DMD Large equally mixed donor pool

Home Help [Feedback] [For Subscribers] [Archive] [Search] [Contents]
QUICK SEARCH:   [advanced]


     

Drug Metabolism and Disposition Fast Forward
First published on January 13, 2006; DOI: 10.1124/dmd.105.007534

0090-9556/06/3405-738-742$20.00
DMD 34:738-742, 2006

This Article
Right arrow Abstract Freely available
Right arrow Full Text (PDF)
Right arrow All Versions of this Article:
dmd.105.007534v1
34/5/738    most recent
Right arrow Submit a response
Right arrow Alert me when this article is cited
Right arrow Alert me when eLetters are posted
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Huang, L.
Right arrow Articles by Grimm, S.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Huang, L.
Right arrow Articles by Grimm, S.

ATP-DEPENDENT TRANSPORT OF ROSUVASTATIN IN MEMBRANE VESICLES EXPRESSING BREAST CANCER RESISTANCE PROTEIN

Liyue Huang1, Yi Wang, and Scott Grimm

Department of Drug Metabolism and Pharmacokinetics, AstraZeneca Pharmaceuticals LP, Wilmington, Delaware

(Received September 20, 2005; accepted December 28, 2005)


   Abstract
Top
 Abstract
Materials and Methods
 Results
 Discussion
 References
 
MDR1/ABCB1, MRP2/ABCC2, and breast cancer resistance protein (BCRP)/ABCG2 are expressed in the liver and intestine and contribute to the disposition of many drugs. Rosuvastatin, a 3-hydroxy-3-methylglutaryl-CoA reductase inhibitor for the treatment of patients with dyslipidemia, is primarily excreted via bile as unchanged drug. The present study was designed to determine whether rosuvastatin is transported by MDR1, MRP2, and BCRP. The apparent permeability value for rosuvastatin across MDR1-Madin-Darby canine kidney cells was low (~8 nm/s), and no directional transport was observed. Rosuvastatin uptake into control Sf9 membranes and membranes expressing MRP2 was similar in the presence or absence of GSH. In contrast, ATP dramatically stimulated rosuvastatin uptake into membranes expressing BCRP, but not control membranes. Rosuvastatin transport occurred into an osmotically sensitive space and was saturable. An Eadie-Hofstee analysis suggested that there were two transport sites in BCRP, with an apparent Km of 10.8 µM for the high affinity site and 307 µM for the low affinity site. These data demonstrate that rosuvastatin is transported efficiently by BCRP and suggest that BCRP plays a significant role in the disposition of rosuvastatin.

A number of ATP-binding cassette transporters (ABC) are expressed at the canalicular membranes of hepatocytes and the brush-border membranes of enterocytes including the P-glycoprotein (Pgp) (MDR1/ABCB1), the multidrug resistance protein 2 (MRP2/ABCC2), and the breast cancer resistance protein (BCRP/ABCG2), which is also known as the mitoxantrone resistance protein (MXR). These efflux transporters have distinct, but overlapping, substrate specificity and have been shown to be involved in biliary excretion of many drugs (Sasabe et al., 1998Go; Jonker et al., 2000Go). MDR1 Pgp, which is overexpressed in resistant tumor cells, transports a wide variety of hydrophobic drugs such as daunorubicin, vinblastine, and loperamide (Hochman et al., 2002Go). MRP2 has been shown to transport many glutathione-S-glucuronide and sulfate conjugates. In addition to conjugates, MRP2 transports some unconjugated organic anions including methotrexate, irinotecan, and ampicillin (Gerk and Vore, 2002Go). The BCRP gene has been cloned and the protein product was found to be a half-transporter, consisting of a single 70-kDa, six-transmembrane peptide. Structurally diverse compounds such as anthracyclines, mitoxantrone, methotrexate, the camptothecins, and estrone-3-sulfate have been identified as BCRP substrates (Sarkadi et al., 2004Go).

Rosuvastatin is a 3-hydroxy-3-methylglutaryl-CoA reductase inhibitor for the treatment of patients with dyslipidemia. The absolute bioavailability of rosuvastatin was estimated as 20%, with an estimated hepatic extraction ratio of 0.63 (Martin et al., 2003aGo). Metabolism of rosuvastatin seems to be a minor route of elimination in humans, and rosuvastatin is mainly excreted into bile as parent. In a human mass balanced study, 90% of the dose was recovered in feces, primarily (92%) as unchanged drug (Martin et al., 2003bGo).

