title: Metabolite identification via LC-SPE-NMR-MS of the in vitro biooxidation products of a lead mGlu5 allosteric antagonist and impact on the improvement of metabolic stability in the series.
abstract: Detailed information on the metabolic fate of lead compounds can be a powerful tool for an informed approach to the stabilization of metabolically labile compounds in the lead optimization phase. The combination of high performance liquid chromatography (HPLC) with nuclear magnetic resonance (NMR) spectroscopy and mass spectrometry (MS) has been used to give comprehensive structural data on metabolites of novel drugs in development. Recently, increased automation and the embedding of on-line solid-phase extraction (SPE) into a integrated LC-SPE-NMR-MS system have improved enormously the detection limits of this approach. The new technology platform allows the analysis of complex mixtures from microsome incubations, combining low material requirements with relatively high throughput. Such characteristics make it possible to thoroughly characterize metabolites of selected compounds at earlier phases along the path to lead identification and clinical candidate selection, thus providing outstanding guidance in the process of eliminating undesired metabolism and detecting active or potentially toxic metabolites. Such an approach was applied at the lead identification stage of a backup program on metabotropic glutamate receptor 5 (mGlu5) allosteric inhibition. The major metabolites of a lead 5-aminothiazole-4-carboxylic acid amide 1 were synthesized and screened, revealing significant in vitro activity and possible involvement in the overall pharmacodynamic behavior of 1. The information collected on the metabolism of the highly active compound 1 was pivotal to the synthesis of related compounds with improved microsomal stability.
authors.affiliation: Pharmaceuticals Division, F. Hoffmann-La Roche AG, 4070 Basel, Switzerland. simona_m.ceccarelli@roche.com
authors.forename: Simona M
authors.initials: SM
authors.lastname: Ceccarelli
authors.forename: Götz
authors.initials: G
authors.lastname: Schlotterbeck
authors.forename: Patrick
authors.initials: P
authors.lastname: Boissin
authors.forename: Martin
authors.initials: M
authors.lastname: Binder
authors.forename: Bernd
authors.initials: B
authors.lastname: Buettelmann
authors.forename: Steven
authors.initials: S
authors.lastname: Hanlon
authors.forename: Georg
authors.initials: G
authors.lastname: Jaeschke
authors.forename: Sabine
authors.initials: S
authors.lastname: Kolczewski
authors.forename: Ernst
authors.initials: E
authors.lastname: Kupfer
authors.forename: Jens-Uwe
authors.initials: JU
authors.lastname: Peters
authors.forename: Richard H P
authors.initials: RH
authors.lastname: Porter
authors.forename: Eric P
authors.initials: EP
authors.lastname: Prinssen
authors.forename: Marianne
authors.initials: M
authors.lastname: Rueher
authors.forename: Iris
authors.initials: I
authors.lastname: Ruf
authors.forename: Will
authors.initials: W
authors.lastname: Spooren
authors.forename: Andreas
authors.initials: A
authors.lastname: Stämpfli
authors.forename: Eric
authors.initials: E
authors.lastname: Vieira
mesh_terms: D000494:Allosteric Regulation; D000631:Aminopyridines / Q000138:chemical synthesis / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D006801:Humans; D020128:Inhibitory Concentration 50; D009682:Magnetic Resonance Spectroscopy / Q000379:methods; D013058:Mass Spectrometry / Q000379:methods; D008862:Microsomes, Liver / Q000737:chemistry / Q000378:metabolism*; D010084:Oxidation-Reduction; D004364:Pharmaceutical Preparations / Q000138:chemical synthesis / Q000378:metabolism*; D064529:Receptor, Metabotropic Glutamate 5; D018094:Receptors, Metabotropic Glutamate / Q000037:antagonists & inhibitors*; D052616:Solid Phase Extraction / Q000379:methods; D013237:Stereoisomerism; D013844:Thiazoles / Q000138:chemical synthesis / Q000378:metabolism*
chemical_list: D000631:Aminopyridines; D004364:Pharmaceutical Preparations; D064529:Receptor, Metabotropic Glutamate 5; D018094:Receptors, Metabotropic Glutamate; D013844:Thiazoles