Olaparib (AZD2281)

Administered via i.p. injection at 10 uL/g of body weight

Breast cancer cell lines including SW620 colon, A2780 ovarian, HCC1937, Hs578T, MDA-MB-231, MDA-MB-436, and T47D

Combining with cediranib, Olaparib is currently in Phase I/II study for treatment of recurrent papillary-serous ovarian, fallopian tube or peritoneal cancer or treatment of recurrent triple-negative breast cancer.

Olaparib (AZD2281, KU0059436) is a selective inhibitor of PARP1 and PARP2 with IC50 of 5 nM and 1 nM, respectively.

Olaparib is one of the first PARP inhibitors.

5 nM, 5 nM, 5 nM, 5 nM, 5 nM, 5 nM

Combining with temozolomide, Olaparib (10 mg/kg, p.o.) significantly suppresses tumor growth in SW620 xenografts. [1] Olaparib shows great response to Brca1-/-, p53-/- mammary tumors (50 mg/kg i.p. per day), while no responses to HR-deficient Ecad-/-, p53-/- mammary tumors. Olaparib even does not show dose-limiting toxicity in tumor-bearing mice. [3] Olaparib has been used to treat with BRCA mutated tumors, such as ovarian, breast and prostate cancers. Moreover, Olaparib shows selectively inhibition to ATM (Ataxia Telangiectasia Mutated)-deficient tumor cells, which indicates to be a potential agent for treating ATM mutant lymphoid tumors. [4]

FlashPlate assay (96-well screening assay), To columns 1 through 10, 1 uL of Olaparib (in DMSO) is added, and 1 uL DMSO only is added to the positive (POS) and negative (NEG) control wells (columns 11 and 12, respectively) of a pretreated FlashPlate. PARP-1 is diluted 1:40 in buffer (buffer B: 10% glycerol (v/v), 25 mM HEPES, 12.5 mM MgCl2, 50 mM KCl, 1 mM DTT, 0.01% NP-40 (v/v), pH 7.6) and 40 uL added to all 96 wells (final PARP-1 concentration in the assay is ca.1 ng/uL). The plate is sealed and shaken at RT for 15 min. Following this, 10 uL of positive reaction mix (0.2 ng/uL of double-stranded oligonucleotide [M3/M4] DNA per well, 5 uM of NAD+ final assay concentration, and 0.075 uCi 3H-NAD+ per well) is added to the appropriate wells (columns 1-11). The negative reaction mix, lacking the DNA oligonucleotide, is added to column 12 (with the mean negative control value used as the background). The plate is resealed and shaken for a further 60 min at RT to allow the reaction to continue. Then, 50 uL of ice-cold acetic acid (30%) is added to each well to stop the reaction, and the plate is sealed and shaken for a further 60 min at RT. Tritiated signal bound to the FlashPlate is then determined in counts per minute (CPM) using the TopCount plate reader.

434, 46

, , Dr David Schrmann from University of Basel, Olaparib (AZD2281)purchased from Selleck, in vivo suppression of PAR formation by the PARP inhibitor AZD2281 upon induction of DNA damage Primary human lung fibroblast cells (MRC-5) were pre-treated with the indicated concentration of the PARP inhibitor AZD2281 for two hours. Oxidative DNA damage was induced by 500 M H2O2 for 10 min and cellular PARP activity was measured by immuno-staining of poly(ADP)-ribose (PAR) (right panels). The in vivo effect of PARP inhibition was compared to cells without DNA damage induction and inhibitor (control) and H2O2-treated cells without inhibitor. Average nuclear PAR staining intensities of more than 50 cells were statistically analysed by Kruskal-Wallis and the post-hoc Dunns Multiple Comparison tests (left panel). Asterisks indicate highly significant (p1%) differences to H2O2-treated cells without PARP inhibitor. Thick horizontal bars mark medians and error bars the interquartile range.

DMSO 87 mg/mL, Water <1 mg/mL, Ethanol <1 mg/mL