Binary and ternary combinations of anti-HIV protease inhibitors: effect on gene expression and functional activity of CYP3A4 and efflux transporters
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Deep Kwatra
Abstract
Background: The purpose of this study is to identify the effect of binary and ternary combinations of anti-HIV protease inhibitors (PIs) on the expression of metabolizing enzyme (CYP3A4) and efflux transporters [multidrug resistance-associated protein 2 (MRP2), P-glycoprotein (P-gp) and breast cancer resistant protein (BCRP)] in a model intestinal cell line (LS-180).
Methods: LS-180 cells were treated with various combinations of PIs (amprenavir, indinavir, saquinavir and lopinavir), and the mRNA expression levels of metabolizing enzyme and efflux transporters were measured using quantitative reverse transcription polymerase chain reaction. The alteration of gene expression was further correlated to the expression of nuclear hormone receptor PXR. Uptake of fluorescent and radioactive substrates was carried out to study the functional activity of these proteins. Cytotoxicity and adenosine triphosphate (ATP) assays were carried out to measure stress responses.
Results: Binary and ternary combinations of PIs appeared to modulate the expression of CYP3A4, MRP2, P-gp and BCRP in a considerable manner. Unlike the individual PIs, their binary combinations showed much greater induction of metabolizing enzyme and efflux proteins. However, such pronounced induction was not observed in the presence of ternary combinations. The observed trend of altered mRNA expression was found to correlate well with the change in expression levels of PXR. The gene expression was found to correlate with activity assays. Lack of cytotoxicity and ATP activity was observed in the treatment samples, suggesting that these alterations in expression levels were probably not stress responses.
Conclusions: In the present study, we demonstrated that combinations of drugs can have serious consequences toward the treatment of HIV infection by altering their bioavailability and disposition.
Acknowledgments
This work was supported by National Institutes of Health grant 1R01 AI071199.
Conflict of interest statement
Authors’ conflict of interest disclosure: The authors stated that there are no conflicts of interest regarding the publication of this article. Research support played no role in thestudy design; in the collection, analysis, and interpretation of data; in the writing of the report; or in the decision to submit the report for publication.
Employment or leadership: None declared.
Honorarium: None declared.
References
1. Pal D, Kwatra D, Minocha M, Paturi DK, Budda B, Mitra AK. Efflux transporters- and cytochrome P-450-mediated interactions between drugs of abuse and antiretrovirals. Life Sci 2011;88:959–71.10.1016/j.lfs.2010.09.012Search in Google Scholar
2. Pal D, Mitra AK. MD- and CYP3A4-mediated drug-drug interactions. J Neuroimmune Pharmacol 2006;1:323–39.10.1007/s11481-006-9034-2Search in Google Scholar
3. Yerly S, Kaiser L, Race E, Bru JP, Clavel F, Perrin L. Transmission of antiretroviral-drug-resistant HIV-1 variants. Lancet 1999;354:729–33.10.1016/S0140-6736(98)12262-6Search in Google Scholar
4. Ford J, Khoo SH, Back DJ. The intracellular pharmacology of antiretroviral protease inhibitors. J Antimicrob Chemother 2004;54:982–90.10.1093/jac/dkh487Search in Google Scholar
5. Turriziani O, Antonelli G, Dianzani F. Cellular factors involved in the induction of resistance of HIV to antiretroviral agents. Int J Antimicrob Agents 2000;16:353–6.10.1016/S0924-8579(00)00264-8Search in Google Scholar
