NU 7026

Kinase assays

DNA-PK activity was measured at 30°C, in a final volume of 40 μl, in buffer containing 25 mm HEPES (pH 7.4), 12.5 mm MgCl2, 50 mm KCl, 1 mm DTT, 10% v/v Glycerol, 0.1% w/v NP-40, and 1 mg of the substrate GST-p53N66 in polypropylene 96-well plates. Varying concentrations of inhibitor were added. After 10 min of incubation, ATP was added to give a final concentration of 50 μm, along with a 30-mer double-stranded DNA oligonucleotide to initiate the reaction. After 1 h with shaking, 150 μl of PBS were added to the reaction, and 5 μl were then transferred to a 96-well opaque white plate containing 45 μl of PBS per well, where the GSTp53N66 substrate was allowed to bind to the wells for 1 h. To detect the phosphorylation event on the serine 15 residue of p53 elicited by DNA-PK, a p53 phosphoserine-15 antibody was used in a basic ELISA procedure.

Cell lines

CHO cell lines V3 transfected with human DNA-PKcs gene

Preparation method

The solubility of this compound in DMSO is <2.81mg/mL. General tips for obtaining a higher concentration: Please warm the tube at 37℃ for 10 minutes and/or shake it in the ultrasonic bath for a while. Stock solution can be stored below -20℃ for several months.

Reacting condition

10 μM

Applications

NU7026 potentiated ionizing radiation cytotoxicity in exponentially growing DNA-PK proficient cells, which can act as a potent radiosensitizer and shows potential as tools for anticancer therapeutic intervention.

Animal models

Female BALB/c mice

Dosage form

four times per day at 100 mg/kg, i.p.

Application

NU7026 shows a significant radiosensitisation effect in BALB/c mice.

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] Veuger, S.J., et al., Radiosensitization and DNA repair inhibition by the combined use of novel inhibitors of DNA-dependent protein kinase and poly(ADP-ribose) polymerase-1. Cancer Res, 2003. 63(18): p. 6008-15.

[2] Nutley, B.P., et al., Preclinical pharmacokinetics and metabolism of a novel prototype DNA-PK inhibitor NU7026. Br J Cancer, 2005. 93(9): p. 1011-8.