OxidatiVe Biotransformation of BMS-690514
Chem. Res. Toxicol., Vol. 24, No. 1, 2011 133
which would lead to loss of the deuterium label from the
hydroxylation site, is also feasible (Scheme 2). Our studies were
not able to distinguish between these two possible pathways.
and tables for the structural characterization of the M1 metabo-
lite of BMS-690514. This material is available free of charge
The availability of deuterated BMS-690514 presents some
interesting opportunities to explore how the ratio of M1 to M37
may be affected in incubations with HLM or with various P450-
enzymes relative to incubations with unlabeled BMS-690514.
Currently, we have not conducted these investigations because
on the chromatographic systems used in these experiments, M37
coelutes with M2, a hydroxylated metabolite with similar mass-
spectrometric characteristics, which would confound the data
interpretation.
Pharmacoactivity of M1 and M37. Both M1 and M37 were
hydroxylated on the pyrrolotriazine nucleus of the parent drug,
which is a core template in a number of effective tyrosine kinase
inhibitors (5). Since the pyrrolotriazine nucleus of M1 underwent
a rearrangement to a pyridotriazine, it was expected that M1
would not retain inhibitory kinase activity. Consistently, in vitro
receptor assays showed that M1 was inactive toward the kinase
activity of EGFR, HER2, and VEGFR2 receptors, with IC50
values >1 µM (8). Moreover, structure-activity studies identi-
fied that the substitution of a methyl group at the 7-position of
the pyrrolotriazine group (position 16 of BMS-690514) led to
substantial loss of inhibitory activity (5), implicating the
importance of this position for the retention of activity. Since
M37 had a hydroxyl group located at this position, it is likely
that M37 is less potent than the parent drug. Since M37 was
not a circulating metabolite in humans, the pharmacological
activity of M37 was not further explored in receptor binding
assays.
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Although M1 was relatively stable at low temperature (<4
°C), it was thermally labile and decomposed into multiple peaks
when kept at ambient temperature overnight (data not shown).
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synthesize M1 and utilize it for further studies, e.g., as a
bioanlatyical reference standard or for toxicity testing. Although
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epoxide or aldehyde intermediates, we observed good extraction
recovery of radioactive components from plasma obtained from
in vivo absorption, distribution, metabolism, and excretion
(ADME) studies with [14C]BMS-690514 (8, 9). In addition, no
glutathione or cysteine adducts were identified in any samples
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trapping experiment with methoxyamine showed that the
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methoxime species, suggesting that the internal trapping of the
aldehyde group leading to the formation of a pyridotriazine ring
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Summary. Metabolites M1 and M37 of BMS-690514 were
two monohydroxylated metabolites of BMS-690514, each
hydroxylated at the same position on the pyrrolotriazine moiety.
During its formation, M1 underwent a novel structural rear-
rangement where the pyrrolotriazine group rearranged into a
pyridotriazine structure. In contrast, the pyrrolotriazine moiety
of M37 remained intact. These two metabolites may be formed
either via a common intermediate or through mechanistically
distinct pathways.
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Supporting Information Available: Synthesis and charac-
terization of [2H]BMS-690514 and [3H]BMS-690514; figures