AtRasentan hydrochloride 是内皮素 A 受体选择性拮抗剂，抑制内皮素 A 受体活性的IC50为 0.0551 nM。

Atrasentan hydrochloride is an antagonist of the endothelin receptor(IC50 = 55.1 μM for ETA).

Atrasentan (0-50 μM) significantly inhibits LNCaP and C4-2b prostate cancer cell growth. ABT-627 in combination with Taxotere elicits a significantly greater loss of viable prostate cancer cells relative to either agent alone and shows a greater degree of down-regulation of the NF-κB DNA binding activity [2]. Atrasentan profoundly induces several CYPs and drug transporters. It is a moderate P-gp inhibitor (IC50: 15.1 μM in P388/dx cells) and a weak BCRP inhibitor (IC50: 59.8 μM in MDCKII-BCRP cells) [3].

Atrasentan (3 mg/kg, p.o.) inhibits the pressor response induced by big endothelin-1 (1 nmol/kg) in pithed rats [1]. Aatrasentan (10 mg/kg, i.p.) inhibited the C4-2b tumor growth within the bone environment to some extent in the SCID-hu model [2].

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].

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].

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].

195733-43-8

C29H39ClN2O6

547.08

(+)-A 127722 hydrochloride;盐酸阿曲生坦;ABT-627 hydrochloride;A-147627 hydrochloride

0.1 M HCL:< 1 mg/mL (insoluble),ultrasonic;warming;adjust pH to 1 with HCl;heat to 60°C
DMSO:30 mg/mL (54.84 mM)
H2O:0.4 mg/mL (0.73 mM),ultrasonic and warming and adjust pH to 4 with HCl and heat to 60°C
Powder: -20°C for 3 years
In solvent: -80°C for 2 years