Olaparib 是一种自噬和线粒体自噬激活剂，是 PARP1/PARP2 的小分子抑制剂，抑制PARP-1和PARP-2的IC50分别为 5 和 1 nM。

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 μM).

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 (AZD2281) 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 μM AZD2281 were approximately 2.1, 1.5, and 1.2 μM in HSC-2 cells, 1.7, 1.4, and 0.98 μM in Ca9-22 cells, and 2.4, 1.5 and 1.3 μM in SAS cells, respectively. The combination of 1 μM cisplatin and 1 μM 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].

Animals bearing SW620 xenografted tumors were treated with AZD2281 (10 mg/kg, po) in combination with TMZ (50 mg/kg, po) once daily for 5 consecutive days, after which the tumors were left to grow out. A considerable inhibition of tumor volumes as compared with that of the TMZ alone group was observed for the TMZ plus AZD2281 combinations [1]. Tumor volumes of control group mice increased during the experimental period. The tumor growth of cisplatin and AZD2281 (25 mg/kg/day, every three days for five treatments) groups significantly decreased compared to the control group, and that of combination group was further decreased [2]. Combination treatment of Calu-6 xenografts with olaparib and fractionated radiotherapy caused significant tumor regression relative to radiotherapy alone. Olaparib alone, when given as single or multiple daily doses, or in combination with fractionated radiotherapy, increased the perfusion of tumor blood vessels [4].

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].

HSC-2, Ca9-22, and SAS oral carcinoma cells were seeded in 24-well plates at a density of 2 × 104 cells/well. After overnight incubation, 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].

Once the tumor diameter had reached 7 mm, the mice were randomly assigned to the following groups: (a) control (200 μL saline); (b) cisplatin (2 mg/kg per body weight, dissolved in 200 μL sterilized water); (c) AZD2281 (25 mg/kg per body weight, dissolved in 200 μL sterilized water); or (d) combination (both cisplatin and AZD2281). The chemicals were administered intraperitoneally every three days, five times. Although AZD2281 is administered orally in the clinic, intraperitoneal injection was recommended by the manufacturer because of easier manipulation and the ethical constraints associated with oral gavage administration to mice. Tumor size and body weight were measured at the time of administration. The tumor volume was calculated using following equation. Tumor volume = verticality × width × height × 0.5236. Three days after the last administration, all surviving mice were sacrificed [2].

763113-22-0

C24H23FN4O3

434.471

奥拉帕尼;AZD2281;KU0059436

H2O:<1 mgml
Ethanol:<1 mgml
DMSO:80 mg/mL (184.1 mM)
Powder: -20°C for 3 years
In solvent: -80°C for 2 years