title: Structure and stereochemistry of the active metabolite of clopidogrel.
abstract: Clopidogrel (SR25990C, PLAVIX) is a potent antiplatelet drug, which has been recently launched and is indicated for the prevention of vascular thrombotic events in patients at risk. Clopidogrel is inactive in vitro, and a hepatic biotransformation is necessary to express the full antiaggregating activity of the drug. Moreover, 2-oxo-clopidogrel has been previously suggested to be the essential key intermediate metabolite from which the active metabolite is formed. In the present paper, we give the evidence of the occurrence of an in vitro active metabolite after incubation of 2-oxo-clopidogrel with human liver microsomes. This metabolite was purified by liquid chromatography, and its structure was studied by a combination of mass spectometry (MS) and NMR experiments. MS results suggested that the active metabolite belongs to a family of eight stereoisomers with the following primary chemical structure: 2-[1-[1-(2-chlorophenyl)-2-methoxy-2-oxoethyl]-4-sulfanyl-3-piperidinylidene]acetic acid. Chiral supercritical fluid chromatography resolved these isomers. However, only one of the eight metabolites retained the biological activity, thus underlining the critical importance of associated absolute configuration. Because of its highly labile character, probably due to a very reactive thiol function, structural elucidation of the active metabolite was performed on the stabilized acrylonitrile derivative. Conjunction of all our results suggested that the active metabolite is of S configuration at C 7 and Z configuration at C 3-C 16 double bound.
authors.affiliation: Sanofi-Synthelabo Recherche, Toulouse cedex, France.
authors.forename: Jean-Marie
authors.initials: JM
authors.lastname: Pereillo
authors.forename: Mohamed
authors.initials: M
authors.lastname: Maftouh
authors.forename: Alain
authors.initials: A
authors.lastname: Andrieu
authors.forename: Marie-Francoise
authors.initials: MF
authors.lastname: Uzabiaga
authors.forename: Olivier
authors.initials: O
authors.lastname: Fedeli
authors.forename: Pierre
authors.initials: P
authors.lastname: Savi
authors.forename: Marc
authors.initials: M
authors.lastname: Pascal
authors.forename: Jean-Marc
authors.initials: JM
authors.lastname: Herbert
authors.forename: Jean-Pierre
authors.initials: JP
authors.lastname: Maffrand
authors.forename: Claudine
authors.initials: C
authors.lastname: Picard
mesh_terms: D000181:Acrylonitrile / Q000378:metabolism; D001711:Biotransformation; D001792:Blood Platelets / Q000187:drug effects / Q000378:metabolism; D000077144:Clopidogrel; D006801:Humans; D066298:In Vitro Techniques; D009682:Magnetic Resonance Spectroscopy; D013058:Mass Spectrometry; D008862:Microsomes, Liver / Q000378:metabolism; D008958:Models, Molecular; D008968:Molecular Conformation; D010975:Platelet Aggregation Inhibitors / Q000737:chemistry* / Q000493:pharmacokinetics*; D013237:Stereoisomerism; D013988:Ticlopidine / Q000031:analogs & derivatives / Q000737:chemistry* / Q000493:pharmacokinetics*
chemical_list: D010975:Platelet Aggregation Inhibitors; D000077144:Clopidogrel; D000181:Acrylonitrile; D013988:Ticlopidine