In vivo and in vitro diclofenac 5-hydroxylation mediated primarily by cytochrome P450 3A enzymes in common marmoset livers genotyped for P450 2C19 variants

Nakanishi, Kazuyuki; Uehara, Shotaro; Kusama, Takashi; Inoue, Takashi; Shimura, Kanami; Kamiya, Yusuke; Murayama, Norie; Shimizu, Makiko; Uno, Yasuhiro; Sasaki, Erika; Yamazaki, Hiroshi

doi:10.1016/j.bcp.2018.04.002

Cited by 16 publications

(10 citation statements)

References 23 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The fraction of diclofenac metabolized to 4′-hydroxydiclofenac by P450 2C9 in the current humanized-liver model was approximately 0.5 (an average of 0.55, based on 0.62 for C max and 0.47 for AUC infinity ). We recently reported that marmoset P450 2C19 (rather than P450 2C9) and P450 3A4 mediated diclofenac 4′- and 5-hydroxylation, respectively, in an almost similar manner both in vitro and in vivo . The reason why diclofenac works as an efficient in vitro human P450 2C9 probe but not as an in vivo human P450 2C9 probe is not currently known.…”

Section: Discussionmentioning

confidence: 99%

“…We recently reported that marmoset P450 2C19 (rather than P450 2C9) and P450 3A4 mediated diclofenac 4′-and 5-hydroxylation, respectively, in an almost similar manner both in vitro and in vivo. 17 The reason why diclofenac works as an efficient in vitro human P450 2C9 probe but not as an in vivo human P450 2C9 probe is not currently known. Presumably this discrepancy results from different contributions of P450 3A to diclofenac 4′-hydroxylation in vitro and in vivo.…”

Section: ■ Discussionmentioning

confidence: 99%

“…S -Warfarin and 7-hydoxywarfarin were determined by a liquid chromatography (LC) system consisting of a fluorescence detector and pump (Shimadzu, Kyoto, Japan) with an analytical octadecylsilane (C 18 ) reversed-phase column (5 μm, 4.6 × 250 mm, Mightysil RP-18 GP 2, Kanto Chemicals, Tokyo, Japan) according to previously reported methods with minor modifications. − Diclofenac, 4′-hydroxydiclofenac, 5-hydroxydiclofenac, and diclofenac acyl-β- d -glucuronide were determined according to previously reported methods with modifications using LC–tandem mass spectrometry. An API 4000 tandem mass spectrometer (AB Sciex, Framingham, MA) was operated in electrospray positive ionization mode and was directly coupled to a 20A System liquid chromatograph (Shimadzu, Kyoto, Japan) equipped with a C 18 column (4.6 × 250 mm, Kanto Chemicals).…”

Section: Methodsmentioning

confidence: 99%

See 2 more Smart Citations

Different Roles of Human Cytochrome P450 2C9 and 3A Enzymes in Diclofenac 4′- and 5-Hydroxylations Mediated by Metabolically Inactivated Human Hepatocytes in Previously Transplanted Chimeric Mice

Miura

Uehara

Shimizu

et al. 2019

Chem. Res. Toxicol.

Self Cite

View full text Add to dashboard Cite

To investigate the respective roles of cytochromes P450 2C9 and 3A in drug oxidation in human livers, the in vivo pharmacokinetics of S-warfarin and diclofenac were analyzed after intravenous administrations in chimeric mice that had been transplanted with human hepatocytes. P450 2C9 was metabolically inactivated in the humanized mice by orally pretreating them with tienilic acid. After intravenous administration of S-warfarin, a significant difference in the concentration–time profiles of the primary metabolite 7-hydroxywarfarin between untreated mice and mice treated with tienilic acid was observed. In contrast, there were no apparent differences in the profiles for S-warfarin between the treated and untreated groups. The mean values of the maximum concentrations (C max) and the areas under the plasma concentration versus time curves (AUCinfinity) for 7-hydroxywarfarin were significantly lower (22 and 16% of the untreated values, respectively) in the treated group. This presumably resulted from suppressed P450 2C9 activity in the primary oxidative metabolism in vivo in the treated group. After diclofenac administration, plasma levels of diclofenac, 5-hydroxydiclofenac, and diclofenac acylglucuronide were roughly similar in pretreated and untreated mice. However, the mean C max and AUCinfinity values for 4′-hydroxydiclofenac were significantly lower (38 and 53% of the untreated group, respectively) in the treated group. The reported value of ∼0.8 for the fraction of S-warfarin metabolized to 7-hydroxywarfarin mediated by P450 2C9 in in vitro systems was similar to the value implied by the present humanized-liver mouse model pretreated with tienilic acid in which the AUC of 7-hydroxywarfarin was reduced by 84%. In contrast, the fractions of diclofenac metabolized to 4′-hydroxydiclofenac in in vitro and in vivo experiments were inconsistent. These results suggested that humanized-liver mice orally treated with tienilic acid might constitute an in vivo model for metabolically inactivated P450 2C9 in human hepatocytes transplanted into chimeric mice. Moreover, diclofenac, a typical in vitro P450 2C9 probe substrate, was cleared differently in vitro and in humanized-liver mice in vivo.

show abstract

Section: Discussionmentioning

confidence: 99%

Section: ■ Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Different Roles of Human Cytochrome P450 2C9 and 3A Enzymes in Diclofenac 4′- and 5-Hydroxylations Mediated by Metabolically Inactivated Human Hepatocytes in Previously Transplanted Chimeric Mice

Miura

Uehara

Shimizu

et al. 2019

Chem. Res. Toxicol.