Statins including rosuvastatin, pitavastatin, cerivastatin, and pravastatin are substrates for OATP-C (OATP1B1), an organic anion-transporting polypeptide expressed exclusively in the basolateral membrane of hepatocytes (Hsiang et al., 1999Go; Shitara et al., 2003Go; Hirano et al., 2004Go; Schneck et al., 2004Go). It has been shown that statins are substrates for multiple efflux transporters. Atorvastatin, cerivastatin, and simvastatin have been reported to be Pgp substrates (Hochman et al., 2004Go; Kivisto et al., 2004Go). Pravastatin seems to be an MRP2 substrate. ATP-dependent transport of pravastatin has been demonstrated in the canalicular membranes isolated from rats, and the biliary excretion of pravastatin is decreased in Mrp2-mutant rats (Yamazaki et al., 1997Go). The transport of pravastatin in the basal-to-apical direction (B->A) is 2.5 times greater than that in the apical-to-basal (A->B) direction across Madin-Darby canine kidney (MDCK) monolayer expressing both OATP-C and MRP2 (Sasaki et al., 2002Go). Recent studies indicate that several statins are also transported by BCRP in vitro (Hirano M et al., 2005Go; Matsushima et al., 2005Go).

The present study was designed to determine whether rosuvastatin is a substrate for MDR1, MRP2, or BCRP. The results indicate that rosuvastatin is transported efficiently by BCRP in membrane vesicles and suggest that BCRP may play a significant role in the disposition of rosuvastatin.


   Materials and Methods
Top
 Abstract
 Materials and Methods
Results
 Discussion
 References
 
Materials. Rosuvastatin was supplied by AstraZeneca Pharmaceuticals LP. GF120918 was obtained from GlaxoSmithKline (Research Triangle Park, NC) under a material transfer agreement. [3H]Estradiol-17-ß-D-glucuronide (E217G) was purchased from PerkinElmer (Boston, MA). [3H]Rosuvastatin was supplied by GE Healthcare (Little Chalfont, Buckinghamshire, UK). Other chemicals were purchased from Sigma-Aldrich (St. Louis, MO). Cell culture reagents were purchased from Invitrogen (Carlsbad, CA). Madin-Darby canine kidney cells transfected with human MDR1 (MDR1-MDCK) were obtained from The Netherlands Cancer Institute (Amsterdam, The Netherlands). Transwells (12-well, 11-mm diameter, 0.4-µm pores) were purchased from Corning Costar (Cambridge, MA)

MRP2-Sf9-VT membranes (membranes isolated from Sf9 cells expressing MRP2; ATP-dependent transport of 200 nM [3H]leukotriene C4 was 20.8 pmol/mg protein/min), CTRL-Sf9 membranes (control membranes isolated from Sf9 cells expressing ß-galactosidase), MXR-M-VT membranes (membranes isolated from mammalian cells expressing BCRP; ATP-dependent transport of 100 µM [3H]methotrexate was 172.6 pmol/mg protein/min), and CTRL-M membranes (control membranes isolated from mammalian cells that have no BCRP expression) were purchased from Solvo Biotechnology (Budapest, Hungary).

Monolayer Efflux Studies. MDR1-MDCK cells were cultured and transport experiments were conducted as described by Polli et al. (2001Go). Briefly, cells were grown in Dulbecco's modified Eagle's medium supplemented with 10% fetal bovine serum in the absence of any antibiotics or selection agents. Cells were seeded onto Transwells at a density of 300,000 cells/cm2, and medium was replaced daily and 2 h before transport experiments were performed. Monolayers were used for transport studies 3 days after seeding. Compounds were dissolved in dimethyl sulfoxide and then diluted in Hanks' balanced transport buffer (pH 7.4) (Mediatech, Herndon, VA). The amount of dimethyl sulfoxide in the final transport solution was less than 0.5% (v/v). Rosuvastatin and reference compounds were tested in triplicate over 60 min in a 37°C shaking water bath.

Rosuvastatin concentrations were analyzed by liquid chromatographytandem mass spectrometry using a Sciex API 3000 with TurboIon spray source (Applied Biosystems/MDS Sciex, South San Francisco, CA) (Schneck et al., 2004Go). Erythromycin and propranolol concentrations were determined by liquid chromatography-tandem mass spectrometry using a Micromass Quattro Ultima (Micromass UK Ltd, Cheshire, UK) equipped with an electrospray ionization interface. The samples were loaded (injection volume 10–20 µl) on columns by means of an HTS Pal autosampler (CTC Analytics AG, Zwingen, Switzerland). Chromatography was performed on columns of Synergi MAX-RP 30 x 2.0 mm, 4 µm (Phenomenex, Torrance, CA) at a flow rate of 1.5 ml/min. The mobile phase consisted of two solvents: 0.1% formic acid in water (A) and 10% methanol in acetonitrile (B). The gradient profile was 0 to 0.3 min 0% B, 0.3 to 1.3 min linear gradient to 95% B, 1.3 to 1.6 min 95% B, and 1.6 to 2 min 0% B.

