Copanlisib (BAY 80-6946) is a phosphoinositide 3-kinase (PI3K) inhibitor with potential antineoplastic activity. Copanlisib inhibits the activation of the PI3K signaling pathway, which may result in inhibition of tumor cell growth and survival in susceptible tumor cell populations. Activation of the PI3K signaling pathway is frequently associated with tumorigenesis and dysregulated PI3K signaling may contribute to tumor resistance to a variety of antineoplastic agents.

PI3Kδ:0.7 nM, PI3Kβ:3.7 nM, PI3Kα:0.5 nM, PI3Kγ:6.4 nM

在KPL4细胞和LPA刺激的PC3细胞中，Copanlisib降低pAKT水平。在一组存在PIK3CA突变和/或HER2过表达的人类癌症细胞系中，Copanlisib展示了抗增殖活性并诱导了凋亡。[1] HER2靶向疗法与Copanlisib的联合应用比单独使用任一疗法更有效地抑制增长，并能在细胞中恢复对trastuzumab和lapatinib的敏感性。[2]

在大鼠KPL4或HCT116肿瘤异种移植模型中，Copanlisib（6 mg/kg，静脉注射）可诱导100%的完全肿瘤消退。在携带Lu7860厄洛替尼耐药、源自患者的非小细胞肺癌和MAXF1398源自患者的腺泡状乳腺肿瘤模型的裸鼠中，Copanlisib（14 mg/kg，静脉注射）同样可抑制肿瘤生长。[1]

Biochemical lipid kinase assays: The effect of BAY 80-6946 on PI3Kα, PI3Kβ, and PI3Kγ activity is measured by the inhibition of 33P incorporation into phosphatidylinositol (PI) in 384-well MaxiSorp? plates coated with 2 μg/well of PI and phosphatidylserine (PS) (1:1 molar ratio). In each PI3K isoform assay, 9 μL of reaction buffer (50 mM MOPSO, pH 7.0, 100 mM NaCl, 4 mM MgCl2, 0.1% BSA) containing 7.5 ng of His-tagged N-terminal truncated p110α or p110β protein, or 25 ng of purified human p110γ protein, is used. The reaction is started by adding 5 μL of a 40-μM ATP solution containing 20 μCi/mL [33>/sup>P]-ATP. After 2 hours incubation at room temperature, the reaction is terminated by addition of 5 μL of a 25-mM EDTA solution. The plates are washed and Ultima Gold? scintillation cocktail (25 μL) is then added. The radioactivity incorporated into the immobilized PI substrate is determined with a BetaPlate Liquid Scintillation Counter.

Cell proliferation over a 72-hour period is determined using the CellTiter-Glo? luminescent cell viability kit. Briefly, cells are plated in separate microtiter plates. Following an overnight incubation at 37oC, luminescence values in the t=0 hour plates are determined. Test compounds diluted in growth medium are added to the t=72 hour plates, and the cells are then incubated for 72 hours at 37oC. Luminescence values are determined with a Wallac 1420 Victor2? 1420 multilabel HTS counter after a 10-minute reaction with CellTiter-Glo? solution. The percentage inhibition of cell growth is calculated by subtracting the luminescence values in the t=0 hour plates from the corresponding values in the t=72 hour plates. Differences in values between drug-treated cells and controls are used to determine the percentage inhibition of cell growth.(Only for Reference)