Novel Pathways Of Ponatinib Disposition Catalysed By CYP1A1 Involving Generation Of Potentially Toxic Metabolites

De Lin, Rumen Kostov, Jeffrey Huang, Colin J Henderson, C Wolf (Lead / Corresponding author)

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Abstract

Ponatinib, a pan-BCR-ABL tyrosine kinase inhibitor for the treatment of chronic myeloid leukemia (CML), causes severe side-effects including vascular occlusions, pancreatitis and liver toxicity, although the underlying mechanisms remain unclear. Modification of critical proteins through reactive metabolites are thought to be responsible for a number of adverse drug reactions. In vitro metabolite screening of ponatinib with human liver microsomes and glutathione revealed unambiguous signals of glutathionedrug (P-GSH) adducts. Further profiling of human P450s indicated that CYP1A1 was the predominant P450 enzyme driving this reaction. P-GSH conjugate formation paralleled the disappearance of hydroxylated ponatinib metabolites, suggesting the initial reaction was epoxide generation. Mouse glutathione S-transferase p1 (mGstp1) further enhanced P-GSH adduct formation in vitro. Ponatinib pharmacokinetics were determined in vivo in WT mice and mice humanised for CYP1A1/2 and treated with the CYP1A1 inducers 2,3,7,8-tetrachlorodibenzodioxin (TCDD) or 3-methylcholanthrene (3-MC). Ponatinib exposure was significantly decreased in treated mice compared to controls (7.7- and 2.2 fold for WT and humanised CYP1A1/2, respectively). Interestingly, the P-GSH conjugate was only found in the feces of CYP1A1-induced mice, not in control animals. Proteinadducts were also identified by LC-MS/MS analysis of mGstp1 tryptic digests. These results indicate not only that CYP1A1 could be involved in ponatinib disposition, which has not been previously reported, but also that electrophilic intermediates resulting from CYP1A1 metabolism in normal tissues may contribute to ponatinib toxicity. These data are consistent with a recent report that CML patients who smoke are at a greater risk of disease progression and premature death. (Lauseker et al., 2017).
Original languageEnglish
Pages (from-to)12-19
Number of pages8
JournalJournal of Pharmacology and Experimental Therapeutics
Volume363
Issue number1
Early online date7 Sep 2017
DOIs
Publication statusPublished - Oct 2017

Fingerprint

Cytochrome P-450 CYP1A1
Poisons
Leukemia, Myelogenous, Chronic, BCR-ABL Positive
Glutathione Transferase
Premature Mortality
Methylcholanthrene
Epoxy Compounds
Liver Microsomes
ponatinib
Drug-Related Side Effects and Adverse Reactions
Feces
Smoke
Pancreatitis
Protein-Tyrosine Kinases
Cytochrome P-450 Enzyme System
Glutathione
Blood Vessels
Disease Progression
Pharmacokinetics
Liver

Cite this

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title = "Novel Pathways Of Ponatinib Disposition Catalysed By CYP1A1 Involving Generation Of Potentially Toxic Metabolites",
abstract = "Ponatinib, a pan-BCR-ABL tyrosine kinase inhibitor for the treatment of chronic myeloid leukemia (CML), causes severe side-effects including vascular occlusions, pancreatitis and liver toxicity, although the underlying mechanisms remain unclear. Modification of critical proteins through reactive metabolites are thought to be responsible for a number of adverse drug reactions. In vitro metabolite screening of ponatinib with human liver microsomes and glutathione revealed unambiguous signals of glutathionedrug (P-GSH) adducts. Further profiling of human P450s indicated that CYP1A1 was the predominant P450 enzyme driving this reaction. P-GSH conjugate formation paralleled the disappearance of hydroxylated ponatinib metabolites, suggesting the initial reaction was epoxide generation. Mouse glutathione S-transferase p1 (mGstp1) further enhanced P-GSH adduct formation in vitro. Ponatinib pharmacokinetics were determined in vivo in WT mice and mice humanised for CYP1A1/2 and treated with the CYP1A1 inducers 2,3,7,8-tetrachlorodibenzodioxin (TCDD) or 3-methylcholanthrene (3-MC). Ponatinib exposure was significantly decreased in treated mice compared to controls (7.7- and 2.2 fold for WT and humanised CYP1A1/2, respectively). Interestingly, the P-GSH conjugate was only found in the feces of CYP1A1-induced mice, not in control animals. Proteinadducts were also identified by LC-MS/MS analysis of mGstp1 tryptic digests. These results indicate not only that CYP1A1 could be involved in ponatinib disposition, which has not been previously reported, but also that electrophilic intermediates resulting from CYP1A1 metabolism in normal tissues may contribute to ponatinib toxicity. These data are consistent with a recent report that CML patients who smoke are at a greater risk of disease progression and premature death. (Lauseker et al., 2017).",
author = "De Lin and Rumen Kostov and Jeffrey Huang and Henderson, {Colin J} and C Wolf",
note = "This work was supported by Cancer Research UK [Grant C4639/A12330]",
year = "2017",
month = "10",
doi = "10.1124/jpet.117.243246",
language = "English",
volume = "363",
pages = "12--19",
journal = "Journal of Pharmacology and Experimental Therapeutics",
issn = "0022-3565",
publisher = "American Society for Pharmacology and Experimental Therapeutics",
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TY - JOUR

