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In vitro functional assay, inhibiting agonist stimulated G-protein activation (measured using GTPγS binding) in CHO membranes expressing the cloned human mu, kappa, and delta receptors. In the present study, opioid ligand binding and GTPγS functional assays were converted to a homogeneous SPA permitting the development of simpler assays with dramatically increased throughput. Optimization in the presence of sodium chloride was designed to bias these assays toward the detection of opioid antagonists. Kb values for compounds with modest binding affinities were not determined. See. Rodgers G., Hubert C., McKinzie J., Suter T., Statnick M., Emmerson P., and Stancato L. Assay Drug Dev. Technol. 1 (2003) 627 for the assay development method
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note
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1H NMR, MS, and one of the following methods for purity verification: HRMS, EA or HPLC. See WO2004026305 (A1) for details of compound preparation.
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