Use of canalicular membrane vesicles (CMVs) from rats, dogs, monkeys and humans to assess drug transport across the canalicular membrane

Shilling, Adam D.; Azam, Farooq; Kao, John Y.; Leung, Louis

doi:10.1016/j.vascn.2005.08.003

Cited by 32 publications

(30 citation statements)

References 28 publications

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“…The study by Zelcer et al (2003), similarly to our data, showed cooperative interaction with lower affinities (K 0.5 ϭ 120 M). A study using human canalicular membrane vesicles showed Michaelis-Menten type kinetics with a K m of 364 M (Shilling et al, 2006). For rat protein classic Michaelis-Menten type kinetics was shown (Borst et al, 2006b).…”

Section: Heré Di-szabó Et Almentioning

confidence: 99%

“…One of them demonstrated a sigmoidal transport with a Hill number of 1.16 (Ninomiya et al, 2005; also reviewed in Borst et al, 2006b), whereas the other group calculated a Hill number of 1.5 (Gerk et al, 2004). The reported K m values range from single digit Ito et al, 2001) through double digit (Borst et al, 2006b) to triple digit numbers (Shilling et al, 2006). Our data are similar to results obtained using rat Mrp2-Sf9 for which MichaelisMenten type kinetics and double digit K m (K m ϭ 16 M) was found (Borst et al, 2006b).…”

Section: Heré Di-szabó Et Almentioning

confidence: 99%

“…However, we have repeated E 2 17␤G transport using MDCKII membranes overexpressing rat Mrp2 and obtained Michaelis-Menten type kinetics (data not shown). Likewise, one of the articles cited (Shilling et al, 2006) used rat canalicular membrane vesicles membranes and observed hyperbolic kinetics.…”

Section: Heré Di-szabó Et Almentioning

confidence: 99%

“…In preclinical studies mostly rodents are used to investigate the pharmacokinetics and toxicity of the compounds. Species specificity studies have been carried out for many MRP2 orthologs (Ninomiya et al, 2005(Ninomiya et al, , 2006Zimmermann et al, 2008;Shilling et al, 2006). However, detailed studies that included membrane as well as cellular experimental systems have only been performed for the human and the mouse protein (Zimmermann et al, 2008).…”

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Multidrug Resistance Protein 2-Mediated Estradiol-17β-d-glucuronide Transport Potentiation: In Vitro-in Vivo Correlation and Species Specificity

et al. 2008

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ABSTRACT:Multidrug resistance protein 2 (MRP2) is a multispecific organic anion transporter expressed at important pharmacological barriers, including the canalicular membrane of hepatocytes. At this location it is involved in the elimination of both endogenous and exogenous waste products, mostly as conjugates, to the bile. Estradiol-17␤-D-glucuronide (E 2 17␤G), a widely studied endogenous substrate of MRP2, was shown earlier to recognize two binding sites of the transporter in vesicular transport assays. MRP2 modulators (substrates and nonsubstrates) potentiate the transport of E 2 17␤G by MRP2. We correlated data obtained from studies of different complexities and investigated the speciesspecific differences between rat and human MRP2-mediated transport. We used vesicular transport assays, sandwich-cultured primary hepatocytes, and in vivo biliary efflux in rats. Our results demonstrate that the rat Mrp2 transporter, unlike the human MRP2, transports E 2 17␤G according to Michaelis-Menten type kinetics. Nevertheless, in the presence of modulator drugs E 2 17␤G transport mediated by the rat transporter also shows cooperative kinetics as potentiation of E 2 17␤G transport was observed in the vesicular transport assay. We also demonstrated that the potentiation exists both in rat and in human hepatocytes and in vivo in rats.

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Section: Heré Di-szabó Et Almentioning

confidence: 99%

Section: Heré Di-szabó Et Almentioning

confidence: 99%

Section: Heré Di-szabó Et Almentioning

confidence: 99%

mentioning

confidence: 99%

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Multidrug Resistance Protein 2-Mediated Estradiol-17β-d-glucuronide Transport Potentiation: In Vitro-in Vivo Correlation and Species Specificity

et al. 2008

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“…Even though the interspecies scaling methods based on physiologically allometric procedures have been successfully applied, particularly into extrapolation of hepatic enzymatic metabolism and urinary excretion (Dedrick et al, 1970;Iwatsubo et al, 1997;Ito et al, 1998), the in vitro or in vivo model for biliary excretion predication is far from being mature (Mahmood and Sahajwalla, 2002). The remarkable interspecies differences in biliary excretion of xenobiotics and drugs/metabolites (Ishizuka et al, 1999;Shilling et al, 2006) may cause significant overestimation of biliary excretion in humans simply by an exponential allometric extrapolation approach (Lave et al, 1999;Pahlman et al, 1999;Ayrton and Morgan, 2001). Therefore, understanding the molecular mechanisms underlying the marked species difference in hepatobiliary elimination of drugs and their metabolites should greatly advance the current allometric scaling models for estimation of human pharmacokinetics.…”

