Hck-IN-1 (compound B9), a diphenylpyrazolo compound, is a selective Nef-dependent Hck inhibitor with IC 50 s of 2.8 μM, >20 μM for Nef:Hck complex and Hck, respectively. Hck-IN-1 is a direct and wide HIV-1 Nef antagonists with an IC 50 of 100-300 nM for wild-type HIV-1 replication. Hck-IN-1 binds pocket residue Asn126 and has anti-retroviral activity In Vitro Hck-IN-1 (compound B9) shows weak activity against other Src-family members in vitro, with IC 50 values >20 μM for c-Src, Lck and Lyn. B9 (1 μM; 8 days) completely inhibits Nef-dependent SFK activation at a concentration of 1.0 μM. Hck-IN-1 (0.1, 0.3, 1, 3 μM) also inhibits Nef-mediated enhancement of HIV-1 infectivity in a concentration-dependent manner in the reporter cell line, TZM-bl. Hck-IN-1 inhibits the replication of all eleven HIV-1 Nef chimeras with IC 50 values of ~ 300 nM in CEM-T4 cells, demonstrating that the compound is broadly active against HIV replication supported by a wide range of HIV-1 Nef proteins. MCE has not independently confirmed the accuracy of these methods. They are for reference only. Western Blot AnalysisCell Line: CEM-T4 cells Concentration: 1 μM Incubation Time: 8 days Result: Completely inhibited Nef-dependent SFK activation at a concentration of 1.0 μM. Form:Solid IC50& Target:HIV-1 Nef 100-300 nM (IC 50 )