| 名稱 | Berzosertib |
| 描述 | Berzosertib (VE-822) is an ATR inhibitor (Ki<0.2 nM) that also inhibits ATM (Ki=34 nM). Berzosertib has antitumor activity and has been used in trials investigating the treatment of ovarian tumors, plasmacytoid tumors of the ovary, solid tumors in adults, advanced solid tumors, and advanced solid tumors. |
| 細(xì)胞實(shí)驗(yàn) | VE-822 is dissolved in DMSO and stored, and then diluted with appropriate media before use[1]. Gemcitabine (10 nM) is added 24 h pre-XRT and is replaced with fresh medium before addition of VE-822. PSN-1 cells are treated with VE-822 (80 nM) for 1 h before, through to 18 h after, XRT (6 Gy). Apoptosis is analyzed 48 h after XRT by flow cytometry using an Annexin V-FITC kit with PI[1]. |
| 激酶實(shí)驗(yàn) | A375 cells are pre-treated with 1 μM JNK-IN-8 for the indicated amounts of time. Remove the medium and wash 3 times with PBS. Resuspend the cell pellet with 1 mL Lysis Buffer (1% NP-40, 1% CHAPS, 25 mM Tris, 150 mM NaCl, Phosphatase Inhibitor Cocktail, and Protease Inhibitor Cocktail). Rotate end-to-end for 30 min at 4°C. Lysates are cleared by centrifugation at 14000 rpm for 15 min in the Eppendorf. The cleared lysates gel filtered into Kinase Buffer (0.1% NP-40, 20 mM HEPES, 150 mM NaCl, Phosphatase Inhibitor Cocktail, Protease Inhibitor Cocktail) using Bio-Rad 10DG colums. The total protein concentration of the gel-filtered lysate should be around 5-15 mg/mL. Cell lysate is labeled with the probe from ActivX at 5 μM for 1 hour. Samples are reduced with DTT, and cysteines are blocked with iodoacetamide and gel filtered to remove excess reagents and exchange the buffer. Add 1 volume of 2X Binding Buffer (2% Triton-100, 1% NP-40, 2 mM EDTA, 2X PBS) and 50 μL streptavidin bead slurry and rotate end-to-end for 2 hours, centrifuge at 7000 rpm for 2 min. Wash 3 times with 1X Binding Buffer and 3 times with PBS. Add 30 μL 1X sample buffer to beads, heat samples at 95°C for 10 min. Run samples on an SDS-PAGE gel at 110V. After transferred, the membrane is immunoblotted with JNK antibody[1]. |
| 體外活性 | 方法: 人胰腺癌細(xì)胞 PSN-1 和 MiaPaCa-2 用 Berzosertib (80 nmol/L) 和 gemcitabine (100 nM)、XRT (6?Gy) 處理，使用 Western Blot 檢測靶點(diǎn)蛋白表達(dá)水平。
結(jié)果: Berzosertib 減少了磷酸化 Ser345-Chk1。Berzosertib 在輻射反應(yīng)中沒有抑制 ATM、Chk2 或 DNA-PK 磷酸化，這進(jìn)一步支持了 Berzosertib 對 ATR 的選擇性。[1]
方法: 骨肉瘤細(xì)胞 MNNG/HOS 和 143B 用 Berzosertib (0-100 μM) 處理 48 h，使用 MTT assay 檢測細(xì)胞活力。
結(jié)果: Berzosterib 導(dǎo)致 MNNG/HOS 和 143B 細(xì)胞存活率呈劑量依賴性下降。[2] |
| 體內(nèi)活性 | 方法: 為檢測體內(nèi)抗腫瘤活性，用 Berzosertib (60 mg/kg，灌胃給藥，每天一次) 和 XRT (6?Gy，一次) 治療攜帶 PSN-1 異種移植物的 Nude 小鼠，給藥六天。
結(jié)果: 單獨(dú)使用 Berzosertib 對腫瘤生長沒有影響，但 XRT 加 Berzosertib 給藥 6 天或 4 天，使單獨(dú)使用 XRT 的 TV600 增加了一倍多。[1] |
| 存儲條件 | Keep away from direct sunlight
Powder: -20°C for 3 years | In solvent: -80°C for 1 year
Shipping with blue ice/Shipping at ambient temperature. |
| 溶解度 | DMSO : 28 mg/mL (60.4 mM), Sonication is recommended.
Ethanol : < 1 mg/mL (insoluble or slightly soluble)
10% DMSO+40% PEG300+5% Tween 80+45% Saline : 2 mg/mL (4.31 mM), Sonication is recommended.
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| 關(guān)鍵字 | VX970 | VX 970 | VE822 | VE 822 | Berzosertib | ATR | ATM |
| 相關(guān)產(chǎn)品 | Schisandrin B | AZD-7648 | ART0380 | VE-821 | Elimusertib | CP-466722 | Tuvusertib | GJ103 sodium salt | GJ071 oxalate | NU6027 | Camonsertib | (Z)-Mirin |
| 相關(guān)庫 | 抗乳腺癌化合物庫 | 經(jīng)典已知活性庫 | 抗癌活性化合物庫 | 已知活性化合物庫 | 激酶抑制劑庫 | 抗衰老化合物庫 | 高選擇性抑制劑庫 | 干細(xì)胞分化化合物庫 | 免疫/炎癥分子化合物庫 | 藥物功能重定位化合物庫 | 抗癌臨床化合物庫 | 抗癌藥物庫 |