HS-173 is an effective PI3Kα inhibitor (IC50: 0.8 nM).

PI3Kα:0.8 nM

通过阻断PI3K/Akt信号,HS-173可使体内肝纤维化进程显著减慢.其还可使小鼠体内血管形成减少.

在体外试验中,通过影响细胞周期分布并激活半胱天冬酶,HS-173可诱导细胞凋亡,并使VEGF诱导的血管生成被阻断。在胰腺癌细胞中,HS-173和索拉非尼联用有协同抗癌效果。在T47D,SK-BR3,和MCF7细胞中,HS-173具有明显的抗增殖这个（IC50：0.6/1.5/7.8 μM）。 HS-173可使癌细胞系(Hep3B和SkBr3)中的PI3K通路被完全抑制。

PI3-Kinase assay: The PI3K assay is performed using the Kinase-Glo Max luminescent kinase assay kit which quantifies the amount of ADP produced by the PI3K reaction. In brief, an active PI3K (100 ng) is preincubated with compound for 5 min in kinase reaction buffer (25 mM MOPS [pH 7.0], 5 mM MgCl2, and 1 mM EGTA) and 10 μg l-α-phosphatidylinositol (PI). Before addition of PI, it is sonicated with sonication buffer (25 mM MOPS [pH 7.0], 1 mM EGTA) in water for 20 min for allowing miscelle formation. Then reaction is started by the addition of 10 μM ATP and ran for 180 min. To terminate kinase reaction, same volume of Kinase-Glo Max buffer is added. After 10 min, the plates are then read on a GloMax plate reader for luminescence detection.

Cell viability is performed by a MTT assay. Briefly, T47D cells are plated in 96-well plates for 24 h. Then, the medium is removed and cells were treated with either DMSO as a control or various concentrations of inhibitors. The final concentration of DMSO in the medium was ≤0.1% (v/v). After the cells are incubated for 48 h, 20 μL MTT solution (5 mg/mL) is added to each well for another 4 h at 37 °C. The formazan crystals that formed are dissolved in DMSO (100 μL/well) by constant shaking for 5 min. The plate is then read on a microplate reader at 540 nm. Three replicate wells are used for each analysis. The median inhibitory concentration (IC50, defined as the drug concentration at which cell growth is inhibited by 50%) is assessed from the dose?response curves. To assess the effect of compounds on cell proliferation, T47D cells are cultured with compound (0.1?100 μM) for 48 h before MTT analysis.(Only for Reference)