Combination of Tenofovir and Emtricitabine plus Efavirenz: In Vitro Modulation of ABC Transporter and Intracellular Drug Accumulation

Bousquet, Laurence; Pruvost, Alain; Guyot, Anne-Cécile; Farinotti, Robert; Mabondzo, Aloı̈se

doi:10.1128/aac.00733-08

Cited by 46 publications

(40 citation statements)

References 56 publications

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“…Traditional mass spectrometric methods identify the NNRTI EFV in negative ionization mode, largely due to the presence of a highly electronegative CF 3 constituent (Table 1) [42]. However, consistent with previously published observations, analyte detection in negative ionization mode significantly impacted overall peak shape and sensitivity of the NRTI TFV [43].…”

Section: 0 Results and Discussionsupporting

confidence: 70%

The development and validation of a method using high-resolution mass spectrometry (HRMS) for the qualitative detection of antiretroviral agents in human blood

Marzinke

Breaud

Parsons

et al. 2014

Clinica Chimica Acta

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Background Antiretroviral drugs are used for the treatment and prevention of HIV infection. Non-adherence to antiretroviral drug regimens can compromise their clinical efficacy and lead to emergence of drug-resistant HIV. Clinical trials evaluating antiretroviral regimens for HIV treatment and prevention can also be compromised by poor adherence and non-disclosed off-study antiretroviral drug use. This report describes the development and validation of a high throughput, qualitative method for the identification of antiretroviral drugs using high-resolution mass spectrometry (HRMS) for the retrospective assessment of off-study antiretroviral drug use and the determination of potential antiretroviral therapy (ART) non-compliance. Methods Serum standards were prepared that contained 15 antiretroviral drugs: 9 protease inhibitors (PIs), 4 nucleotide/nucleoside reverse transcriptase inhibitors (NRTIs), and 2 nonnucleoside/nucleotide reverse transcriptase inhibitors (NNRTIs). Analytical separation was achieved on a Hypersil Gold PFP (100 × 3 mm) column and the eluent was analyzed using the Thermo Exactive Orbitrap mass spectrometer (Exactive-MS) operated in full scan mode. Limit of identification, signal intensity precision, retention time analysis, selectivity, and carryover studies were conducted. Concordance with liquid chromatographic-tandem mass spectrometric (LC-MS/MS) methods was evaluated using remnant plasma samples from a clinical trial. Results The limit of identification ranged from 5-10 ng/ml for 14 drugs (9 PIs, 1 NNRTI, 4 NRTIs) and was 150 ng/ml for 1 NNRTI. Precision studies with high and low control mixtures revealed signal intensity coefficients of variation of 3.0-27.5%. The Exactive-MS method was selective for the compounds of interest. Overall, concordance ranged from 89.1%-100% for the screening of antiretroviral drugs in clinical plasma specimens as compared to LC-MS/MS methods. Conclusion Using the Exactive-MS, we developed and validated a highly selective, robust method for the multiplexed detection of 15 antiretroviral drugs.

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Section: 0 Results and Discussionsupporting

confidence: 70%

The development and validation of a method using high-resolution mass spectrometry (HRMS) for the qualitative detection of antiretroviral agents in human blood

Marzinke

Breaud

Parsons

et al. 2014

Clinica Chimica Acta

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“…Whereas it is unequivocal that efavirenz can induce ABC transporter expression and function in vitro in LS180 cells (35,39,40), data for PBMCs are contradictory (8,9), and there are two studies (one in humans and one in rats) indicating that efavirenz does not provoke changes in intestinal P-gp expression in vivo (6,26). Although our data indicate a slight decrease in P-gp expression in PBMCs, these changes did not translate to changes in P-gp function.…”

Section: Discussionmentioning

confidence: 99%

No Evidence for Induction of ABC Transporters in Peripheral Blood Mononuclear Cells in Humans after 14 Days of Efavirenz Treatment

