| 名稱 | Atrasentan hydrochloride |
| 描述 | Atrasentan hydrochloride (ABT-627 hydrochloride) is an antagonist of the endothelin receptor(IC50 = 55.1 μM for ETA). |
| 細胞實驗 | 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]. |
| 激酶實驗 | 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]. |
| 動物實驗 | 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 | Atrasentan（0-50 μM）顯著抑制LNCaP和C4-2b前列腺癌細胞生長。ABT-627與Taxotere聯(lián)合使用，相比單獨使用任一化合物，可顯著降低前列腺癌活細胞的數(shù)量，并更大程度地下調(diào)NF-κB DNA結(jié)合活性[2]。Atrasentan顯著誘導(dǎo)多種CYPs和藥物轉(zhuǎn)運體的表達。它是一種中等強度的P-gp抑制劑（IC50：15.1 μM，在P388/dx細胞中）和一種弱BCRP抑制劑（IC50：59.8 μM，在MDCKII-BCRP細胞中）[3]。 |
| 體內(nèi)活性 | Atrasentan（3 mg/kg, p.o.）通過抑制大型內(nèi)皮素-1（1 nmol/kg）誘導(dǎo)的加壓反應(yīng)，在去髓大鼠中顯示出效果[1]。此外，Atrasentan（10 mg/kg, i.p.）在SCID-hu模型中對C4-2b腫瘤在骨環(huán)境中的生長有一定程度的抑制作用[2]。 |
| 存儲條件 | Powder: -20°C for 3 years | In solvent: -80°C for 1 year
Shipping with blue ice/Shipping at ambient temperature. |
| 溶解度 | H2O : 0.4 mg/mL (0.73 mM), when pH is adjusted to 4 with HCl. Sonication and heating to 60℃ are recommended.
DMSO : 30 mg/mL (54.84 mM), Sonication is recommended.
0.1 M HCl : < 1 mg/mL (insoluble)
10% DMSO+40% PEG300+5% Tween 80+45% Saline : 2 mg/mL (3.66 mM), Sonication is recommended.
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| 關(guān)鍵字 | Inhibitor | inhibit | ET-A | EndothelinReceptor | Endothelin Receptor | Atrasentan hydrochloride | Atrasentan | ABT-627 Hydrochloride | ABT627 Hydrochloride | ABT-627 | ABT627 | ABT 627 Hydrochloride | ABT 627 | A-147627 Hydrochloride | A147627 Hydrochloride | A-147627 | A147627 | A-127722 Hydrochloride | A127722 Hydrochloride | A 147627 Hydrochloride | A 147627 | A 127722 Hydrochloride | (+)-A 127722 |
| 相關(guān)產(chǎn)品 | Clazosentan | BQ-788 | Aprocitentan | SB-209670 | Edonentan | Sparsentan | Macitentan | BMS 182874 | BMS 182874 hydrochloride | Bosentan | Ambrisentan | Sitaxsentan sodium |
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