Olaparib, Inhibitor

Olaparib (AZD2281, Ku-0059436)

Cat. Num. AAA575811

• Purity: 99.88%
• Synonyms: Olaparib; Olaparib (AZD2281; Ku-0059436); inhibitor

• Purity/Purification: 99.88%
• Form/Format: POWDER

• CAS: 763113-22-0
• Formula: C₂₄H₂₃FN₄O₃
• Solubility: DMSO: 80 mg/ml (184.1 mM); Ethanol: <1mg/ml; Water: <1 mg/ml; (< 1 mg/ml refers to the product slightly soluble or insoluble)
• Receptor (IC50): PARP1: 5 nM (cell free); PARP2: 1 nM (cell free)
• Smiles: Fc1ccc(Cc2n[nH]c(=O)c3ccccc23)cc1C(=O)N1CCN(CC1)C(=O)C1CC1
• Analytical Data: *NMR: 1H NMR spectrum of sample consistents with structure of the compound.; * HPLC: sample tested with high performance liquid chromatography, purity 100.00%.
• In vitro Activity: Following the establishment of a growth-inhibition curve for MMS on SW620 cells, the addition of the PARP-1 inhibitor at nM concentrations dose-dependently increased the effectiveness of MMS. Olaparib was applied to SW620 cell lysates at a similar concentration range and identified the IC50 for PARP-1 inhibition to be around 6 nM and the total ablation of PARP-1 activity to be at concentrations of 30-100 nM [1]. Concentrations of cisplatin capable of suppressing cell survival by 90% in the presence of 0, 1 or 3 nM AZD2281 were approximately 2.1, 1.5, and 1.2 nM in HSC-2 cells, 1.7, 1.4, and 0.98 nM in Ca9-22 cells, and 2.4, 1.5 and 1.3 nM in SAS cells, respectively. The combination of 1 nM cisplatin and 1 nM AZD2281 attenuated growth rates compared with single treatments of either cisplatin or AZD2281 [2]. The largest difference in drug sensitivity between BRCA2-deficient KB2P cells (IC50: 91 nmol/L) and BRCA2-proficient KP cells (IC50: 8,135 nmol/L) was observed for AZD2281 [3].
• Kinase Assay: This assay determined the ability of test compounds to inhibit PARP-1 enzyme activity. The method that was used was as reported. We measured PARP-2 activity inhibition by using a variation of the PARP-1 assay in which PARP-2 protein (recombinant) was bound down by a PARP-2 specific antibody in a 96-well white-walled plate. PARP-2 activity was measured following 3H-NAD+ DNA additions. After washing, scintillant was added to measure 3 H-incorporated ribosylations. For tankyrase-1, an AlphaScreen assay was developed in which HIS-tagged recombinant TANK-1 protein was incubated with biotinylated NAD+ in a 384-well ProxiPlate assay. Alpha beads were added to bind the HIS and biotin tags to create a proximity signal, whereas the inhibition of TANK-1 activity was directly proportional to the loss of this signal. All experiments were repeated at least three times [1].
• Cell Assay: HSC-2, Ca9-22, and SAS oral carcinoma cells were seeded in 24-well plates at a density of 2 × 104 cells/well. After overnightincubation, the culture medium was replaced with fresh medium containing various concentrations of PARP inhibitor AZD228 or cisplatin. After 24 h of treatment, the number of viable cells was assessed using an MTT assay as reported previously. Briefly, one-tenth of the fluid volume of 5 mg/mL MTT in RPMI-1640 medium was added to each well, followed by incubation for 4 h at 37 °C. After incubation, the medium was carefully removed and an adequate volume of 0.1 N HCl in isopropanol was added to each well and the resultant formazan crystals was dissolved. Absorbance was determined at 570 nm by microplate reader in 96-well assay plates. All experiments were performed in triplicate [2].; Cell line: Breast cancer cell lines including SW620 colon, MBS127101 ovarian.
• Preparation and Storage: 2 years -80°C in solvent; 3 years -20°C powder.

Structure

Related Product Information for Olaparib, inhibitor

Olaparib is a small molecule inhibitor of PARP1/PARP2 (IC50: 5/1 nM) but is less effective against the PARP tankyrase-1 (IC50: 1.5 nM).