| 名稱(chēng) | Atrasentan |
| 描述 | Atrasentan (ABT-627) is an endothelin receptor antagonist that can inhibit the activity of ETA (IC50 value is 0.0551 nM). |
| 細(xì)胞實(shí)驗(yàn) | All three prostate cancer cell lines (LNCaP, C4-2b, and PC-3 cells) are seeded at a density of 3 × 10^3 cells per well in 96-well microtiter culture plates. After overnight incubation, the medium is removed and replaced with a fresh medium containing different concentrations of ABT-627 (0-50 μM) diluted from a 10-mM stock. After 72 h of incubation with the drug, 20 μL of MTT solution (5 mg/mL in PBS) is added to each well and incubated further for 2 h. Upon termination, the supernatant is aspirated and the MTT formazan formed by metabolically viable cells is dissolved in isopropanol (100 μL). The plates are mixed for 30 min on a gyratory shaker, and the absorbance is measured at 595 nm on a plate reader [2]. |
| 激酶實(shí)驗(yàn) | Cells are incubated and treated with Atrasentan. They are then washed twice with PBS and lysed in ice-cold lysis buffer [20 mM Tris (pH 7.4), 150 mM NaCl, 1% Triton X-100, 1 mM EDTA, 1 mM EGTA, 2.5 mM sodium PPi, 1 mM β-glycerophosphate, 1 mM sodium orthovanadate, 1 μg/mL leupeptin, and 1 mM PMSF]. The extracts are centrifuged to remove cellular debris, and the protein content of the supernatants is determined using the bicinchoninic acid (BCA) protein assay reagent. Proteins (150 μg) are incubated with gentle rocking at 4°C overnight with immobilized Akt antibody cross-linked to agarose hydrazide beads. After the Akt is selectively immunoprecipitated from the cell lysates, the immunoprecipitated products are washed twice with lysis buffer and twice with kinase assay buffer [25 mM Tris (pH 7.5), 10 mM MgCl2, 5 mM β-glycerol phosphate, 0.1 mM sodium orthovanadate, 2 mM DTT] and then resuspended in 40 μL of kinase assay buffer containing 200 μM ATP and 1 μg GSK-3α/β fusion protein. The kinase assay reaction is allowed to proceed at 30°C for 30 min and stopped by the addition of Lamelli SDS sample buffer. Reaction products are resolved by 10% SDS-PAGE, followed by Western blotting with antiphosphorylated GSK-3α/β antibody. For analysis of the total amount of Akt, 40 μg of protein from the lysate samples are resolved by 10% SDS-PAGE, followed by Western blotting with anti-Akt antibody [2]. |
| 動(dòng)物實(shí)驗(yàn) | YM598 (0.3, 1, and 3 mg/kg), atrasentan (0.3, 1, and 3 mg/kg), or 0.5% methylcellulose as vehicle is orally administered to rats with a dosing cannula. The dosing volume of the test substances and vehicle is set at 5 mL/kg. Approximately 20 min after administration of compounds, the rats are anesthetized with sodium pentobarbital, and then pithed and ventilated 30 min after dosing. Approximately 1 h after oral administration of compounds, big endothelin-1 (1 nmol/kg) is intravenously administered, and blood pressure is measured. In these two experiments, the dose of test compound that causes 50% inhibition (ID50) of the big endothelin-1-induced increase in diastolic blood pressure is determined by linear regression analysis [1]. |
| 體外活性 | 方法：Atrasentan (ABT-627)（ 10μM,24小時(shí)） 處理PPC-1-ET 系列細(xì)胞，觀察細(xì)胞生長(zhǎng)情況。
結(jié)果：Atrasentan 處理顯著增加了凋亡細(xì)胞的數(shù)量 。[2] |
| 體內(nèi)活性 | 方法：Atrasentan （ABT-627）（20 mg/kg ，腹膜內(nèi)注射） 處理人腫瘤異種移植模型 HT29小鼠，觀察Atrasentan對(duì)腫瘤缺氧的影響。
結(jié)果：Atrasentan 可以明顯減少腫瘤缺氧。[1]
方法：?jiǎn)为?dú)Atrasentan（20 mg/kg，口服，每天）和單獨(dú)多西他賽（5 mg / kg，腹腔注射，每3天一次）；ABT-627（20 mg/kg，口服每天）和多西他賽（5 mg/kg，腹腔注射，每 3 天一次）治療腫瘤異種移植模型小鼠，觀察小鼠體內(nèi)腫瘤生長(zhǎng)。
結(jié)果：用 Atrasentan + 多西他賽聯(lián)合治療小鼠的腫瘤負(fù)荷和生長(zhǎng)速率顯著低于單獨(dú)使用Atrasentan或多西他賽治療的小鼠。[2] |
| 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 | DMSO : 9 mg/mL (17.63 mM)
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| 關(guān)鍵字 | A127722 | Endothelin Receptor | ABT627 | A 147627 | Inhibitor | inhibit | ABT 627 | A147627 | A 127722 | Atrasentan | A-127722 |
| 相關(guān)產(chǎn)品 | Clazosentan | Aprocitentan | Edonentan | Sparsentan | Macitentan | Sulfisoxazole | BMS 182874 | BMS 182874 hydrochloride | Ambrisentan | Bosentan | Sitaxsentan sodium | Sitaxsentan |
| 相關(guān)庫(kù) | 抑制劑庫(kù) | 經(jīng)典已知活性庫(kù) | 已知活性化合物庫(kù) | GPCR靶點(diǎn)分子庫(kù) | 膜蛋白靶向化合物庫(kù) | 藥物功能重定位化合物庫(kù) | 抗癌藥物庫(kù) |