Molecule

Description

S/GSK1349572 (GSK1349572) is a two-metal-binding HIV integrase inhibitor with IC50 of 2.7 nM.

Targets

IC₅₀

In Vitro

S/GSK1349572 shows the potent inhibitory effect on nine clinical isolates from integrase inhibitor-na?ve HIV-2-infected patients with EC50 ranging from 0.2 nM -1.4 nM. ^[1] In vitro, S/GSK1349572 inhibits recombinant HIV-1 integrase-catalyzed strand transfer with IC50 of 2.7 nM. Furthermore, S/GSK1349572 potently inhibits HIV replication in cells such as peripheral blood mononuclear cells (PBMCs), MT-4 cells and CIP4 cells infected with a self-inactivating PHIV lentiviral vector with EC50 of 0,51 nM, 0.71 nM and 2.2 nM, respectively. ^[2] In vitro, S/GSK1349572 exhibits potent activity against five different nonnucleoside reverse transcription inhibitor--resistant or nucleoside reverse transcription inhibitor--resistant viruses with EC50 ranging from 1.3 nM -2.1 nM. Similarly to that against wild-type virus, S/GSK1349572 shows equivalent activity against two protease inhibitor-resistant viruses with EC50 of 0.36 nM and 0.37 nM, respectively. ^[2]

In Vivo

Clinical Trials

S/GSK1349572 (GSK1349572) is currently in Phase III clinical trials in HIV-1 Infected Antiretroviral Naive Adult Subjects.

Features

S/GSK1349572 (GSK1349572) is a next-generation and two-metal-binding HIV integrase strand transfer inhibitor.

Description

In MT-4 cells, S/GSK1349572 shows synergistic antiretroviral activity against HIV in combination with nonnucleoside reverse transcription (RT) inhibitors (efavirenz and nevirapine), nucleoside RT inhibitors (stavudine and abacavir) or HIV protease inhibitors (lopinavir and amprenavir). ^[2] GSK1349572 plus Abacavir/Lamivudine fixed-dose combination therapy is currently in Phase III clinical trials in HIV-1 Infected Antiretroviral Therapy Naive Adult Subjects.

In vitro strand transfer assay

The inhibitory potencies of S/GSK1349572 and other INIs are measured in a strand transfer assay using recombinant HIV integrase. A complex of integrase and biotinylated preprocessed donor DNA-streptavidin-coated Acintillation proximity assay (SPA) beads is 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 minutes at 37 °C. These beads are spun down and preincubated with diluted INIs for 60 minutes at 37 °C. Then a ₃H-labeled target DNA substrate is added to give a final concentration of 7 nM substrate, and the strand transfer reaction mixture was incubated at 37 °C for 25 to 45 minutes, which allows for a linear increase in the strand transfer of donor DNA to radiolabeled target DNA. The signal is read using a Wallac MicroBeta scintillation plate reader.

Cell Lines

MT-4

Concentrations

0 to 10 μM

Incubation Time

4 days or 5 days

Methods

MT-4 cells growing exponentially at a density of 500000 or 600000 /mL are infected with HIV-1 strain IIIB at a viral multiplicity of infection of 0.001 or a 50% tissue culture infective dose of 4 to 10. The cells are then aliquoted to 96-well plates in the presence of varying concentrations of S/GSK1349572. After incubation for 4 or 5 days, antiviral activity is determined by a cell viability assay that either measured bioluminescence with a CellTiter-Glo luminescent reagent or measured absorbance at 560 and 690 nm using the yellow tetrazolium MTT reagent [3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyltetrazolium bromide].