The apparent permeability (Papp) values were calculated using the equation Papp = (1/AC0) x (dQ/dt), where A is the membrane surface area, C0 is the donor drug concentration at t = 0, and dQ/dt is the amount of drug transported within a given time period. Flux ratio = Papp (B->A)/Papp (A->B).

Rosuvastatin Transport in Membrane Vesicles. ATP-dependent transport of [3H]rosuvastatin or [3H]E217G, a known substrate for MRP2, was measured using a rapid filtration technique (Huang et al., 2000Go). Briefly, membranes were quickly thawed at 37°C. Transport was initiated by adding membranes (30–66 µg) to the preincubated buffer (50 mM Tris, 50 mM mannitol, and 10 mM MgCl2, pH 7.5) containing 5 mM ATP or AMP and [3H]rosuvastatin or [3H]E217G. Transport was terminated by the addition of 2.5 ml of ice-cold transport buffer, and vesicle-associated [3H]rosuvastatin was separated from free [3H]rosuvastatin by rapid filtration through a GF/F filter (Whatman, Florham Park, NJ). Filters were further washed twice with 2.5 ml of ice-cold buffer and assayed for radioactivity in 10 ml of liquid scintillation cocktail. Because nonspecific binding to the filter was relatively minimal, the data were not corrected for the binding to the filter unless stated otherwise in the figure legend. ATP-dependent transport was determined as the difference in uptake between 5 mM ATP and 5 mM AMP.

Statistics. Statistical differences (p < 0.05) were determined by one-way analysis of variance, followed by a multiple comparison test or unpaired Student's t test. Prism 3.0 software (GraphPad Software Inc., San Diego, CA) was used to produce the best-fitting curves.


   Results
Top
 Abstract
 Materials and Methods
 Results
Discussion
 References
 
Pgp-Mediated Transport of Rosuvastatin. Rosuvastatin transport across MDR1-MDCK monolayers was measured at concentrations of 10 and 50 µM in the presence or absence of 2 µM GF120918, a potent inhibitor for both Pgp and BCRP. Rosuvastatin transport in both B->A and A->B directions was similar, with a Papp value of ~8 nm/s, and was not affected by GF120918. In contrast, transport of the Pgp substrate erythromycin (10 µM) in the B->A direction was 35-fold greater than that in the A->B direction (Table 1).


View this table:
[in this window]
[in a new window]
  		TABLE 1 Transport of rosuvastatin across monolayers of MDR1-MDCK cells in the presence and absence of 2 µM GF120918

Values are the means ± S.D. of triplicate determinations from a single experiment.

 

MRP2-Mediated Transport of Rosuvastatin. No significant ATP-dependent transport of rosuvastatin was observed in either CTRL-Sf9-(control) or MRP2-Sf9-membranes in the absence of glutathione (GSH) (Fig. 1A). Significant ATP-dependent transport of rosuvastatin was observed in the presence of 5 mM GSH in both MRP2- and control-membrane vesicles (Fig. 1B). However, ATP-dependent transport of rosuvastatin between control and MRP2-Sf9-membranes was not significantly different (p > 0.05).


Figure 1
View larger version (20K):
[in this window]
[in a new window]
  		FIG. 1. Transport of rosuvastatin in MRP2- or CTRL-Sf9-membrane vesicles. Transport was initiated by adding membranes (50 µg) to the preincubated mixtures containing 5 mM ATP or AMP and [3H]rosuvastatin. A, in the absence of GSH; B, in the presence of 5 mM GSH. Values are the means ± S.D. of triplicate or duplicate determinations from a single experiment.

 
BCRP-Mediated Transport of Rosuvastatin. ATP dramatically stimulated rosuvastatin uptake in MXR-M-VT membranes expressing high levels of BCRP in a time-dependent manner (Fig. 2). An initial uptake rate of ATP-dependent transport of rosuvastatin (5 µM) was too rapid to be determined accurately, and the transport at 30 s was 239.7 pmol/mg protein. In contrast, no ATP-dependent transport of rosuvastatin was observed in CTRL-M membranes that did not express BCRP. The difference in the uptake between ATP and AMP was sensitive to extravesicular osmolarity. As the concentration of extravesicular sucrose increased, rosuvastatin uptake decreased in the presence of ATP (p < 0.05), whereas the uptake did not change significantly in the presence of AMP (p > 0.05) (Fig. 3).