6. Patel J, Buddha B, Dey S, Pal D, Mitra AK. In vitro interaction of the HIV protease inhibitor ritonavir with herbal constituents: changes in P-gp and CYP3A4 activity. Am J Ther 2004;11:262–77.10.1097/01.mjt.0000101827.94820.22Search in Google Scholar
7. Schinkel AH, Jonker JW. Mammalian drug efflux transporters of the ATP binding cassette (ABC) family: an overview. Adv Drug Deliv Rev 2003;55:3–29.10.1016/S0169-409X(02)00169-2Search in Google Scholar
8. Huisman MT, Smit JW, Schinkel AH. Significance of P-glycoprotein for the pharmacology and clinical use of HIV protease inhibitors. AIDS 2000;14:237–42.10.1097/00002030-200002180-00005Search in Google Scholar PubMed
9. Maffeo A, Bellomi F, Solimeo I, Bambacioni F, Scagnolari C, De Pisa F, et al. P-glycoprotein expression affects the intracellular concentration and antiviral activity of the protease inhibitor saquinavir in a T cell line. New Microbiol 2004;27:119–26.Search in Google Scholar
10. Luetkemeyer AF, Havlir DV, Currier JS. Complications of HIV disease and antiretroviral therapy. Top Antivir Med 2012;20: 48–60.Search in Google Scholar
11. Fan J, Maeng HJ, Du Y, Kwan D, Pang KS. Transport of 5,5-diphenylbarbituric acid and its precursors and their effect on P-gp, MRP2 and CYP3A4 in Caco-2 and LS180 cells. Eur J Pharm Sci 2011;42:19–29.10.1016/j.ejps.2010.10.001Search in Google Scholar PubMed
12. Gupta A, Mugundu GM, Desai PB, Thummel KE, Unadkat JD. Intestinal human colon adenocarcinoma cell line LS180 is an excellent model to study pregnane X receptor, but not constitutive androstane receptor, mediated CYP3A4 and multidrug resistance transporter 1 induction: studies with anti-human immunodeficiency virus protease inhibitors. Drug Metab Dispos 2008;36:1172–80.10.1124/dmd.107.018689Search in Google Scholar PubMed
13. Kwatra D, Budda B, Vadlapudi AD, Vadlapatla RK, Pal D, Mitra AK. Transfected MDCK cell line with enhanced expression of CYP3A4 and P-glycoprotein as a model to study their role in drug transport and metabolism. Mol Pharm 2012;9:1877–86.10.1021/mp200487hSearch in Google Scholar PubMed PubMed Central
14. Kwatra D, Vadlapatla RK, Vadlapudi AD, Pal D, Mitra AK. Interaction of gatifloxacin with efflux transporters: a possible mechanism for drug resistance. Int J Pharm 2010;395:114–21.10.1016/j.ijpharm.2010.05.027Search in Google Scholar PubMed PubMed Central
15. Hariharan S, Gunda S, Mishra GP, Pal D, Mitra AK. Enhanced corneal absorption of erythromycin by modulating P-glycoprotein and MRP mediated efflux with corticosteroids. Pharm Res 2009;26:1270–82.10.1007/s11095-008-9741-xSearch in Google Scholar PubMed PubMed Central
16. Dey S, Gunda S, Mitra AK. Pharmacokinetics of erythromycin in rabbit corneas after single-dose infusion: role of P-glycoprotein as a barrier to in vivo ocular drug absorption. J Pharmacol Exp Ther 2004;311:246–55.10.1124/jpet.104.069583Search in Google Scholar PubMed
17. Dixit V, Hariparsad N, Li F, Desai P, Thummel KE, Unadkat JD. Cytochrome P450 enzymes and transporters induced by anti-human immunodeficiency virus protease inhibitors in human hepatocytes: implications for predicting clinical drug interactions. Drug Metab Dispos 2007;35:1853–9.10.1124/dmd.107.016089Search in Google Scholar PubMed
18. Pfrunder A, Gutmann H, Beglinger C, Drewe J. Gene expression of CYP3A4, ABC-transporters (MDR1 and MRP1-MRP5) and hPXR in three different human colon carcinoma cell lines. J Pharm Pharmacol 2003;55:59–66.10.1111/j.2042-7158.2003.tb02434.xSearch in Google Scholar PubMed
19. Svard J, Spiers JP, Mulcahy F, Hennessy M. Nuclear receptor-mediated induction of CYP450 by antiretrovirals: functional consequences of NR1I2 (PXR) polymorphisms and differential prevalence in Whites and Sub-Saharan Africans. J Acquir Immune Defic Syndr 2010;55:536–49.10.1097/QAI.0b013e3181f52f0cSearch in Google Scholar PubMed