Self Cite

View full text Add to dashboard Cite

show abstract

“…Paclitaxel 6α-hydroxylation, diclofenac 4′-hydroxylation, S-and R,S-warfarin 7-hydroxylation, and omeprazole 5-hydroxylation activities of recombinant dog CYP2C proteins were determined as described previously (Uno et al, 2011;Uehara et al, 2015;Uehara et al, 2016;Nakanishi et al, 2018;Uno et al, 2022a). Briefly, typical reaction mixtures (0.20 mL) contained dog liver microsomes (0.50 mg protein/mL) or recombinant protein (5.0 pmol), an NADPH-generating system, and substrate (10 or 100 μM) in 50 mM potassium phosphate buffer (pH 7.4).…”

Section: Enzymatic Characterization Of Dog Cyp2csmentioning

confidence: 99%

“…Briefly, typical reaction mixtures (0.20 mL) contained dog liver microsomes (0.50 mg protein/mL) or recombinant protein (5.0 pmol), an NADPH-generating system, and substrate (10 or 100 μM) in 50 mM potassium phosphate buffer (pH 7.4). The lower substrate concentrations were adopted according to the known values of the Michaelis-Menten constant (K m ) for human CYP2C8/9/19 enzymes (Uehara et al, 2015;Uehara et al, 2016;Nakanishi et al, 2018).…”

Section: Enzymatic Characterization Of Dog Cyp2csmentioning

confidence: 99%

Novel Cytochrome P450 2C94 Functionally Metabolizes Diclofenac and Omeprazole in Dogs

et al. 2023

View full text Add to dashboard Cite

Cytochromes P450 (P450s or CYPs) are important drug-metabolizing enzymes. Because dogs are frequently used in drug metabolism studies, knowledge of dog CYP2C enzymes is essential because, in humans, these enzymes are abundant and play major roles in liver and intestine. The present study identified and characterized novel dog CYP2C94 along with previously identified dog CYP2C21 and CYP2C41. Dog CYP2C21, CYP2C41, and CYP2C94 cDNAs, respectively, contained open reading frames of 490, 489, and 496 amino acids and shared high sequence identities (70%, 75%, and 58%) with human CYP2Cs. Dog CYP2C94 mRNA was preferentially expressed in liver, just as dog CYP2C21 and CYP2C41 mRNAs were. In dog liver, CYP2C21 mRNA was the most abundant, followed by CYP2C94 and CYP2C41 mRNAs. Moreover, the hepatic expressions of all three dog CYP2C mRNAs varied in four individual dogs, two of which did not express CYP2C41 mRNA. The three dog CYP2C genes had similar gene structures, and CYP2C94, although located on the same chromosome, was in a genomic region far from the gene cluster containing CYP2C21 and CYP2C41.Metabolic assays with recombinant proteins showed that dog CYP2C94, along with CYP2C21 and CYP2C41, efficiently catalyzed oxidations of diclofenac, warfarin, and/or omeprazole, indicating that dog CYP2C94 is a functional enzyme. Novel dog CYP2C94 is expressed abundantly in liver and encodes a functional enzyme that metabolizes human CYP2C substrates; it is, therefore, likely responsible for drug clearances in dogs.

show abstract

Identification of emerging contaminants and their transformation products in a moving bed biofilm reactor (MBBR)–based drinking water treatment plant around River Yamuna in India

Tak

Tiwari

Vellanki

2020

Environ Monit Assess

View full text Add to dashboard Cite

In vivo and in vitro diclofenac 5-hydroxylation mediated primarily by cytochrome P450 3A enzymes in common marmoset livers genotyped for P450 2C19 variants

Cited by 16 publications

References 23 publications

Different Roles of Human Cytochrome P450 2C9 and 3A Enzymes in Diclofenac 4′- and 5-Hydroxylations Mediated by Metabolically Inactivated Human Hepatocytes in Previously Transplanted Chimeric Mice

Different Roles of Human Cytochrome P450 2C9 and 3A Enzymes in Diclofenac 4′- and 5-Hydroxylations Mediated by Metabolically Inactivated Human Hepatocytes in Previously Transplanted Chimeric Mice

Novel Cytochrome P450 2C94 Functionally Metabolizes Diclofenac and Omeprazole in Dogs

Identification of emerging contaminants and their transformation products in a moving bed biofilm reactor (MBBR)–based drinking water treatment plant around River Yamuna in India

Contact Info

Product

Resources

About