Figure 2
View larger version (13K):
[in this window]
[in a new window]
  		FIG. 2. Time course of rosuvastatin (5 µM) transport in MXR-M-VT (BCRP) and CTRL-M membrane vesicles. Transport was initiated by adding membranes (35 µg for MXR-M-VT, 60 µg for CTRL-M) to the preincubated mixtures containing 5 mM ATP or AMP and [3H]rosuvastatin. Data were corrected for the nonspecific binding of rosuvastatin to the filter, and the values are the means ± S.D. of triplicate determinations from a single experiment.

 

Figure 3
View larger version (12K):
[in this window]
[in a new window]
  		FIG. 3. Effect of osmolarity on ATP-dependent transport of rosuvastatin (25 µM) in MXR-M-VT membrane vesicles. Uptake was measured at 2 min in the presence of 5 mM ATP or AMP after prior incubation (37°C, 10 min) of the vesicles in the buffer containing the indicated sucrose concentrations. Transport was initiated by adding a mixture containing [3H]rosuvastatin and ATP or AMP to the preincubated membranes. The values are the means ± S.D. of triplicate determinations from a single experiment. *, p < 0.05.

 
Concentrations of rosuvastatin ranging from 0.2 to 1000 µM were used to determine whether BCRP-mediated transport of rosuvastatin was saturable (Fig. 4). An Eadie-Hofstee analysis suggested that there were two binding sites in BCRP for rosuvastatin, with an apparent Km of 10.8 ± 1.1 µM for the high affinity site, and 307 ± 89.4 µM for the low affinity site.


Figure 4
View larger version (16K):
[in this window]
[in a new window]
  		FIG. 4. Concentration dependence of ATP-dependent transport of rosuvastatin in MXR-M-VT membrane vesicles. ATP-dependent transport was determined as the difference in the uptake between 5 mM ATP and AMP at 30 s. An Eadie-Hofstee plot is shown in the inset. The values are the combined results from two independent experiments.

 
GF120918 (2 µM), an inhibitor of Pgp and BCRP, inhibited ATP-dependent transport of rosuvastatin by 89% (p < 0.05) (Fig. 5). Pravastatin up to 250 µM inhibited rosuvastatin transport by only 30%.


Figure 5
View larger version (17K):
[in this window]
[in a new window]
  		FIG. 5. Effect of GF120918 and pravastatin on ATP-dependent transport of rosuvastatin (1 µM) in MXR-M-VT-membrane vesicles. Transport was measured at 30 s and was initiated by adding [3H]rosuvastatin and ATP or AMP to the preincubated mixtures containing membranes and test compounds. Data are expressed as percentage of control (control values were 75.3 ± 5.7 or 63.2 ± 7.3 pmol/mg protein) and represent means ± S.D. of triplicate determinations from a single experiment. *, p < 0.05.

 

   Discussion
Top
 Abstract
 Materials and Methods
 Results
 Discussion
References
 
The purpose of this study was to identify and characterize ATP-dependent transporters for rosuvastatin. Our results provide the direct evidence that rosuvastatin is a substrate for BCRP. Rosuvastatin transport in MXR-M-VT membranes was dramatically stimulated by ATP and was saturable, with an apparent Km of 10.8 µM for the high affinity binding or transport site. This value is comparable to the Ki (15.4 ± 2.4 µM) for inhibiting [3H]estrone-3-sulfate uptake by human BCRP-membrane vesicles (Hirano et al., 2005Go), suggesting that rosuvastatin and estrone-3-sulfate may share the same transport or binding site. The uptake in the presence of ATP, but not in the presence of AMP, was decreased as the intravesicular space was decreased by increasing extravesicular sucrose concentration (osmolarity). These data indicate that rosuvastatin transport occurred into the intravesicular space, and the observed difference in uptake between ATP and AMP was mainly due to transport, not binding (Vore et al., 1996Go). Western blot analysis showed that high levels of BCRP, moderate levels of MRP4 and MRP5, very low levels of MRP1, and undetectable levels of Pgp, MRP2, and MRP3 were expressed in MXR-M-VT; and total MRPs expressed in MXR-M-VT were much lower than that in control membranes (personal communication with the chief scientist Dr. Peter Krajsci in Solvo Biotechnology). Moreover, rosuvastatin transport in MXR-M-VT membranes was inhibited by GF120918. Thus, the observed ATP-dependent transport in MXR-M-VT membranes is probably mediated by BCRP. In an attempt to determine whether pravastatin is also transported by BCRP, the ability of pravastatin to inhibit BCRP-mediated transport of rosuvastatin was evaluated. However, pravastatin up to 250 µM inhibited rosuvastatin transport (1 µM) by only 30%, suggesting that pravastatin is not transported by BCRP, or possibly binds to a different site. These data suggest that the transporter mechanisms for the first-pass and biliary excretion between pravastatin and rosuvastatin may be different.

