title: AL-3138 antagonizes FP prostanoid receptor-mediated inositol phosphates generation: comparison with some purported FP antagonists.
abstract: The aim of this study was to pharmacologically characterize the antagonist properties of a novel prostaglandin F2alpha (PGF2alpha) analogue (11-deoxy-16-fluoro PGF2alpha; AL-3138) using a variety of second-messenger assays of prostaglandin receptor subtypes. A detailed comparison was made between AL-3138 and some purported FP receptor antagonists such as PGF2alpha dimethylamine, PGF2alpha dimethylamide, glibenclamide and phloretin using the FP receptor-mediated phosphoinositide turnover assay in A7r5 rat thoracic aorta smooth muscle cells and mouse Swiss 3T3 fibroblasts. The potency and efficacy of AL-3138 as an FP receptor agonist were: EC50 = 72.2 +/- 17.9 nM (Emax = 37%) (n = 3) in A7r5 cells and EC50 = 20.5 +/- 2.8 nM (Emax = 33%) (n = 5) in 3T3 cells. Being a partial agonist, the antagonist potency of AL-3138 against fluprostenol in A7r5 cells was determined to be: Ki = 296 +/- 17 nM (n = 3) and Kb = 182 +/- 44 nM (n = 5) (-log Kb = 6.79 +/- 0.1). AL-3138 exhibited very minimal or no antagonistic effects at EP2, EP4, DP and TP prostaglandin receptors. Both PGF2alpha dimethylamide and PGF2alpha dimethylamine were inactive as FP receptor antagonists, whereas phloretin and glibenclamide were very weak and had -log Kb values of 5.28 +/- 0.09 (n = 3) and 3.58 +/- 0.32 (n = 3), respectively. However, phloretin antagonized functional responses of EP2 and DP prostanoid receptors, and also the V1-vasopressin receptor. AL-3138 competed for [3H]PGF2alpha binding to FP receptors with a relatively high affinity (IC50high = 312 +/- 95 nM) matching its functional antagonist potency. In conclusion, AL-3138 is a more potent and selective FP receptor antagonist than glibenclamide, phloretin, PGF2alpha dimethylamide and PGF2alpha dimethylamine and is therefore a unique and novel pharmacological tool to help characterize FP receptor-mediated functions.
authors.affiliation: Molecular Pharmacology Unit, Alcon Research Ltd, Fort Worth, TX 76134, USA. naj.sharif@alconlabs.com
authors.forename: N A
authors.initials: NA
authors.lastname: Sharif
authors.forename: J Y
authors.initials: JY
authors.lastname: Crider
authors.forename: T L
authors.initials: TL
authors.lastname: Davis
mesh_terms: D016475:3T3 Cells; D000818:Animals; D001667:Binding, Competitive / Q000187:drug effects; D016466:CHO Cells; D002417:Cattle; D002460:Cell Line; D002461:Cell Line, Transformed; D003338:Corpus Luteum / Q000378:metabolism; D006224:Cricetinae; D000242:Cyclic AMP / Q000378:metabolism; D015237:Dinoprost / Q000031:analogs & derivatives* / Q000235:genetics* / Q000378:metabolism / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D005260:Female; D005905:Glyburide / Q000494:pharmacology; D006801:Humans; D007295:Inositol Phosphates / Q000378:metabolism*; D008566:Membranes / Q000378:metabolism; D051379:Mice; D010693:Phloretin / Q000494:pharmacology; D011448:Prostaglandin Antagonists / Q000494:pharmacology*; D011461:Prostaglandins F, Synthetic / Q000494:pharmacology; D011869:Radioligand Assay; D011982:Receptors, Prostaglandin / Q000819:agonists / Q000037:antagonists & inhibitors* / Q000502:physiology
chemical_list: C422825:AL-3138; D007295:Inositol Phosphates; D011448:Prostaglandin Antagonists; D011461:Prostaglandins F, Synthetic; D011982:Receptors, Prostaglandin; C034671:prostaglandin F2alpha receptor; C121227:AL 8810; C006326:fluprostenol; C032516:prostaglandin F2alpha N-dimethylamide; D015237:Dinoprost; D000242:Cyclic AMP; D010693:Phloretin; D005905:Glyburide