title: 1- and 3-hydroxylations, in addition to 4-hydroxylation, of debrisoquine are catalyzed by cytochrome P450 2D6 in humans.
abstract: Twenty-one healthy Swedish Caucasian volunteers, representing different groups with 0-13 functional cytochrome P450 (CYP) 2D6 genes, were given a single oral dose of 20 mg of debrisoquine. The hypothesis of further oxidation of the main metabolite, (S)-4-hydroxydebrisoquine, in subjects with multiple CYP2D6 genes was tested by screening the 0-8-hr urine samples for dihydroxylated metabolites of debrisoquine with protonated molecular ions at m/z 208, using LC/MS. Three peaks were detected in a subject with 13 functional CYP2D6 genes. One compound was identified as dihydroxylated debrisoquine (presumably with hydroxylation at position 4 plus one of the positions in the aromatic ring). This metabolite had not been previously demonstrated in humans and was detected only in this subject. The other two compounds, which were measurable in various amounts in all subjects investigated, were identified as 2-(guanidinomethyl)phenylacetic acid and 2-(guanidinoethyl)benzoic acid. They had been previously detected in the urine of humans, dogs, and rats. They were distinguished by acid-catalyzed deuterium exchange of the hydrogens at the alpha-position, with respect to the carboxylic acid group, of the former but not the latter acid. The acids are formed by 3- and 1-hydroxylation of debrisoquine, respectively, followed by ring opening to aldehydes, which are further oxidized to acids. Strong Spearman rank correlations between debrisoquine products of 1- or 3-hydroxydebrisoquine and debrisoquine/4-hydroxydebrisoquine ratios (rS = 0.97 and rS = 0.96, respectively), using the intensity of the peaks of the reconstructed ion-current chromatograms, clearly showed that both hydroxylation steps are catalyzed by CYP2D6. Because reference compounds for the two acids were not available, the absolute quantities could not be determined.
authors.affiliation: Department of Medical Laboratory Sciences and Technology, Division of Clinical Pharmacology, Karolinska Institute, Huddinge University Hospital, Huddinge, Sweden.
authors.forename: B
authors.initials: B
authors.lastname: Eiermann
authors.forename: P O
authors.initials: PO
authors.lastname: Edlund
authors.forename: A
authors.initials: A
authors.lastname: Tjernberg
authors.forename: P
authors.initials: P
authors.lastname: Dalén
authors.forename: M L
authors.initials: ML
authors.lastname: Dahl
authors.forename: L
authors.initials: L
authors.lastname: Bertilsson
mesh_terms: D000959:Antihypertensive Agents / Q000493:pharmacokinetics* / Q000652:urine; D002384:Catalysis; D002853:Chromatography, Liquid; D019389:Cytochrome P-450 CYP2D6 / Q000378:metabolism*; D003647:Debrisoquin / Q000493:pharmacokinetics* / Q000652:urine; D006801:Humans; D006900:Hydroxylation; D013058:Mass Spectrometry; D012016:Reference Values
chemical_list: D000959:Antihypertensive Agents; D019389:Cytochrome P-450 CYP2D6; D003647:Debrisoquin