Establishment and Characterization of the Transformants Stably-Expressing MDR1 Derived from Various Animal Species in LLC-PK1

Takeuchi, Toshiyuki; Yoshitomi, Sumie; Higuchi, Tomoaki; Ikemoto, Keiko; Niwa, Shin‐Ichi; Ebihara, Takuya; Katoh, Miki; Yokoi, Tsuyoshi; Asahi, Satoru

doi:10.1007/s11095-006-0285-7

Cited by 122 publications

(103 citation statements)

References 31 publications

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“…The Food and Drug Administration considers these techniques to be a reference tool for testing the transport of new chemical entities by Pgp, as species-to-species extrapolation of the interactions of compounds with P-gp is hazardous in vivo (10). The correlation between the interaction of many compounds in vitro with human MDR1 and mouse Mdr1a is poor (9,23), and recently published data suggest that there are differences between rodents and humans in the transporter profiles of the brain capillary endothelial cells forming the BBB (24). Thus, a specific human model is needed to clarify the P-gp substrate status of the drugs, so as to prevent any drug-drug interaction that could occur in clinical practice and variability in the brain distribution due to variations in transporter expression.…”

Section: Discussionmentioning

confidence: 99%

“…Most studies on the carrier-mediated transport of radiotracers across the BBB are performed on rodents using chemical inhibition or physical disruption of the P-gp or Bcrp transporter genes (8). However, some differences in the spectra of ABC substrates of the human and rodent isoforms of Pgp indicate that models of human ABCs should be used (9,10). A few studies have been done on humans, but they require expensive, time-consuming experimental protocols using cyclosporine A or newer-generation P-gp inhibitors (11,12).…”

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confidence: 99%

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Transport of Selected PET Radiotracers by Human P-Glycoprotein (ABCB1) and Breast Cancer Resistance Protein (ABCG2): An In Vitro Screening

Tournier¹,

Valette²,

Peyronneau³

et al. 2011

J Nucl Med

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Radiolabeled compounds used for brain imaging with PET must readily cross the blood-brain barrier (BBB) to reach their target. Efflux transporters at the BBB-P-glycoprotein (P-gp) and the breast cancer resistance protein (BCRP)-could limit their uptake by the brain. The assays were performed using the nonradioactive form of each compound (ultraviolet high-performance liquid chromatography analysis) and, when available, the 18 F-labeled analogs (g-counting). Results: Befloxatone appeared to be transported solely by BCRP. Loperamide, verapamil, and diprenorphine were the only P-gp substrates. Other ligands were transported by neither P-gp nor BCRP. Conclusion: The present method can readily be used to screen new-compound transport by Pgp or BCRP, even before any radiolabeling. Compounds that were previously thought to be transported by P-gp in rodents, such as p-MPPF, WAY-100635, and flumazenil, cannot be considered substrates of human P-gp. The impact of BCRP and P-gp at the BBB on the transport of befloxatone and diprenorphine in vivo remains to be evaluated with PET.

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Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

Transport of Selected PET Radiotracers by Human P-Glycoprotein (ABCB1) and Breast Cancer Resistance Protein (ABCG2): An In Vitro Screening

Tournier¹,

Valette²,

Peyronneau³

et al. 2011

J Nucl Med

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“…Although the fact that permeability of P-gp substrates to the rat small intestine was predicted from in vitro study using cell lines expressing human P-gp may come under intense scrutiny, several reports have shown the good correlation between the functional activity of human MDR1 and that of rat mdr1a/1b. Takeuchi et al (2006) established the LLC-PK1 cell lines, which stably express various MDR1 (derived from human, monkey, canine, rat, and mouse) to investigate species differences in P-gpmediated efflux activity. When the permeability ratios (BL to AP/AP to BL) of P-gp substrate drugs were compared among these cell lines, a good correlation was observed between human and rat MDR1 (both MDR1a and 1b).…”

Section: Discussionmentioning

confidence: 99%

Scaling of in Vitro Membrane Permeability to Predict P-glycoprotein-Mediated Drug Absorption in Vivo

