Comparison of drug transporter gene expression and functionality in Caco-2 cells from 10 different laboratories

Hayeshi, Rose; Hilgendorf, Constanze; Artursson, Per; Augustijns, Patrick; Brodin, Birger; Dehertogh, Pascale; Fisher, Karen A.; Fossati, Lina; Hovenkamp, E.; Korjamo, Timo; Masungi, Chantal; Maubon, Nathalie; Mols, Raf; Müllertz, Anette; Mönkkönen, Jukka; O’Driscoll, Caitriona M.; Oppers-Tiemissen, H.M.; Ragnarsson, Eva; Rooseboom, Martijn; Ungell, Anna‐Lena

doi:10.1016/j.ejps.2008.08.004

Cited by 230 publications

(207 citation statements)

References 36 publications

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“…5 This had resulted in improvements in permeability, as assessed in a Caco-2 assay (apical pH =6.5; basolateral pH = 7.4). 8 This was consistent with a reduction in hydrogen bond donor count, but had led to an unacceptable loss in potency for the three molecular matched pairs ( Figure 1). Notably, methylation of R4 appeared to have the least impact on Caco-2 permeability, with more significant changes being observed at R1 and R7.…”

Section: Body Textsupporting

confidence: 60%

“…By contrast, 9-substitution resulted in an increase in both potency and permeability. Methyl substitution (8)(9)(10) showed similar SAR in terms of potency, with 6-and 8-substitution resulting in lower potencies relative to the un-substituted core. As before, 9-substitution resulted in increased potency and an improvement in terms of Ligand Lipophilicity Efficiency (LLE) 13 of +1.3 relative to the un-substituted core.…”

Section: Body Textmentioning

confidence: 99%

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Achieving improved permeability by hydrogen bond donor modulation in a series of MGAT2 inhibitors

et al. 2013

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Section: Body Textsupporting

confidence: 60%

Section: Body Textmentioning

confidence: 99%

Achieving improved permeability by hydrogen bond donor modulation in a series of MGAT2 inhibitors

et al. 2013

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“…They found a decreased efflux ratio for DRV at a concentration of 100 M (efflux ratio was 3.7) and stated that there was only a limited impact of efflux transporters on the intestinal absorption of DRV. This difference in results may be due to the interlaboratory differences in P-gp expression in the Caco-2 cell line (Hayeshi et al, 2008).…”

Section: Resultsmentioning

confidence: 81%

In Situ Intestinal Perfusion in Knockout Mice Demonstrates Inhibition of Intestinal P-Glycoprotein by Ritonavir Causing Increased Darunavir Absorption

Holmstock

Mols²,

Annaert³

et al. 2010

Drug Metab Dispos

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ABSTRACT:Darunavir is a second-generation protease inhibitor designed to have antiviral efficacy against HIV-1 with multiple resistance mutations to protease inhibitors. It is always coadministered with a subtherapeutic dose of ritonavir. It has been shown that darunavir and ritonavir are substrates of P-glycoprotein (P-gp). We explored the contribution of P-gp to the transport characteristics of darunavir (up to 100 M) using Caco-2 monolayers and the recently developed in situ intestinal perfusion technique using wild-type and mdr1a/1b(؊/؊) mice. We observed that, in vitro, P-gp has a modulatory effect on the absorption of darunavir, even at a concentration of 100 M (efflux ratio ‫؍‬ 25). Simulated intestinal fluids partially inhibited P-gp functionality, which was further inhibited by adding the P-gp inhibitors verapamil, 6-[(2S,4R,6E)-4-methyl-2-(methylamino)-3-oxo-6-octenoic acid]cyclosporine D (PSC833), N-(4-[2-(1,2,3,4-tetrahydro-6,7-dimethoxy-2-isoquinolinyl) ethyl]-phenyl)-9,10-dihydro-5-methoxy-9-oxo-4-acridine carboxamide (GF120918), or ritonavir. Using the in situ intestinal perfusion technique, we demonstrated that coperfusion with ritonavir resulted in a similar apparent permeability coefficient to that observed using P-gp knockout mice, which was 2.7-fold higher than in control mice. We conclude that, in mice, even at a relevant intraluminal concentration of darunavir, P-gp has a modulatory effect on the absorption of darunavir. However, this P-gp-mediated darunavir transport is inhibited when it is combined with ritonavir. IntroductionThe combination of multiple antiretroviral drugs with different modes of action (highly active antiretroviral therapy) has contributed to a spectacular progress in the fight against HIV (Hammer et al., 2008). The standard regimen for newly diagnosed patients usually includes two nucleoside reverse transcriptase inhibitors combined with either a protease inhibitor [boosted with ritonavir (RTV)] or a non-nucleoside reverse transcriptase inhibitor. Darunavir [(DRV) Prezista; Tibotec Pharmaceuticals, Cork, Ireland] is a second-generation protease inhibitor (PI) designed to have antiviral efficacy against HIV-1 with multiple resistance mutations to protease inhibitors (Busse and Penzak, 2007;Tremblay, 2008). RTV is always coadministered in a subtherapeutic dose as a pharmacokinetic booster of DRV.In vitro and in vivo studies have demonstrated that all HIV PI are high-affinity substrates for P-glycoprotein [(P-gp) multidrug resistance 1 (MDR1)], which can significantly influence the disposition of antiretroviral drugs (Kis et al., 2009). P-gp is widely distributed with a high level of expression in the small intestine, liver, kidney, and brain (Ho and Kim, 2005). Presently, however, there is still uncertainty about the role of intestinal P-gp in vivo. It is not sure whether P-gp plays a pivotal role as a biochemical barrier for compounds at concentrations relevant for the intraluminal environment, because of a possible saturation of the efflux system. In addition, it has b...