The Eadie-Hofstee analysis suggested that there were two binding or transport sites for rosuvastatin in MXR-M-VT membranes. Although multiple binding sites in BCRP have not been reported previously, and the presence of unknown transporters for rosuvastatin in these membranes cannot be excluded, our results are consistent with the findings of multiple binding sites in other ABC transporters. It has been proposed that MDR1 has three binding sites, including two transport sites and an allosteric site. Pgp-mediated drug transport was stimulated by prazosin and progesterone (Shapiro et al., 1999Go). Similarly, two transport or binding sites have been proposed for MRP2. Zelcer et al. (2003Go) postulated two MRP2 transport sites with positive cooperativity to explain the ability of sulfinpyrazone and indomethacin to stimulate MRP2-mediated transport.

Based on the fact that rosuvastatin is an organic anion and that pravastatin is an MRP2 substrate, we had speculated that rosuvastatin was an MRP2 substrate. However, there was no ATP-dependent transport of rosuvastatin in MRP2-Sf9-membranes or control membranes in the absence of GSH, although significant ATP-dependent transport of E217G, a known MRP2 substrate, was clearly observed. Uptake of E217G measured over 5 min was 839 ± 64 and 146 ± 4 (pmol/mg protein) in the presence of 5 mM ATP and AMP, respectively. The high expression level of MRP2 in these membranes, but not in control membranes, was confirmed by Western blot analysis (data not shown). It has been shown that GSH stimulates MRP-mediated transport of vincristine (Loe et al., 1998Go). In the presence of 5 mM GSH, significant ATP-dependent transport of rosuvastatin was observed in both control and MRP2-Sf9-VT membranes. However, there was no significant difference in ATP-dependent transport of rosuvastatin between control and MRP2-Sf9-VT membranes, suggesting that endogenous transporters in control Sf9 cells transport rosuvastatin in the presence of GSH. These data do not support our speculation that rosuvastatin is an MRP2 substrate. However, the possibility that MRP2 transports rosuvastatin in the presence of GSH cannot be excluded. It is also possible that the expression levels of endogenous MRP-like proteins in Sf9 cells expressing MRP2 are lower than that in control cells, and MRP2-mediated transport is masked by the high background transport in control membranes.

The role of Pgp in rosuvastatin efflux was assessed using MDR1-MDCK cells, a cell line widely used to identify Pgp substrates. No significant directional transport of rosuvastatin across MDR1-MDCK cells was observed. The apparent permeability of rosuvastatin was low, and was similar to that of mannitol, a marker for paracellular diffusion, suggesting that rosuvastatin may not be able to penetrate into cells without the activity of uptake transporters. The lack of directional transport of rosuvastatin across MDR1-MDCK cells may reflect its inability to access Pgp rather than it not being transported by Pgp. Therefore, rosuvastatin uptake into membrane vesicles isolated from wild-type and MDR1-MDCK cells was further examined. Marginal ATP-dependent transport of rosuvastatin was observed in membranes isolated from both wild-type and MDR1-MDCK cells. The ATP-dependent transport at 1 min was 11 and 9 (pmol/mg protein) for wild-type and MDR1-MDCK membranes, respectively (p > 0.05) (data not shown). These data do not support the idea that rosuvastatin is a Pgp substrate.

Recently, Tiberg et al. (2004Go) reported that rosuvastatin transport in the serosal to the mucosal (secretory) direction was higher than that in the mucosal to the serosal (absorptive) direction across rat intestinal segments mounted in Ussing chambers, with the highest efflux in the ileum. The efflux ratios were 8.2, 7.2, 15, and 1.7 across duodenum, jejunum, ileum, and colon segments, respectively, which is consistent with the expression pattern of Bcrp in rat intestine (Tanaka et al., 2005Go). Bcrp mRNA levels are high throughout the intestinal tract, with the highest levels in the ileum, which differs from the limited expression of Mrp2 in proximal segments of rat small intestine (Mottino et al., 2000Go). Mrp2 protein is present mainly in brush-border membranes of the proximal segments and gradually decreases from jejunum to the distal ileum. It has been shown that BCRP plays an important role in biliary excretion and oral absorption of topotecan (Jonker et al., 2000Go). The present data clearly demonstrated that rosuvastatin is a substrate for BCRP, and this transporter probably contributes to its biliary excretion. However, it is not known whether BCRP plays a significant role in limiting oral absorption of rosuvastatin. Rosuvastatin has poor permeability, and its flux across cell monolayers in vitro is mainly through the paracellular pathway. If this is the case in vivo, rosuvastatin may not be able to access BCRP for efflux without the involvement of uptake transporters. Recently, it was found that OATP-B is expressed at the apical membrane of human intestinal epithelial cells and may involve pH-dependent absorption of pravastatin (Kobayashi et al., 2003Go). An uptake transport system for rosuvastatin at the apical membranes of the intestines may exist and, therefore, BCRP may limit its oral absorption.

