Design and Synthesis of Selective, High-Affinity Inhibitors of Human Cytochrome P450 2J2.

Laboratoire de Chimie et Biochimie Pharmacologiques et Toxicologiques, UMR 8601 CNRS, Universite Paris 5 René Descartes, 45 Rue des Saints-Pères, 75270 Paris Cedex 06, France.
Bioorganic & Medicinal Chemistry Letters (Impact Factor: 2.42). 06/2006; 16(10):2777-80. DOI: 10.1016/j.bmcl.2006.02.004
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The active site topology, substrate specificity, and biological roles of the human cytochrome P450 CYP2J2, which is mainly expressed in the cardiovascular system, are poorly known even though recent data suggest that it could be a novel biomarker and potential target for therapy of human cancer. This paper reports a first series of high-affinity, selective CYP2J2 inhibitors that are related to terfenadine, with K(i) values as low as 160nM, that should be useful tools to determine the biological roles of CYP2J2.

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    • "A specific substrate/inhibitor pair for CYP2J2 may reveal a role for CYP2J2 in drug metabolism that may be underestimated, especially in extrahepatic tissues. CYP2J2-selective inhibition has been studied previously , and several compounds mostly related to the backbone structure of terfenadine have been identified (Lafite et al., 2006, 2007; Chen et al., 2009). However, selectivity of these agents against most This work was supported by the National Institutes of Health National Heart, Lung, and Blood Institute [Grant R01-HL096706]; and the National Institutes of Health National Institute of General Medical Sciences [P01-GM32165]. "
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    ABSTRACT: CYP2J2, an arachidonic acid epoxygenase, is recognized for its role in the first-pass metabolism of astemizole and ebastine. To fully assess the role of CYP2J2 in drug metabolism, a selective substrate and potent specific chemical inhibitor are essential. In this study, we report amiodarone 4-hydoxylation as a specific CYP2J2-catalyzed reaction with no CYP3A4, or other drug-metabolizing enzyme, involvement. Amiodarone 4-hydroxylation enabled the determination of liver relative activity factor and intersystem extrapolation factor for CYP2J2. Amiodarone 4-hydroxylation correlated with astemizole O-demethylation but not with CYP2J2 protein content in a sample of human liver microsomes. To identify a specific CYP2J2 inhibitor, 138 drugs were screened using terfenadine and astemizole as probe substrates with recombinant CYP2J2. Forty-two drugs inhibited CYP2J2 activity by ≥50% at 30 μM, but inhibition was substrate-dependent. Of these, danazol was a potent inhibitor of both hydroxylation of terfenadine (IC(50) = 77 nM) and O-demethylation of astemizole (K(i) = 20 nM), and inhibition was mostly competitive. Danazol inhibited CYP2C9, CYP2C8, and CYP2D6 with IC(50) values of 1.44, 1.95, and 2.74 μM, respectively. Amiodarone or astemizole were included in a seven-probe cocktail for cytochrome P450 (P450) drug-interaction screening potential, and astemizole demonstrated a better profile because it did not appreciably interact with other P450 probes. Thus, danazol, amiodarone, and astemizole will facilitate the ability to determine the metabolic role of CYP2J2 in hepatic and extrahepatic tissues.
    Drug metabolism and disposition: the biological fate of chemicals 02/2012; 40(5):943-51. DOI:10.1124/dmd.111.043505 · 3.25 Impact Factor
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    • "The design and synthesis of high-affinity and selective CYP2J2 inhibitors derived from terfenadone, a derivative of the drug terfenadine, has been described in detail by Lafite et al. (2006). We synthesized an additional novel hydrochloride salt compound , 1-[4-(vinyl) phenyl]-4-[4-(diphenyl-hydroxymethyl)-piperidinyl]-butanone hydrochloride, labeled as compound 26 (C26) (Chen et al., 2009). "
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    ABSTRACT: Cytochrome P450 2J2 (CYP2J2) epoxygenase converts arachidonic acid to four regioisomeric epoxyeicosatrienoic acids (EETs) that exert multiple biological effects in the cardiovascular system and in various human solid cancers. However, it is unknown whether this enzyme is expressed or plays any role in malignant hematological diseases. In this study, we found strong and highly selective CYP2J2 expression in five human-derived malignant hematological cell lines and in leukemia cells from peripheral blood and bone marrow in 36 of 42 patients (86%) with malignant hematologic diseases. Furthermore, increased levels of EETs were detected in urine and blood samples from these patients. Addition of exogenous EET or CYP2J2 overexpression in cultured human-derived malignant hematologic cell lines markedly accelerated proliferation and attenuated apoptosis. Addition of the selective CYP2J2 inhibitor compound 26 (C26; 1-[4-(vinyl) phenyl]-4-[4-(diphenyl-hydroxymethyl)-piperidinyl]-butanone hydrochloride) inhibited cell proliferation and increased apoptosis, an effect that was significantly reversed by EET. CYP2J2 overexpression and exogenous EET activated AMP-activated protein kinase, c-Jun NH(2)-terminal kinase, and phosphatidylinositol 3-kinase/Akt signaling pathways, and increased epidermal growth factor receptor phosphorylation levels. CYP2J2 overexpression also enhanced malignant xenograft growth, which was efficiently inhibited by oral administration of C26 in Tie2-CYP2J2 transgenic mice and in severe combined immunodeficiency (SCID) xenograft mice. Together, these results suggest that CYP2J2 plays a key role in the pathogenesis of human hematologic malignant diseases. Selective inhibition of CYP2J2 may be a promising therapeutic strategy for these conditions.
    Journal of Pharmacology and Experimental Therapeutics 10/2010; 336(2):344-55. DOI:10.1124/jpet.110.174805 · 3.97 Impact Factor
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    ABSTRACT: Although selective inhibitors of cytochrome P450 enzymes can be used to determine relative contributions of the enzymes to xenobiotic metabolism, characterization of CYP2J2 in drug metabolism is more challenging due to lack of selective, well-characterized inhibitors. Thus, selectivity of hydroxyebastine, which has high affinity for recombinant CYP2J2, was studied. The IC50 value of hydroxyebastine in CYP2J2-mediated astemizole O-demethylation activity was lower than that of its structural analog, terfenadine alcohol. Terfenadine alcohol inhibited several other P450 activities, such as CYP2D6, more potently than CYP2J2, and is thus not suitable as a CYP2J2-selective inhibitor. Inhibitory potential values of hydroxyebastine in CYP2J2-catalyzed astemizole O-demethylation, tolbutamide hydroxylation (CYP2C9), S-mephenytoin hydroxylation (CYP2C19), and dextromethorphan O-demethylation (CYP2D6) were 0.45, 2.74, 10.22, and 3.83 μM, respectively. The inhibitory potential of other P450 enzymes, such as CYP1A2, CYP2B6, CYP2E1, and CYP3A, was negligible. Although hydroxyebastine was a relatively potent inhibitor of CYP2J2, it provided a selectivity of only > 6-fold (CYP2J2 vs. other P450s). However, hydroxyebastine can serve as a relatively selective inhibitor of CYP2J2 and can be used to characterize the contribution of CYP2J2 to xenobiotic metabolism due to the lack of a more specific inhibitor.
    Journal of the Korean Society for Applied Biological Chemistry 10/2011; 54(5). DOI:10.1007/BF03253143 · 0.69 Impact Factor
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