CYP3A4-transfected Caco-2 cells as a tool for understanding biochemical absorption barriers: Studies with sirolimus and midazolam

被引:91
作者
Cummins, CL [1 ]
Jacobsen, W [1 ]
Christians, U [1 ]
Benet, LZ [1 ]
机构
[1] Univ Calif San Francisco, Dept Biopharmaceut Sci, San Francisco, CA 94143 USA
关键词
D O I
10.1124/jpet.103.058065
中图分类号
R9 [药学];
学科分类号
1007 ;
摘要
CYP3A4-transfected Caco-2 cells were used as an in vitro system to predict the importance of drug metabolism and transport on overall drug absorption. We examined the transport and metabolism of two drugs; midazolam, an anesthetic agent and CYP3A4 substrate, and sirolimus, an immunosuppressant and a dual CYP3A4/P-glycoprotein (P-gp) substrate, in the presence of cyclosporine (CsA, a CYP3A4/P-gp inhibitor) or N-{4-[2-(1,2,3,4-tetrahydro-6,7-dimethoxy-2-isoquinolinyl)-ethyl]-phenyl}-9,10-dihydro-5-methoxy-9-oxo-4-acridine carboxamine (GG918) (an inhibitor of P-gp and not CYP3A4). All major CYP3A4 metabolites were formed in the cells (1-OH > 4- OH midazolam and 39-O-desmethyl > 12-OH > 11- OH sirolimus), consistent with results from human liver microsomes. There was no bidirectional transport of midazolam across CYP3A4-transfected Caco-2 cells, whereas there was a 2.5-fold net efflux of sirolimus (1 muM) that disappeared in the presence of CsA or GG918. No change in the absorption rate or extraction ratio (ER) for midazolam was observed when P-gp was inhibited with GG918. Addition of GG918 had a modest impact on the absorption rate and ER for sirolimus (increased 58% and decreased 25%, respectively), whereas a 6.1-fold increase in the absorption rate and a 75% decrease in the ER were found when sirolimus was combined with CsA. Although both midazolam and sirolimus metabolites were preferentially excreted to the apical compartment, only sirolimus metabolites were transported by P-gp as determined from inhibition studies with GG918. Using CYP3A4-transfected Caco-2 cells we determined that, in contrast to P-gp, CYP3A4 is the major factor limiting sirolimus absorption. The integration of CYP3A4 and P-gp into a combined in vitro system was critical to unveil the relative importance of each biochemical barrier.
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页码:143 / 155
页数:13
相关论文
共 28 条
[1]   Biochemical, cellular, and pharmacological aspects of the multidrug transporter [J].
Ambudkar, SV ;
Dey, S ;
Hrycyna, CA ;
Ramachandra, M ;
Pastan, I ;
Gottesman, MM .
ANNUAL REVIEW OF PHARMACOLOGY AND TOXICOLOGY, 1999, 39 :361-398
[2]  
ARCECI RJ, 1992, BLOOD, V80, P1528
[3]   Intestinal drug absorption and metabolism in cell cultures: Caco-2 and beyond [J].
Artursson, P ;
Borchardt, RT .
PHARMACEUTICAL RESEARCH, 1997, 14 (12) :1655-1658
[4]   CORRELATION BETWEEN ORAL-DRUG ABSORPTION IN HUMANS AND APPARENT DRUG PERMEABILITY COEFFICIENTS IN HUMAN INTESTINAL EPITHELIAL (CACO-2) CELLS [J].
ARTURSSON, P ;
KARLSSON, J .
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1991, 175 (03) :880-885
[5]  
Benet L.Z., 1996, GOODMAN GILMANS PHAR, V3, P27
[6]   Intestinal drug metabolism and antitransport processes: A potential paradigm shift in oral drug delivery [J].
Benet, LZ ;
Wu, CY ;
Hebert, MF ;
Wacher, VJ .
JOURNAL OF CONTROLLED RELEASE, 1996, 39 (2-3) :139-143
[7]   Automated, fast and sensitive quantification of drugs in blood by liquid chromatography-mass spectrometry with on-line extraction: immunosuppressants [J].
Christians, U ;
Jacobsen, W ;
Serkova, N ;
Benet, LZ ;
Vidal, C ;
Sewing, KF ;
Manns, MP ;
Kirchner, GI .
JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES, 2000, 748 (01) :41-53
[8]  
Crowe A, 1999, DRUG METAB DISPOS, V27, P627
[9]   In vitro and in situ absorption of SDZ-RAD using a human intestinal cell line (Caco-2) and a single pass perfusion model in rats:: Comparison with rapamycin [J].
Crowe, A ;
Lemaire, M .
PHARMACEUTICAL RESEARCH, 1998, 15 (11) :1666-1672
[10]   Unmasking the dynamic interplay between intestinal P-glycoprotein and CYP3A4 [J].
Cummins, CL ;
Jacobsen, W ;
Benet, LZ .
JOURNAL OF PHARMACOLOGY AND EXPERIMENTAL THERAPEUTICS, 2002, 300 (03) :1036-1045