| 名稱 | Tazemetostat |
| 描述 | Tazemetostat (EPZ6438) is a histone methyltransferase EZH2 inhibitor (IC50=11 nM) that is orally active, selective, and SAM-competitive. Tazemetostat exhibits antitumor activity and may be used for the treatment of epithelioid sarcoma/follicular lymphoma. |
| 細(xì)胞實(shí)驗(yàn) | For the adherent cell line proliferation assays, plating densities for each cell line are determined based on growth curves (measured by ATP content) and density over a 7-d time course. On the day before compound treatment, cells are plated in either 96-well plates in triplicate (for the day 0–7 time course) or 6-well plates (for replating on day 7 for the remainder of the time course). On day 0, cells are either untreated, DMSO-treated, or treated with EPZ-6438 starting at 10 μM and decreasing in either threefold or fourfold dilutions. Plates are read on day 0, day 4, and day 7 using Cell Titer Glo, with compound/media being replenished on day 4. On day 7, the six-well plates are trypsinized, centrifuged, and resuspended in fresh media for counting by Vi-Cell. Cells from each treatment are replated at the original density in 96-well plates in triplicate. Cells are allowed to adhere to the plate overnight, and cells are treated as on day 0. On days 7, 11, and 14, plates are read using Cell Titer Glo, with compound/media being replenished on day 11. Averages of triplicates are used to plot proliferation over the time course, and calculate IC50 values. For cell cycle and apoptosis, G401 and RD cells are plated in 15-cm dishes in duplicate at a density of 1 × 106 cells per plate. Cells are incubated with EPZ-6438 at 1 μM, in a total of 25 mL, over a course of 14 d, with cells being split back to original plating density on day 4, 7, and 11. Cell cycle analysis and TUNEL assay are performed using a Guava flow cytometer, following the manufacturer's protocol.(Only for Reference) |
| 激酶實(shí)驗(yàn) | Biochemical Methods: EPZ-6438 is incubated for 30 min with 40 μL per well of 5 nM PRC2 (final assay concentration in 50 μL is 4 nM ) in 1X assay buffer (20 mM Bicine [pH 7.6], 0.002% Tween-20, 0.005% Bovine Skin Gelatin and 0.5 mM DTT). 10 μL per well of substrate mix comprising assay buffer 3 H-SAM, unlabeled SAM, and peptide representing histone H3 residues 21-44 containing C-terminal biotin (appended to a C-terminal amide-capped lysine) are added to initiate the reaction (both substrates are present in the final reaction mixture at their respective Km values, an assay format referred to as ‘‘balanced conditions''. The final concentrations of substrates and methylation state of the substrate peptide are indicated for each enzyme Reactions are incubated for 90 min at room temperature and quenched with 10 μL per well of 600 μM unlabeled SAM, Then transferred to a 384-well flashplate and washed after 30 min. |
| 體外活性 | 方法：滑膜肉瘤細(xì)胞 Fuji 和 HS-SY-II 用 Tazemetostat (0.039-20 μmol/L) 處理 14 天，使用 CellTiter-Glo Luminescent Cell Viability Assay 檢測(cè)細(xì)胞活力。
結(jié)果：Tazemetostat 處理 Fuji 和 HS-SY-II 細(xì)胞增殖的濃度依賴性降低，IC50 值分別為 0.15 μmol/L 和 0.52 μmol/L。[1]
方法：人腎癌細(xì)胞 G401 和人惡性胚胎橫紋肌瘤細(xì)胞 RD 用 Tazemetostat (0.006-1100 nM) 處理 4 天，使用 Western Blot 方法檢測(cè)靶點(diǎn)蛋白表達(dá)水平。
結(jié)果：Tazemetostat 處理導(dǎo)致整體 H3K27Me3 水平的濃度依賴性降低。[2] |
| 體內(nèi)活性 | 方法：為檢測(cè)體內(nèi)抗腫瘤活性，將 Tazemetostat (125-500 mg/kg，0.5% NaCMC plus 0.1% Tween 80 in water) 口服給藥給攜帶人腎癌腫瘤 G401 的 SCID 小鼠，每天兩次，持續(xù)二十八天。
結(jié)果：Tazemetostat 導(dǎo)致 SMARCB1突變型 MRT 異種移植物完全和持續(xù)消退。[1]
方法：為研究在蛛網(wǎng)膜下腔出血 (SAH) 誘導(dǎo)的神經(jīng)炎癥中的作用，將 Tazemetostat (1-9 mg/kg) 腹腔注射給血管內(nèi)穿孔法誘導(dǎo)蛛網(wǎng)膜下腔出血的大鼠模型。
結(jié)果：Tazemetostat 對(duì) EZH2 的抑制通過 H3K27me3/SOCS3/TRAF6/NF-κB 信號(hào)通路減輕了大鼠 SAH 后的神經(jīng)炎癥。[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 | 10% DMSO+40% PEG300+5% Tween 80+45% Saline : 3.3 mg/mL (5.76 mM), Please add co-solvents sequentially, clarifying the solution as much as possible before adding the next one. Dissolve by heating and/or sonication if necessary. Working solution is recommended to be prepared and used immediately.
DMSO : 29.2 mg/mL (50.98 mM), Sonication and heating are recommended.
H2O : < 1 mg/mL (insoluble or slightly soluble)
Ethanol : < 1 mg/mL (insoluble or slightly soluble)
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| 關(guān)鍵字 | epigenetic | inhibit | Tazemetostat | Histone Methyltransferase | EPZ-6438 | Inhibitor | E 7438 | EPZ 6438 | E7438 | EZH1 | cancer | EZH2 | PRC2 |
| 相關(guān)產(chǎn)品 | Tazemetostat hydrobromide | XY1 | MRTX-1719 | MAK683 | GSK126 | iso-Azalansta | EPZ015666 | UNC 0631 | AMI-1 free acid | MS37452 | MAK-683 hydrochloride | Piribedil |
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