title: Species difference in glucuronidation formation kinetics with a selective mTOR inhibitor.
abstract: The mammalian target of rapamycin (mTOR) is a protein kinase that shows key involvement in age-related disease and promises to be a target for treatment of cancer. In the present study, the elimination of potent ATP-competitive mTOR inhibitor 3-(6-amino-2-methylpyrimidin-4-yl)-N-(1H-pyrazol-3-yl)imidazo[1,2-b]pyridazin-2-amine (compound 1) is studied in bile duct-cannulated rats, and the metabolism of compound 1 in liver microsomes is compared across species. Compound 1 was shown to undergo extensive N-glucuronidation in bile duct-catheterized rats. N-glucuronides were detected on positions N1 (M2) and N2 (M1) of the pyrazole moiety as well as on the primary amine (M3). All three N-glucuronide metabolites were detected in liver microsomes of the rat, dog, and human, while primary amine glucuronidation was not detected in cynomolgus monkey. In addition, N1- and N2-glucuronidation showed strong species selectivity in vitro, with rat, dog, and human favoring N2-glucuronidation and monkey favoring N1-glucuronide formation. Formation of M1 in monkey liver microsomes also followed sigmoidal kinetics, singling out monkey as unique among the species with regard to compound 1 N-glucuronidation. In this respect, monkeys might not always be the best animal model for N-glucuronidation of uridine diphosphate glucuronosyltransferase (UGT) 1A9 or UGT1A1 substrates in humans. The impact of N-glucuronidation of compound 1 could be more pronounced in higher species such as monkey and human, leading to high clearance in these species. While compound 1 shows promise as a candidate for investigating the impact of pan-mTOR inhibition in vivo, opportunities may exist through medicinal chemistry efforts to reduce metabolic liability with the goal of improving systemic exposure.
authors.affiliation: Departments of Pharmacokinetics and Drug Metabolism (L.M.B., J.L., A.C., Z.Z., Y.T.) and Discovery Research (P.K.), Amgen, Inc., Cambridge, Massachusetts.
authors.forename: Loren M
authors.initials: LM
authors.lastname: Berry
authors.forename: Jingzhou
authors.initials: J
authors.lastname: Liu
authors.forename: Adria
authors.initials: A
authors.lastname: Colletti
authors.forename: Paul
authors.initials: P
authors.lastname: Krolikowski
authors.forename: Zhiyang
authors.initials: Z
authors.lastname: Zhao
authors.forename: Yohannes
authors.initials: Y
authors.lastname: Teffera
mesh_terms: D000818:Animals; D001711:Biotransformation; D002851:Chromatography, High Pressure Liquid; D004285:Dogs; D005260:Female; D020719:Glucuronides / Q000378:metabolism*; D006574:Heterocyclic Compounds, 2-Ring / Q000378:metabolism* / Q000494:pharmacology; D006801:Humans; D007700:Kinetics; D008252:Macaca fascicularis; D009682:Magnetic Resonance Spectroscopy; D008297:Male; D008862:Microsomes, Liver / Q000187:drug effects / Q000201:enzymology*; D011743:Pyrimidines / Q000378:metabolism* / Q000494:pharmacology; D051381:Rats; D017207:Rats, Sprague-Dawley; D013045:Species Specificity; D058570:TOR Serine-Threonine Kinases / Q000037:antagonists & inhibitors*; D053719:Tandem Mass Spectrometry
chemical_list: C000592573:3-(6-amino-2-methylpyrimidin-4-yl)-N-(1H-pyrazol-3-yl)imidazo(1,2-b)pyridazin-2-amine; D020719:Glucuronides; D006574:Heterocyclic Compounds, 2-Ring; D011743:Pyrimidines; D058570:TOR Serine-Threonine Kinases