In summary, the present studies demonstrate that rosuvastatin is transported efficiently by BCRP in vitro. These data suggest that BCRP may play a significant role in the disposition of rosuvastatin.


   Acknowledgments
 
We thank Robert Williams and Julie Zalikowski for bioanalytical support.


   Footnotes
 
Article, publication date, and citation information can be found at http://dmd.aspetjournals.org.

doi:10.1124/dmd.105.007534.

ABBREVIATIONS: ABC, ATP-binding cassette transporter; BCRP, breast cancer resistance protein; E217G, estradiol-17ß-D-glucuronide; GSH, glutathione; MDR1, multidrug resistance protein 1; MDR1-MDCK, Madin-Darby canine kidney cells transfected with the human MDR1 gene; MRP2, multidrug resistance protein 2; MXR, mitoxantrone resistance protein; Papp, apparent permeability; Pgp, P-glycoprotein; OATP, organic anion-transporting polypeptide; GF120918, 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; CTRL, control; A->B apical to basolateral; B->A, basolateral to apical.

1 Current affiliation: Amgen, Cambridge, Massachusetts. Back

Address correspondence to: Dr. Yi Wang, Department of Drug Metabolism and Pharmacokinetics, AstraZeneca Pharmaceuticals LP, Wilmington, DE 19850. E-mail: yi.wang{at}astrazeneca.com


   References
Top
 Abstract
 Materials and Methods
 Results
 Discussion
 References
 


Gerk PM and Vore M (2002) Regulation of expression of the multidrug resistance-associated protein 2 (MRP2) and its role in drug disposition. J Pharmacol Exp Ther 302: 407–415.[Abstract/Free Full Text]

Hirano M, Maeda K, Matsushima S, Nozaki Y, Kusuhara H, and Sugiyama Y (2005) Involvement of BCRP (ABCG2) in the biliary excretion of pitavastatin. Mol Pharmacol 68: 800–807.[Abstract/Free Full Text]

Hirano M, Maeda K, Shitara Y, and Sugiyama Y (2004) Contribution of OATP2 (OATP1B1) and OATP8 (OATP1B3) to the hepatic uptake of pitavastatin in humans. J Pharmacol Exp Ther 311: 139–146.[Abstract/Free Full Text]

Hochman JH, Pudvah N, Qiu J, Yamazaki M, Tang C, Lin JH, and Prueksaritanont T (2004) Interactions of human P-glycoprotein with simvastatin, simvastatin acid and atorvastatin. Pharm Res (NY) 21: 1686–1691.

Hochman JH, Yamazaki M, Ohe T, and Lin JH (2002) Evaluation of drug interactions with P-glycoprotein in drug discovery: in vitro assessment of the potential for drug-drug interactions with P-glycoprotein. Curr Drug Metab 3: 257–273.[CrossRef][Medline]

Hsiang B, Zhu Y, Wang Z, Wu Y, Sasseville V, Yang WP, and Kirchgessner TG (1999) A novel human hepatic organic anion transporting polypeptide (OATP2): identification of a liver specific human organic anion transporting polypeptide and identification of rat and human hydroxymethylglutaryl-CoA reductase inhibitor transporters. J Biol Chem 274: 37161–37168.[Abstract/Free Full Text]

Huang L, Smit JW, Meijer DKE, and Vore M (2000) Mrp2 is essential for estradiol-17ß(ß-D-glucuronide)-induced cholestasis in rats. Hepatology 32: 66–72.[CrossRef][Medline]

Jonker JW, Smit JW, Brinkhuis RF, Maliepaard M, Beijnen JH, Schellens JHM, and Schinkel AH (2000) Role of breast cancer resistance protein in the bioavailability and fetal penetration of topotecan. J Natl Cancer Inst 92: 1651–1656.[Abstract/Free Full Text]

