title: Identification of nonpeptidic urotensin II receptor antagonists by virtual screening based on a pharmacophore model derived from structure-activity relationships and nuclear magnetic resonance studies on urotensin II.
abstract: The vasoactive cyclic 11-amino acid peptide urotensin II (U-II) has recently been discovered as the endogenous ligand of the orphan G-protein-coupled receptor GPR14. As U-II might be involved in the regulation of cardiovascular homeostasis and pathology, a nonpeptidic GPR14/U-II antagonist is of considerable basic and therapeutic interest. We have performed structure-activity relationship studies on U-II by investigating 25 peptide analogues to mobilize intracellular calcium in GPR14-transfected CHO cells, demonstrating that only the side chains of the residues Trp-7, Lys-8, and Tyr-9 are required for receptor recognition and activation. The solution structure of U-II derived by nuclear magnetic resonance has served as a structural template for a three-dimensional three point pharmacophore query for the virtual screening of the Aventis compound repository for nonpeptidic U-II receptor antagonists. Highly active lead compounds of six different scaffold classes could be identified, antagonizing the biological activity of U-II in vitro. The most potent compound identified by the virtual screening approach, 1-(3-carbamimidoyl-benzyl)-4-methyl-1H-indole-2-carboxylic acid (naphthalen-1-ylmethyl)amide, reveals an IC(50) of 400 nM in a functional fluorometric imaging plate reader assay and constitutes a promising lead.
authors.affiliation: Lead Generation Chemistry, Aventis Pharma Germany GmbH, Industriepark Hoechst, D-65926 Frankfurt am Main, Germany.
authors.forename: Stefanie
authors.initials: S
authors.lastname: Flohr
authors.forename: Michael
authors.initials: M
authors.lastname: Kurz
authors.forename: Evi
authors.initials: E
authors.lastname: Kostenis
authors.forename: Alexandre
authors.initials: A
authors.lastname: Brkovich
authors.forename: Alain
authors.initials: A
authors.lastname: Fournier
authors.forename: Thomas
authors.initials: T
authors.lastname: Klabunde
mesh_terms: D019943:Amino Acid Substitution; D000818:Animals; D016466:CHO Cells; D002118:Calcium / Q000378:metabolism; D020650:Combinatorial Chemistry Techniques; D006224:Cricetinae; D016208:Databases, Factual; D005470:Fluorometry; D006801:Humans; D007211:Indoles / Q000138:chemical synthesis* / Q000737:chemistry / Q000494:pharmacology; D008024:Ligands; D009682:Magnetic Resonance Spectroscopy; D008958:Models, Molecular; D009281:Naphthalenes / Q000138:chemical synthesis* / Q000737:chemistry / Q000494:pharmacology; D010446:Peptide Fragments / Q000138:chemical synthesis* / Q000737:chemistry / Q000494:pharmacology; D011956:Receptors, Cell Surface / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000378:metabolism; D043562:Receptors, G-Protein-Coupled; D013329:Structure-Activity Relationship; D014162:Transfection; D014579:Urotensins / Q000737:chemistry*
chemical_list: C453979:1-(3-carbamimidoyl-benzyl)-4-methyl-1H-indole-2-carboxylic acid(naphthalen-1-ylmethyl)amide; D007211:Indoles; D008024:Ligands; D009281:Naphthalenes; D010446:Peptide Fragments; D011956:Receptors, Cell Surface; D043562:Receptors, G-Protein-Coupled; C121697:UTS2R protein, human; D014579:Urotensins; C029677:urotensin II; D002118:Calcium