Comparison of Two Immortalized Human Cell Lines to Study Nuclear Receptor-Mediated CYP3A4 Induction

Harmsen, Stefan; Koster, Andries S.; Beijnen, Jos H.; Schellens, Jan H.M.; Meijerman, Irma

doi:10.1124/dmd.107.017335

Cited by 54 publications

(34 citation statements)

References 35 publications

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“…The screening was performed in the intestinal cell line LS180, because previous comparison of the CYP3A4 inducibility in both LS180 and other widely used cell lines such as Caco-2 [14] and HepG2 [15] had revealed that LS180 cells represent a better model to study CYP3A4 induction.…”

Section: Discussionmentioning

confidence: 99%

Nuclear receptor mediated induction of cytochrome P450 3A4 by anticancer drugs: a key role for the pregnane X receptor

Harmsen

Meijerman

Beijnen

et al. 2008

Cancer Chemother Pharmacol

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Purpose Induction of cytochrome P450 (CYP) 3A4, an enzyme that is involved in the biotransformation of more than 50% of all drugs, by xenobiotics is an important cause of pharmacokinetic drug-drug interactions in oncology. In addition to rifampicin and hyperforin, the anticancer drug paclitaxel has also been shown to be an inducer of CYP3A4 via activation of the pregnane X receptor (PXR). We therefore screened 18 widely used anticancer drugs for their ability to activate PXR-mediated CYP3A4 induction. Methods A CYP3A4 reporter gene assay was employed to identify PXR agonists among the eighteen anticancer drugs. Subsequently CYP3A4 mRNA and protein expression following treatment with these PXR agonists was assessed. Finally, the eVect of pre-treatment with these agents on the 1'-hydroxylation of midazolam (a speciWc CYP3A4 probe) was determined. Results Paclitaxel, erlotinib, tamoxifen, ifosfamide, Xutamide and docetaxel are able to activate PXR, while only strong PXR activation leads to signiWcant induction of CYP3A4 activity. Conclusions The identiWed PXR agonists may have the propensity to cause clinically relevant drug-drug interactions as a result of CYP3A4 induction.

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Section: Discussionmentioning

confidence: 99%

Nuclear receptor mediated induction of cytochrome P450 3A4 by anticancer drugs: a key role for the pregnane X receptor

Harmsen

Meijerman

Beijnen

et al. 2008

Cancer Chemother Pharmacol

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“…LS180 cells were used in the present study because it was previously demonstrated that these cells are a more suitable cell line to study the induction of CYP3A4 compared with the more frequently used cell line HepG2 (Harmsen et al, 2008). Of all tested CAM, Echinacea and vitamin B 12 were shown to moderately inhibit PXRmediated CYP3A4 induction by rifampicin.…”

Section: Discussionmentioning

confidence: 99%

“…One of the major transcriptional regulators of CYP3A4 is the pregnane X receptor (PXR) (Pascussi et al, 2003;Harmsen et al, 2008). After activation of PXR by ligand binding, PXR forms a heterodimer with retinoid X receptor in the nucleus.…”

Section: Introductionmentioning

confidence: 99%

Milk Thistle’s Active Components Silybin and Isosilybin: Novel Inhibitors of PXR-Mediated CYP3A4 Induction

et al. 2013

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Because cancer is often treated with combination therapy, unexpected pharmacological effects can occur because of drug-drug interactions. Several drugs are able to cause upregulation or downregulation of drug transporters or cytochrome P450 enzymes, particularly CYP3A4. Induction of CYP3A4 may result in decreased plasma levels and therapeutic efficacy of anticancer drugs. Since the pregnane X receptor (PXR) is one of the major transcriptional regulators of CYP3A4, PXR antagonists can possibly prevent CYP3A4 induction. Currently, a limited number of PXR antagonists are available. Some of these antagonists, such as sulphoraphane and coumestrol, belong to the so-called complementary and alternative medicines (CAM). Therefore, the aim was to determine the potential of selected CAM (b-carotene, Echinacea purpurea, garlic, Ginkgo biloba, ginseng, grape seed, green tea, milk thistle, saw palmetto, valerian, St. John's Wort, and vitamins B 6 , B 12 , and C) to inhibit PXR-mediated CYP3A4 induction at the transcriptional level, using a reporter gene assay and a real-time polymerase chain reaction assay in LS180 colon adenocarcinoma cells. Furthermore, computational molecular docking and a LanthaScreen time-resolved fluorescence resonance energy transfer (TR-FRET) PXR competitive binding assay were performed to explore whether the inhibiting CAM components interact with PXR. The results demonstrated that milk thistle is a strong inhibitor of PXR-mediated CYP3A4 induction. The components of milk thistle responsible for this effect were identified as silybin and isosilybin. Furthermore, computational molecular docking revealed a strong interaction between both silybin and isosilybin and PXR, which was confirmed in the TR-FRET PXR assay. In conclusion, silybin and isosilybin might be suitable candidates to design potent PXR antagonists to prevent drug-drug interactions via CYP3A4 in cancer patients.

