<i>In Vitro</i>Pre- and Post-Exposure Prophylaxis Using HIV Inhibitors as Microbicides Against Cell-Free or Cell-Associated HIV-1 Infection

Terrazas-Aranda, Katty; Herrewege, Yven Van; Lewi, Paul; Roey, Jens Van; Vanham, Guido

doi:10.1177/095632020701800304

Cited by 19 publications

(27 citation statements)

References 45 publications

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“…Since then researchers from various fields have actively been developing technologies to combat the rapid spread of HIV. These strategies include pre-exposure prophylactics (28,41) and microbicides (14,31) designed to topically deliver single or combination antiviral agents. Currently, microbicide delivery systems under development include gels, rings, films, and suppositories.…”

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confidence: 99%

Vaginal Microbicide Gel for Delivery of IQP-0528, a Pyrimidinedione Analog with a Dual Mechanism of Action against HIV-1

Mahalingam

Simmons

Ugaonkar

et al. 2011

Antimicrob Agents Chemother

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Pyrimidinediones, a novel class of compounds, have previously been shown to possess antiviral activity at nanomolar concentrations. One member of this class of compounds, IQP-0528, was selected as the lead molecule for formulation development owing to its stability at physiologically relevant conditions, wide therapeutic window, and antiviral activity in the nanomolar range. Here, we report the development of two vaginal gels-3.0% hydroxyethyl cellulose (HEC) formulation and a 0.65% Carbopol formulation-for the sustained delivery of IQP-0528. Stability studies under accelerated conditions confirmed the chemical stability of IQP-0528 and mechanical stability of the gel formulation for 3 months. In vitro release studies revealed that diffusion-controlled release of IQP-0528 occurred over 6 h, with an initial lag time of approximately 1 h. Based on the drug release profile, the 3.0% HEC gel was selected as the lead formulation for safety and activity evaluations. The in vitro and ex vivo safety evaluations showed no significant loss in cell viability or significant inflammatory response after treatment with a 3.0% HEC gel containing 0.25% IQP-0528. In an in vitro HIV-1 entry inhibition assay, the lead formulation showed an 50% effective concentration of 0.14 g/ml for gel in culture media, which corresponds to ϳ0.001 M IQP-0528. The antiviral activity was further confirmed by using polarized cervical explants, in which the formulation showed complete protection against HIV infection. In summary, these results are encouraging and warrant further evaluation of IQP-0528 gel formulations in in vivo models, as well as the development of alternative formulations for the delivery of IQP-0528 as a microbicide.

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confidence: 99%

Vaginal Microbicide Gel for Delivery of IQP-0528, a Pyrimidinedione Analog with a Dual Mechanism of Action against HIV-1

Mahalingam

Simmons

Ugaonkar

et al. 2011

Antimicrob Agents Chemother

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“…As expected, no firefly luciferase activity was measured in the CCR5-negative X4M reporter cell line. Clearly, all antiretrovirals not only block viral spread to the target cells, but also inhibit viral spread The TCID 50 of BaL-infected effector cells (i.e., resting PBMC/CD4 ϩ T cells, activated PBMC/CD4 ϩ T cells, and macrophages) was determined in a 7-day coculture with R5M target cells. Firefly luciferase was used to assess target cell infection, and the TCID 50 was calculated using the method of Reed and Muench, expressed as the number of cells needed for 50% infection and plotted on the left y axis (black symbols).…”

Section: Resultsmentioning

confidence: 99%

“…TCID 50 determination of cell-free HIV and HIV-infected effector cells. The 50% tissue culture infective dose (TCID 50 ) was determined for cell-free BaL, as well as for BaL-infected effector cells (i.e., resting PBMC, activated PBMC, and macrophages) using the R5M cell line as HIV target cells. The FL reporter enzyme is transcribed and expressed under the influence of Tat-long terminal repeat (LTR) interaction and is therefore restricted to infected R5M cells.…”

Section: Methodsmentioning

confidence: 99%

“…Percentages of viral inhibition were calculated compared to untreated control wells and plotted against the compound concentrations. Nonlinear regression analysis was used to calculate the 50% effective concentration (EC 50 ) based on at least three independent measurements and using GraphPad Prism version 5.03 for Windows (GraphPad Software, San Diego, CA). Significant differences were calculated using a Z test with Bonferroni correction.…”

