| 名稱 | Exatecan Mesylate |
| 描述 | Exatecan Mesylate (DX8951f) is a DNA topoisomerase I inhibitor (IC50: 2.2 μM, 0.975 μg/mL). |
| 細胞實驗 | Growth inhibition experiments are carried out in 96-well flat-bottomed microplates, and the amount of viable cells at the end of the incubation is determined by MTT assay. Thus, 500-20000 cells/well in 150 μL of medium are plated and grown for 24 h (P388, CCRF-CEM and K562 cells for 4h), the drug (including Exatecan Mesylate, in 150 μL medium/well), or the medium alone as a control, is added, and the cells are cultured for an additional 3 days. After the addition of MTT (20 μL/well, 5 mg/mL in phosphate-buffered saline), the plates are incubated for 4 h and centrifuged at 800 g for 5 min, then the medium is removed and the blue dye formed is dissolved in 150 μL of DMSO. the absorbance is measured at 540 nm using a Microplate Reader [1]. |
| 激酶實驗 | Cells (5×10^6) are lysed with SDS buffer (10 mM HEPES, 2 mM orthovanadate, 10 mM NaF, 10 mM pyrophosphate, 1 mM PMSF, 10 μg/mL leupeptin, 10% 2-mercaptoethanol, 10% glycerol,8% SDS, 42 mM Tris-HCl, 0.002% bromophenol blue, pH 7.4). Protein in the whole-cell lysates is separated in 7.5% polyacrylamide gel and blotted onto the nitrocellulose membrane. The membrane is treated with anti-Topo I human antibody and subsequently, with horseradish peroxidase-conjugated protein A. The Topo I-specific band is detected with ECL reagents. To obtain a nuclear extract, cells (5×10^7) are washed with ice-cold buffer (2 mM K2HPO4, 5 mM MgCl2, 150 mM NaCl, 1 mM EGTA, 0.1 mM dithiothreitol), resuspended in buffer containing 0.35% Triton-X100 and PMSF and then incubated on ice for 10 min. The resulting lysates are centrifuged, and precipitates are then incubated with buffer containing 0.35 M NaCl for 1 hr at 4°C. After centrifugation (18,000g, 10 min), the protein concentration of the supernatant (nuclear extract) is determined using a protein assay kit. The same amount of nuclear protein is analyzed by Western blotting analysis using the anti-Topo I antibody [2]. |
| 動物實驗 | At 3 weeks after BxPC-3-GFP and MIA-PaCa-2-GFP orthotopic implantation, mice are randomized into five different groups of 5 mice each for treatment purposes. Group 1 serves as the negative control and does not receive any treatment. Groups 2 and 3 are treated with Exatecan Mesylate at 25 and 15 mg/kg/dose, respectively. Groups 4 and 5 receive gemcitabine treatments at 300 and 150 mg/kg/dose, respectively. At 6 weeks after BxPC-3-GFP orthotopic implantation, mice are randomized into three different groups of 20 mice each for treatment purposes. Group 1 serves as the negative control and does not receive any treatment. Group 2 is treated with 25 mg/kg/dose Exatecan Mesylate and group 3 receives 300 mg/kg/dose gemcitabine. Dosing for both drugs is performed once a week for 3 weeks, discontinued for 2 weeks, and then continued for another 3 weeks. In both early and late cancer models, primary tumor size and body weights are measured once a week. Tumor volumes are calculated using the formula a × b2 × 0.5, where a and b represent the larger and smaller diameters, respectively. At the termination of the studies, mice are sacrificed and explored. Final tumor weights and direct GFP images of primary tumor and metastases are recorded for each mouse. The tumor growth IR is calculated using the formula IR (%) = (1 ? TWt/TWc) × 100, where TWt and TWc are the mean tumor weight of treated and control groups, respectively [3]. |
| 體外活性 | Exatecan Mesylate (DX-8951f) 顯著抑制多種癌癥細胞系的增殖，對乳腺癌細胞、結腸癌細胞、胃癌細胞和肺癌細胞的平均GI50分別為2.02 ng/mL、2.92 ng/mL、1.53 ng/mL和0.877 ng/mL [1]。Exatecan 對PC-6和PC-6/SN2-5細胞展示出細胞毒性活動，其平均GI50分別為0.186 ng/mL和0.395 ng/mL。此外，Exatecan Mesylate (34 nM) 在PC-6和PC-6/SN2-5細胞中穩(wěn)定DNA-TopoI復合體 [2]。 |
| 體內(nèi)活性 | Exatecan Mesylate（3.325-50 mg/kg，靜脈注射）在攜帶腫瘤細胞的小鼠模型中展示出抗腫瘤活性，未觀察到毒性死亡[1]。Exatecan Mesylate（15, 25 mg/kg，靜脈注射）在MIA-PaCa-2早期模型及BxPC-3早期模型中，顯著抑制MIA-PaCa和BxPC-3原發(fā)腫瘤生長。在BxPC-3晚期癌癥模型中，Exatecan Mesylate（15, 25 mg/kg，靜脈注射）還顯著抑制了BxPC-3的淋巴轉移，并完全消除了肺轉移[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 | DMSO : 8 mg/mL (15.05 mM), Sonication is recommended.
H2O : 6 mg/mL (11.29 mM), Sonication and heating are recommended.
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| 關鍵字 | Exatecan | Exatecan Mesylate | Topoisomerase | Inhibitor | inhibit |
| 相關產(chǎn)品 | Prulifloxacin | Norfloxacin | Ciprofloxacin | Berberine chloride | Etoposide | Dexamethasone acetate | Ciprofloxacin monohydrochloride | EIDD-1931 | Paclitaxel | Methotrexate |
| 相關庫 | 抑制劑庫 | 抗乳腺癌化合物庫 | 抗癌活性化合物庫 | 經(jīng)典已知活性庫 | 已知活性化合物庫 | 抗衰老化合物庫 | 抗卵巢癌化合物庫 | 藥物功能重定位化合物庫 | 抗癌臨床化合物庫 | 抗癌藥物庫 |