| 名稱 | Lexibulin |
| 描述 | Lexibulin (CYT-997)(CYT-997) is a potent tubulin polymerization inhibitor (IC50: 10-100 nM, in Y cell lines). Lexibulin blocks the formation of the mitotic spindle and leading to cell cycle arrest at the G2/M phase; this may result in disruption of the tumor vasculature and tumor blood flow, and tumor cell death. |
| 細胞實驗 | Cells are exposed to various concentrations of CYT997 for ~72 hours. Cell proliferation is assessed with either the Alamar blue or MTT assays. For MTT assays, 5 mg/mL of MTT is added to all wells, plates are incubated for 6 hours at 37 °C, and then lysis buffer is added (10% SDS in 0.01 N HCl) and absorbance is measured at 620 nm in a BMG Technologies Lumistar or Polarstar plate reader. For Alamar blue assays, Alamar blue (10 μL/well) is added to each well and the plates are incubated at 37 °C for 4 hours. The fluorescence is then measured using a fluorescence plate reader with an excitation filter at 544 nm and an emission filter at 590 nm. For cell cycle analysis, cells are fixed and permeabilized with 70% ethanol in PBS and incubated at 4 °C overnight. RNase-treated samples (10 μg RNase/mL for 20 minutes at 37 °C) are stained with propidium iodide (5 μg/mL) at 4 °C for a minimum of 10 minutes. Cell cycle variables are determined by fluorescence-activated cell sorting (FACS) analysis using a Beckman-Coulter Quanta SC MPL system and analyzed using CXP Software. For apoptosis analysis, cells are detached and collected. Annexin staining is done using the Vybrant Apoptosis Assay Kit. Cells are stored on ice and analyzed on a Beckman Coulter Quanta MPL within 1 hour of preparation. Annexin V–positive cells are determined using two-channel analysis.(Only for Reference) |
| 激酶實驗 | Turbidimetric assay for tubulin polymerization: The effect of CYT997 on microtubule polymerization is determined using conventional turbidimetric assay using bovine neuronal tubulin in which the assembly of microtubules is monitored by an increase in absorbance at 340 nm. Increasing concentrations of CYT997 is added to 100 μL of tubulin/GTP/glycerol. Turbidimetric assays of microtubule assembly is done by incubating bovine microtubule protein in cuvettes at 37 °C in a thermostatically controlled spectrophotometer measuring the change in absorbance at 340 nm over time in PEM buffer [80 nM PIPES (pH 6.9), 2 mM MgCl2, 0.5 mM EGTA, and 5% glycerol]. |
| 體外活性 | CYT997（7.5 mg/kg,i.p.）給藥肝轉(zhuǎn)移,6 h時明顯降低血流,效果與陽性對照CA4P（100 mg/kg）的作用程度相近.與體外抗骨髓瘤活性一致,CYT997（15 mg/kg/day）處理激進型系統(tǒng)性骨髓性白血病小鼠模型,可使小鼠壽命延長.與靜脈注射半衰期(1.5小時)相比,CYT997口服處理大鼠的半衰期(2.5 小時)稍長, 絕對口服生物有效性為50%-70%.與紫杉醇相比,CYT997口服給藥PC3移植瘤小鼠,對腫瘤生長的抑制作用更強,該作用呈劑量依賴性.CYT997對攜帶小鼠乳腺癌4T1細胞的同位模型也有效,該模型對Paclitaxel治療是有一定抗性的. |
| 體內(nèi)活性 | CYT997對16種癌細胞均有毒性（IC50：9/101 nM,HepG2和KHOS/NP細胞）。CYT997對HCT15細胞有較好作用（IC50：52 nM）,其具有多耐藥機制Pgp(MDR+)。處理A549細胞24 h,CYT997（1 μM）對微管快速重組有誘導(dǎo)作用,包括現(xiàn)有的微管網(wǎng)絡(luò)遭破壞,及菌斑處細胞質(zhì)微管蛋白累積,還顯著改變細胞形態(tài),包括粘附細胞丟失,細胞減少。通過抑制微管聚合,CYT997可使細胞周期停在G2-M分界處,且促使磷酸化Bcl-2增多,還可使cyclin B1表達、caspase-3激活及PARP產(chǎn)生增高。CYT997處理1小時后,快速可逆的提高HUVEC單層膜通透性（IC50：80 nM）。與CYT997破壞細胞微管蛋白一致,其有效抑制增殖,誘導(dǎo)細胞周期停滯,且誘導(dǎo)人骨髓細胞系和原代MM細胞凋亡。 |
| 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 : 16 mg/mL (36.8 mM)
DMSO : 80 mg/mL (184.1 mM)
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| 關(guān)鍵字 | inhibit | Reactive Oxygen Species | Apoptosis | CYT 997 | Microtubule/Tubulin | CYT997 | Inhibitor | Lexibulin |
| 相關(guān)產(chǎn)品 | L-Glutamic acid | Sodium 2-oxopropanoate | Metronidazole | 5-Fluorouracil | Dextran sulfate sodium salt (MW 4500-5500) | Stavudine | Tributyrin | Imeglimin hydrochloride | L-Ascorbic acid | Acetylcysteine | Salicylic acid | Sodium 4-phenylbutyrate |
| 相關(guān)庫 | 抑制劑庫 | 經(jīng)典已知活性庫 | 抗癌活性化合物庫 | 已知活性化合物庫 | 抗衰老化合物庫 | 抗肝癌化合物庫 | 藥物功能重定位化合物庫 | 微管靶向化合物庫 | 抗癌臨床化合物庫 | 抗癌藥物庫 |