Liquid chromatography-mass spectrometry and liquid chromatography-NMR characterization of in vitro metabolites of a potent and irreversible peptidomimetic inhibitor of rhinovirus 3C protease.
In vitro metabolism of AG7088 [trans-(4S,2'R,5'S,3"'S)-4-[2'-4-(4-fluorobenzyl)-6'-methyl-5'-[(5"-methylisoxazole-3"-carbonylamino]-4-oxoheptanoylamino]-5-(2"'-oxopyrrolidin-3-"'-yl)pent-2-enoic acid ethyl ester] was studied in liver microsomes isolated from mice, rats, rabbits, dogs, monkeys, and humans. The structures of the metabolites were characterized by liquid chromatography (LC)-tandem mass spectrometry and LC-NMR methods. Hydrolysis of the ethyl ester to produce metabolite M4 (AG7185) is the predominant pathway in all species, with the greatest activity observed in rodents and rabbits, followed by monkeys, dogs, and humans. Several hydroxylation products were identified as minor metabolites, including diastereomers M1 and M2, with a hydroxy group at the P1-lactam moiety, and M3, with a hydroxy group at the methyl position of the methylisoxazole ring. Rodent and rabbit liver microsomes formed almost exclusively the acid metabolite M4 (AG7185), with very little hydroxylated metabolites, whereas monkey liver microsomes formed more secondary metabolites (i.e., acid analogs of the hydroxylated metabolites). The overall metabolic profile of AG7088 formed in dog liver microsomes closely resembled that of human liver microsomes; therefore, this species may be the most appropriate animal model relative to humans for exposure to AG7088 and its metabolites.
Zhang
K E
KE
Department of Developmental Pharmacology, Agouron Pharmaceuticals, Incorporated, A Pfizer Company, San Diego, California, USA.
Hee
B
B
Lee
C A
CA
Liang
B
B
Potts
B C
BC
D000086042:3C Viral Proteases; D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003546:Cysteine Endopeptidases / Q000187:drug effects*; D004285:Dogs; D000882:Haplorhini; D006801:Humans; D066298:In Vitro Techniques; D007555:Isoxazoles / Q000737:chemistry / Q000494:pharmacology*; D009682:Magnetic Resonance Spectroscopy / Q000379:methods*; D008297:Male; D013058:Mass Spectrometry / Q000379:methods*; D018716:Molecular Mimicry; D015394:Molecular Structure; D010455:Peptides / Q000494:pharmacology*; D010649:Phenylalanine / Q000031:analogs & derivatives; D011480:Protease Inhibitors / Q000737:chemistry / Q000494:pharmacology*; D011760:Pyrrolidinones / Q000737:chemistry / Q000494:pharmacology*; D014633:Valine / Q000031:analogs & derivatives; D014764:Viral Proteins
D007555:Isoxazoles; D010455:Peptides; D011480:Protease Inhibitors; D011760:Pyrrolidinones; D014764:Viral Proteins; D010649:Phenylalanine; D003546:Cysteine Endopeptidases; D000086042:3C Viral Proteases; D014633:Valine; C118874:rupintrivir