Olaparib (AZD2281, Ku-0059436)

Catalog No.S1060

Olaparib (AZD2281, KU0059436) is a selective inhibitor of PARP1/2 with IC50 of 5 nM/1 nM in cell-free assays, 300-times less effective against tankyrase-1. Phase 3.

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Olaparib (AZD2281, Ku-0059436) Chemical Structure

Olaparib (AZD2281, Ku-0059436) Chemical Structure
Molecular Weight: 434.46

Validation & Quality Control

Cited by 73 publications:

13 customer reviews :

Quality Control & MSDS

Related Compound Libraries

Olaparib (AZD2281, Ku-0059436) is available in the following compound libraries:

PARP Inhibitors with Unique Features

  • Selective PARP Inhibitors

    AG-14361 PARP1-selective, Ki<5 nM. UPF 1069 PARP2-selective, IC50=0.3 μM.

  • Most Potent PARP Inhibitor

    MK-4827 (Niraparib) PARP1, IC50=3.8 nM; PARP2, IC50=2.1 nM.

  • PARP Inhibitor in Clinical Trial

    Iniparib (BSI-201) Phase III for solid tumors.

  • Newest PARP Inhibitor

    BMN 673 Novel PARP inhibitor with IC50 of 0.58 nM. Also a potent inhibitor of PARP-2, but does not inhibit PARG and highly sensitive to PTEN mutation. Novel PARP inhibitor with IC50

Product Information

  • Compare PARP Inhibitors
    Compare PARP Products
  • Research Area
  • Inhibition Profile
  • Olaparib (AZD2281, Ku-0059436) Mechanism

Product Description

Biological Activity

Description Olaparib (AZD2281, KU0059436) is a selective inhibitor of PARP1/2 with IC50 of 5 nM/1 nM in cell-free assays, 300-times less effective against tankyrase-1. Phase 3.
Targets PARP2 [1]
(Cell-free assay)
PARP1 [1]
(Cell-free assay)
Tankyrase-1 [1]
(Cell-free assay)
IC50 1 nM 5 nM 1.5 μM
In vitro Olaparib would act against BRCA1 or BRCA2 mutations. Olaparib is not sensitive to tankyrase-1 (IC50 >1 μM). Olaparib could ablate the PARP-1 activity at concentrations of 30-100 nM in SW620 cells. Olaparib is hypersensitive to BRCA1-deficient cell lines (MDA-MB-463 and HCC1937), compared with BRCA1- and BRCA2-proficient cell lines (Hs578T, MDA-MB-231, and T47D). [1] Olaparib is strongly sensitive to KB2P cells due to suppression of base excision repair by PARP inhibition, which may result in the conversion of single-strand breaks to double-strand breaks during DNA replication, thus activating BRCA2-dependent recombination pathways. [2]
Cell Data
Cell LinesAssay TypeConcentrationIncubation TimeFormulationActivity DescriptionPMID
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KP6.3NXnX[IZuT3Kxd4ToJGlvcGmkaYTpc44hSXO|YYm=M4nkT|Qh\A>?MXfJR|UxRTFyLkSyPEDPxE1iMWKxPFU2QTZzMx?=
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Mouse H2AX−/− ES CellsMUnDfZRwfG:6aXOgRZN{[Xl?NWXSVYxVOi53IN88US=>NETSb5EzOCCqMoDaV4lodmmoaXPhcpRtgSCrbnjpZol1eyClZXzsJJN2en[rdnHsMl60NlM{PTV2OEm=
Mouse ATM−/− ES CellsNY[3cmhtS3m2b4TvfIlkKEG|c3H5NHz1OWMzNjVizszNNUHFcmVOOjBiaB?=NF;lRnBUcWewaX\pZ4FvfGy7IHnubIljcXS|IHPlcIwhe3W{dnn2ZYw>M1zzU|I{OzV3NEi5
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PC-9PTEN−NYPHfWR1T3Kxd4ToJGlvcGmkaYTpc44hSXO|YYm=M3[4clIxKM7:TR?=NITGZoEyPDRiaB?=M3fjfmlEPTB;Nj61NkDPxE1?NFLFZWszOzJ|OUiwPS=>
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BT20M{TNRmdzd3e2aDDJcohq[mm2aX;uJGF{e2G7MU[1JIRigQ>?NVHQVodbUUN3ME23Mlch|ryPM1S5UlI{PzZyNEm2
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... Click to View More Cell Line Experimental Data

In vivo 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]
Features A potent PARP inhibitor (currently in late stage clinical trials).

