Stable isotope- and mass spectrometry-based metabolomics as tools in drug metabolism: A study expanding tempol pharmacology

Fei Li, Xiaoyan Pang, Kristopher W. Krausz, Changtao Jiang, Chi Chen, John A. Cook, Murali C. Krishna, James B. Mitchell, Frank J. Gonzalez, Andrew David Patterson

Research output: Contribution to journalArticle

28 Citations (Scopus)

Abstract

The application of mass spectrometry-based metabolomics in the field of drug metabolism has yielded important insights not only into the metabolic routes of drugs but has provided unbiased, global perspectives of the endogenous metabolome that can be useful for identifying biomarkers associated with mechanism of action, efficacy, and toxicity. In this report, a stable isotope- and mass spectrometry-based metabolomics approach that captures both drug metabolism and changes in the endogenous metabolome in a single experiment is described. Here the antioxidant drug tempol (4-hydroxy-2,2,6,6- tetramethylpiperidine-N-oxyl) was chosen because its mechanism of action is not completely understood and its metabolic fate has not been studied extensively. Furthermore, its small size (MW = 172.2) and chemical composition (C 9H18NO2) make it challenging to distinguish from endogenous metabolites. In this study, mice were dosed with tempol or deuterated tempol (C9D17HNO2) and their urine was profiled using ultraperformance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry. Principal component analysis of the urinary metabolomics data generated a Y-shaped scatter plot containing drug metabolites (protonated and deuterated) that were clearly distinct from the endogenous metabolites. Ten tempol drug metabolites, including eight novel metabolites, were identified. Phase II metabolism was the major metabolic pathway of tempol in vivo, including glucuronidation and glucosidation. Urinary endogenous metabolites significantly elevated by tempol treatment included 2,8-dihydroxyquinoline (8.0-fold, P < 0.05) and 2,8-dihydroxyquinoline- β-d-glucuronide (6.8-fold, P < 0.05). Urinary endogenous metabolites significantly attenuated by tempol treatment including pantothenic acid (1.3-fold, P < 0.05) and isobutrylcarnitine (5.3-fold, P < 0.01). This study underscores the power of a stable isotope- and mass spectrometry-based metabolomics in expanding the view of drug pharmacology.

Original languageEnglish (US)
Pages (from-to)1369-1376
Number of pages8
JournalJournal of Proteome Research
Volume12
Issue number3
DOIs
StatePublished - Mar 11 2013

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Metabolomics
Metabolites
Metabolism
Isotopes
Mass spectrometry
Mass Spectrometry
Pharmacology
Pharmaceutical Preparations
Metabolome
Pantothenic Acid
Glucuronides
Liquid chromatography
Biomarkers
Metabolic Networks and Pathways
tempol
Principal Component Analysis
Liquid Chromatography
Principal component analysis
Toxicity
Antioxidants

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Chemistry(all)

Cite this

Li, Fei ; Pang, Xiaoyan ; Krausz, Kristopher W. ; Jiang, Changtao ; Chen, Chi ; Cook, John A. ; Krishna, Murali C. ; Mitchell, James B. ; Gonzalez, Frank J. ; Patterson, Andrew David. / Stable isotope- and mass spectrometry-based metabolomics as tools in drug metabolism : A study expanding tempol pharmacology. In: Journal of Proteome Research. 2013 ; Vol. 12, No. 3. pp. 1369-1376.
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abstract = "The application of mass spectrometry-based metabolomics in the field of drug metabolism has yielded important insights not only into the metabolic routes of drugs but has provided unbiased, global perspectives of the endogenous metabolome that can be useful for identifying biomarkers associated with mechanism of action, efficacy, and toxicity. In this report, a stable isotope- and mass spectrometry-based metabolomics approach that captures both drug metabolism and changes in the endogenous metabolome in a single experiment is described. Here the antioxidant drug tempol (4-hydroxy-2,2,6,6- tetramethylpiperidine-N-oxyl) was chosen because its mechanism of action is not completely understood and its metabolic fate has not been studied extensively. Furthermore, its small size (MW = 172.2) and chemical composition (C 9H18NO2) make it challenging to distinguish from endogenous metabolites. In this study, mice were dosed with tempol or deuterated tempol (C9D17HNO2) and their urine was profiled using ultraperformance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry. Principal component analysis of the urinary metabolomics data generated a Y-shaped scatter plot containing drug metabolites (protonated and deuterated) that were clearly distinct from the endogenous metabolites. Ten tempol drug metabolites, including eight novel metabolites, were identified. Phase II metabolism was the major metabolic pathway of tempol in vivo, including glucuronidation and glucosidation. Urinary endogenous metabolites significantly elevated by tempol treatment included 2,8-dihydroxyquinoline (8.0-fold, P < 0.05) and 2,8-dihydroxyquinoline- β-d-glucuronide (6.8-fold, P < 0.05). Urinary endogenous metabolites significantly attenuated by tempol treatment including pantothenic acid (1.3-fold, P < 0.05) and isobutrylcarnitine (5.3-fold, P < 0.01). This study underscores the power of a stable isotope- and mass spectrometry-based metabolomics in expanding the view of drug pharmacology.",
author = "Fei Li and Xiaoyan Pang and Krausz, {Kristopher W.} and Changtao Jiang and Chi Chen and Cook, {John A.} and Krishna, {Murali C.} and Mitchell, {James B.} and Gonzalez, {Frank J.} and Patterson, {Andrew David}",
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Li, F, Pang, X, Krausz, KW, Jiang, C, Chen, C, Cook, JA, Krishna, MC, Mitchell, JB, Gonzalez, FJ & Patterson, AD 2013, 'Stable isotope- and mass spectrometry-based metabolomics as tools in drug metabolism: A study expanding tempol pharmacology', Journal of Proteome Research, vol. 12, no. 3, pp. 1369-1376. https://doi.org/10.1021/pr301023x