Section: Methodsmentioning

confidence: 99%

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In Vitro Activities of Candidate Microbicides against Cell-Associated HIV

Selhorst

Grupping

Bourlet

et al. 2012

Antimicrob Agents Chemother

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Most research on HIV transmission and microbicides focuses on the inhibition of cell-free virus (CFV) present in genital secretions. However, an effective microbicide should also block the transmission of cell-associated virus (CAV) originating from seminal T cells and macrophages. Because inhibition of CAV remains controversial, especially for viral entry inhibitors, we developed a novel in vitro assay to evaluate the activities of different classes of candidate microbicides against cell-free HIV and HIVinfected leukocytes (i.e., resting peripheral blood mononuclear cells [PBMC], activated PBMC, and monocyte-derived macrophages). The assay is based on two CD4 ؉ CXCR4 ؉ T-cell lines (R5MaRBLE and X4MaRBLE) that both contain a firefly luciferase reporter gene but differ in the expression of the CCR5 coreceptor. Consequently, the quantification of the luciferase activities and the Gag p24 concentrations in cocultures of R5-tropic HIV-infected leukocytes with each cell line separately allowed us to discriminate between the infection of the cell lines (i.e., target cells), the ongoing infection in the HIV-infected leukocytes (i.e., effector cells), and the total infection of the coculture (i.e., effector plus target cells). All 14 antiretrovirals tested were able to block target cell infection by all three sources of CAV, although a small decrease in activity (2-to 18-fold) was observed for all entry inhibitors. On the other hand, the production of Gag p24 by the infected effector cells could be blocked only by protease inhibitors. Overall, these results show that entry and protease inhibitors are eligible drug classes for inclusion in future combination microbicides.

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“…In adults, emtricitabine recommended dose is 200 mg once a day (QD) [2]. Both in vitro [3] and in vivo [4] testing demonstrated that emtricitabine presents enough potential to be tested in the prevention of HIV-1, either alone or in combination [5].…”

Section: Introductionmentioning

confidence: 99%

High-Performance Liquid Chromatographic Method for Analysis of Emtricitabine in Rat Plasma: Method Development, Validation and Application to a Pharmacokinetic Study

Singh

Pai

2013

ISRN Chromatography

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A new reverse phase liquid chromatographic method for the investigation of emtricitabine in rat plasma was developed after oral administration to Wistar rats. The desired chromatographic separation was achieved on Phenomenex C 18 column (250 mm × 4.6 mm I.D., 5 m) column, under isocratic conditions using UV detection at 280 nm. The optimized mobile phase consisted of a mixture of 10 mM potassium dihydrogen phosphate buffer-(adjusted to pH 6.8) methanol-2% acetic acid in a ratio of (73 : 25 : 2, v/v/v) at a flow rate of 1 mL min −1 . The system was found to produce sharp and well-resolved peaks for emtricitabine with retention time of 5.78 min. The linear regression analysis for the calibration curves showed a good linear correlation over the concentration range 0.050-3.0 g mL −1 , with determination coefficients, R 2 , exceeding 0.9970. The limits of detection (LOD) and quantitation (LOQ) were found to be 0.016 g mL −1 and 0.049 g mL −1 , respectively. The method was successfully applied for the pharmacokinetic in rats. Emtricitabine concentration in plasma reached ( max ) was 1.357 g mL −1 about 2 h after oral administration of 15 mg/kg/rat. The AUC 0−24 was 12.175 g mL −1 * h and the apparent elimination half-life (t 1/2 ) was 8.153 h. This method was found to be suitable for examining emtricitabine concentration in rats, after oral administration of emtricitabine in a single dose.

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In VitroPre- and Post-Exposure Prophylaxis Using HIV Inhibitors as Microbicides Against Cell-Free or Cell-Associated HIV-1 Infection

Cited by 19 publications

References 45 publications

Vaginal Microbicide Gel for Delivery of IQP-0528, a Pyrimidinedione Analog with a Dual Mechanism of Action against HIV-1

Vaginal Microbicide Gel for Delivery of IQP-0528, a Pyrimidinedione Analog with a Dual Mechanism of Action against HIV-1

In Vitro Activities of Candidate Microbicides against Cell-Associated HIV

High-Performance Liquid Chromatographic Method for Analysis of Emtricitabine in Rat Plasma: Method Development, Validation and Application to a Pharmacokinetic Study

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