The Design, Synthesis And Potential Utility Of Fluorescence Probes That Target Dfg-Out Conformation Of P38 Alpha For High Throughput Screening Binding Assay

The design, synthesis and utility of fluorescence probes that bind to the DFG-out conformation of p38 alpha kinase are described. Probes that demonstrate good affinity for p38 alpha, have been identified and one of the probes, PF-04438255, has been successfully used in an high throughput screening (HTS) assay to identify two novel non-classical p38 alpha inhibitors. In addition, a cascade activity assay was utilized to validate the selective binding of these non-classical kinase inhibitors to the unactive form of the enzyme.