Effects of Bergamottin on Human and Monkey Drug-Metabolizing Enzymes in Primary Cultured Hepatocytes

Wen, Yuan H.; Sahi, Jasminder; Urda, Ellen; Kulkarni, Shaila; Rose, Kelly; Zheng, Xianxian; Sinclair, Jacqueline F.; Cai, Hongbo; Strom, Stephen C.; Kostrubsky, Vsevolod E.

doi:10.1124/dmd.30.9.977

Cited by 42 publications

(31 citation statements)

References 32 publications

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“…Testosterone metabolism and EROD activity on cultured hepatocytes were conducted as described by Kostrubsky, et al 40 and Wen et al 41 , respectively. Briefly, hepatocytes were isolated from control, non-transplanted mice or those showing repopulation with human hepatocytes using a standard col-lagenase perfusion protocol 42 .…”

Section: Drug Metabolizing Enzymes In Cultured Hepatocytesmentioning

confidence: 99%

“…Cells were then plated in HMM media (hepatocyte maintenance media; Lonza, USA) at a density of 1.5 million cells per well in collagen I -coated 6 well plates. Cells were cultured for 48 h without inducers and then for 48 additional hours in the presence of 1 mM Phenobarbital, 10 μM rifampicin or 25 μ M beta-naphthoflavone as previously described 40,41 . Following the induction period, cells were washed 3 times with PBS to remove inducers, and media was replaced with fresh media (without inducers).…”

Section: Drug Metabolizing Enzymes In Cultured Hepatocytesmentioning

confidence: 99%

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Robust expansion of human hepatocytes in Fah−/−/Rag2−/−/Il2rg−/− mice

et al. 2007

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Mice that could be highly repopulated with human hepatocytes would have many potential uses in drug development and research applications. The best available model of liver humanization, the uroplasminogen-activator transgenic model, has major practical limitations. To provide a broadly useful hepatic xenorepopulation system, we generated severely immunodeficient, fumarylacetoacetate hydrolase (Fah)-deficient mice. After pretreatment with a urokinaseexpressing adenovirus, these animals could be highly engrafted (up to 90%) with human hepatocytes from multiple sources, including liver biopsies. Furthermore, human cells could be serially transplanted from primary donors and repopulate the liver for at least four sequential rounds. The expanded cells displayed typical human drug metabolism. This system provides a robust platform to produce high-quality human hepatocytes for tissue culture. It may also be useful for testing the toxicity of drug metabolites and for evaluating pathogens dependent on human liver cells for replication.The liver is the principal site for the metabolism of xenobiotic compounds, including medical drugs. Because many hepatic enzymes are species specific, it is necessary to evaluate the metabolism of candidate pharmaceuticals using cultured primary human hepatocytes 1,2 . Today, hepatocytes are isolated primarily from cadaveric organs and then shipped to the location where testing will be performed. The condition (viability and state of differentiation) of hepatocytes from cadaveric sources is highly variable, and many cell preparations are of marginal quality. The availability of high-quality human hepatocytes is further hampered by the fact that they cannot be substantially expanded in tissue culture 3,4 . Hepatocytes from readily available mammalian species, such as the mouse, are not suitable for drug testing because they have a different complement of metabolic enzymes and

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Section: Drug Metabolizing Enzymes In Cultured Hepatocytesmentioning

confidence: 99%

Section: Drug Metabolizing Enzymes In Cultured Hepatocytesmentioning

confidence: 99%

Robust expansion of human hepatocytes in Fah−/−/Rag2−/−/Il2rg−/− mice

et al. 2007

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“…RNA was extracted with Trizol reagent using the method recommended by the manufacturer (Invitrogen, Carlsbad, CA). The microarray was fabricated and used as described previously (Kane et al, 2001;Wen et al, 2002). Briefly, three oligonucleotides per gene were designed and amino-modified 50mer oligos were spotted onto SuModic slides using a Molecular Dynamic Gen III robotic spotter (Amersham Biosciences).…”

Section: Methodsmentioning

confidence: 99%

Comparative Effects of Thiazolidinediones on in Vitro P450 Enzyme Induction and Inhibition

Sahi¹,

Black²,

Hamilton³

et al. 2003

Drug Metab Dispos

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102

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This article is available online at http://dmd.aspetjournals.org ABSTRACT:Rosiglitazone and pioglitazone are thiazolidinediones used for treatment of noninsulin-dependent diabetes mellitus. These compounds, along with troglitazone, were evaluated for the ability to induce cytochrome P450 enzymes (P450) in primary human hepatocyte cultures and to inhibit P450 in human microsomes. In induction studies, all three thiazolidinediones caused a dose-dependent increase in CYP3A4 activity and immunoreactive protein.While troglitazone was the most potent, rosiglitazone and pioglitazone generally exceeded troglitazone in absolute CYP3A4 activity achieved at concentrations >10 M. A comparable concentration-dependent increase in CYP2B6 immunoreactive protein was observed with all three thiazolidinediones. Microarray analysis revealed rifampin > troglitazone > pioglitazone > rosiglitazone in terms of CYP3A4 mRNA induction potential with 10 M compound. Inhibition studies conducted for CYP1A2, CYP2C8, CYP2C9, CYP2C19, CYP2D6, CYP3A4, CYP2A6, and CYP2E1 showed troglitazone to be the most nonselective and potent inhibitor followed by rosiglitazone and pioglitazone. In vitro, the thiazolidinediones were strong inhibitors of CYP2C8, with K i values between 1.7 and 5.6 M, and of CYP3A4, with K i values between 1.6 and 11.8 M.Troglitazone, in addition, inhibited CYP2C9 (K i 0.6 M). Although the inhibitory effects of the thiazolidinediones have not been demonstrated clinically, our results suggest there is potential for interactions with CYP2C8 substrates. This is the first report of in vitro induction of P450 enzymes by rosiglitazone and pioglitazone. While only the induction of CYP3A4 by troglitazone has been demonstrated in vivo, these results suggest that other thiazolidinediones may have the potential to cause clinically significant drug interactions at sufficiently high doses.

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“…Bergamottin (BG) and 6Ј,7Ј-dihydroxybergamottin (DHBG), the two most abundant furanocoumarins present in grapefruit, have been characterized as reversible inhibitors and as mechanism-based inactivators of CYP3A4 (Schmiedlin-Ren et al, 1997;He et al, 1998;Guo et al, 2000;Ohnishi et al, 2000;Tassaneeyakul et al, 2000). Subsequently, studies in primary cultured hepatocytes and more clinically related research with BG and DHBG were pursued (Wen et al, 2002;Goosen et al, 2004;Kakar et al, 2004;Paine et al, 2004Paine et al, , 2006. Accelerated degradation of CYP3A4 after covalent modification of the apoprotein by BG and DHBG was confirmed to play a role in the grapefruit juice effect by several in vivo studies (Goosen et al, 2004;Kakar et al, 2004;Paine et al, 2006).…”

Section: Introductionmentioning

confidence: 99%