Shirasaka¹,

Masaoka²,

Kataoka³

et al. 2008

Drug Metab Dispos

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ABSTRACT:In a previous study, the concentration-dependent permeability of P-glycoprotein (P-gp) substrate drugs, quinidine, verapamil, and vinblastine, in several cell monolayers with different levels of P-gp expression was analyzed kinetically to obtain fundamental parameters for P-gp-mediated transport, V max and K m(app) values. Both V max and K m(app) values of each drug were found to show linear correlations with the expression level of P-gp. These findings imply the possibility of estimating the V max and K m(app) values of P-gp substrate drugs in the in vivo intestinal membrane on the basis of the P-gp expression level. In the present study, concentrationdependent drug permeability to the rat small intestines (upper jejunum and ileum) was simulated on the basis of V max and K m(app) values of each drug estimated from the P-gp expression level in the rat small intestines. To validate the predictability of these procedures, drug permeability in the rat small intestines was measured by the in situ single-pass perfusion method. It was confirmed that simulated permeability of each drug in the rat jejunum and ileum corresponded well with permeability measured by the in situ single-pass perfusion method. This study clearly demonstrated the potential to estimate the permeability of P-gp substrate drugs in the human intestine from its P-gp expression level and thus the possibility to predict the oral absorption of those drugs.P-glycoprotein (P-gp), one of the most important efflux transporters in the field of biopharmacy, is an ATP-binding cassette (ABC) transporter encoded by the multidrug resistance 1 gene (MDR1/ABCB1). P-gp is expressed not only in cancer cells but also in many normal tissues (Loo and Clarke, 1999). So far, many reports have described the effects of P-gp on the pharmacokinetic profiles of clinically important drugs, by denaturing absorption, distribution, and excretion. For example, P-gp located in the apical domain of the enterocytes of the gastrointestinal tract limits the uptake and absorption of its substrate drugs after oral administration (Hunter and Hirst, 1997;Meijer et al., 1997;Jonker et al., 1999). Because of the great impact of P-gp on the pharmacokinetic profiles of a variety of drugs, various in vitro experimental techniques are now used to identify the compounds subjected to P-gp-mediated efflux (Kim et al., 1998;Polli et al., 1999Polli et al., , 2001Susanto and Benet, 2002; Shirasaka et al., 2006a,b). However, it is still difficult to quantitatively predict the effect of P-gp on in vivo intestinal absorption of such drugs.In a previous study, we measured the apical (AP) to basal (BL) absorptive permeability of P-gp substrate drugs in several cultured cell monolayers with different expression levels of P-gp (Shirasaka et al., 2008). In all cell monolayers, AP to BL permeability of P-gp substrate drugs showed a sigmoid-type relation to their donor (AP) concentration and reached a maximal value at the higher concentration range because of the saturation of P-gp-mediated efflux. Con...

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“…The LLC-PK1 cells expressing rat Mdr1a and human MDR1 were cultured using a slight modification of a method reported previously (Takeuchi et al, 2006). In brief, cells were incubated in complete culture medium consisting of M199 (Invitrogen, Carlsbad, CA) or Dulbecco's modified Eagle's medium (Invitrogen) with 20% fetal bovine serum (Invitrogen), 500 g/ml G418, and 150 ng/ml colchicine.…”

Section: Methodsmentioning

confidence: 99%

Quantitative Investigation of the Impact of P-Glycoprotein Inhibition on Drug Transport across Blood-Brain Barrier in Rats

et al. 2010

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ABSTRACT:The magnitude of P-glycoprotein [(P-gp)/multidrug resistance protein 1 (MDR1)]-mediated drug-drug interaction (DDI) at the blood-brain barrier (BBB) in rats was estimated by in vitro-in vivo correlation (IVIVC). In in vitro studies, rat Mdr1a-expressing LLC-PK1 cells were examined for the evaluation of P-gp inhibitory activity using digoxin as a P-gp probe substrate. The in vitro K i value was calculated using a modified corrected flux ratio that reflects the P-gp function. In in vivo studies, digoxin with or without P-gp inhibitors was administered to rats by constant intravenous infusion to evaluate the effect of P-gp inhibition on digoxin transport to the brain under steady-state conditions. In the presence of elacridar, the brain-to-plasma concentration ratio (K p,brain ) of digoxin was approximately 14 times the control value. However, no significant change in the K p,brain was observed

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Establishment and Characterization of the Transformants Stably-Expressing MDR1 Derived from Various Animal Species in LLC-PK1

Abstract: Results in the present study indicate that all MDR1 stably expressing cells have efflux activity for various P-gp substrates, and that interspecies differences and similarities of the P-gp substrate efflux activity may exist.

Cited by 122 publications

References 31 publications

Transport of Selected PET Radiotracers by Human P-Glycoprotein (ABCB1) and Breast Cancer Resistance Protein (ABCG2): An In Vitro Screening

Transport of Selected PET Radiotracers by Human P-Glycoprotein (ABCB1) and Breast Cancer Resistance Protein (ABCG2): An In Vitro Screening

Scaling of in Vitro Membrane Permeability to Predict P-glycoprotein-Mediated Drug Absorption in Vivo

Quantitative Investigation of the Impact of P-Glycoprotein Inhibition on Drug Transport across Blood-Brain Barrier in Rats

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