BAY 80-6946 (Copanlisib)3X 积分

Biochemical lipid kinase assays

The effect of BAY 80-6946 on PI3Kα, PI3Kβ and PI3Kγ activity was 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 was used. The reaction was started by adding 5 μL of 40 μM ATP solution containing 20 μCi/mL [γ-33P]-ATP. After 2-hr incubation at room temperature, the reaction was terminated by addition of 5 μL of 25 mM EDTA solution. The plates were washed and Ultima Gold scintillation cocktail (25 μL) was then added. The radioactivity incorporated into the immobilized PI substrate was determined with a BetaPlate Liquid Scintillation Counter.

Cell lines

A panel of cancer cell lines

Preparation method

The solubility of this compound in DMSO is limited. General tips for obtaining a higher concentration: Please warm the tube at 37 °C for 10 minutes and/or shake it in the ultrasonic bath for a while. Stock solution can be stored below - 20 °C for several months.

Reacting condition

5 μM; 72 hrs

Applications

BAY 80-6946 showed significant anti-proliferative activity in a series of cancers cells exhibiting constitutively activated PI3K signaling. Several breast cancer, endometrial cancer and hematologic tumor cell lines were extremely sensitive to BAY 80-6946 (IC50 values < 10 nM).

Animal models

A rat KPL4 tumor xenograft model

Dosage form

0.5 ~ 6 mg/kg; i.v.; every 2 days for a total of 5 doses starting on day 14 after tumor cell implantation

Applications

On day 25 (i.e. 3 days after the last dose), BAY 80-6946 at doses of 0.5, 1, 3 and 6 mg/kg showed TGI rates of 77%, 84%, 99% and 100%, respectively. In addition, BAY 80-6946 at doses of 3 and 6 mg/kg resulted in complete tumor regression.

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]. Liu N, Rowley BR, Bull CO, Schneider C, Haegebarth A, Schatz CA, Fracasso PR, Wilkie DP, Hentemann M, Wilhelm SM, Scott WJ, Mumberg D, Ziegelbauer K. BAY 80-6946 is a highly selective intravenous PI3K inhibitor with potent p110α and p110δ activities in tumor cell lines and xenograft models. Mol Cancer Ther. 2013 Nov;12(11):2319-30.