S/GSK1349572

Inhibitory activities

The inhibitory potencies of S/GSK1349572 and was measured in a strand transfer assay using recombinant HIV integrase as previously described. A complex of integrase and biotinylated preprocessed donor DNA-streptavidin-coated Acintillation proximity assay (SPA) beads was formed by incubating 2 μM purified recombinant integrase with 0.66 μM biotinylated donor DNA-4 mg/ml streptavidin-coated SPA beads in 25 mM sodium morpholinepropanesulfonic acid (MOPS) (pH 7.2), 23 mM NaCl, and 10 mM MgCl2 for 5 min at 37℃. These beads were spun down and preincubated with diluted S/GSK1349572 for 60 min at 37℃. Then a 3H-labeled target DNA substrate was added to give a final concentration of 7 nM substrate, and the strand transfer reaction mixture was incubated at 37℃ for 25 to 45 min, which allowed for a linear increase in the strand transfer of donor DNA to radiolabeled target DNA. The signal was read using a Wallac MicroBeta scintillation plate reader.

Cell lines

MT-4 cells infected with HIV-1 strain IIIB; PBMCs.

Preparation method

Soluble in DMSO > 10 mM. General tips for obtaining a higher concentration: Please warm the tube at 37℃ for 10 minutes and/or shake it in the ultrasonic bath for a while. Stock solution can be stored below -20℃ for several months.

Reaction Conditions

0.16, 0.8, 4, 20 nM; 4 or 5 days.

Applications

In MT-4 cells, S/GSK1349572 inhibits HIV-1 with EC50 value of 0.71 nM. Also, S/GSK1349572 inhibits HIV-1 in PBMCs and in the PHIV assay with EC50 values of 0.51 and 2.2 nM, respectively. In proliferating IM-9, U-937, MT-4, and Molt-4 cells, S/GSK1349572 exhibits 50% cytotoxic concentrations (CC50) of 4.8, 7.0, 14 and 15 μM, respectively. In MT-4 cells infected with HIV-1 NL432, S/GSK1349572 inhibits viral replication.

Animal models

C57BL/6 mice

Dosage form

2.7 mg/kg/day; two weeks; administrated orally.

Applications

In C57BL/6 mice, S/GSK1349572 significantly increases serum creatinine, which is consistent with integrase inhibitors competitively inhibiting creatinine secretion.

Other notes

Please test the solubility of all compounds indoor, and the actual solubility may slightly differ with the theoretical value. This is caused by an experimental system error and it is normal.

References:

[1]. Kobayashi M, Yoshinaga T, Seki T, et al. In Vitro antiretroviral properties of S/GSK1349572, a next-generation HIV integrase inhibitor. Antimicrob Agents Chemother, 2011, 55(2): 813-821.

[2]. Eadon MT, Zhang H, Skaar TC, et al. A two-week regimen of high-dose integrase inhibitors does not cause nephrotoxicity in mice. Antivir Chem Chemother, 2015.