| 名稱 | NU 7026 |
| 描述 | NU 7026 (DNA-PK Inhibitor II) is an effective DNA-PK inhibitor (IC50: 0.23 μM, in cell-free assays), 60-fold selective for DNA-PK than PI3K and no inhibition against both ATM and ATR. |
| 細胞實驗 | NU7026 is dissolved in DMSO and stored, and then diluted with appropriate media before use[2]. I83 cells are plated in RPMI 1640 medium with 10% FBS (1.5×105 cells/mL) and treated with vehicle (DMSO), 5 μM CLB, CLB IC50, 10 μM NU7026, or the combination of both drugs for 0, 6, 24, and 48 h. Cell cycle distribution, apoptosis, DNA-PK phosphorylation, and γH2AX determination are determined, and they are expressed as a percentage of cells in each phase of the cycle. DNA content is analyzed with a FACSCalibur flow cytometer equipped with CellQuest software[2]. |
| 激酶實驗 | Mammalian DNA-PK (500 ng/μL) is isolated from HeLa cell nuclear extract after chromatography using Q-Sepharose, S-Sepharose, and Heparin agarose. DNA-PK (250 ng) activity is measured at 30°C, in a final volume of 40 μL, in buffer containing 25 mM HEPES (pH 7.4), 12.5 mM MgCl2, 50 mM KCl, 1 mM DTT, 10% v/v Glycerol, 0.1% w/v NP-40, and 1 mg of the substrate GST-p53N66 (the NH2-terminal 66 amino acid residues of human wild-type p53 fused to glutathione S-transferase) in polypropylene 96-well plates. To the assay mix, varying concentrations of inhibitor (in DMSO at a final concentration of 1% v/v) are added. After 10 min of incubation, ATP is added to give a final concentration of 50 μM, along with a 30-mer double-stranded DNA oligonucleotide (final concentration of 0.5 ng/mL), to initiate the reaction. After 1 h with shaking, 150 μL of PBS are added to the reaction, and 5 μL are then transferred to a 96-well opaque white plate containing 45 μL of PBS per well, where the GSTp53N66 substrate is allowed to bind to the wells for 1 h. To detect the phosphorylation event on the serine 15 residue of p53 elicited by DNA-PK, a p53 phosphoserine-15 antibody is used in a basic ELISA procedure. An antirabbit horseradish peroxidase-conjugated secondary antibody is then used in the ELISA before the addition of chemiluminescence reagent to detect the signal as measured by chemiluminescent counting via a TopCount NXT[1]. |
| 體外活性 | NU7026（20 mg/kg,i.p.或p.o.）的生物利用度分別為20 和15%.在小鼠體內(nèi),NU7026（20 mg/kg, i.v.）具有快速的血漿清除率為0.108/h. |
| 體內(nèi)活性 | NU7026對V3YAC細胞系中的DNA DSB修復(fù)有抑制作用（56%）。在CLL細胞系(I83)和初級CLL-淋巴細胞中,NU7026（<10 μM）與苯丁酸氮芥具有協(xié)同的細胞毒活性。在K562細胞中,NU7026（10 μM）增強正定霉素,阿霉素,伊達比星,胺苯吖啶,依托泊苷和米托蒽醌抑制生長的作用,PF50值范圍大約為19(對mAMSA)到2(對伊達比星)。NU7026（10 μM）也會增強白血病細胞中依托泊苷的生長抑制作用（PF50：10.53）。NU7026（10 μM）增強K562細胞中依托泊苷誘導的細胞周期G2期阻滯。在CH1人卵巢癌細胞中,NU7026（10 μM）暴露4小時結(jié)合3 Gy輻射對明顯的電波敏化作用是必需的。NU7026（10 μM）增加I83細胞中苯丁酸氮芥誘導的G(2)/M期阻滯。在I83細胞中,NU7026（10 μM）增強整個細胞周期中苯丁酸氮芥誘導的γH2AX。NU7026（10 μM）增加I83細胞系中苯丁酸氮芥誘導的細胞凋亡。NU7026（55 μM）導致p53缺失的MEFs中明顯的端粒融合誘導,并導致p53和連接酶IV雙重缺失的MEFs中端粒融合更少。 |
| 存儲條件 | Powder: -20°C for 3 years | In solvent: -80°C for 1 year | Shipping with blue ice. |
| 溶解度 | DMSO : 2.8 mg/mL (10 mM)
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| 關(guān)鍵字 | DNA-dependent protein kinase | inhibit | Apoptosis | NU 7026 | DNA-PK | Inhibitor | LY-293646 | NU-7026 | LY 293646 |
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