T1 - Novel Pathways Of Ponatinib Disposition Catalysed By CYP1A1 Involving Generation Of Potentially Toxic Metabolites

AU - Lin, De

AU - Kostov, Rumen

AU - Huang, Jeffrey

AU - Henderson, Colin J

AU - Wolf, C

N1 - This work was supported by Cancer Research UK [Grant C4639/A12330]

PY - 2017/10

Y1 - 2017/10

N2 - Ponatinib, a pan-BCR-ABL tyrosine kinase inhibitor for the treatment of chronic myeloid leukemia (CML), causes severe side-effects including vascular occlusions, pancreatitis and liver toxicity, although the underlying mechanisms remain unclear. Modification of critical proteins through reactive metabolites are thought to be responsible for a number of adverse drug reactions. In vitro metabolite screening of ponatinib with human liver microsomes and glutathione revealed unambiguous signals of glutathionedrug (P-GSH) adducts. Further profiling of human P450s indicated that CYP1A1 was the predominant P450 enzyme driving this reaction. P-GSH conjugate formation paralleled the disappearance of hydroxylated ponatinib metabolites, suggesting the initial reaction was epoxide generation. Mouse glutathione S-transferase p1 (mGstp1) further enhanced P-GSH adduct formation in vitro. Ponatinib pharmacokinetics were determined in vivo in WT mice and mice humanised for CYP1A1/2 and treated with the CYP1A1 inducers 2,3,7,8-tetrachlorodibenzodioxin (TCDD) or 3-methylcholanthrene (3-MC). Ponatinib exposure was significantly decreased in treated mice compared to controls (7.7- and 2.2 fold for WT and humanised CYP1A1/2, respectively). Interestingly, the P-GSH conjugate was only found in the feces of CYP1A1-induced mice, not in control animals. Proteinadducts were also identified by LC-MS/MS analysis of mGstp1 tryptic digests. These results indicate not only that CYP1A1 could be involved in ponatinib disposition, which has not been previously reported, but also that electrophilic intermediates resulting from CYP1A1 metabolism in normal tissues may contribute to ponatinib toxicity. These data are consistent with a recent report that CML patients who smoke are at a greater risk of disease progression and premature death. (Lauseker et al., 2017).

AB - Ponatinib, a pan-BCR-ABL tyrosine kinase inhibitor for the treatment of chronic myeloid leukemia (CML), causes severe side-effects including vascular occlusions, pancreatitis and liver toxicity, although the underlying mechanisms remain unclear. Modification of critical proteins through reactive metabolites are thought to be responsible for a number of adverse drug reactions. In vitro metabolite screening of ponatinib with human liver microsomes and glutathione revealed unambiguous signals of glutathionedrug (P-GSH) adducts. Further profiling of human P450s indicated that CYP1A1 was the predominant P450 enzyme driving this reaction. P-GSH conjugate formation paralleled the disappearance of hydroxylated ponatinib metabolites, suggesting the initial reaction was epoxide generation. Mouse glutathione S-transferase p1 (mGstp1) further enhanced P-GSH adduct formation in vitro. Ponatinib pharmacokinetics were determined in vivo in WT mice and mice humanised for CYP1A1/2 and treated with the CYP1A1 inducers 2,3,7,8-tetrachlorodibenzodioxin (TCDD) or 3-methylcholanthrene (3-MC). Ponatinib exposure was significantly decreased in treated mice compared to controls (7.7- and 2.2 fold for WT and humanised CYP1A1/2, respectively). Interestingly, the P-GSH conjugate was only found in the feces of CYP1A1-induced mice, not in control animals. Proteinadducts were also identified by LC-MS/MS analysis of mGstp1 tryptic digests. These results indicate not only that CYP1A1 could be involved in ponatinib disposition, which has not been previously reported, but also that electrophilic intermediates resulting from CYP1A1 metabolism in normal tissues may contribute to ponatinib toxicity. These data are consistent with a recent report that CML patients who smoke are at a greater risk of disease progression and premature death. (Lauseker et al., 2017).

U2 - 10.1124/jpet.117.243246

DO - 10.1124/jpet.117.243246

M3 - Article

C2 - 28882992

VL - 363

SP - 12

EP - 19

JO - Journal of Pharmacology and Experimental Therapeutics

JF - Journal of Pharmacology and Experimental Therapeutics

SN - 0022-3565

IS - 1

ER -