Burhenne

Matthee

Pasáková

et al. 2010

Antimicrob Agents Chemother

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Intracellular concentrations of antiretroviral drugs in peripheral blood mononuclear cells (PBMCs) are an important determinant of therapeutic success. In vitro data indicate that efavirenz induces several ATP-binding cassette (ABC) transporters, and pharmacogenetic studies found an association between ABCB1(C3435T) and efavirenz exposure and between this polymorphism and improved virological outcomes. We therefore aimed to clarify whether efavirenz also induces ABC transporters in vivo in PBMCs and whether intracellular concentrations might be altered after induction. Twelve healthy individuals received multiple oral doses of efavirenz over 14 days (400 mg once daily). Blood samples were drawn on study days 1 (single dose) and 14 (multiple dose), and efavirenz concentrations were analyzed by liquid chromatography-tandem mass spectrometry. Expression of P glycoprotein (P-gp) and of the multidrug resistance-associated proteins 1 and 2 as well as P-gp activity was analyzed in PBMCs on day 1 and day 14 using real-time reverse transcription-PCR (RT-PCR) and rhodamine 123 efflux. Although a clear autoinduction could be confirmed by a significant decrease of efavirenz exposure from day 1 to day 14, efavirenz did not change expression of the ABC transporters or P-gp activity in PBMCs. Moreover, intracellular concentrations of efavirenz were 1.3-to 1.8-fold higher than the corresponding plasma concentrations, and the intracellular/plasma concentration ratio remained constant during the treatment and did not correlate with ABC transporter expression or function. In conclusion, our study confirmed that intracellular concentrations of efavirenz are independent from these efflux transporters and demonstrated for the first time that the transporters are not induced in PBMCs in vivo after 2 weeks of treatment with efavirenz.

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“…Like for their analysis in plasma (see, for example, PMPA in Nirogi et al, 2008), simple gradients starting with a low percentage of MeOH or ACN in ammonium acetate or formate buffers have been applied (Lynch, Eisenberg, & Kernan, 2001;Kim et al, 2004Kim et al, , 2005Bousquet et al, 2009). Although these methods are relatively uncomplicated, the lack of retention for DPs and TPs restricts their applicability.…”

Section: Separation and Detectionmentioning

confidence: 98%

Mass spectrometry in the quantitative analysis of therapeutic intracellular nucleotide analogs

Jansen

Rosing

Schellens

et al. 2011

Mass Spectrometry Reviews

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Nucleoside analogs are widely used in anti-cancer, anti-(retro)viral, and immunosuppressive therapy. Nucleosides are prodrugs that require intracellular activation to mono-, di-, and finally triphosphates. Monitoring of these intracellular nucleotides is important to understand their pharmacology. The relatively involatile salts and ion-pairing agents traditionally used for the separation of these ionic analytes limit the applicability of mass spectrometry (MS) for detection. Both indirect and direct methods have been developed to circumvent this apparent incompatibility. Indirect methods consist of de-phosphorylation of the nucleotides into nucleosides before the actual analysis. Various direct approaches have been developed, ranging from the use of relatively volatile or very low levels of regular ion-pairing agents, hydrophilic interaction chromatography (HILIC), weak anion-exchange, or porous graphitic carbon columns to capillary electrophoresis and matrix-assisted light desorption--time of flight (MALDI-TOF) MS. In this review we present an overview of the publications describing the quantitative analysis of therapeutic intracellular nucleotide analogs using MS. The focus is on the different approaches for their direct analysis. We conclude that despite the technical hurdles, several useful MS-compatible chromatographic approaches have been developed, enabling the use of the excellent selectivity and sensitivity of MS for the quantitative analysis of intracellular nucleotides.

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Combination of Tenofovir and Emtricitabine plus Efavirenz: In Vitro Modulation of ABC Transporter and Intracellular Drug Accumulation

Cited by 46 publications

References 56 publications

The development and validation of a method using high-resolution mass spectrometry (HRMS) for the qualitative detection of antiretroviral agents in human blood

The development and validation of a method using high-resolution mass spectrometry (HRMS) for the qualitative detection of antiretroviral agents in human blood

No Evidence for Induction of ABC Transporters in Peripheral Blood Mononuclear Cells in Humans after 14 Days of Efavirenz Treatment

Mass spectrometry in the quantitative analysis of therapeutic intracellular nucleotide analogs

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