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“…Transporter expression profiling of the Caco-2 cell populations used in our lab along with those obtained from other labs revealed that our Caco-2 cells (as well as those of most other labs tested) express high levels of OATP2B1 with absent or very low levels of the other OATP isoforms (Hayeshi et al, 2008). Previous work has shown that OATP2B1 is predominantly responsible for the apical accumulation of E3S in Caco-2 cells (Sai et al, 2006).…”

Section: Cgamf and E3s Accumulation In Caco-2 Cellsmentioning

confidence: 89%

Interaction of HIV protease inhibitors with OATP1B1, 1B3, and 2B1

et al. 2010

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The effects of human immunodeficiency virus (HIV) protease inhibitors (PI) on the accumulation of the fluorescent bile salt analogue cholyl-glycylamido-fluorescein (CGamF) were determined in organic anion transporting polypeptide (OATP)-1B1 and -1B3-expressing Chinese hamster ovary (CHO) cells. In addition, interaction studies in Caco-2 monolayers, known only to express the OATP2B1 isoform, were conducted using the established OATP substrate estrone 3-sulfate (E3S), since no CGamF accumulation was observed in Caco-2 monolayers. CGamF appeared an excellent substrate for the OATP1B subfamily, with net accumulation clearance values of 7.8 and 142 microl min(-1) mg(-1) protein in OATP1B1 and OATP1B3-transfected cells, respectively. K(i)-values reflecting inhibition of CGamF accumulation by HIV PI correlated well between OATP1B1 and OATP1B3-expressing cells. Lopinavir was the most potent inhibitor (K(i) = 0.5-1.4 microM) of OATP1B-mediated CGamF accumulation compared with atazanavir, darunavir, ritonavir, and saquinavir (K(i) between 1.4 and 3.3 microM). Inhibitory profiles towards OATP2B1-mediated E3S accumulation were different with only indinavir, saquinavir, and ritonavir showing substantial effects. In conclusion, OATP1B3 appears to be a major transport mechanism mediating sodium-independent CGamF accumulation in human liver, and CGamF could be used as a probe substrate for in vitro drug interaction studies. The remarkably potent inhibition of OATP1B1 by lopinavir may explain some clinically relevant drug interactions between lopinavir and OATP1B substrates such as fexofenadine.

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Comparison of drug transporter gene expression and functionality in Caco-2 cells from 10 different laboratories

Cited by 230 publications

References 36 publications

Achieving improved permeability by hydrogen bond donor modulation in a series of MGAT2 inhibitors

Achieving improved permeability by hydrogen bond donor modulation in a series of MGAT2 inhibitors

In Situ Intestinal Perfusion in Knockout Mice Demonstrates Inhibition of Intestinal P-Glycoprotein by Ritonavir Causing Increased Darunavir Absorption

Interaction of HIV protease inhibitors with OATP1B1, 1B3, and 2B1

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