| 名稱(chēng) | PD0166285 |
| 描述 | PD0166285 is a potent Wee1 and Chk1 inhibitor with activity at nanomolar concentrations.PD0166285 is a novel G2 checkpoint abrogator. |
| 細(xì)胞實(shí)驗(yàn) | B16 cells (1 × 105 cells in 100 mm-dishes) are maintained in medium overnight. In addition, the cells are treated with (0, 0.5, 1.0, or 2.0 μM) PD0166285 (including DMSO vehicle) for indicated times. The cells are washed twice with phosphate-buffered saline (PBS). Next, the cells are trypsinized, so the cell numbers in each dish are determined by using a computed cell-counter (Sysmex CDA-500) according to manufacturer's recommendation. (Only for Reference) |
| 激酶實(shí)驗(yàn) | Wee1 Mass Screening: Wee1 mass screening is performed using Amersham's p34cdc2 kinase SPA kit with some modifications. Briefly, 45–60 nM full-length Wee1 kinase is incubated with 25 μM compounds, 20 μM ATP, and 122–441 nM Cdc2/cyclin B in a final volume of 50 μl of enzyme dilution buffer [50 mM Tris (pH 8.0), 10 mM NaCl, 10 mM MgCl2, 1 mM DTT, and 0.1 mM Na3VO4]. After 30 min incubation at 30°C, 30 μl of [33P]ATP containing kinase buffer [67 mM Tris (pH 8.0), 40 mM NaCl, 13 mM MgCl2, 1 mM DTT, and 0.13 mM Na3VO4] containing 1 μM biotinylated peptide, and 0.25 μCi of [γ-33P]ATP is added to the reaction and incubated for another 30 min at 30°C. The reaction is stopped by adding 200 μl of stop buffer [50 μM ATP, 5 mM EDTA, 0.1% Triton X-100, and 1.25 mg/ml SPA beads in PBS]. After centrifugation at 2400 rpm for 15 min, the plate is counted with Wallac's Microbeta counter. |
| 動(dòng)物實(shí)驗(yàn) | E98 tumor fragments (8 mm3) were grafted subcutaneously in the flank of Balb/C nude mice (n = 10). For the experiments with the orthotopic tumors, U251 mg and E98 cells were transduced to express Fluc and mCherry, to generate U251-FM and E98-FM cells. One million cells were injected intracranially. Starting at 12 days after tumor inoculation, mice received the WEE1 inhibitor PD0166285 via intraperitoneal injections (500 μl of a 20 μM solution diluted in NaCl) or vehicle for 5 consecutive days. On days 10–15, mice were irradiated with a single dose of irradiation. |
| 體外活性 | PD0166285通過(guò)納摩爾濃度抑制Wee1活性，使G2/M檢查點(diǎn)失效，誘導(dǎo)提前細(xì)胞分裂。在細(xì)胞水平上，0.5 μM PD0166285顯著抑制七種測(cè)試癌細(xì)胞系中的輻射誘導(dǎo)的Cdc2在Tyr-15和Thr-14的磷酸化。PD0166285通過(guò)廢除G2檢查點(diǎn)來(lái)殺死癌細(xì)胞。PD0166285不抑制Cdc2/cyclin B，而是以更高濃度（3433 nM）抑制Chk1激酶。用抑制劑治療細(xì)胞與微管穩(wěn)定和cyclin D轉(zhuǎn)錄減少有關(guān)。因此，PD0166285可能是一種有潛力的抗癌療法[1][2]。 |
| 體內(nèi)活性 | PD0166285在0.5 μM濃度下，能夠抑制所有測(cè)試細(xì)胞系中的Cdc2Y15/T14磷酸化，這一效果與細(xì)胞的p53狀態(tài)無(wú)關(guān)[1]。此外，通過(guò)藥理學(xué)上靶向WEE1的PD0166285能夠增強(qiáng)U251-FM GBM腫瘤對(duì)體內(nèi)IR的敏感性[3]。 |
| 存儲(chǔ)條件 | Powder: -20°C for 3 years | In solvent: -80°C for 1 year | Shipping with blue ice. |
| 溶解度 | Ethanol : 93 mg/mL (181.5 mM)
DMSO : 93 mg/mL (181.5 mM)
H2O : < 1 mg/mL (insoluble or slightly soluble)
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| 關(guān)鍵字 | PD 166285 | Wee1 | Inhibitor | inhibit | PD0166285 | Apoptosis | PD166285 | PD-0166285 | PD 0166285 |
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| 相關(guān)庫(kù) | 抑制劑庫(kù) | 經(jīng)典已知活性庫(kù) | 抗癌化合物庫(kù) | 已知活性化合物庫(kù) | 細(xì)胞周期化合物庫(kù) | 激酶抑制劑庫(kù) | 細(xì)胞凋亡化合物庫(kù) | 抗衰老化合物庫(kù) | NO PAINS 化合物庫(kù) | 表型篩選靶點(diǎn)鑒定庫(kù) |