title: Identification and structural elucidation of in vitro metabolites of atazanavir by HPLC and tandem mass spectrometry.
abstract: Atazanavir (marketed as Reyataz®) is an important member of the human immunodeficiency virus protease inhibitor class. LC-UV-MS(n) experiments were designed to identify metabolites of atazanavir after incubations in human hepatocytes. Five major (M1-M5) and seven minor (M7-M12) metabolites were identified. The most abundant metabolite, M1, was formed by a mono-oxidation on the t-butyl group at the non-prime side. The second most abundant metabolite, M2, was also a mono-oxidation product, which has not yet been definitively identified. Metabolites, M3 and M4, were structural isomers, which were apparently formed by oxidative carbamate hydrolysis. The structure of M5 comprises the non-prime side of atazanavir which contains a pyridinyl-benzyl group. Metabolite M6a was formed by the cleavage of the pyridinyl-benzyl side chain, as evidenced by the formation of the corresponding metabolic product, the pyridinyl-benzoic acid (M6b). Mono-oxidation also occurred on the pyridinyl-benzyl group to produce the low abundance metabolite M8. Oxidation of the terminal methyl groups produced M9 and M10, respectively, which have low chemical stability. Trace-level metabolites of di-oxidations, M11 and M12, were also detected, but the complexity of the molecule precluded identification of the second oxidation site. To our knowledge, metabolites M6b and M8 have not been reported.
authors.affiliation: Concert Pharmaceuticals, Inc., Lexington, MA 02421, USA. ccheng@concertpharma.com
authors.forename: Changfu
authors.initials: C
authors.lastname: Cheng
authors.forename: Richard
authors.initials: R
authors.lastname: Gallegos
authors.forename: Gary
authors.initials: G
authors.lastname: Bridson
authors.forename: Lijun
authors.initials: L
authors.lastname: Wu
authors.forename: Scott
authors.initials: S
authors.lastname: Harbeson
authors.forename: Robert
authors.initials: R
authors.lastname: Zelle
authors.forename: Roger
authors.initials: R
authors.lastname: Tung
mesh_terms: D000069446:Atazanavir Sulfate; D002478:Cells, Cultured; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005260:Female; D022781:Hepatocytes / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D007477:Ions / Q000032:analysis / Q000737:chemistry / Q000378:metabolism; D008297:Male; D009842:Oligopeptides / Q000032:analysis / Q000737:chemistry* / Q000378:metabolism; D011725:Pyridines / Q000032:analysis / Q000737:chemistry* / Q000378:metabolism; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D007477:Ions; D009842:Oligopeptides; D011725:Pyridines; D000069446:Atazanavir Sulfate