Maraviroc

Maraviroc (UK-427857, Selzentry) belongs to small molecule drugs and is a C-C chemokine receptor 5 antagonist with oral activity and selectivity. This compound is used for the treatment of HIV-1 infection and has also shown anti-inflammatory, neuroprotective, and potential antitumor synergistic effects in research.

MIP-1β:7.2 nM, RANTES:5.2 nM, HOS cells:2 nM, HeLa P4 cells:0.2 nM, MIP-1α:3.3 nM, R5 HIV-1 (Ba-L):6.4 nM, HIV-1 (Ba-L):1.1 nM (in PM-1 cells), CHO cells:1.4 nM, TZM-bl cells:0.0002 µM

Methods: TZM-bl cells were used to incubate serially diluted Maraviroc with HIV-1 Ba-L strain for 48 hours, followed by detection via luciferase assay.
Results: The EC50 value of Maraviroc was 0.015 μM, with no activity against CXCR4-tropic strains, indicating its specific CCR5 antagonism. [1]

Methods: A humanized NOD/scid-IL-2Rγc null mouse model was used, with HIV-1ADA infection established via intraperitoneal injection. Maraviroc was administered via intraperitoneal injection at a dose of 120 mg/kg twice daily for 3 weeks, with DMSO:PBS mixture as solvent.
Results: Maraviroc significantly reduced viral loads in blood and brain tissue, alleviated immunosuppression, decreased Aβ production and Tau protein phosphorylation in the brain, and protected the blood-brain barrier and neurons. [2]

Inhibition of chemokine binding to CCR5: Binding of 125I-labeled MIP-1α, MIP-1β, and RANTES to CCR5 is measured using intact HEK-293 cells stably expressing the receptor or membrane preparations thereof. Briefly, cells are resuspended in binding buffer (50 mM HEPES containing 1 mM CaCl2, 5 mM MgCl2, and 0.5% bovine serum albumin [BSA] and adjusted to pH 7.4) to a density of 2 × 106 cells/ml. For membrane preparations, phosphate-buffered saline (PBS)-washed cells are resuspended in lysis buffer (20 mM HEPES, 1 mM CaCl2, 1 tablet COMPLETE per 50 mL, pH 7.4) prior to homogenization in a Polytron hand-held homogenizer, ultracentrifugation (40,000× g for 30 min), and resuspension in binding buffer to a protein concentration of 0.25 mg/mL (12.5 μg of membrane protein is used in each well of a 96-well plate). 125I-radiolabeled MIP-1α, MIP-1β, and RANTES are prepared and diluted in binding buffer to a final concentration of 400 pM in the assay. Maraviroc dilutions are added to each well to a final volume of 100 μL, the assay plates incubate for 1 hour, and the contents filter through preblocked and washed Unifilter plates which are counted following overnight drying.

Drug susceptibility assays are performed in 24-well tissue culture plates. Duplicate eight-point dilution series of Maraviroc are prepared in DMSO and medium to yield a final DMSO concentration of 0.1% (vol/vol) in the assay. PHA-stimulated PBMC or PM-1 cells are infected with virus for 1 hour at 37 °C. Cells are subsequently washed once, and 3.6 × 105 PBMC or 2.0 × 105 PM-1 cells are added to each well of assay plates containing diluted Maraviroc. Plates are incubated for 5 days (lab-adapted strains) or 7 days (primary isolates) at 37 °C in a humidified 5% CO2 (vol/vol) atmosphere.(Only for Reference)

DMSO: 255 mg/mL (496.43 mM), Sonication is recommended.

Ethanol: 51.4 mg/mL (100.06 mM), Sonication is recommended.

10% DMSO+40% PEG300+5% Tween 80+45% Saline: 5 mg/mL (9.73 mM), Sonication is recommended.