Measurement of plasma and intracellular concentrations of raltegravir in patients with HIV infection

Sandkovsky, Uriel; Swindells, Susan; Robbins, Brian L.; Nelson, Sarah R.; Acosta, Edward P.; Fletcher, Courtney V.

doi:10.1097/qad.0b013e328359a978

Cited by 10 publications

(9 citation statements)

References 8 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Here, we report that processing samples through oil resulted in higher cell-associated drug concentrations compared with standard techniques utilizing multiple washes or even processing samples on ice [8,11,12]. We did observe similar amounts of antiretrovirals present in samples washed once with HBSS and samples centrifuged through oil.…”

Section: Discussionmentioning

confidence: 58%

“…Low passage cells were cultured in RPMI + 10% fetal calf serum (Life Technologies, Grand Island, NY, USA) at 37°C with 5% CO 2 CEM.ss cells were plated in T-75 flasks at an initial concentration of 4 × 10 5 cells/ml. Raltegravir (RAL), a gift from Merck & Co. Inc. (Rahway, NJ, USA) was added to the flasks with a final concentration of 3 μM to approximate average peak plasma concentrations [11]. In additional experiments cells were treated with a combination of 5 nM of the HIV-1 NNRTI efavirenz (EFV) as well as the HIV-1 protease inhibitors 0.5 nM atazanavir (ATV), and 1.25 nM darunavir (DRV), lopinavir (LPV) and ritonavir (RTV) simultaneously.…”

Section: Methodsmentioning

confidence: 99%

“…Cell-associated RAL sample extracts were analyzed using a validated method published previously [11,12]. Briefly, methanol extracts were centrifuged to remove cellular debris.…”

Section: Methodsmentioning

confidence: 99%

“…The assay standard curve has a range from 0.00562 fmol/μl to 22.5 fmol/μl and is linear over this range. HBSS wash samples were treated using a plasma sample protocol as published [11]. An aliquot of sample was spiked with 13C6 RAL, and extracted with methanol.…”

Section: Methodsmentioning

confidence: 99%

See 3 more Smart Citations

A Rapid Spin Through Oil Results in Higher Cell-Associated Concentrations of Antiretrovirals Compared with Conventional Cell Washing

et al. 2015

View full text Add to dashboard Cite

Background Determination of cell-associated antiretroviral drug concentrations is necessary for research into reservoirs of HIV. Variation exists in cell-associated drug concentrations among research groups. One cause for this may be washing cells during processing. We explored spinning cells through oil to minimize this variability. Methods & results Raltegravir, atazanavir, darunavir, efavirenz, lopinavir and ritonavir concentrations were assessed in CEM.ss T cells washed with HBSS and oil-spun cells. Oil-spun cells had significantly higher concentrations for all drugs compared with samples washed with HBSS. Conclusion The decline in cell-associated drug concentrations with saline washes compared with a single spin through oil shows the utility of a spin through oil. Oil centrifugation results in high cell-associated drug concentrations, and can be done in a fast, efficient manner.

show abstract

Section: Discussionmentioning

confidence: 58%

Section: Methodsmentioning

confidence: 99%

“…Cell-associated RAL sample extracts were analyzed using a validated method published previously [11,12]. Briefly, methanol extracts were centrifuged to remove cellular debris.…”

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 2 more Smart Citations

A Rapid Spin Through Oil Results in Higher Cell-Associated Concentrations of Antiretrovirals Compared with Conventional Cell Washing

et al. 2015

View full text Add to dashboard Cite

show abstract

“…One possible explanation for this could be that raltegravir is concentrated within cells, so lower extracellular (plasma) concentrations are not correlated with high intracellular concentrations. However, in our and other studies, raltegravir concentrations in PBMCs are strongly correlated with plasma concentrations and do not show any accumulation of the drug inside the cells that could explain the lack of correlation between plasma concentrations and efficacy (22,24,25). It is likely that 100% of measured drug concentrations do not need to be above the IC 95 to achieve PrEP efficacy.…”

Section: Discussionmentioning

confidence: 70%

Correlation between Plasma, Intracellular, and Cervical Tissue Levels of Raltegravir at Steady-State Dosing in Healthy Women

