Skip Navigation LinksHome > August 15, 2005 - Volume 39 - Issue 5 > Atazanavir Inhibits P-Glycoprotein and Multidrug Resistance-...
JAIDS Journal of Acquired Immune Deficiency Syndromes:
Letters to the Editor

Atazanavir Inhibits P-Glycoprotein and Multidrug Resistance-Associated Protein Efflux Activity

Lucia, Mothanje Barbara MD, PhD*; Golotta, Caterina MD†; Rutella, Sergio MD, PhD*; Rastrelli, Elena MD*; Savarino, Andrea MD*; Cauda, Roberto MD, PhD*

Free Access
Article Outline
Collapse Box

Author Information

*Department of Infectious Diseases, Catholic University, Rome, Italy, †Department of Hematology, Catholic University, Rome, Italy

To the Editor:

P-glycoprotein (P-gp) and the multidrug resistance-associated protein (MRP) are the best studied energy-dependent adenosine triphosphate (ATP)-binding cassette (ABC) drug efflux pumps.1,2 P-gp and MRP function as major transporters for the HIV-protease inhibitors (PIs),3,4 and for this reason, their expression and function are determinants of intracellular concentrations of these agents.5 A variety of cell types directly involved in HIV pathobiology express both P-gp and MRP (eg, peripheral blood lymphocytes [PBLs], bone marrow stem cells).6,7 Among the PIs, ritonavir (RTV) is the most potent inhibitor of P-gp and MRP efflux, although all the PIs are substrates for P-gp and MRP-1 and, to some degree, also inhibitors,3,8,9 whereas they do not significantly induce P-gp/MRP expression in normal PBLs.10,11 Atazanavir (ATV) is a potent novel azapeptide inhibitor of the HIV-1 protease with a pharmacokinetic profile permitting once-daily administration and a lower degree of dyslipidemia.12 The objective of this study was to analyze the effect of ATV on P-gp and MRP function in normal PBLs as well as in hemopoietic stem cells (HSCs) from human umbilical cord blood cells (UBCs), which provide a powerful in vitro model of normal human hematopoiesis.13 Cells were coincubated with the following HIV PIs: ATV (Bristol-Myers Squibb), RTV (Abbott Laboratories), saquinavir (SQV; Roche Products), and indinavir (IDV; Merck Sharp & Dohme). PBLs were collected by 5 healthy volunteers. UBCs were obtained from 3 informed mothers undergoing cesarean delivery of full-term infants in accordance with hospital ethical regulations. Mononuclear cells were separated by Ficoll-Hypaque density centrifugation, washed, and subsequently incubated with Miltenyi MultiSort anti-CD34 MicroBeads (Miltenyi Biotec; 100 μL/108 cells); after extensive washing, they were processed using a VarioMACS magnetic device to remove non-CD34+ cells, as previously described.14 CD34+ cells were eluted after removal of unwanted unbound cells by discontinuation of the magnetic fields in the VarioMACS device. All CD34+ immunoselected samples had CD34 cell purity greater than 95%. P-gp and MRP activity was evaluated using functional assays for rhodamine 123 (Rh123; Sigma Chemicals)15 and carboxyfluorescein diacetate (CF; Molecular Probes, Eugene, OR),16 respectively, with or without ATV (0.2-10 μM), RTV (3 μM), SQV (3 μM), and IDV (3 μM), as previously reported.8,17 The anthracycline compound doxorubicin (DOX; Adria Laboratories, Columbus, OH), a common P-gp/MRP naturally fluorescent substrate,18 was used for HSC functional analysis. A human acute lymphoblastic leukemia cell line (CEM) and its vinblastine-resistant subline (CEM/VBL100), kindly provided by Dr. M. Cianfriglia (Istituto Superiore di Sanità, Rome, Italy), were used as negative and positive controls, respectively, for P-gp expression and function. Results were expressed as percentages of dim and bright cells in the case of bimodality of the fluorescence during dye efflux or as mean fluorescence intensity (MFI) when fluorescence distribution in a cell population was Gaussian.8,17 P-gp phenotype was detected by using the specific monoclonal antibody MRK-16 (10 μg/mL; Kamiya Biomedicals, Seattle, WA), as previously reported.8

