title: Urinary biomarkers of trimethoprim bioactivation in vivo following therapeutic dosing in children.
abstract: The antimicrobial trimethoprim-sulfamethoxazole (TMP-SMX) is widely used for the treatment of skin and soft-tissue infections in the outpatient setting. Despite its therapeutic benefits, TMP-SMX has been associated with a number of adverse drug reactions, which have been primarily attributed to the formation of reactive metabolites from SMX. Recently, in vitro experiments have demonstrated that TMP may form reactive intermediates as well. However, evidence of TMP bioactivation in patients has not yet been demonstrated. In this study, we performed in vitro trapping experiments with N-acetyl-l-cysteine (NAC) to determine stable markers of reactive TMP intermediates, focusing on eight potential markers (NAC-TMP adducts), some of which were previously identified in vitro. We developed a specific and sensitive assay involving liquid chromatography followed by tandem mass spectrometry for measurement of these adducts in human liver microsomal samples and expanded the methodology toward the detection of these analytes in human urine. Urine samples from four patients receiving TMP-SMX treatment were analyzed, and all samples demonstrated the presence of six NAC-TMP adducts, which were also detected in vitro. These adducts are consistent with the formation of imino-quinone-methide and para-quinone-methide reactive intermediates in vivo. As a result, the TMP component of TMP-SMX should be considered as well when evaluating adverse drug reactions to TMP-SMX.
authors.affiliation: Division of Clinical Pharmacology and Therapeutic Innovation, Children's Mercy Hospitals and Clinics, and ‡Department of Pediatrics, School of Medicine, University of Missouri Kansas City , 2401 Gillham Road, Kansas City, Missouri 64108, United States.
authors.forename: Leon
authors.initials: L
authors.lastname: van Haandel
authors.forename: Jennifer L
authors.initials: JL
authors.lastname: Goldman
authors.forename: Robin E
authors.initials: RE
authors.lastname: Pearce
authors.forename: J Steven
authors.initials: JS
authors.lastname: Leeder
mesh_terms: D000111:Acetylcysteine / Q000378:metabolism; D000890:Anti-Infective Agents / Q000493:pharmacokinetics* / Q000494:pharmacology / Q000652:urine; D015415:Biomarkers / Q000652:urine; D001711:Biotransformation; D002648:Child; D002675:Child, Preschool; D002853:Chromatography, Liquid; D006801:Humans; D008862:Microsomes, Liver / Q000378:metabolism; D053719:Tandem Mass Spectrometry; D014295:Trimethoprim / Q000493:pharmacokinetics* / Q000494:pharmacology / Q000652:urine; D015662:Trimethoprim, Sulfamethoxazole Drug Combination / Q000493:pharmacokinetics* / Q000652:urine
chemical_list: D000890:Anti-Infective Agents; D015415:Biomarkers; D015662:Trimethoprim, Sulfamethoxazole Drug Combination; D014295:Trimethoprim; D000111:Acetylcysteine