Mitchell

Roemer

Nkwopara

et al. 2014

Antimicrob Agents Chemother

Self Cite

View full text Add to dashboard Cite

Raltegravir is an antiretroviral with potential value for preexposure prophylaxis (PrEP) against HIV, but the intracellular pharmacokinetics in genital tissue have not been described. In this study, healthy, HIV-uninfected nonpregnant women took 400 mg of raltegravir twice daily for 22 days. On day 8, 15, and 22, blood was collected 0, 4, 6, 8, and 12 h and cervical biopsy specimens taken 0, 6, and 12 h after raltegravir dosing. Plasma and intracellular raltegravir concentrations in peripheral blood mononuclear cells (PBMC) and cervical tissue were measured by tandem mass spectrometry. Linear mixed effects models evaluated correlations between different sample types, as well as differences in concentration between phases of the menstrual cycle. Ten women were enrolled: 9 completed all three visits and 1 completed two visits. The age (mean ؎ standard deviation) of participants was 30 ؎ 8 years. Trough plasma concentrations of raltegravir 12 h after a directly observed dose were above the HIV 95% inhibitory concentration (IC 95 ) of 33 nM (14.6 ng/ml) in 96% of measurements, compared to 67% of PBMC and 89% of cervical tissue trough values. Across all measurements, only 2% (3/135) of plasma values fell below the IC 95 , compared to 10% (13/135) for PBMC and 6% (5/81) for cervical tissue. There was no impact of menstrual phase on raltegravir concentrations. In conclusion, cervical tissue raltegravir concentrations were no greater than plasma concentrations, and ϳ10% of all cervical tissue trough values were below the IC 95 , making the current twice-daily formulation of raltegravir impractical for PrEP. P reexposure prophylaxis (PrEP) using antiretroviral medication has proven effective for infants and adults to prevent HIV acquisition. Studies have shown significant reduction in risk of HIV acquisition in high-risk infants born to HIV-infected women (1, 2), men who have sex with men (3), and HIV-1 serodiscordant heterosexual partners (4). The ideal agent for PrEP should have a long half-life and high concentrations in the genital tract at the site of viral exposure, should be nontoxic, and should not be part of first-line HIV regimens. Tenofovir was chosen as a potential PrEP agent in part because it fulfills almost all of these criteria (5). However, tenofovir is part of the first-line HIV treatment regimen in many parts of the world (6), raising concerns about the impact of development of drug resistance if HIV transmission occurs while taking PrEP.Raltegravir is an HIV integrase inhibitor which blocks strand transfer of viral DNA and integration into the host's genome and has minimal adverse effects (7). Genital fluid concentrations of raltegravir are higher than plasma concentrations, an attractive quality for a PrEP agent (8, 9). However, no studies have examined intracellular tissue concentrations of raltegravir in the genital tract, where the level of the drug would be critical to ensuring efficacy (10). Raltegravir is a substrate for P glycoprotein transporter, which is expressed in cervical tissue (11) and...

show abstract

Mucosal tissue pharmacokinetics of the integrase inhibitor raltegravir in a humanized mouse model: Implications for HIV pre-exposure prophylaxis

Veselinović

Yang²,

Sykes³

et al. 2016

Virology

View full text Add to dashboard Cite

Orally administered anti-retroviral drugs show considerable promise for HIV/AIDS pre-exposure prophylaxis (PrEP). For the success of these strategies, pharmacokinetic (PK) data defining the optimal concentration of the drug needed for protection in relevant mucosal exposure sites is essential. Here we employed a humanized mouse model to derive comprehensive PK data on the HIV integrase inhibitor raltegravir (RAL), a leading PrEP drug candidate. Under steady state conditions following oral dosing, plasma and multiple mucosal tissues were sampled simultaneously. RAL exhibited higher drug exposure in mucosal tissues relative to that in plasma with one log higher exposure in vaginal and rectal tissue and two logs higher exposure in intestinal mucosa reflecting the trends seen in the human studies. These data demonstrate the suitability of RAL for HIV PrEP and validate the utility of humanized mouse models for deriving important preclinical PK-PD data.

show abstract

Measurement of plasma and intracellular concentrations of raltegravir in patients with HIV infection

Cited by 10 publications

References 8 publications

A Rapid Spin Through Oil Results in Higher Cell-Associated Concentrations of Antiretrovirals Compared with Conventional Cell Washing

A Rapid Spin Through Oil Results in Higher Cell-Associated Concentrations of Antiretrovirals Compared with Conventional Cell Washing

Correlation between Plasma, Intracellular, and Cervical Tissue Levels of Raltegravir at Steady-State Dosing in Healthy Women

Mucosal tissue pharmacokinetics of the integrase inhibitor raltegravir in a humanized mouse model: Implications for HIV pre-exposure prophylaxis

Contact Info

Product

Resources

About