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confidence: 99%

Absolute Difference of Hepatobiliary Transporter Multidrug Resistance-Associated Protein (MRP2/Mrp2) in Liver Tissues and Isolated Hepatocytes from Rat, Dog, Monkey, and Human

Li¹,

Zhang²,

Hua³

et al. 2008

Drug Metab Dispos

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ABSTRACT:We previously reported that hepatobiliary transporter multidrug resistance-associated protein (MRP2/Mrp2) is considered to be the major cause of the interspecies differences detected by efflux of fluorescent substrates in isolated hepatocytes. In the present study, the interspecies differences of MRP2/Mrp2 were first evaluated by quantitative real-time polymerase chain reaction and Western blotting. The mRNA levels were able to distinguish the difference among species with a rank order comparable with the corresponding activities observed, whereas the extents of the differences remained unknown. The cross-reactions of MRP2/Mrp2 protein of different species with anti-human MRP2 polyclonal antibody were found by Western blotting. However, because of the unknown binding affinity of antibody to MRP2/Mrp2 protein across species and lack of purified MRP2/Mrp2 proteins for calibration, the immunoblotting assay was excluded from the absolute quantification of MRP2/Mrp2 protein for multiple species. By using our newly developed liquid chromatography-tandem mass spectrometry quantification method, we were able to measure the absolute amount of MRP2/Mrp2 in liver tissues and isolated hepatocytes across species. Freshly isolated hepatocytes conserved MRP2/ Mrp2 protein levels that are comparable with those in the liver tissues. The amount of Mrp2 in rat liver was approximately 10-fold higher than that in other species. Moreover, a significant loss of Mrp2 protein in the membrane fraction of rat cryopreserved hepatocytes was observed. Thus, the absolute differences of MRP2/ Mrp2 levels in various species were determined, for the first time, by direct quantification. The results could potentially fill the translational gaps of in vitro/in vivo or preclinical species to human extrapolation of hepatobiliary elimination mediated by MRP2/ Mrp2.Xenobiotics and their metabolites are generally eliminated and detoxified by phase I and phase II enzymatic metabolism, by phase III transporter-mediated drug efflux to bile, or by both mechanisms. The excretion of drugs by hepatocytes into bile is one of the primary elimination routes for xenobiotics and the conjugate metabolites (Arias et al., 1993). In each stage of drug discovery, accurate prediction of human pharmacokinetics for a potential drug candidate is of great value (Mahmood, 1999). Even though the interspecies scaling methods based on physiologically allometric procedures have been successfully applied, particularly into extrapolation of hepatic enzymatic metabolism and urinary excretion (Dedrick et al., 1970;Iwatsubo et al., 1997;Ito et al., 1998), the in vitro or in vivo model for biliary excretion predication is far from being mature (Mahmood and Sahajwalla, 2002). The remarkable interspecies differences in biliary excretion of xenobiotics and drugs/metabolites (Ishizuka et al., 1999;Shilling et al., 2006) may cause significant overestimation of biliary excretion in humans simply by an exponential allometric extrapolation approach (Lave et al., 1999;Pahlman et al., ...

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Introduction to Drug Transporters

Leung¹,

Oganesian²

2008

Drug Metabolism Handbook

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Use of canalicular membrane vesicles (CMVs) from rats, dogs, monkeys and humans to assess drug transport across the canalicular membrane

Cited by 32 publications

References 28 publications

Multidrug Resistance Protein 2-Mediated Estradiol-17β-d-glucuronide Transport Potentiation: In Vitro-in Vivo Correlation and Species Specificity

Multidrug Resistance Protein 2-Mediated Estradiol-17β-d-glucuronide Transport Potentiation: In Vitro-in Vivo Correlation and Species Specificity

Absolute Difference of Hepatobiliary Transporter Multidrug Resistance-Associated Protein (MRP2/Mrp2) in Liver Tissues and Isolated Hepatocytes from Rat, Dog, Monkey, and Human

Introduction to Drug Transporters

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