| 名稱 | Olaparib |
| 描述 | Olaparib (KU0059436) is a small molecule inhibitor of PARP1/PARP2 (IC50=5/1 nM), with weak inhibitory activity against PARP tankyrase-1 (IC50=1.5 μM), and is selective and orally active. Olaparib exhibits autophagy and mitochondrial autophagy activation activity. |
| 細(xì)胞實驗 | 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]. |
| 激酶實驗 | 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]. |
| 動物實驗 | 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]. |
| 體外活性 | 方法：人子宮頸癌細(xì)胞 SiHa 和 ME180 用 Olaparib (5-10 μM) 和 cisplatin (1-30 μM) 處理 72 h，使用 MTT 方法檢測細(xì)胞生長抑制情況。
結(jié)果：Olaparib 和 cisplatin 共同處理比用單一藥物處理的細(xì)胞顯示出顯著的細(xì)胞生長抑制作用。[1]
方法：人子宮內(nèi)膜癌細(xì)胞 HEC-6 和 HEC-6-PTEN 用 Olaparib (10 μM) 處理 72 h，使用 Flow Cytometry 方法分析細(xì)胞周期情況。
結(jié)果：Olaparib 誘導(dǎo) HEC-6 和 HEC-6-PTEN 細(xì)胞顯著增加了亞 G1 群體。[2]
方法：雞淋巴瘤細(xì)胞 DT40 用 Olaparib (0.01-10 μM) 處理 30 min，使用 Western Blot 方法檢測靶點蛋白表達(dá)水平。
結(jié)果：Olaparib 劑量依賴性抑制 PARylation 的表達(dá)水平，抑制 PARP 的活化。[3] |
| 體內(nèi)活性 | 方法：為檢測體內(nèi)抗腫瘤活性，將 Olaparib (10 mg/kg) 和 TMZ (50 mg/kg) 口服給藥給攜帶人結(jié)直腸癌腫瘤 SW620 的小鼠，每天一次，持續(xù)五天。
結(jié)果：與單獨(dú) TMZ 組相比，TMZ 加 Olaparib 組合治療組觀察到腫瘤體積的顯著抑制。[4]
方法：為研究 Olaparib 對哮喘的治療作用，將 Olaparib (1-10 mg/kg) 腹腔注射給基于 OVA 的哮喘 C57BL/6 小鼠模型，每天一次，持續(xù)三天。
結(jié)果：Olaparib 能顯著減少氣道嗜酸性粒細(xì)胞增多、粘液產(chǎn)生和高反應(yīng)性。Olaparib 的保護(hù)作用與抑制 Th2 細(xì)胞因子 eotaxin、IL-4、IL-5、IL-6、IL-13 和 M-CSF 以及卵清蛋白特異性 IgE 有關(guān)，同時增加 Th1 細(xì)胞因子 IFN-γ。Olaparib 有可能成為人類哮喘臨床試驗的候選藥物。[5] |
| 存儲條件 | Powder: -20°C for 3 years | In solvent: -80°C for 1 year | Shipping with blue ice. |
| 溶解度 | H2O : < 1 mg/mL (insoluble or slightly soluble)
DMSO : 50 mg/mL (115.09 mM)
Ethanol : < 1 mg/mL (insoluble or slightly soluble)
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| 關(guān)鍵字 | PARP | Mitochondrial Autophagy | inhibit | Autophagy | orally | Inhibitor | PARP1 | Mitophagy | poly ADP ribose polymerase | PARP2 | KU 0059436 | Olaparib | AZD-2281 | AZD 2281 | KU-0059436 |
| 相關(guān)產(chǎn)品 | Guanidine hydrochloride | Naringin | Taurine | Gefitinib | Hydroxychloroquine | Curcumin | Stavudine | Paeonol | Sodium 4-phenylbutyrate |
| 相關(guān)庫 | 抑制劑庫 | 抗癌上市藥物庫 | 抗癌活性化合物庫 | 已知活性化合物庫 | EMA 上市藥物庫 | 抗衰老化合物庫 | FDA 上市藥物庫 | 藥物功能重定位化合物庫 | 抗癌臨床化合物庫 | 抗癌藥物庫 |