptor Antagonists - isolation & purification; Serotonin 5-HT3 Receptor Antagonists - pharmacology; Xenopus laevis
• ISSN: 1521-0103
• DOI: 10.1124/jpet.115.230011

Citral, eucalyptol, and linalool are widely used as flavorings, fragrances, and cosmetics. Here, we examined their effects on electrophysiological and binding properties of human 5-HT3 receptors expressed in Xenopus oocytes and human embryonic kidney 293 cells, respectively. Data were analyzed using nonlinear mixed-effects modeling to account for random variance in the peak current response between oocytes. The oils caused an insurmountable inhibition of 5-HT-evoked currents (citral IC50 = 120 µM; eucalyptol = 258 µM; linalool = 141 µM) and did not compete with fluorescently labeled granisetron, suggesting a noncompetitive mechanism of action. Inhibition was not use-dependent but required a 30-second preapplication. Compound washout caused a slow (∼180 seconds) but complete recovery. Coapplication of the oils with bilobalide or diltiazem indicated they did not bind at the same locations as these channel blockers. Homology modeling and ligand docking predicted binding to a transmembrane cavity at the interface of adjacent subunits. Liquid chromatography coupled to mass spectrometry showed that an essential oil extracted from Lippia alba contained 75.9% citral. This inhibited expressed 5-HT3 receptors (IC50 = 45 µg ml(-1)) and smooth muscle contractions in rat trachea (IC50 = 200 µg ml(-1)) and guinea pig ileum (IC50 = 20 µg ml(-1)), providing a possible mechanistic explanation for why this oil has been used to treat gastrointestinal and respiratory ailments. These results demonstrate that citral, eucalyptol, and linalool inhibit 5-HT3 receptors, and their binding to a conserved cavity suggests a valuable target for novel allosteric modulators.