20. Dumond JB, Vourvahis M, Rezk NL, Patterson KB, Tien HC, White N, et al. A phenotype-genotype approach to predicting CYP450 and P-glycoprotein drug interactions with the mixed inhibitor/inducer tipranavir/ritonavir. Clin Pharmacol Ther 2010;87:735–42.10.1038/clpt.2009.253Search in Google Scholar PubMed PubMed Central
21. Harmsen S, Meijerman I, Beijnen JH, Schellens JH. The role of nuclear receptors in pharmacokinetic drug-drug interactions in oncology. Cancer Treat Rev 2007;33:369–80.10.1016/j.ctrv.2007.02.003Search in Google Scholar PubMed
22. Burk O, Arnold KA, Geick A, Tegude H, Eichelbaum M. A role for constitutive androstane receptor in the regulation of human intestinal MDR1 expression. Biol Chem 2005;386:503–13.10.1515/BC.2005.060Search in Google Scholar PubMed
23. Maher JM, Cheng X, Slitt AL, Dieter MZ, Klaassen CD. Induction of the multidrug resistance-associated protein family of transporters by chemical activators of receptor-mediated pathways in mouse liver. Drug Metab Dispos 2005;33:956–62.10.1124/dmd.105.003798Search in Google Scholar
24. Dou W, Mukherjee S, Li H, Venkatesh M, Wang H, Kortagere S, et al. Alleviation of gut inflammation by Cdx2/Pxr pathway in a mouse model of chemical colitis. PloS One 2012;7:e36075.10.1371/journal.pone.0036075Search in Google Scholar
25. Ho EA, Piquette-Miller M. Regulation of multidrug resistance by pro-inflammatory cytokines. Curr Cancer Drug Targets 2006;6:295–311.10.2174/156800906777441753Search in Google Scholar
26. Fairchild CR, Cowan KH. Keynote address: multidrug resistance: a pleiotropic response to cytotoxic drugs. Int J Radiat Oncol Biol Phys 1991;20:361–7.10.1016/0360-3016(91)90121-JSearch in Google Scholar
27. Luo S, Pal D, Shah SJ, Kwatra D, Paturi KD, Mitra AK. Effect of HEPES buffer on the uptake and transport of P-glycoprotein substrates and large neutral amino acids. Mol Pharm 2010;7:412–20.10.1021/mp900193eSearch in Google Scholar PubMed PubMed Central
28. Khandelwal A, Krasowski MD, Reschly EJ, Sinz MW, Swaan PW, Ekins S. Machine learning methods and docking for predicting human pregnane X receptor activation. Chem Res Toxicol 2008;21:1457–67.10.1021/tx800102eSearch in Google Scholar PubMed PubMed Central
29. Ekins S, Chang C, Mani S, Krasowski MD, Reschly EJ, Iyer M, et al. Human pregnane X receptor antagonists and agonists define molecular requirements for different binding sites. Mol Pharmacol 2007;72:592–603.10.1124/mol.107.038398Search in Google Scholar PubMed
30. Li H, Redinbo MR, Venkatesh M, Ekins S, Chaudhry A, Bloch N, et al. Novel yeast-based strategy unveils antagonist binding regions on the nuclear xenobiotic receptor PXR. J Biol Chem 2013;288:13655–68.10.1074/jbc.M113.455485Search in Google Scholar PubMed PubMed Central
©2014 by Walter de Gruyter Berlin/Boston
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Articles in the same Issue
- Frontmatter
- Editorial
- Importance of ABC transporters in different tissues
- Review
- Pharmacogenetic potential biomarkers for carbamazepine adverse drug reactions and clinical response
- Reviews in Population Pharmacogenomics
- The pharmacogenetics of drug metabolizing enzymes in the Lebanese population
- Original Articles
- Characterization of ABC transporters in human skin
- Binary and ternary combinations of anti-HIV protease inhibitors: effect on gene expression and functional activity of CYP3A4 and efflux transporters
- Raloxifene pharmacodynamics is influenced by genetic variants in the RANKL/RANK/OPG system and in the Wnt signaling pathway
- The effect of AST-120 on the single-dose pharmacokinetics of metoprolol extended-release tablets in healthy subjects
- Case Report
- Liver enzyme abnormalities during antipsychotic treatment: a case report of risperidone-associated hepatotoxicity
