| 名稱 | PD0166285 |
| 描述 | PD0166285 is a potent Wee1 and Chk1 inhibitor with activity at nanomolar concentrations.PD0166285 is a novel G2 checkpoint abrogator. |
| 細胞實驗 | 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) |
| 激酶實驗 | 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. |
| 動物實驗 | 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通過納摩爾濃度抑制Wee1活性，使G2/M檢查點失效，誘導提前細胞分裂。在細胞水平上，0.5 μM PD0166285顯著抑制七種測試癌細胞系中的輻射誘導的Cdc2在Tyr-15和Thr-14的磷酸化。PD0166285通過廢除G2檢查點來殺死癌細胞。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，能夠抑制所有測試細胞系中的Cdc2Y15/T14磷酸化，這一效果與細胞的p53狀態(tài)無關(guān)[1]。此外，通過藥理學上靶向WEE1的PD0166285能夠增強U251-FM GBM腫瘤對體內(nèi)IR的敏感性[3]。 |
| 存儲條件 | Powder: -20°C for 3 years | In solvent: -80°C for 1 year | Shipping with blue ice. |
| 溶解度 | Ethanol : 93 mg/mL (181.49 mM), Sonication is recommended.
DMSO : 93 mg/mL (181.49 mM), Sonication is recommended.
H2O : < 1 mg/mL (insoluble or slightly soluble)
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| 關(guān)鍵字 | Wee1 | PD166285 | PD-0166285 | PD0166285 | PD 166285 | PD 0166285 | Myt1 | Inhibitor | inhibit | Chk1 | Apoptosis |
| 相關(guān)產(chǎn)品 | L-Glutamic acid | Cysteamine hydrochloride | Alginic acid | Flubendazole | 5-Fluorouracil | Stavudine | Dextran sulfate sodium salt (MW 4500-5500) | Tributyrin | Myricetin | L-Ascorbic acid sodium salt | L-Ascorbic acid | Sodium 4-phenylbutyrate |
| 相關(guān)庫 | 抑制劑庫 | 經(jīng)典已知活性庫 | 抗癌化合物庫 | 已知活性化合物庫 | 細胞周期化合物庫 | 激酶抑制劑庫 | 細胞凋亡化合物庫 | 抗衰老化合物庫 | NO PAINS 化合物庫 | 表型篩選靶點鑒定庫 |