Protocol(Only for Reference)

Kinase Assay:

[1]

FlashPlate assay (96-well screening assay) To columns 1 through 10, 1 μL of Olaparib (in DMSO) is added, and 1 μL 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 μL added to all 96 wells (final PARP-1 concentration in the assay is ~1 ng/μL). The plate is sealed and shaken at RT for 15 min. Following this, 10 μL of positive reaction mix (0.2 ng/μL of double-stranded oligonucleotide [M3/M4] DNA per well, 5 μM of NAD+ final assay concentration, and 0.075 μCi 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 μL 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.
In vitro isolated enzyme assay PARP-2 activity inhibition uses a variation of the PARP-1 assay in which PARP-2 protein (recombinant) is bound down by a PARP-2 specific antibody in a 96-well white-walled plate. PARP-2 activity is measured following 3H-NAD+ DNA additions. After washing, scintillant is added to measure 3H-incorporated ribosylations.

Cell Assay:

[1]

Cell lines Breast cancer cell lines including SW620 colon, A2780 ovarian, HCC1937, Hs578T, MDA-MB-231, MDA-MB-436, and T47D
Concentrations 1-300 nM
Incubation Time 7-14 days
Method

The cytotoxicity of Olaparib is measured by clonogenic assay. Olaparib is dissolved in DMSO and diluted by culture media before use. The cells are seeded in six well plates and left to attach overnight. Then Olaparib is added at various concentrations and the cells are incubated for 7-14 days. After that the surviving colonies are counted for calculating the IC50.

Animal Study:

[3]

Animal Models Brca1-/-;p53-/- mammary tumors are generated in K14cre;Brca1F/F;p53F/F mice.
Formulation 50 mg/mL stocks in DMSO with 10% 2-hydroxyl-propyl-β-cyclodextrine/PBS
Dosages 50 mg/kg
Administration Administered via i.p. injection at 10 μL/g of body weight

Conversion of different model animals based on BSA (Value based on data from FDA Draft Guidelines)

SpeciesMouseRatRabbitGuinea pigHamsterDog
Weight (kg)0.020.151.80.40.0810
Body Surface Area (m2)0.0070.0250.150.050.020.5
Km factor36128520
Animal A (mg/kg) = Animal B (mg/kg) multiplied by  Animal B Km
Animal A Km

For example, to modify the dose of resveratrol used for a mouse (22.4 mg/kg) to a dose based on the BSA for a rat, multiply 22.4 mg/kg by the Km factor for a mouse and then divide by the Km factor for a rat. This calculation results in a rat equivalent dose for resveratrol of 11.2 mg/kg.

Rat dose (mg/kg) = mouse dose (22.4 mg/kg) ×  mouse Km(3)  = 11.2 mg/kg
rat Km(6)
1

References

[1] Menear KA, et al. J Med Chem, 2008, 51(20), 6581-6591.

[2] Evers B, et al, Clin Cancer Res, 2008, 14(12), 3916-3925.

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Clinical Trial Information( data from http://clinicaltrials.gov, updated on 2016-07-23)

NCT Number Recruitment Conditions Sponsor
/Collaborators
Start Date Phases
NCT02822157 Not yet recruiting Ovarian Epithelial Cancer Universitaire Ziekenhuizen Leuven|AstraZeneca August 2016 Phase 2
NCT02810743 Not yet recruiting Breast Cancer The Netherlands Cancer Institute August 2016 Phase 3
NCT02489006 Not yet recruiting Ovarian Cancer|Fallopian Tube Cancer|Neoadjuvant Treatment|Debulking Surgical Procedures University Health Network, Toronto July 2016 Phase 2
NCT02392676 Withdrawn Platinum Sensitive Relapsed Ovarian Cancer AstraZeneca|Myriad Genetic Laboratories, Inc. July 2016 Phase 3
NCT02546661 Recruiting Muscle Invasive Bladder Cancer AstraZeneca June 2016 Phase 1