We investigated whether ATV alone or in combination with other PIs (ie, RTV, SQV) inhibits P-gp and MRP function in normal PBLs. Rh123 retention was studied in the absence and presence of increasing ATV concentrations corresponding to its steady-state pharmacokinetic profile.19 ATV increased Rh123 retention in a concentration-dependent manner, effectively blocking efflux at 10 μM (Fig. 1). We also observed an augmented inhibitory effect between ATV and RTV or SQV (see Fig. 1). ATV also significantly inhibited MRP efflux function (data not shown) as well as DOX efflux by HSCs (see Fig. 1). P-gp expression of drug-selected CEM/VBL100 was higher than that of the corresponding parental cells (3% CEM/P-gp+, 100% CEM/VBL100/P-gp+), and higher doses of ATV were required to inhibit Rh123 efflux (CEM-MFI: efflux 169, ATV-10 μM: efflux 207, ATV-50 μM: efflux 389; CEM/VBL100-MFI: efflux 131, ATV-10 μM: efflux 190, ATV-50 μM: efflux 295, ATV-100 μM: efflux 411), suggesting that ATV may be a possible P-gp/MRP substrate and competitively inhibit P-gp/MRP function as a drug efflux pump.

Figure 1
Figure 1
Image Tools

Although PIs represent a clear advance in the management of HIV disease, drug resistance, side effects, and drug interactions are common. To understand and predict pharmacokinetics and drug interactions, it is essential to determine the interaction of these agents with drug transporters of the ABC family that have been shown to control the intracellular concentrations of PIs.5 ATV is a new once-daily administered PI with excellent anti-HIV activity that is metabolized by cytochrome CYP3A substrate.12,19 It is not known, however, whether ATV interacts with drug transporter proteins because it differs from the peptidomimetic PIs (eg, ritonavir, nelfinavir) by its C-2 symmetric chemical structure.19 Our results indicate that ATV is a possible substrate for P-gp and MRP in HIV-target cells such as normal PBLs and, like other PIs, competitively inhibits their efflux activity in a dose-dependent manner. The concentrations of ATV used here are comparable to those achieved clinically.19 We also observed an increased inhibitory effect of ATV in combination with RTV and SQV that follows the hierarchy ATV+RTV > ATV+SQV, which is most likely attributable to the higher inhibitory potential shown by ATV compared with the older PIs. These data support a recent observation that suggests a predominant effect of RTV on CYP3A4 metabolism and of ATV on ABC transporter activity when ATV, RTV, and SQV are coadministered.20 In addition, we observed a significant ATV-mediated inhibition of the efflux of the anthracycline compound DOX by HSCs from human UBCs. Because P-gp and MRP transport a broad range of anticancer agents (ie, anthracyclines, Vinca alkaloids, taxanes),21 our results help to explain the higher hematologic toxicity induced by the combination of PI-containing highly active antiretroviral therapy (HAART) plus chemotherapy in patients with AIDS-related neoplasms receiving MDR-related cytotoxic agents.22,23 The modulator effect exerted by PIs when used in conjunction with anticancer drugs may enhance the cytotoxicity of chemotherapy drugs as a result of reduced drug efflux and increased intracellular concentration.

Mothanje Barbara Lucia, MD, PhD*

Caterina Golotta, MD†

Sergio Rutella, MD, PhD*

Elena Rastrelli, MD*

Andrea Savarino, MD*

Roberto Cauda, MD, PhD*

*Department of Infectious Diseases Catholic University Rome, Italy †Department of Hematology Catholic University Rome, Italy

Back to Top | Article Outline

REFERENCES

1. Loo TW, Clarke DM. Merk Frosst Award Lecture 1998. Molecular dissection of the human multidrug resistance P-glycoprotein. Biochem Cell Biol. 1999;77:11-23.

