Atypical kinetics of cytochrome P450 2J2: Epoxidation of arachidonic acid and reversible inhibition by xenobiotic inhibitors

Leow, Jacqueline Wen Hui; Verma, Ravi Kumar; Lim, Amos Boon Hao; Fan, Hao; Chan, Eric Chun Yong

doi:10.1016/j.ejps.2021.105889

Cited by 11 publications

(10 citation statements)

References 108 publications

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“…Moreover, our previous study on AA metabolism based on a CYP2J2 homology model, derived from the crystal structure of CYP2R1, suggested the possibility of two AA substrates binding concomitantly to its orthosteric binding site and alternative binding site (Leow et al, 2021). The observation corroborated the comparable large active site within the binding pocket to that of CYP3A4, which allows for the accommodation of bulky molecules and binding of more than one molecule (Lee et al, 2010).…”

Section: Introductionsupporting

confidence: 52%

“…Our collective findings provided compelling evidences that the dysregulation of CYP2J2-mediated AA-EETs metabolism affects both the viability and normal beating function of cardiomyocytes. As inhibition of CYP2J2 is substratedependent (Leow et al, 2021), AA was additionally adopted as the physiologically relevant substrate.…”

Section: Discussionmentioning

confidence: 99%

“…Quantification of all drugs and metabolites was performed on Agilent 1290 Infinity UHPLC coupled with QTRAP 3500 MS/MS. Analytical method was modified based on our previous studies (Cheong et al, 2019;Leow et al, 2021;. In brief, chromatographic separation was achieved on a Luna Omega Polar C 18 UHPLC column (1.6 μM, 2.1×100 mm, Phenomenex, Torrance, CA).…”

Section: Lc-ms/ms Methodsmentioning

confidence: 99%

“…Prism were directly adopted to analyze the inhibition of rivaroxaban metabolism by osimertinib. In view of the atypical substrate inhibition kinetics observed for AA (Leow et al, 2021), AA-osimertinib data was fitted using our previously modified reversible inhibition model with substrate inhibition kinetics (Equation 8).…”

Section: Analysis Of Reversible Inhibition Kinetics and Evaluation Of...mentioning

confidence: 99%

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Inhibition of Cytochrome P450 2J2-Mediated Metabolism of Rivaroxaban and Arachidonic Acid by Ibrutinib and Osimertinib

Wang

Chan

2022

Drug Metab Dispos

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Covalent tyrosine kinase inhibitors (TKIs) ibrutinib and osimertinib are associated with cardiac arrhythmia. The interactions between these TKIs with cytochrome P450 2J2 (CYP2J2) that is highly expressed in human heart are unknown. In vitro metabolism experiments were performed to characterize CYP2J2-mediated metabolism of ibrutinib and osimertinib. Unbound distribution coefficient (K puu ) for both TKIs was determined in AC16 cardiomyocytes. In vitro reversible and time-dependent CYP2J2 inhibition experiments were conducted with exogenous and endogenous substrates, namely rivaroxaban and arachidonic acid (AA), respectively, where kinetic parameters were estimated via one-site and multisite kinetic modeling. Ibrutinib was efficiently metabolized by CYP2J2 to a hydroxylated metabolite, M35, following substrate inhibition kinetics. Osimertinib is not a substrate of CYP2J2. Both TKIs depicted K puu values above 1 and equipotently inhibited CYP2J2-mediated hydroxylation of rivaroxaban in a concentration-dependent manner without time-dependency. The mode of reversible inhibition of CYP2J2-mediated metabolism of rivaroxaban and AA by osimertinib was described by Michaelis-Menten kinetics, while a two-site kinetic model recapitulated the atypical inhibitory kinetics of ibrutinib assuming multiple substratebinding domains within the CYP2J2 active site. The inhibition of ibrutinib and osimertinib on cardiac AA metabolism could be clinically significant considering the preferable distribution of both TKIs to cardiomyocytes with R cut-off values of 1.160 and 1.026, respectively. The dysregulation of CYP2J2-mediated metabolism of AA to cardioprotective epoxyeicosatrienoic acids by ibrutinib and osimertinib serves as a novel mechanism for TKI-induced cardiac arrhythmia. Mechanistic characterization of This article has not been copyedited and formatted. The final version may differ from this version.

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Section: Introductionsupporting

confidence: 52%

Section: Discussionmentioning

confidence: 99%

Section: Lc-ms/ms Methodsmentioning

confidence: 99%

Section: Analysis Of Reversible Inhibition Kinetics and Evaluation Of...mentioning

confidence: 99%

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Inhibition of Cytochrome P450 2J2-Mediated Metabolism of Rivaroxaban and Arachidonic Acid by Ibrutinib and Osimertinib

Wang

Chan

2022

Drug Metab Dispos

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“…Recent publications have discussed the advancement of atypical Michaelis-Menten kinetics for a variety of CYP enzymes, including several enzymes involved in the secondary metabolism cascade for vitamin D. This includes CYP3A4 [Arendse et al, 2013] and CYP2J2 [Leow and Chan, 2019;Leow et al, 2021]. In particular, CYP2J2 shares 72.5% sequence similarity with CYP2R1, so it is not unreasonable to assume that CYP2R1 may also exhibit atypical Michaelis-Menten kinetics at varying sufficiency levels.…”