Kivisto KT, Zukunft J, Hofmann U, Niemi M, Rekersbrink S, Schneider S, Luippold G, Schwab M, Eichelbaum M, and Fromm MF (2004) Characterisation of cerivastatin as a P-glycoprotein substrate: studies in P-glycoprotein-expressing cell monolayers and mdr1a/b knock-out mice. Naunyn-Schmiedeberg's Arch Pharmacol 370: 124–130.[Medline]

Kobayashi D, Nozawa T, Imai K, Nezu J, Tsuji A, and Tamai I (2003) Involvement of human organic anion transporting polypeptide OATP-B (SLC21A9) in pH-dependent transport across intestinal apical membrane. J Pharmacol Exp Ther 306: 703–708.[Abstract/Free Full Text]

Loe DW, Deeley RG, and Cole SP (1998) Characterization of vincristine transport by the M(r) 190,000 multidrug resistance protein (MRP): evidence for cotransport with reduced glutathione. Cancer Res 58: 5130–5136.[Abstract/Free Full Text]

Martin PD, Warwick MJ, Dane AL, Brindley C, and Short T (2003a) Absolute oral bioavailability of rosuvastatin in healthy white adult male volunteers. Clin Ther 25: 2553–2563.[CrossRef][Medline]

Martin PD, Warwick MJ, Dane AL, Hill SJ, Giles PB, Phillips PJ, and Lenz E (2003b) Metabolism, excretion and pharmacokinetics of rosuvastatin in healthy adult volunteers. Clin Ther 25: 2822–2835.[CrossRef][Medline]

Matsushima S, Maeda K, Kondo C, Hirano M, Sasaki M, Suzuki H, and Sugiyama Y (2005) Identification of the hepatic efflux transporters of organic anions using double transfected human organic anion-transporting polypeptide 1B1 (OATP1B1)/multidrug resistance-associated protein 2, OATP1B1/multidrug resistance 1, and OATP1B1/breast cancer resistance protein. J Pharmacol Exp Ther 314: 1059–1067.[Abstract/Free Full Text]

Mottino AD, Hoffman T, Jennes L, and Vore M (2000) Expression and localization of multidrug resistant protein mrp2 in rat small intestine. J Pharmacol Exp Ther 293: 717–723.[Abstract/Free Full Text]

Polli JW, Wring SA, Humphreys JE, Huang L, Morgan JB, Webster LO, and Serabjit-Singh CS (2001) Rational use of in vitro P-glycoprotein assays in drug discovery. J Pharmacol Exp Ther 299: 620–628.[Abstract/Free Full Text]

Sarkadi B, Ozvegy-Laczka C, Nemet K, and Varadi A (2004) ABCG2—a transporter for all seasons. FEBS Lett 567: 116–120.[CrossRef][Medline]

Sasabe H, Tsuji A, and Sugiyama Y (1998) Carrier-mediated mechanism for the biliary excretion of the quinolone antibiotic grepafloxacin and its glucuronide in rats. J Pharmacol Exp Ther 284: 1033–1039.[Abstract/Free Full Text]

Sasaki M, Suzuki H, Ito K, Abe T, and Sugiyama Y (2002) Transcellular transport of organic anions across a double-transfected Madin-Darby canine kidney cell monolayer expressing both human organic anion-transporting polypeptide (OATP2/SLC21A6) and multidrug resistance-associated protein 2 (MRP2/ABCC2). J Biol Chem 277: 6497–6503.[Abstract/Free Full Text]

Schneck DW, Birmingham BK, Zalikowski JA, Mitchell PD, Wang Y, Martin PD, Lasseter KC, Brown CD, Windass AS, and Raza A (2004) The effect of gemfibrozil on the pharmacokinetics of rosuvastatin. Clin Pharmacol Ther 75: 455–463.[CrossRef][Medline]

Shapiro AB, Fox K, Lam P, and Ling V (1999) Stimulation of P-glycoprotein-mediated drug transport by prazosin and progesterone. Evidence for a third drug-binding site. Eur J Biochem 259: 841–850.[Medline]

Shitara Y, Itoh T, Sato H, Li AP, and Sugiyama Y (2003) Inhibition of transporter-mediated hepatic uptake as a mechanism for drug-drug interaction between cerivastatin and cyclosporin A. J Pharmacol Exp Ther 304: 610–616.[Abstract/Free Full Text]

Tanaka Y, Slitt AL, Leazer TM, Maher JM, and Klaassen CD (2005) Tissue distribution and hormonal regulation of the breast cancer resistance protein (Bcrp/Abcg2) in rats and mice. Biochem Biophys Res Commun 326: 181–187.[CrossRef][Medline]

Tiberg M, Dudley A, and Ungell A (2004) Regional and pH dependent permeability of rosuvastatin. AAPS J 6 No. 4, abstract T2202.