2. Borst P, Evers R, Kool M, et al. A family of drug transporters: the multidrug resistance-associated proteins. J Natl Cancer Inst. 2000;92:1295-1302.

3. Lee CG, Gottesman MM, Cardarelli CO, et al. HIV-1 protease inhibitors are substrates for the MDR1 multidrug transporter. Biochemistry. 1998;37:3594-3601.

4. Olson PD, Scadden DT, D'Aquila RT, et al. The protease inhibitor ritonavir inhibits the functional activity of the multidrug resistance related-protein 1 (MRP-1). AIDS. 2002;16:1743-1747.

5. Jones K, Hoggard PG, Sales SD, et al. Differences in the intracellular accumulation of HIV protease inhibitors in vitro and the effect of active transport. AIDS. 2001;15:675-681.

6. Drach D, Zhao S, Drach J, et al. Subpopulations of normal peripheral blood and bone marrow cells express a functional multidrug resistance. Blood. 1992;80:2729-2734.

7. Legrand O, Perrot JY, Tang R, et al. Expression of the multidrug resistance-associated protein (MRP) mRNA and protein in normal peripheral blood and bone marrow haemopoietic cells. Br J Haematol. 1996;94:23-33.

8. Lucia MB, Rutella S, Leone G, et al. HIV-protease inhibitors contribute to P-glycoprotein efflux function defect in peripheral blood lymphocytes from HIV-positive patients receiving HAART. J Acquir Immune Defic Syndr. 2001;27:321-330.

9. Srinivas R, Middlemas D, Flynn P, et al. Human immunodeficiency virus protease serve as substrates for multidrug transporter MDR1 and MRP1 but retain antiviral efficacy in cell lines expressing these transporters. Antimicrob Agents Chemother. 1998;42:3157-3162.

10. Lucia MB, Rutella S, Leone G, et al. In vitro and in vivo modulation of MDR1/P-glycoprotein in HIV-infected patients administered highly active antiretroviral therapy and liposomal doxorubicin. J Acquir Immune Defic Syndr. 2002;30:369-378.

11. Lucia MB, Savarino A, Straface E, et al. Role of lymphocyte multidrug resistance associated protein 1 (MRP1) in HIV infection: expression, function, and consequences of inhibition. J Acquir Immune Defic Syndr. (In press).

12. Havlir DV, O'Marro SD. Atazanavir: new option for treatment of HIV infection. Clin Infect Dis. 2004;38:1599-1604.

13. Leglise MC, Darodes de Tailly P, Vignot JL, et al. A cellular model for drug interactions on hematopoiesis: the use of human umbilical cord blood progenitors as a model for the study of drug-related myelosuppression of normal hematopoiesis. Cell Biol Toxicol. 1996;12:39-53.

14. Rutella S, Bonanno G, Marone M, et al. Identification of a novel subpopulation of human cord blood CD34-CD133-CD7-CD45+ lineage-cells capable of lymphoid/cell differentiation after in vitro exposure to IL-15. J Immunol. 2003;15:2977-2988.

15. Lee JS, Paull K, Alvarez M, et al. Rhodamine efflux patterns predict P-glycoprotein substrates in the National Cancer Institute drug screen. Mol Pharmacol. 1994;46:627-638.

16. Laupeze B, Amiout L, Courtois L, et al. Use of the anionic dye carboxy-2′-dichlorofluorescein for sensitive flow cytometric detection of multidrug resistance-associated protein activity. Int J Oncol. 1999;15:571-576.

17. Savarino A, Lucia MB, Rastrelli E, et al. Anti-HIV effects of chloroquine. Inhibition of viral particle glycosylation and synergism with protease inhibitors. J Acquir Immune Defic Syndr. 2004;35:223-232.

18. Harbottle A, Daly AK, Campbell C. Role of glutathione s-transferase P1, P-glycoprotein and multidrug resistance-associated protein 1 in acquired doxorubicin resistance. Int J Cancer. 2001;92:777-783.