Stable isotope- and mass spectrometry-based metabolomics as tools in drug metabolism : A study expanding tempol pharmacology. / Li, Fei; Pang, Xiaoyan; Krausz, Kristopher W.; Jiang, Changtao; Chen, Chi; Cook, John A.; Krishna, Murali C.; Mitchell, James B.; Gonzalez, Frank J.; Patterson, Andrew David.

In: Journal of Proteome Research, Vol. 12, No. 3, 11.03.2013, p. 1369-1376.

Research output: Contribution to journalArticle

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T1 - Stable isotope- and mass spectrometry-based metabolomics as tools in drug metabolism

T2 - A study expanding tempol pharmacology

AU - Li, Fei

AU - Pang, Xiaoyan

AU - Krausz, Kristopher W.

AU - Jiang, Changtao

AU - Chen, Chi

AU - Cook, John A.

AU - Krishna, Murali C.

AU - Mitchell, James B.

AU - Gonzalez, Frank J.

AU - Patterson, Andrew David

PY - 2013/3/11

Y1 - 2013/3/11

N2 - The application of mass spectrometry-based metabolomics in the field of drug metabolism has yielded important insights not only into the metabolic routes of drugs but has provided unbiased, global perspectives of the endogenous metabolome that can be useful for identifying biomarkers associated with mechanism of action, efficacy, and toxicity. In this report, a stable isotope- and mass spectrometry-based metabolomics approach that captures both drug metabolism and changes in the endogenous metabolome in a single experiment is described. Here the antioxidant drug tempol (4-hydroxy-2,2,6,6- tetramethylpiperidine-N-oxyl) was chosen because its mechanism of action is not completely understood and its metabolic fate has not been studied extensively. Furthermore, its small size (MW = 172.2) and chemical composition (C 9H18NO2) make it challenging to distinguish from endogenous metabolites. In this study, mice were dosed with tempol or deuterated tempol (C9D17HNO2) and their urine was profiled using ultraperformance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry. Principal component analysis of the urinary metabolomics data generated a Y-shaped scatter plot containing drug metabolites (protonated and deuterated) that were clearly distinct from the endogenous metabolites. Ten tempol drug metabolites, including eight novel metabolites, were identified. Phase II metabolism was the major metabolic pathway of tempol in vivo, including glucuronidation and glucosidation. Urinary endogenous metabolites significantly elevated by tempol treatment included 2,8-dihydroxyquinoline (8.0-fold, P < 0.05) and 2,8-dihydroxyquinoline- β-d-glucuronide (6.8-fold, P < 0.05). Urinary endogenous metabolites significantly attenuated by tempol treatment including pantothenic acid (1.3-fold, P < 0.05) and isobutrylcarnitine (5.3-fold, P < 0.01). This study underscores the power of a stable isotope- and mass spectrometry-based metabolomics in expanding the view of drug pharmacology.

AB - The application of mass spectrometry-based metabolomics in the field of drug metabolism has yielded important insights not only into the metabolic routes of drugs but has provided unbiased, global perspectives of the endogenous metabolome that can be useful for identifying biomarkers associated with mechanism of action, efficacy, and toxicity. In this report, a stable isotope- and mass spectrometry-based metabolomics approach that captures both drug metabolism and changes in the endogenous metabolome in a single experiment is described. Here the antioxidant drug tempol (4-hydroxy-2,2,6,6- tetramethylpiperidine-N-oxyl) was chosen because its mechanism of action is not completely understood and its metabolic fate has not been studied extensively. Furthermore, its small size (MW = 172.2) and chemical composition (C 9H18NO2) make it challenging to distinguish from endogenous metabolites. In this study, mice were dosed with tempol or deuterated tempol (C9D17HNO2) and their urine was profiled using ultraperformance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry. Principal component analysis of the urinary metabolomics data generated a Y-shaped scatter plot containing drug metabolites (protonated and deuterated) that were clearly distinct from the endogenous metabolites. Ten tempol drug metabolites, including eight novel metabolites, were identified. Phase II metabolism was the major metabolic pathway of tempol in vivo, including glucuronidation and glucosidation. Urinary endogenous metabolites significantly elevated by tempol treatment included 2,8-dihydroxyquinoline (8.0-fold, P < 0.05) and 2,8-dihydroxyquinoline- β-d-glucuronide (6.8-fold, P < 0.05). Urinary endogenous metabolites significantly attenuated by tempol treatment including pantothenic acid (1.3-fold, P < 0.05) and isobutrylcarnitine (5.3-fold, P < 0.01). This study underscores the power of a stable isotope- and mass spectrometry-based metabolomics in expanding the view of drug pharmacology.

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