Vore M, Hoffman T, and Gosland M (1996) ATP-dependent transport of beta-estradiol 17-(beta-D-glucuronide) in rat canalicular membrane vesicles. Am J Physiol 271: G791–G798.[Medline]

Yamazaki M, Akiyama S, Ni'inuma K, Nishigaki R, and Sugiyama Y (1997) Biliary excretion of pravastatin in rats: contribution of the excretion pathway mediated by canalicular multi-specific organic anion transporter (cMOAT). Drug Metab Dispos 25: 1123–1129.[Abstract/Free Full Text]

Zelcer N, Huisman MT, Reid G, Wielinga P, Breedveld P, Kuil A, Knipscheer P, Schellens JH, Schinkel AH, and Borst (2003) Evidence for two interacting ligand binding sites in human multidrug resistance protein 2 (ATP binding cassette C2). J Biol Chem 278: 23538–23544.[Abstract/Free Full Text]


This article has been cited by other articles:


Home page
 J Clin PharmacolHome page
T. Zhu, W. M. Awni, B. Hosmane, M. T. Kelly, D. J. Sleep, J. C. Stolzenbach, K. Wan, T. O. Chira, and R. S. Pradhan
ABT-335, the Choline Salt of Fenofibric Acid, Does Not Have a Clinically Significant Pharmacokinetic Interaction With Rosuvastatin in Humans
J. Clin. Pharmacol., January 1, 2009; 49(1): 63 - 71.
[Abstract] [Full Text] [PDF]

Home page
 Drug Metab. Dispos.Home page
S. Kitamura, K. Maeda, Y. Wang, and Y. Sugiyama
Involvement of Multiple Transporters in the Hepatobiliary Transport of Rosuvastatin
Drug Metab. Dispos., October 1, 2008; 36(10): 2014 - 2023.
[Abstract] [Full Text] [PDF]

Home page
 Mol. Pharmacol.Home page
A. Verhulst, R. Sayer, M. E. De Broe, P. C. D'Haese, and C. D. A. Brown
Human Proximal Tubular Epithelium Actively Secretes but Does Not Retain Rosuvastatin
Mol. Pharmacol., October 1, 2008; 74(4): 1084 - 1091.
[Abstract] [Full Text] [PDF]

Home page
 Drug Metab. Dispos.Home page
C. Chen, J. L. Stock, X. Liu, J. Shi, J. W. Van Deusen, D. A. DiMattia, R. G. Dullea, and S. M. de Morais
Utility of a Novel Oatp1b2 Knockout Mouse Model for Evaluating the Role of Oatp1b2 in the Hepatic Uptake of Model Compounds
Drug Metab. Dispos., September 1, 2008; 36(9): 1840 - 1845.
[Abstract] [Full Text] [PDF]

Home page
 J. Pharmacol. Exp. Ther.Home page
K. Abe, A. S. Bridges, W. Yue, and K. L. R. Brouwer
In Vitro Biliary Clearance of Angiotensin II Receptor Blockers and 3-Hydroxy-3-methylglutaryl-Coenzyme A Reductase Inhibitors in Sandwich-Cultured Rat Hepatocytes: Comparison with in Vivo Biliary Clearance
J. Pharmacol. Exp. Ther., September 1, 2008; 326(3): 983 - 990.
[Abstract] [Full Text] [PDF]

Home page
 J Clin PharmacolHome page
B. L. Urquhart, R. G. Tirona, and R. B. Kim
Nuclear Receptors and the Regulation of Drug-Metabolizing Enzymes and Drug Transporters: Implications for Interindividual Variability in Response to Drugs
J. Clin. Pharmacol., May 1, 2007; 47(5): 566 - 578.
[Abstract] [Full Text] [PDF]

This Article
Right arrow Abstract Freely available
Right arrow Full Text (PDF)
Right arrow All Versions of this Article:
dmd.105.007534v1
34/5/738    most recent
Right arrow Submit a response
Right arrow Alert me when this article is cited
Right arrow Alert me when eLetters are posted
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Huang, L.
Right arrow Articles by Grimm, S.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Huang, L.
Right arrow Articles by Grimm, S.

Home Help [Feedback] [For Subscribers] [Archive] [Search] [Contents]
All ASPET Journals Molecular Pharmacology Pharmacological Reviews
Molecular Interventions Drug Metabolism and Disposition