AZD0156 , an effective and specific inhibitors of ATM kinase, is potential antineoplastic activities and chemo-/radio-sensitizing.

ATM:0.58 nM.

AZD0156 effectively blocks the kinase activity of ATM and its associated signaling pathways, inhibiting the activation of DNA damage checkpoints. This disruption in DNA damage repair promotes apoptosis in tumor cells and results in cell death, particularly in those with an overexpression of ATM[1].

AZD0156 is dissolved in 100% DMSO. HT29 cells are seeded into 384 well assay plates at a density of 6000 cells/well in 40 μL EMEM medium containing 1% L glutamine and 10% FBS and allowed to adhere overnight. The following morning compound of Formula (I) in 100% DMSO is added to assay plates by acoustic dispensing. After lh incubation at 37°C and 5% C02,?40 nL of 3 mM 4NQO in 100% DMSO is added to all wells by acoustic dispensing, except minimum control wells which are left untreated with 4NQO to generate a null response control. Plates are returned to the incubator for a further lh. Then cells are fixed by adding 20 μL of 3.7% formaldehyde in PBS solution and incubating for 20 mins at r.t. Then 20 μL of 0.1% Triton XI 00 in PBS is added and incubated for 10 minutes at r.t., to permeabalise cells. Then the plates are washed once with 50 μL/well PBS, using a Biotek EL405 plate washer.