19. Goldsmith DR, Perry CM. Atazanavir. Drugs. 2003;63:1679-1693.

20. Boffito M, Kurowski M, Kruse G, et al. Atazanavir enhances saquinavir hard-gel concentrations in a ritonavir-boosted once-daily regimen. AIDS. 2004;18:1291-1297.

21. Fischer G, Lum B, Sikic BJ. Clinical studies with modulators of multidrug resistance. In: Fischer G, Sikic B, eds. Drug Resistance Clinical Oncology and Hematology. Philadelphia: WB Saunders; 1995:363-382.

22. Spina M, Gabarre J, Rossi G, et al. Stanford V regimen and concomitant HAART in 59 patients with Hodgkin disease and HIV infection. Blood. 2002;100:1984-1988.

23. Bower M, McCall-Peat N, Ryan N, et al. Protease inhibitors potentiate chemotherapy induced neutropenia. Blood. 2004;104:2943-2946.

Cited By:

This article has been cited 17 time(s).

Pharmaceutical Research
Solid lipid nanoparticles enhance the delivery of the HIV protease inhibitor, atazanavir, by a human brain endothelial cell line
Chattopadhyay, N; Zastre, J; Wong, HL; Wu, XY; Bendayan, R
Pharmaceutical Research, 25(): 2262-2271.
10.1007/s11095-008-9615-2
CrossRef
Drugs of Today
Atazanavir/ritonavir: A review of its use in HIV therapy
von Hentig, N
Drugs of Today, 44(2): 103-132.
10.1358/dot.2008.44.2.1137107
CrossRef
Enfermedades Infecciosas Y Microbiologia Clinica
P-glycoprotein and human immunodeficiency virus infection
Peralta, G; Sanchez, MB; Echevarria, S; Valdizan, EM; Armijo, JA
Enfermedades Infecciosas Y Microbiologia Clinica, 26(3): 150-159.

International Review of Cell and Molecular Biology, Vol 280
Impact of Atp-Binding Cassette Transporters on Human Immunodeficiency Virus Therapy
Weiss, J; Haefeli, WE
International Review of Cell and Molecular Biology, Vol 280, 280(): 219-279.
10.1016/S1937-6448(10)80005-X
CrossRef
Antiviral Therapy
Synergistic inhibition of protease-inhibitor-resistant HIV type 1 by saquinavir in combination with atazanavir or lopinavir
Dam, E; Lebel-Binay, S; Rochas, S; Thibaut, L; Faudon, JL; Thomas, CM; Essioux, L; Hill, A; Schutz, M; Clavel, F
Antiviral Therapy, 12(3): 371-380.

Current Hiv Research
Response of feline immunodeficiency virus (FIV) to tipranavir may provide new clues for development of broad-based inhibitors of retroviral proteases acting on drug-resistant HIV-1
Norelli, S; El Daker, S; D'Ostilio, D; Mele, F; Mancini, F; Taglia, F; Ruggieri, A; Ciccozzi, M; Cauda, R; Ciervo, A; Barreca, ML; Pistello, M; Bendinelli, M; Savarino, A
Current Hiv Research, 6(4): 306-317.

Journal of Neuroscience Research
Up-regulation of P-glycoprotein by HIV Protease Inhibitors in a Human Brain Microvessel Endothelial Cell Line
Zastre, JA; Chan, GNY; Ronaldson, PT; Ramaswamy, M; Couraud, PO; Romero, IA; Weksler, B; Bendayan, M; Bendayan, R
Journal of Neuroscience Research, 87(4): 1023-1036.
10.1002/jnr.21898
CrossRef
Cancer Biology & Therapy
CIAPIN1 confers multidrug resistance by upregulating the expression of MDR-1 and MRP-1 in gastric cancer cells
Hao, ZM; Li, XH; Qiao, TD; Du, R; Hong, L; Fan, DM
Cancer Biology & Therapy, 5(3): 261-266.