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Chemical Information

Download Olaparib (AZD2281, Ku-0059436) SDF
Molecular Weight (MW) 434.46
Formula

C24H23FN4O3

CAS No. 763113-22-0
Storage 3 years -20℃powder
6 months-80℃in solvent
Synonyms N/A
Solubility (25°C) * In vitro DMSO 86 mg/mL warming (197.94 mM)
Water 0.002 mg/mL (0.0 mM)
Ethanol <1 mg/mL (<1 mM)
In vivo 4% DMSO+30% PEG 300+ddH2O 5mg/mL
* <1 mg/ml means slightly soluble or insoluble.
* Please note that Selleck tests the solubility of all compounds in-house, and the actual solubility may differ slightly from published values. This is normal and is due to slight batch-to-batch variations.
Chemical Name 4-(3-(1-(cyclopropanecarbonyl)piperazine-4-carbonyl)-4-fluorobenzyl)phthalazin-1(2H)-one

Customer Product Validation(13)


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Rating
Source Hepatology 2012 55, 1840-1851. Olaparib (AZD2281, Ku-0059436) purchased from Selleck
Method Western Blot
Cell Lines SNU398/SNU449 cells
Concentrations 3 uM
Incubation Time 48 h
Results The results showed that coadministration of SAHA and olaparib resulted in a synergistic increase in the levels of the cleaved caspase-3 and cleaved PARP in SNU-398 cells as compared with SAHA or olaparib treatment alone. In contrast, the same effect in SNU-449 cells was only modest.

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Rating
Source Hepatology 2012 55, 1840-1851. Olaparib (AZD2281, Ku-0059436) purchased from Selleck
Method Western Blot
Cell Lines SNU398/SNU449 cells
Concentrations 3 uM
Incubation Time 48 h
Results Cotreatment with SAHA and olaparib synergistically down-regulated the protein levels of checkpoint protein 1 (Chk1), Chk2, and fanconi anemia group D2 protein (FANCD2) in SNU-398 but not SNU-449 Cells.

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Rating
Source Hepatology 2012 55, 1840-1851. Olaparib (AZD2281, Ku-0059436) purchased from Selleck
Method Apoptosis assay
Cell Lines SNU398/SNU449 cells
Concentrations 3 uM
Incubation Time 48 h
Results Results showed that coadministration of SAHA and olaparib significantly increased apoptosis (12.8%) in SNU-398 cells compared with treatment with individual agent alone, whereas only a minor effect was observed in SNU-449 cells.

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Source Cancer Res 2014 74(21), 5948-54. Olaparib (AZD2281, Ku-0059436) purchased from Selleck
Method Western Blotting
Cell Lines A549 cells
Concentrations 1 uM
Incubation Time 2 h
Results To determine if the PTX/GMX synergy is mediated through PARP, cells were treated with PTX and GMX in the presence or absence of olaparib, a small molecule PARP inhibitor or following siRNA knockdown of PARP-1. Treatment with olaparib was sufficient to inhibit PTXinduced PAR and NAD+ depletion. It was also sufficient to partially restore NAD+ levels in the PTX/GMX treated cells above the threshold level such that viability was also restored from 22% to 68%. SiRNA knockdown of PARP-1 completely inhibited PTX-mediated PAR and partially restored viability to the PTX/GMX treated cells, demonstrating that the synergy is mediated in part through PARP-1. Of note, by eliminating the PARP substrate, NAD+, GMX functionally mimics a PARP inhibitor and prevents PTX-induced PAR (no PAR occurs following GMX treatment alone as expected. GMX does not prevent the induction of double strand DNA breaks (DSB) by PTX as measured by γ-H2AX levels, and thus does not interfere with the DNA damaging aspects of PTX cytotoxicity or the cell’s demand for NAD+.

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Source EMBO Mol Med 2012 4, 1087-1096. Olaparib (AZD2281, Ku-0059436) purchased from Selleck
Method Western Blot
Cell Lines CAL51 cells
Concentrations 0-10 uM
Incubation Time 48 h
Results As expected, H2AX phosphorylation could be detected by Western blotting when cells were exposed to 10 uM olaparib (Fig 5C). Whilst FK866 did not in itself induce H2AX phosphorylation, combining FK866 with olaparib caused detectable gH2AX as measured by Western blotting at concentrations of olaparib as low as 0.01 uM (Fig C).

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Rating
Source J Biol Chem 2011 286, 12157-12165. Olaparib (AZD2281, Ku-0059436) purchased from Selleck
Method Clonogenic Survival Assay
Cell Lines HCT116+3+5 cells, HT29 cells
Concentrations 2 µM
Incubation Time
Results As shown in Fig. A, MSH3-deficient G5 cells were more sensitive to olaparib than the MSH3-restored G5 cell line. These data clearly support the role of MSH3 in DSB repairs in CRC cells. Moreover, the combination of oxaliplatin and olaparib exhibited a synergistic effect in cytotoxicity in the MSH3-deficient G5 cells compared with the parental HCT116+3+5 cells. We also confirmed this effect in two other colon cancer cell lines HT29 (Fig. B) and SW480 (data not shown) in a transient knockdown system using MSH3 siRNA.

