title: An innovative method for rapid characterisation of phospholipase C activity: SB242,084 competitively antagonises 5-HT2C receptor-mediated [3H]phosphatidylinositol depletion.
abstract: 6-chloro-5-methyl-1-[6-(2-methylpyridin-3-yloxy) pyridin-3-ylcarbamoyl] indoline (SB242,084) is a novel, selective 5-HT(2C) receptor antagonist, but its actions at these sites have been little characterised at the cellular level. We employed a rapid and innovative approach to investigate its functional activity at phospholipase C (PLC)-coupled human 5-HT(2C) receptors expressed in CHO cells. PLC activity was determined as a decrease in the [3H]phosphatidylinositol ([3H]PI) content of cell membranes. Serotonin (5-HT) stimulated [3H]PI depletion (pEC50=8.74), and SB242,084, like mesulergine, completely reversed this action of 5-HT (pK(B)=9.25 and 9.01, respectively). Further, in Schild analysis, SB242,084 behaved as a high affinity competitive antagonist, inducing a parallel, rightward displacement of the 5-HT stimulation isotherm without loss of maximum efficacy. The pA2 of 9.50 was similar to its binding affinity (pKi=9.38). SB242,084 also displayed antagonist properties when PLC activity was examined by conventional determination of [3H]inositol phosphate generation. Employing this parameter, the potency of SB242,084 (pK(B)=9.21) and that of mesulergine (pK(B)=9.06) closely resembled those determined by [3H]PI depletion. In conclusion, determination of [3H]PI depletion constitutes a useful and novel technique to characterise agonist and antagonist properties of ligands at PLC-coupled receptors.
authors.affiliation: Psychopharmacology Department, Institut de Recherches Servier, Croissy-sur-Seine (Paris), France.
authors.forename: D
authors.initials: D
authors.lastname: Cussac
authors.forename: A
authors.initials: A
authors.lastname: Newman-Tancredi
authors.forename: Y
authors.initials: Y
authors.lastname: Quentric
authors.forename: M J
authors.initials: MJ
authors.lastname: Millan
mesh_terms: D000631:Aminopyridines / Q000494:pharmacology*; D000818:Animals; D016466:CHO Cells; D006224:Cricetinae; D004873:Ergolines / Q000494:pharmacology; D006801:Humans; D007211:Indoles / Q000494:pharmacology*; D010716:Phosphatidylinositols / Q000378:metabolism*; D044405:Receptor, Serotonin, 5-HT2C; D011985:Receptors, Serotonin / Q000187:drug effects* / Q000502:physiology; D012701:Serotonin / Q000494:pharmacology; D012702:Serotonin Antagonists / Q000494:pharmacology*; D010738:Type C Phospholipases / Q000378:metabolism*
chemical_list: C107820:6-chloro-5-methyl-1-((2-(2-methylpyrid-3-yloxy)pyrid-5-yl)carbamoyl)indoline; D000631:Aminopyridines; D004873:Ergolines; D007211:Indoles; D010716:Phosphatidylinositols; D044405:Receptor, Serotonin, 5-HT2C; D011985:Receptors, Serotonin; D012702:Serotonin Antagonists; D012701:Serotonin; D010738:Type C Phospholipases; C031649:mesulergine