Antimicrobial Agents and Chemotherapy
Pharmacokinetics of saquinavir, atazanavir, and ritonavir in a twice-daily boosted double-protease inhibitor regimen
von Hentig, N; Muller, A; Rottmann, C; Wolf, T; Lutz, T; Klauke, S; Kurowski, M; Oertel, B; Dauer, B; Harder, S; Staszewski, S
Antimicrobial Agents and Chemotherapy, 51(4): 1431-1439.
10.1128/AAC.00854-06
CrossRef
Journal of Antimicrobial Chemotherapy
Interindividual variability in the effect of atazanavir and saquinavir on the expression of lymphocyte P-glycoprotein
Chinn, LW; Gow, JM; Tse, MM; Becker, SL; Kroetz, DL
Journal of Antimicrobial Chemotherapy, 60(1): 61-67.
10.1093/jac/dkm135
CrossRef
Pharmacogenomics Journal
Overview of the pharmacogenetics of HIV therapy
Rodriguez-Novoa, S; Barreiro, P; Jimenez-Nacher, I; Soriano, V
Pharmacogenomics Journal, 6(4): 234-245.
10.1038/sj.tpj.6500374
CrossRef
Antimicrobial Agents and Chemotherapy
Quantitative immunoassay to measure plasma and intracellular atazanavir levels: Analysis of drug accumulation in cultured T cells
Roucairol, C; Azoulay, S; Nevers, MC; Creminon, C; Lavrut, T; Garraffo, R; Grassi, J; Burger, A; Duval, D
Antimicrobial Agents and Chemotherapy, 51(2): 405-411.
10.1128/AAC.00730-06
CrossRef
Hiv Medicine
Atazanavir and lopinavir with ritonavir alone or in combination: analysis of pharmacokinetic interaction and predictors of drug exposure
Di Giambenedetto, S; De Luca, A; Villani, P; Bacarelli, A; Ragazzoni, E; Regazzi, M; Cauda, R; Navarra, P
Hiv Medicine, 9(4): 239-245.
10.1111/j.1468-1293.2008.00555.x
CrossRef
AIDS Research and Human Retroviruses
Comparison of ABC transporter modulation by atazanavir in lymphocytes and human brain endothelial cells: ABC transporters are involved in the atazanavir-limited passage across an in vitro human model of the blood-brain barrier
Bousquet, L; Roucairol, C; Hembury, A; Nevers, MC; Creminon, C; Farinotti, R; Mabondzo, A
AIDS Research and Human Retroviruses, 24(9): 1147-1154.
10.1089/aid.2007.0022
CrossRef
Trends in Parasitology
HIV and malaria co-infection: interactions and consequences of chemotherapy
Skinner-Adams, TS; McCarthy, JS; Gardiner, DL; Andrews, KT
Trends in Parasitology, 24(6): 264-271.
10.1016/j.pt.2008.03.008
CrossRef
AIDS
Genetic factors influencing atazanavir plasma concentrations and the risk of severe hyperbilirubinemia
Rodríguez-Nóvoa, S; Martín-Carbonero, L; Barreiro, P; González-Pardo, G; Jiménez-Nácher, I; González-Lahoz, J; Soriano, V
AIDS, 21(1): 41-46.
10.1097/QAD.0b013e328011d7c1
PDF (99) | CrossRef
AIDS
Atazanavir and lopinavir/ritonavir: pharmacokinetics, safety and efficacy of a promising double-boosted protease inhibitor regimen
Ribera, E; Azuaje, C; Lopez, RM; Diaz, M; Feijoo, M; Pou, L; Crespo, M; Curran, A; Ocaña, I; Pahissa, A
AIDS, 20(8): 1131-1139.
10.1097/01.aids.0000226953.56976.ad
PDF (199) | CrossRef
Back to Top | Article Outline

© 2005 Lippincott Williams & Wilkins, Inc.

Login