Section: Discussionmentioning

confidence: 99%

Physiologically Based Pharmacokinetic Modeling of Vitamin D₃ and Metabolites in Vitamin D–Insufficient Patients

et al. 2022

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A physiologically based pharmacokinetic (PBPK) model of vitamin D 3 and metabolites [25(OH)D 3 , 1,25(OH) 2 D 3 , and 24,25(OH) 2 D 3 ] is presented. In this study, patients with 25(OH)D 3 plasma concentrations below 30 ng/ml were studied after a single dose of 5,000 I.U. (125 µg) cholecalciferol, provided with 5,000 I.U. daily cholecalciferol supplementation until vitamin D replete (25(OH)D 3 plasma concentrations above 30 ng/ml), and had serial plasma samples were collected at each phase for 14 days. Total concentrations of vitamin D 3 and metabolites were measured by ultra-high performance liquid chromatography tandem mass spectrometry. A ninecompartment PBPK model was built using MATLAB to represent the triphasic study nature (insufficient, replenishing, sufficient). Stimulatory and inhibitory effect of 1,25(OH) 2 D 3 were incorporated by fold-changes in the primary metabolic enzymes CYP27B1 and CYP24A1, respectively. Incorporation of dynamic adipose partition coefficients for vitamin D 3 and 25(OH)D 3 and variable enzymatic reactions aided in model fitting.Measures of model predictions agreed well with data from metabolites, with 97%, 88%, and 98% of the data for 25(OH)D 3 , 24,25(OH) 2 D 3 , and 1,25(OH) 2 D 3 , respectively, within 2-fold of unity (fold error values between 0.5 and 2.0). Bootstrapping was performed and optimized parameters were reported with 95% confidence intervals.This PBPK model could be a useful tool for understanding the connections between vitamin D and its metabolites under a variety of clinical situations.

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Understanding Cytochrome P450 Enzyme Substrate Inhibition and Prospects for Elimination Strategies

Zhang,

Wang

et al. 2024

ChemBioChem

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Cytochrome P450 (CYP450) enzymes, which are widely distributed and pivotal in various biochemical reactions, catalyze diverse processes such as hydroxylation, epoxidation, dehydrogenation, dealkylation, nitrification, and bond formation. These enzymes have been applied in drug metabolism, antibiotic production, bioremediation, and fine chemical synthesis. Recent research revealed that CYP450 catalytic kinetics deviated from the classic Michaelis–Menten model. A notable substrate inhibition phenomenon that affects the catalytic efficiency of CYP450 at high substrate concentrations was identified. However, the substrate inhibition of various reactions catalyzed by CYP450 enzymes have not been comprehensively reviewed. This review describes CYP450 substrate inhibition examples and atypical Michaelis–Menten kinetic models, and provides insight into mechanisms of these enzymes. We also reviewed 3D structure and dynamics of CYP450 with substrate binding. Outline methods for alleviating substrate inhibition in CYP450 and other enzymes, including traditional fermentation approaches and protein engineering modifications. The comprehensive analysis presented in this study lays the foundation for enhancing the catalytic efficiency of CYP450 by deregulating substrate inhibition.

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Atypical kinetics of cytochrome P450 2J2: Epoxidation of arachidonic acid and reversible inhibition by xenobiotic inhibitors

Cited by 11 publications

References 108 publications

Inhibition of Cytochrome P450 2J2-Mediated Metabolism of Rivaroxaban and Arachidonic Acid by Ibrutinib and Osimertinib

Inhibition of Cytochrome P450 2J2-Mediated Metabolism of Rivaroxaban and Arachidonic Acid by Ibrutinib and Osimertinib

Physiologically Based Pharmacokinetic Modeling of Vitamin D₃ and Metabolites in Vitamin D–Insufficient Patients

Understanding Cytochrome P450 Enzyme Substrate Inhibition and Prospects for Elimination Strategies

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Atypical kinetics of cytochrome P450 2J2: Epoxidation of arachidonic acid and reversible inhibition by xenobiotic inhibitors

Cited by 11 publications

References 108 publications

Inhibition of Cytochrome P450 2J2-Mediated Metabolism of Rivaroxaban and Arachidonic Acid by Ibrutinib and Osimertinib

Inhibition of Cytochrome P450 2J2-Mediated Metabolism of Rivaroxaban and Arachidonic Acid by Ibrutinib and Osimertinib

Physiologically Based Pharmacokinetic Modeling of Vitamin D3 and Metabolites in Vitamin D–Insufficient Patients

Understanding Cytochrome P450 Enzyme Substrate Inhibition and Prospects for Elimination Strategies

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Product

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Physiologically Based Pharmacokinetic Modeling of Vitamin D₃ and Metabolites in Vitamin D–Insufficient Patients