AZD2281

Biological Activity

AZD 2281 (Olaparib, KU-0059436) is a  novel, potent orally active PARP (poly ADP-ribose polymerase) inhibitorwith IC50 of 5 and 1 nM for PARP-1and PARP-2  respectively. AZD 2281 (Olaparib, KU-0059436) acts as an inhibitor of the enzyme Poly ADP ribose polymerase (PARP) and is one of the first PARP inhibitors.It induces cancer specific synthetic lethality in homologous recombination repair defective cells, including BRCA-deficient tumors.

Customer Product Validations & Biological Datas

	Source	Int J Oncol (2015). Figure 4. AZD2281
	Method	apoptosis assay
	Cell Lines	HCC‑1428 cells
	Concentrations	2 μM
	Incubation Time	4 day
	Results	Inhibition of autophagy by knocking down ATG5 also partially inhibited AZD2281-induced apoptosis, suggesting that autophagy contributes to AZD2281-induced cell death in BRCA mutated breast cancer cells.

	Source	Int J Oncol (2015). Figure 1. AZD2281
	Method	Transmission electron microscopy
	Cell Lines	BRCA1 or BRCA2 mutant breast cancer cell lines
	Concentrations	2 μM
	Incubation Time	4 day
	Results	The growth inhibition effect of AZD2281 was significantly higher in the BRCA wild-type cell lines with BRCA1 allelic loss than in the BRCA wild-type cell line without BRCA1 allelic loss.

Protocol

Cell Experiment
Cell lines	SW620 colorectal cell line
Preparation method	Potentiation of MMS Cytotoxicity by 47 Determined by the Use of Sulforhodamine B Cell Growth Assays. SW620 cells were seeded in 96-well plates and were left to attach overnight. Cells were preincubated with vehicle control (DMSO) or with a single concentration of KU-0059436 (1, 3, 10, 30, 100 or 300 nM) for 1 h before the addition of increasing concentrations of MMS. Cells were incubated in the presence of each drug combination for 4 days before cell growth was quantified by the use of an SRB assay.44 Data were calculated from triplicate wells as the mean percentage of cell growth relative to KU-0059436-only wells, and (SE and IC50 were calculated by the use of XL-FIT 4 software. SW620 cells showed <24% growth inhibition (>76% cell growth) when only KU-0059436 was used at concentrations below 300 nM (data not shown).
Concentrations	1, 3, 10, 100 and 300 nM
Incubation time	4 days

Animal Experiment
Animal models	mouse bearing SW620 xenografted tumors
Formulation	methylcellulose/PBS
Dosages	10 mg/kg once daily for 5 consecutive days
Administration	orally

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

Species	Mouse	Rat	Rabbit	Guinea pig	Hamster	Dog
Weight (kg)	0.02	0.15	1.8	0.4	0.08	10
Body Surface Area (m2)	0.007	0.025	0.15	0.05	0.02	0.5
Km factor	3	6	12	8	5	20

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 K_m factor for a mouse and then divide by the K_m factor for a rat. This calculation results in a rat equivalent dose for resveratrol of 11.2 mg/kg.

Chemical Information

References

Synthetic lethality of PARP and NAMPT inhibition in triple-negative breast cancer cells.
Bajrami et al. EMBO Mol Med. 2012 Aug 30. PMID: 22933245.

Clinical trials and future potential of targeted therapy for ovarian cancer.
Itamochi et al. Int J Clin Oncol. 2012 Aug 28. PMID: 22926640.

PI3K inhibition impairs BRCA1/2 expression and sensitizes BRCA proficient triple negative breast cancer to PARP inhibition.
Ibrahim et al. Cancer Discov. 2012 Aug 24. PMID: 22915752.

Cross-platform pathway-based analysis identifies markers of response to the PARP inhibitor olaparib.
Daemen et al. Breast Cancer Res Treat. 2012 Sep;135(2):505-17. PMID: 22875744.

Olaparib , PARP1 inhibitor in ovarian cancer.
Marchetti et al. Expert Opin Investig Drugs. 2012 Jul 13. PMID: 22788971.