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Source J Exp Clin Cancer Res 2013 32(1), 95. Olaparib (AZD2281, Ku-0059436) purchased from Selleck
Method flow cytometry
Cell Lines MCF7-ATMi and MCF7 cells
Concentrations 0-5 uM
Incubation Time 48 h
Results MCF7-ATMi cells are more sensitive than MCF7-ctr cells to olaparib.

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Rating
Source Nucl Med Commun 2011 32, 1046-51. Olaparib (AZD2281, Ku-0059436) purchased from Selleck
Method Cell growth assay
Cell Lines human Burkitt lymphoma cells
Concentrations 500 nmol/L
Incubation Time 0-5 d
Results A volume of 500 nmol/l ABT-888 and AZD-2281 showed a moderate intrinsc effect on cell proliferation on days 3–5 (Fig. a). ABT-888 and AZD-2281 showed a significant ( P < 0.05) reduction in lymphoma cell growth on days 2–5 (Fig. b–d).

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Source Nucl Med Commun 2011 32, 1046-51. Olaparib (AZD2281, Ku-0059436) purchased from Selleck
Method PARP activity assay
Cell Lines Raji lymphocyte tumor cells
Concentrations 500 nmol/L
Incubation Time 24 h
Results

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Rating
Source Nucl Med Commun 2011 32, 1046–1051 . Olaparib (AZD2281, Ku-0059436) purchased from Selleck
Method Laser confocal microscopy/fluorescent H2AX Antibody Staining
Cell Lines Epstein–Barr virus-infected Raji lymphocyte tumor cells
Concentrations 500 nM
Incubation Time 2 h
Results The Fluor-647 anti-H2A.X-phosphorylated (Ser139) antibody stained significantly more double-stranded breaks in cells treated with ABT-888 and AZD-2281 compared with controls at 8 and 12 Gy (P<0.05).

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Rating
Source Medicine (Baltimore) 2014 93(28), e294. Olaparib (AZD2281, Ku-0059436) purchased from Selleck
Method
Cell Lines SW480, SW480-XRCC2/RNAi cells
Concentrations 1, 10, 50 uM
Incubation Time 2 h
Results 96-hour treatment with 1 mM olaparib, SW480-XRCC2 cell viability was greater than that of SW480 cells, as was that of SW480 cells compared to SW480-XRCC2/RNAi cells (A; P =0.011; P = 0.001, respectively). Surprisingly, following treatment with 10 mM olaparib, the SW480-XRCC2 cell viability was lowest among the three cell lines (B; P ?0.024). However, cell viability began to decline after 72 hours following treatment with 50 mM olaparib (C). These results suggest that CRC cell sensitivity to olaparib increases progressively with the increased XRCC2 expression when the olaparib concentration is within a certain range.

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Rating
Source 2010 Dr. David Schrmann from University of Base. Olaparib (AZD2281, Ku-0059436) purchased from Selleck
Method Kruskal-Wallis and the post-hoc Dunn’s Multiple Comparison tests, immuno-staining
Cell Lines Primary human lung fibroblast cells (MRC-5)
Concentrations 0-100 nM
Incubation Time 2 h
Results 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 µM 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 Dunn’s Multiple Comparison tests (left panel). Asterisks indicate highly significant (p<1%) differences to H2O2-treated cells without PARP inhibitor. Thick horizontal bars mark medians and error bars the interquartile range.

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Rating
Source 2010 Dr. Xiangbing Meng of University of Iowa. Olaparib (AZD2281, Ku-0059436) purchased from Selleck
Method WST-1 method
Cell Lines Hec50/Ishikawa/SKOV3/Caov3/PA-1 cell lines
Concentrations 0-12000 nM
Incubation Time 3 d
Results Effect of AZD 2281 on the viability of endometrial cancer cell line Hec50 and Ishikawa and ovarian cancer cell line SKOV3,Caov3 and PA-1 was detected by WST-1 method after 3 days treatment.

Tech Support

Answers to questions you may have can be found in the inhibitor handling instructions. Topics include how to prepare stock solutions, how to store inhibitors, and issues that need special attention for cell-based assays and animal experiments.

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