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Assay Method Information

Assay Name:  ITC Assay
Description:  The synthesized compounds of Formula I may be screened by one-dimensional 1H nuclear magnetic resonance spectroscopy (1D-1H NMR) binding assays against Bcl-XL. Active compounds in 1D-1H NMR binding assays are then selected and evaluated in the Isothermal Titration Calorimetry assays (ITC), cell viability assays and competitive fluorescence polarization assays (FPA). A group of compounds of Formula I display high binding affinity for Bcl-XL in these assays. The most potent compounds induce significant chemical shift changes in the active site methyl groups (region between 0.38 and 0.42 ppm) in the one-dimensional 1H-NMR spectra of Bcl-XL and also have an IC50 value in the FP displacement assays, which is more effective than Apogossypol. To confirm results of the NMR binding data and the FP assays, binding affinity of the compounds of Formula I for Bcl-XL using ITC assay were tested.
Affinity data for this assay
 

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Last update November 1, 2007
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