Atrasentan, Inhibitor

Atrasentan

Cat. Num. AAA5764510

• Purity: 98.00%
• Synonyms: Atrasentan; ABT-627; A-147627; inhibitor

• Purity/Purification: 98.00%
• Form/Format: Solid. Powder

• Solubility: DMSO: Soluble; (<1mg/ refers to the product slightly soluble or insoluble)
• Formula: C29H38N2O6
• SMILES: CCCCN(CCCC)C(CN(C[C@@H]([C@H]1C(O)=O)c(cc2)cc3c2OCO3)[C@H]1c(cc1)ccc1OC)=O
• CAS Number: 173937-91-2
• Cell Research: All three prostate cancer cell lines (LNCaP, C4-2b, and PC-3 cells) are seeded at a density of 3 x 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 (0-50uM) diluted from a 10-mM stock. After 72 h of incubation with the drug, 20uL of MTT solution (5mg/mL in PBS) is added to each well and incubated further for 2 hours. Upon termination, the supernatant is aspirated and the MTT formazan formed by metabolically viable cells is dissolved in isopropanol (100 uL). The plates are mixed for 30 min on a gyratory shaker, and the absorbance is measured at 595nM on a plate reader [2].
• Kinase Assay: Cells are incubated and treated with Atrasentan. They are then washed twice with PBS and lysed in ice-cold lysis buffer [20mM Tris (pH 7.4), 150mM NaCl, 1% Triton X-100, 1mM EDTA, 1mM EGTA, 2.5mM sodium PPi, 1mM beta-glycerophosphate, 1mM sodium orthovanadate, 1ug/mL leupeptin, and 1mM 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 ug) are incubated with gentle rocking at 4 degree 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 [25mM Tris (pH 7.5), 10mM MgCl2, 5mM beta-glycerol phosphate, 0.1mM sodium orthovanadate, 2mM DTT] and then resuspended in 40uL of kinase assay buffer containing 200uM ATP and 1 ug GSK-3alpha/beta fusion protein. The kinase assay reaction is allowed to proceed at 30 degree 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-3alpha/beta antibody. For analysis of the total amount of Akt, 40 ug of protein from the lysate samples are resolved by 10% SDS-PAGE, followed by Western blotting with anti-Akt antibody [2].
• In Vitro: Atrasentan (3mg/kg, p.o.) inhibits the pressor response induced by big endothelin-1 (1nMol/kg) in pithed rats [1]. Aatrasentan (10mg/kg, i.p.) inhibited the C4-2b tumor growth within the bone environment to some extent in the SCID-hu model [2].
• Preparation and Storage: Store at -20 degree C for 3 years powder; -80 degree C for 2 years in solvent

Related Product Information for Atrasentan, inhibitor

Atrasentan (ABT-627) is an endothelin receptor antagonist (IC50: 0.0551nM for ETA).

Product Categories/Family for Atrasentan, inhibitor

GPCR/G Protein; Endothelin Receptor

References

Yuyama H, et al. Superiority of YM598 over atrasentan as a selective endothelin ETA receptor antagonist. Eur J Pharmacol. 2004 Sep 13;498(1-3):171-7.
Yuyama H, et al. Superiority of YM598 over atrasentan as a selective endothelin ETA receptor antagonist. Eur J Pharmacol. 2004 Sep 13;498(1-3):171-7.
Banerjee S, et al. In vitro and in vivo molecular evidence for better therapeutic efficacy of ABT-627 and taxotere combination in prostate cancer. Cancer Res. 2007 Apr 15;67(8):3818-26.
Banerjee S, et al. In vitro and in vivo molecular evidence for better therapeutic efficacy of ABT-627 and taxotere combination in prostate cancer. Cancer Res. 2007 Apr 15;67(8):3818-26.
Weiss J, et al. Interaction potential of the endothelin-A receptor antagonist atrasentan with drug transporters and drug-metabolising enzymes assessed in vitro. Cancer Chemother Pharmacol. 2011 Oct;68(4):1093-8.
Weiss J, et al. Interaction potential of the endothelin-A receptor antagonist atrasentan with drug transporters and drug-metabolising enzymes assessed in vitro. Cancer Chemother Pharmacol. 2011 Oct;68(4):1093-8.