Biotransformation of a GABAA receptor partial agonist in sprague-dawley rats and cynomolgus monkeys: identification of two unique N-carbamoyl metabolites.
The absorption, metabolism, and excretion of N-[3-fluoro-4-[2-(propylamino)ethoxy]phenyl]-4,5,6,7-tetrahydro-4-oxo-1H-indole-3-carboxamide monomethanesulfonate (1), a GABAA receptor partial agonist potentially useful in treating generalized anxiety disorder, have been evaluated in both Sprague-Dawley rats and cynomolgus monkeys using [14C]1. In both species, mass balance was achieved within 48 h postdose, with the majority of drug-related material excreted within the feces; the clearance of 1 in each species had both metabolic and renal components. In addition to the metabolites produced by aliphatic hydroxylation and/or N-dealkylation of 1, two unique metabolites were detected: a putative carbamic acid (M7) in rat plasma and monkey bile, and an N-carbamoyl glucuronide (M8) in both rat and monkey bile. Metabolite M8 was structurally deciphered by liquid chromatographytandem mass spectrometry and NMR, and was readily generated in vitro upon incubation of [14C]1 with rat liver microsomes fortified with uridine 5'-diphosphoglucuronic acid trisodium salt and alamethicin under a CO2 atmosphere. Treatment of M8 with beta-glucuronidase afforded 1 directly. The presence of M8 in bile and its notable absence from other matrices suggests the enterohepatic cycling of 1 via M8. Although the structure of M7 was not elucidated unequivocally due to its inability to be formed in vitro and its minimal absolute quantities in limited biological matrices, data herein clearly support its structural rationalization. Furthermore, since M7 is the precursor of M8, detection of M8 is indirect evidence of its existence. It is proposed that M7 arises from an equilibrium between 1 and dissolved CO2-equivalents both in vivo and in vitro, similar to carbamino bonds observed in hemoglobin and certain amino acids, respectively.
Shaffer
Christopher L
CL
Department of pharmacokinetics, Pharmacodynamics and Metabolism, Pfizer Global Research and Development, Groton/New London Laboratories, Pfizer Inc., Eastern Point Road, MS 4075, Groton, CT 06340, USA. Christopher.L.Shaffer@pfizer.com
Gunduz
Mithat
M
O'Connell
Thomas N
TN
Obach
R Scott
RS
Yee
Shiyin
S
D000818:Animals; D019540:Area Under Curve; D001646:Bile / Q000378:metabolism; D002219:Carbamates / Q000097:blood / Q000378:metabolism / Q000652:urine; D002250:Carbon Radioisotopes; D002853:Chromatography, Liquid; D005243:Feces / Q000737:chemistry; D005260:Female; D018755:GABA Agonists / Q000097:blood / Q000493:pharmacokinetics* / Q000652:urine; D058785:GABA-A Receptor Agonists; D020719:Glucuronides / Q000097:blood / Q000378:metabolism / Q000652:urine; D007211:Indoles / Q000097:blood / Q000493:pharmacokinetics* / Q000652:urine; D008252:Macaca fascicularis; D009682:Magnetic Resonance Spectroscopy; D008297:Male; D008862:Microsomes, Liver / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley
D002219:Carbamates; D002250:Carbon Radioisotopes; D018755:GABA Agonists; D058785:GABA-A Receptor Agonists; D020719:Glucuronides; D007211:Indoles; C512159:N-(3-fluoro-4-(2-(propylamino)ethoxy)phenyl)-4,5,6,7-tetrahydro-4-oxo-1H-indole-3-carboxamide; C070766:carbamic acid