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“…Among them, the HepG2 cell line has been well characterized and is widely used throughout drug metabolism and toxicology testing. HepG2 cells have demonstrated robust induction response to CYP1A inducers; however, due to the low basal expression of enzymes and little to no induction response to known CYP3A4 inducers, HepG2 cells are not considered an appropriate model to study drug-drug interactions (Westerink and Schoonen, 2007;Harmsen et al, 2008). Gómez-Lechón et al (2001) performed an extensive analysis of the biotransformation properties of BC2 cells by measuring basal and inducible expression of Phase I and II enzymes.…”

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confidence: 99%

In Vitro and in Vivo Induction of Cytochrome P450: A Survey of the Current Practices and Recommendations: A Pharmaceutical Research and Manufacturers of America Perspective

Chu¹,

Einolf²,

Evers³

et al. 2009

Drug Metab Dispos

153

122

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ABSTRACT:Cytochrome P450 (P450) induction is one of the factors that can affect the pharmacokinetics of a drug molecule upon multiple dosing, and it can result in pharmacokinetic drug-drug interactions with coadministered drugs causing potential therapeutic failures. In recent years, various in vitro assays have been developed and used routinely to assess the potential for drug-drug interactions due to P450 induction. There is a desire from the pharmaceutical industry and regulatory agencies to harmonize assay methodologies, data interpretation, and the design of clinical drug-drug interaction studies. In this article, a team of 10 scientists from nine Pharmaceutical Research and Manufacturers of America (PhRMA) member companies conducted an anonymous survey among PhRMA companies to query current practices with regards to the conduct of in vitro induction assays, data interpretation, and clinical induction study practices. The results of the survey are presented in this article, along with reviews of current methodologies of in vitro assays and in vivo studies, including modeling efforts in this area. A consensus recommendation regarding common practices for the conduct of P450 induction studies is included.The convergence of recent advances in molecular biology and genomics, higher throughput chemical synthesis, and automated highthroughput in vitro enzyme and cell-based assays to assess biological activity has led to shorter lead identification and optimization times in drug research. However, these breakthroughs have not yet translated into success in drug development. Surveys suggest that in the last several years, the number of New Drug Applications submitted to regulatory agencies has declined and that the poor success rate during drug development is to some extent due to late-stage failures. It is important that novel and innovative approaches are used for assessing the risks and benefits of new molecular entities (NMEs) early in the research and development life cycle to minimize late-stage attrition.Based on a survey conducted in 2004, the major factors for compound attrition during clinical development are lack of efficacy, toxicity, and safety, and suboptimal pharmacokinetics and/or bioavailability, with the remaining failures due to financial and/or commercial reasons (Kola and Landis, 2004). In that survey, one notable observation was the reduction of compound attrition due to pharmacokinetic and/or bioavailability issues from pre-1991 to the period be- ABBREVIATIONS: NME, new molecular entity; P450, cytochrome P450; DDI, drug-drug interaction; RIF, rifampicin; PhRMA, Pharmaceutical Research and Manufacturers of America; AhR, aryl hydrocarbon receptor; CAR, constitutive androstane receptor; PXR, pregnane X receptor; UGT, uridine diphosphate glucuronosyl transferase; FXR, farnesyl X receptor; PPAR, peroxisome proliferator-activated receptor; VDR, vitamin D receptor; Nrf2, nuclear factor erythroid 2-related factor 2; LBD, ligand binding domain; hPXR, human PXR; SR12813, tetra-ethyl 2-(3,5-di-tertbutyl...

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Comparison of Two Immortalized Human Cell Lines to Study Nuclear Receptor-Mediated CYP3A4 Induction

Cited by 54 publications

References 35 publications

Nuclear receptor mediated induction of cytochrome P450 3A4 by anticancer drugs: a key role for the pregnane X receptor

Nuclear receptor mediated induction of cytochrome P450 3A4 by anticancer drugs: a key role for the pregnane X receptor

Milk Thistle’s Active Components Silybin and Isosilybin: Novel Inhibitors of PXR-Mediated CYP3A4 Induction

In Vitro and in Vivo Induction of Cytochrome P450: A Survey of the Current Practices and Recommendations: A Pharmaceutical Research and Manufacturers of America Perspective

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