For synthesis of KAM-derivatized resin, packed NHS-activated sepharose 4 fast flow resin (volume = 2 mL; GE Healthcare) was washed with anhydrous DMSO (3 × 10 mL). To the washed NHS-activated sepharose resin was added 0.5 mM KAM in anhydrous DMSO (8 mL; 2 μmol compound/mL of resin), followed by the addition of triethylamine (30 μL). The reaction mixture was vortexed to mix and pelleted by centrifugation (100 x g, 2 min). An aliquot of the supernatant (50 μL) was saved for LC/MS analysis. The reaction mixture was allowed incubate overnight at room temp with end-over-end rotating agitation. On the following day, the reaction mixture was pelleted by centrifugation (100 x g, 2 min). An aliquot of the supernatant (50 μL) was saved for LC/MS analysis. Completion of coupling was inferred by loss of starting material following LC/MS analysis. 2-(2-Aminoethoxy)ethanol (100 μL; Sigma-Aldrich) was added to the reaction mixture, vortexed, and incubate overnight at room temp with end-over-end agitation. The KAM-derivatized resin was then washed with anhydrous DMSO (3 × 10 mL) and 95% EtOH (3 × 10 mL).
For synthesis of imatinib-derivatized resin, a similar protocol was followed as described above except that final concentration of compound on the bead was 0.25 μmol compound/mL.
For synthesis of dasatinib-derivatized resin, the protocol for the KAM-derivatized resin was followed.
For synthesis of imatinib-derivatized resin, a similar protocol was followed as described above except that final concentration of compound on the bead was 0.25 μmol compound/mL.
For synthesis of dasatinib-derivatized resin, the protocol for the KAM-derivatized resin was followed.